3.2.1.4: cellulase
This is an abbreviated version!
For detailed information about cellulase, go to the full flat file.
Word Map on EC 3.2.1.4
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3.2.1.4
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cellulose
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trichoderma
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biomass
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reesei
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xylanase
-
cellulolytic
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saccharification
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lignocellulosic
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cellobiose
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straw
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lignin
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polysaccharide
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aspergillus
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glycoside
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beta-glucosidase
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corn
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biofuels
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pectinase
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xylan
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hemicellulose
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bagasse
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penicillium
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endoglucanases
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bioethanol
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xylose
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amylase
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polygalacturonase
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sugarcane
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carbohydrate-binding
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feedstock
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viride
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glucans
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thermocellum
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bran
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carboxymethylcellulose
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microcrystalline
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exoglucanase
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cellulose-binding
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laccase
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biorefinery
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rumen
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biotechnology
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analysis
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novozyme
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silage
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endoxylanase
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compost
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industry
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textile industry
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food industry
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biofuel production
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agriculture
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deconstruct
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xylobiose
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diagnostics
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harzianum
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degradation
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chrysosporium
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synthesis
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detergent
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environmental protection
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xyloglucans
- 3.2.1.4
- cellulose
- trichoderma
- biomass
- reesei
- xylanase
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cellulolytic
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saccharification
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lignocellulosic
- cellobiose
- straw
- lignin
- polysaccharide
- aspergillus
- glycoside
- beta-glucosidase
- corn
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biofuels
- pectinase
- xylan
- hemicellulose
- bagasse
- penicillium
- endoglucanases
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bioethanol
- xylose
- amylase
- polygalacturonase
- sugarcane
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carbohydrate-binding
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feedstock
- viride
- glucans
- thermocellum
- bran
- carboxymethylcellulose
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microcrystalline
- exoglucanase
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cellulose-binding
- laccase
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biorefinery
- rumen
- biotechnology
- analysis
-
novozyme
-
silage
- endoxylanase
-
compost
- industry
- textile industry
- food industry
- biofuel production
- agriculture
-
deconstruct
- xylobiose
- diagnostics
- harzianum
- degradation
- chrysosporium
- synthesis
- detergent
- environmental protection
- xyloglucans
Reaction
Synonyms
(1->4)-beta-D-glucan 4-glucanohydrolase, 1,4-beta-D-endoglucanase, 1,4-beta-D-glucan-4-glucanohydrolase, 168cel5, 9.5 cellulase, Abscission cellulase, AEG, Ag-EGase III, AgCMCase, alkali cellulase, Alkaline cellulase, AnCel5A, AtCel5, ATEG_07420, avicelase, BC-EG70a, Bc22Cel, BCE1, BcsZ, beta-1,4-endoglucan hydrolase, beta-1,4-endoglucanase, beta-1,4-glucanase, Bgl7A, bifunctional endoglucanase/xylanase, BlCel9, BP_Cel9A, Bx-ENG-1, Bx-ENG-2, Bx-ENG-3, C4endoII, carbomethyl cellulase, Carboxymethyl cellulase, Carboxymethyl-cellulase, carboxymethylcellulase, Cat 1, Caylase, CBH45-1, cbh6A, CBHI, CBHII, CcCel6C, CEL1, Cel1 EGase, Cel12A, Cel1753, Cel28a, Cel44A, Cel45A, Cel48A, Cel5, Cel5A, cel5B, Cel5E, Cel6A, Cel6A (E2), Cel6B, Cel6C, CEL7, Cel7A, Cel7B, Cel8, Cel8A, Cel8M, Cel8Y, Cel9A, CEL9A-50, CEL9A-65, Cel9A-68, CEL9A-82, Cel9A-90, Cel9B, CEL9C1, Cel9K, Cel9M, Cel9Q, CelA, CelB, CelC2 cellulase, CelCM3, CelDR, CelE, CelF, Celf_1230, Celf_3184, CelG, CelG endoglucanase, CelI15, cell-bound bacterial cellulase, CelL15, CelL73, cellic Ctec2, cellobiohydrolase, cellobiohydrolase I, celluase A, Celluclast, celludextrinase, Cellulase, cellulase 12A, cellulase A, cellulase A 3, cellulase Cel48F, cellulase Cel9A, cellulase Cel9M, cellulase CelC2, cellulase CelE, cellulase CM3, Cellulase E1, Cellulase E2, Cellulase E4, Cellulase E5, cellulase EGX, cellulase II, cellulase III, cellulase K, Cellulase SS, cellulase T, Cellulase V1, cellulase Xf818, cellulases I, cellulases III, cellulosin AP, Cellulysin, CelP, celS, CelStrep, celVA, CelX, CenA, CenC, CfCel6A, CfCel6C, CfEG3a, CHU_1280, CHU_2103, CjCel9A, Clocel_2741, CMCase, CMCase-I, CMCax, CMcellulase, Csac_1076, Csac_1078, CSCMCase, ctCel9D-Cel44A, CTendo45, ctendo7, CtGH5, Cthe_0435, CTHT_0045780, CX-cellulase, CyPB, DCC85_10145, DK-85, Dockerin type 1, Dtur_0671, E1 endoglucanase, Econase, EfPh, EG I, EG III, EG1, EG12, EG2, EG25, EG271, EG28, EG3, EG35, EG44, EG47, EG51, Eg5a, EG60, EGA, EGase, EGase II, EGB, EGC, EGCCA, EGCCC, EGCCD, EGCCF, EGCCG, EGD, EGE, EGF, egGH45, EGH, EGI, EGII, EGII/Cel5A, EGIV, EGL, EGL 1, Egl-257, Egl1, Egl499, Egl5a, EglA, eglB, EglC, EGLII, EglS, EGM, EGPf, EGPh, EGSS, EgV, EGX, EGY, EGZ, endo-1,4-B-glucanase, endo-1,4-beta-D-glucanase, endo-1,4-beta-glucanase, endo-1,4-beta-glucanase 1, endo-1,4-beta-glucanase 2, endo-1,4-beta-glucanase E1, endo-1,4-beta-glucanase V1, endo-beta-1,3-1,4-glucanase, endo-beta-1,4-glucanase, endo-beta-1,4-glucanase 1, endo-beta-1,4-glucanase 2, endo-beta-1,4-glucanase CMCax, endo-beta-1,4-glucanase EG27, endo-beta-1,4-glucanase EG45, endo-beta-D-1,4-glucanohydrolase, endo-beta-glucanase, endo-glucanase, ENDO1, ENDO2, endocellulase, endocellulase E1, endocellulases I, endocellulases II, endocellulases III, endocellulases IV, endogenous beta-1,4-endoglucanase, endogenous cellulase, endoglucanase, endoglucanase 1, endoglucanase 35, endoglucanase 47, endoglucanase CBP105, endoglucanase Cel 12A, endoglucanase Cel 5A, endoglucanase Cel 7B, endoglucanase Cel5A, endoglucanase Cel6A, endoglucanase D, endoglucanase EG-I, endoglucanase EG25, endoglucanase EG28, endoglucanase EG44, endoglucanase EG47, endoglucanase EG51, endoglucanase EG60, endoglucanase H, endoglucanase II, endoglucanase IIa, endoglucanase IV, endoglucanase L, endoglucanase M, endoglucanase V, endoglucanase Y, endolytic cellulase, EngA, EngH, EngL, EngM, engXCA, EngY, EngZ, family 7 cellobiohydrolase, FI-CMCASE, FnCel5A, FpCel45, Fpcel45a, GE40, GE40 endoglucanase, GH12 endo-1,4-beta-glucanase, GH124 endoglucanase, GH45 endoglucanase, gh45-1, GH5 cellulase, GH5 endoglucanase, GH6 endoglucanase, GH7 endoglucanase, GH9 termite cellulase, Glu1, Glu2, glycoside hydrolase family 9 endoglucanase, GtGH45, Lp-egl-1, manganese dependent endoglucanase, Maxazyme, Meicelase, mesophilic endoglucanase, mgCel6A, More, MtGH45, nmGH45, Onozuka R10, pancellase SS, PaPopCel1, PF0854, PH1171, PttCel9A, RCE1, RCE2, Roth 3056, RtGH124, Rucel5B, Sl-cel7, Sl-cel9C1, SnEG54, SoCel5, ssgluc, SSO1354, SSO1354 enzyme, SSO1354 protein, SSO1949, SSO2534, STCE1, Sumizyme, T12-GE40, TC Serva, TeEG-I, TeEgl5A, TfCel9A, ThEG, theme C glycoside hydrolase family 9 endo-beta-glucanase, Thermoactive cellulase, thermostable carboxymethyl cellulase, THITE_2110957, TM_1525, TM_1751, umcel5G, umcel9y-1, Vul_Cel5A
ECTree
3.2.1.4 cellulaseAdvanced search results
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results (Engineering
Engineering on EC 3.2.1.4 - cellulase
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
F194A
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site-directed mutagenesis, the mutant shows 2fold increased activity compared to wild-type enzyme
E289V
I62T/L79I/A93T/S308P/I370V/L374P/M416V/F472I/I484V/W494R
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S40 mutant, DNA shuffling. Higher hydrolytic activities than the wild-type enzyme
K120E/D272H/S283G/S308P/L374P
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M1-23 mutant, random mutation. Higher hydrolytic activities than the wild-type enzyme
K120E/S283G/S308P/L374P
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M1 mutant, random mutation. Higher hydrolytic activities than the wild-type enzyme, shows 1.25fold increase in activity
N39D/K120E/N175H/V255A/S308P/L386S/K398R
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S75 mutant, DNA shuffling. Higher hydrolytic activities than the wild-type enzyme
T32I/N39D/K120E/S248G/S283G/S308P/R314G/I370N/L374P/N403D/N451D/S467N
-
S78 mutant, DNA shuffling. Higher hydrolytic activities than the wild-type enzyme
V74A/K120E/D272G/K337E/S355P/D459G/K479E/K482E/K491N
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M44 mutant, random mutation. Higher hydrolytic activities than the wild-type enzyme, shows 1.56fold increase in activity
V74A/K120E/D272G/K337E/S355P/T449I/D459G/K479E/K482E/D488N/K491N
-
M44-11 mutant, random mutation. Higher hydrolytic activities than the wild-type enzyme
K120E/D272H/S283G/S308P/L374P
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M1-23 mutant, random mutation. Higher hydrolytic activities than the wild-type enzyme
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K120E/S283G/S308P/L374P
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M1 mutant, random mutation. Higher hydrolytic activities than the wild-type enzyme, shows 1.25fold increase in activity
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V74A/K120E/D272G/K337E/S355P/D459G/K479E/K482E/K491N
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M44 mutant, random mutation. Higher hydrolytic activities than the wild-type enzyme, shows 1.56fold increase in activity
-
V74A/K120E/D272G/K337E/S355P/T449I/D459G/K479E/K482E/D488N/K491N
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M44-11 mutant, random mutation. Higher hydrolytic activities than the wild-type enzyme
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DELTA1-90
expressed as a soluble protein in Pichia pastoris, the wild-type enzyme is anchored to membrane
A211D
-
site-directed mutagenesis, the mutant shows reduced activity at different pH values compared to the wild-type enzyme
A253S
-
site-directed mutagenesis, the mutant shows reduced activity at different pH values compared to the wild-type enzyme
D216C
-
replacement of Asp with cysteinesulfinate by combination of site-directed mutagenesis and chemical modification, the substituted cysteinyl residue is oxidized to cysteine sulfinic acid with hydrogen peroxide, the resulting protein product retains its native structure, almost inactive mutant
D216N
-
site-directed mutagenesis, the mutant shows reduced activity and a shift in pH dependence compared to the wild-type enzyme
D252C
-
site-directed mutagenesis, substitution of the catalytic acid residue Asp252, almost inactive mutant
D287C
-
site-directed mutagenesis, the mutant shows reduced activity at different pH values compared to the wild-type enzyme
D287E
-
site-directed mutagenesis, the mutant shows reduced activity at different pH values compared to the wild-type enzyme
D287N
-
site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0
D392A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
D392C
-
replacement of Asp with cysteinesulfinate by combination of site-directed mutagenesis and chemical modification, the substituted cysteinyl residue is oxidized to cysteine sulfinic acid with hydrogen peroxide, the resulting protein product retains its native structure. Oxidation of the Asp392Cys mutant enzyme restores 52% of wild-type activity when assessed at pH 7.5. The replacement of Asp392 with cysteine sulfinate induced an acidic shift in the pH profile of the enzyme such that this enzyme derivative is more active than wild-type CenA below pH 5.5
D392N
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
D392S
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
E368A
-
site-directed mutagenesis, the mutant shows reduced activity at pH values of pH 7 and pH 9, but increased activity at pH 5, compared to the wild-type enzyme
E368A/E407A
-
site-directed mutagenesis, the double mutant shows decreased activity at pH 5.0 compared to the wild-type enzyme
E407A
-
site-directed mutagenesis, the mutant shows reduced activity at pH values of pH 7 and pH 9, but increased activity at pH 5, compared to the wild-type enzyme
K292A
-
site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0
K292A/E407A
-
site-directed mutagenesis, the double mutant shows increased activity at pH 5.0 compared to the wild-type enzyme
L387P
-
site-directed mutagenesis, the mutant shows reduced activity at pH values of pH 7 and pH 9, but increased activity at pH 5, compared to the wild-type enzyme
N320C
-
site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0
N320D
-
site-directed mutagenesis, the mutant shows highly reduced activity at different pH values compared to the wild-type enzyme
N320E
-
site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0
N360D
-
site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0
N360H
-
site-directed mutagenesis, the mutant shows reduced activity at different pH values compared to the wild-type enzyme
N360K
-
site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0
N360R
-
site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0
Q256C
-
site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0
Q256D
-
site-directed mutagenesis, the mutant shows reduced activity at different pH values compared to the wild-type enzyme
Q256E
-
site-directed mutagenesis, the mutant shows reduced activity at different pH values compared to the wild-type enzyme
S319A
-
site-directed mutagenesis, the mutant shows reduced activity at different pH values compared to the wild-type enzyme
S319D
-
site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0
S319H
-
site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0
S319K
-
site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0
S319R
-
site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0
Y321A
-
site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0
Y321F
-
site-directed mutagenesis, the mutant shows reduced activity at pH values of pH 7 and pH 9, but increased activity at pH 5, compared to the wild-type enzyme
Y321F/E407A
-
site-directed mutagenesis, the double mutant shows increased activity at pH 5.0 compared to the wild-type enzyme
Y321H
-
site-directed mutagenesis, the mutant shows highly reduced activity at different pH values compared to the wild-type enzyme
Y321K
-
site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0
Y321R
-
site-directed mutagenesis, the mutant shows highly reduced activity at different pH values compared to the wild-type enzyme
K94R/S365P
optimal temperature of K94R/S365P is increased by 7.5°C compared to wild-type. K94R/S365P retains 78.3% relative activity at 70°C, while the wild-type retains 5.8%. K94R/S365P shows 45.1fold higher activity than the wild-type at 70°C and 3.1fold higher activity at 42.5°C, which is the optimal temperature ofthe wild type. K94R/S365P is stimulated in 2.5fold lower concentration of CaCl2 and displays delayed aggregation temperature in the presence of CaCl2 compared to the wild type. In long-term hydrolysis, K94R/S365P reduces the newly released reducing sugars after 12 h reaction
K94R/S365P
-
optimal temperature of K94R/S365P is increased by 7.5°C compared to wild-type. K94R/S365P retains 78.3% relative activity at 70°C, while the wild-type retains 5.8%. K94R/S365P shows 45.1fold higher activity than the wild-type at 70°C and 3.1fold higher activity at 42.5°C, which is the optimal temperature ofthe wild type. K94R/S365P is stimulated in 2.5fold lower concentration of CaCl2 and displays delayed aggregation temperature in the presence of CaCl2 compared to the wild type. In long-term hydrolysis, K94R/S365P reduces the newly released reducing sugars after 12 h reaction
-
G117S
1.6fold increase in activity, mutation might directly affect the substratebinding affinity
V9A/K353E
1.4fold increase in activity, mutation might directly affect the substratebinding affinity
D117N
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
N95D
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
D117N
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
-
N95D
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
-
N92D
replaxement of the general base in the catalytic mechanism, drastic decrease in activity. Unlike Asn92, residue Asp92 is mobile
T130I/S134Q/M259I/A241E/S183T/Q307R/A309S
temperature required to reduce the initial activity by 50% within 120 min is increased by 4.4 degrees compared to wild-type
T130I/S134Q/M259I/A241E/S183T/Q307R/A309S/T112S/N125K/Q204K/F206Y/I365
temperature required to reduce the initial activity by 50% within 120 min is increased by 5.4 degrees compared to wild-type, specific activity is the same as wild-type
C106A/C159A
kcat/KM for p-nitrophenyl cellobiose is 1.3fold higher than wild-type value. Activity towards carboxymethyl cellulose is increased by 1.7fold
C106A/C159A/C372A/C412A
kcat/KM for p-nitrophenyl cellobiose is 1.4fold higher than wild-type value. Activity towards carboxymethyl cellulose is increased by 2.1fold
C106S
melting temperature of the mutant enzyme is 2°C lower than the wild-type enzyme
C159A
kcat/KM for p-nitrophenyl cellobiose is 1.8fold lower than wild-type value
C372/AC412A
kcat/KM for p-nitrophenyl cellobiose is 2.9fold higher than wild-type value. Activity towards carboxymethyl cellulose is increased by 1.6fold
D385N
activity towards carboxymethyl cellulose is 29.9% of wild-type activity
DELTAQ1-G5
activity towards carboxymethyl cellulose is 135.6% of wild-type activity. kcat/Km for p-nitrophenyl cellobiose is 2.3fold higher than wild-type value. Thermostability is not significantly influenced
E163A
kcat/KM for p-nitrophenyl cellobiose is 6fold lower than wild-type value
E201A
E201Q
activity towards carboxymethyl cellulose is 1.12% of wild-type activity. kcat/Km for p-nitrophenyl cellobiose is 43fold lower than wild-type value
E342A
E342Q
activity towards carboxymethyl cellulose is 0.01% of wild-type activity
G158A
kcat/KM for p-nitrophenyl cellobiose is 2fold lower than wild-type value
H155A
kcat/KM for p-nitrophenyl cellobiose is 140fold lower than wild-type value
H161A
kcat/KM for p-nitrophenyl cellobiose is neatrly identical to wild-type value
H297A
activity towards carboxymethyl cellulose is 0.08% of wild-type activity
H297N
activity towards carboxymethyl cellulose is 1.31% of wild-type activity. pH-optimum is 7.0, compared to 5.5-6 for wild-type enzyme
I157A
kcat/KM for p-nitrophenyl cellobiose is nearly identical to wild-type value
I162A
kcat/KM for p-nitrophenyl cellobiose is 140fold lower than wild-type value
N200A
activity towards carboxymethyl cellulose is 5.43% of wild-type activity
P164A
kcat/KM for p-nitrophenyl cellobiose is 6fold lower than wild-type value
P74C
melting temperature of the mutant enzyme is 2°C lower than the wild-type enzyme
P74C/C106S
melting temperature of the mutant enzyme is 2°C lower than the wild-type enzyme
Q306A
25% of the activity with avicel as compared to wild-type enzyme
R102A
activity towards carboxymethyl cellulose is 0.67% of wild-type activity
R156A
kcat/KM for p-nitrophenyl cellobiose is 10fold lower than wild-type value
T160A
kcat/KM for p-nitrophenyl cellobiose is nearly identical to wild-type value
W377A
W82A
75% of the activity with avicel as compared to wild-type enzyme
Y299A
activity towards carboxymethyl cellulose is 0.21% of wild-type activity
Y299F
C106S
-
melting temperature of the mutant enzyme is 2°C lower than the wild-type enzyme
-
P74C
-
melting temperature of the mutant enzyme is 2°C lower than the wild-type enzyme
-
P74C/C106S
-
melting temperature of the mutant enzyme is 2°C lower than the wild-type enzyme
-
Q306A
-
25% of the activity with avicel as compared to wild-type enzyme
-
A3V/A6Q/T7K/A8P/N10T/E18K/P22D/P58T/Y60L/N157A/D181N/E183Q
mutant is more active and stable than wild-type Cel7A or Trichoderma reesei Cel7A in aqueous ionic liquids solutions, i.e. up to 43% (w/w) 1,3-dimethylimdazolium dimethylphosphate and 20% (w/w) 1-ethyl-3-methylimidazolium acetate
A6L/A8E/N10V/P58E/T59S/Y60L/L73V/G80A/V84I/S87N/S89D/K92T/L105V/L108M/Q109E/N220T/V222F/S301K/I308V/S311G/N312K/Q316N/P317S/N318E/D320T/I321W/T325G/T438N/G439P/T440P/P441G/S442G/H471M
mutant is more active and stable than wild-type Cel7A or Trichoderma reesei Cel7A in aqueousionic liquids solutions, i.e. up to 43% (w/w) 1,3-dimethylimdazolium dimethylphosphate and 20% (w/w) 1-ethyl-3-methylimidazolium acetate. Increase in melting temperature of 1.9-3.9°C compared to wild-type
G266C/D320C
introduction of an additional disulfide bridge, decrease in activity
G4C/A72C
introduction of an additional disulfide bridge, improves thermostability
G4C/A72C/N54C/P191C/T243C/A375C
introduction of 3 additional disulfide bridges, improves thermostability
N54C/P191C
introduction of an additional disulfide bridge, improves thermostability
Q190C/I200C
introduction of an additional disulfide bridge, decrease in activity
T243C/A375C
introduction of an additional disulfide bridge, improves thermostability
A170S
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
A87S
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
G91A
-
site-directed mutagenesis, the mutant has 3-4fold higher activity towards carboxymethyl cellulose than the wild type enzyme
G91A/K429A
-
site-directed mutagenesis, the double mutant has 7-13fold higher activity towards carboxymethyl cellulose than the wild type enzyme, the mutations show synegistic effects
G91A/Y97W
-
site-directed mutagenesis, the double mutant has 7-13fold higher activity towards carboxymethyl cellulose than the wild type enzyme, the mutations show synegistic effects
G91A/Y97W/K429A
-
site-directed mutagenesis, the triple mutant has 7-13fold higher activity towards carboxymethyl cellulose than the wild type enzyme, the mutations show synegistic effects
G91I
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
I347V
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
K429A
-
site-directed mutagenesis, the mutant has 3-4fold higher activity towards carboxymethyl cellulose than the wild type enzyme
L103I
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
N245S
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
N38D
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Q42N/K43N
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
S173A
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
S90D
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Y329F
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Y97F
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Y97W
-
site-directed mutagenesis, the mutant has 3-4fold higher activity towards carboxymethyl cellulose than the wild type enzyme
Y97W/K429A
-
site-directed mutagenesis, the double mutant has 7-13fold higher activity towards carboxymethyl cellulose than the wild type enzyme, the mutations show synegistic effects
E410Q
-
mutation totally inactivates carboxymethyl cellulase activity of the protein
G145D/N207K
random mutagenesis, the mutant shows increased activity with carboxymethyl cellulose compared to the wild-type enzyme
G263C/R307H
random mutagenesis, the mutant shows increased activity with carboxymethyl cellulose compared to the wild-type enzyme
P228R
random mutagenesis, the mutant shows increased activity with carboxymethyl cellulose compared to the wild-type enzyme
T157I/G251D/V259D
random mutagenesis, the mutant shows increased activity with carboxymethyl cellulose compared to the wild-type enzyme
T67N/D142E/S218N/V242D/D330E
random mutagenesis, the mutant shows increased activity with carboxymethyl cellulose compared to the wild-type enzyme
N194A
-
mutation in potential N-glycosylation site, no notable effect on the enzyme thermostability. Slight shift of pH optimum 4.5 for wild-type to 5.0 and 32-35% increase in the specific activity against carboxymethylcellulose and barley beta-glucan
N19A
-
mutation in potential N-glycosylation site, no notable effect on the enzyme thermostability but 26% decrease in the specific activity against carboxymethylcellulose and 12% against barley beta-glucan
N42A
-
mutation in potential N-glycosylation site, no notable effect on the enzyme thermostability. Slight shift of pH optimum 4.5 for wild-type to 5.0 and 12-13% increase in the specific activity against carboxymethylcellulose and barley beta-glucan
S127C/A165C
site-directed mutagenesis, introduction of a disulfide bond into mutant DSB2, the mutant enzyme displays 15-21% increased specific activity against carboxymethylcellulose and beta-glucan, and increased thermostability compared to wild-type enzyme EGLII
Y171C/L201C
site-directed mutagenesis, introduction of a disulfide bond into mutant DSB3, the mutant enzyme displays 15-21% increased specific activity against carboxymethylcellulose and beta-glucan, and increased thermostability compared to wild-type enzyme EGLII
S127C/A165C
-
site-directed mutagenesis, introduction of a disulfide bond into mutant DSB2, the mutant enzyme displays 15-21% increased specific activity against carboxymethylcellulose and beta-glucan, and increased thermostability compared to wild-type enzyme EGLII
-
Y171C/L201C
-
site-directed mutagenesis, introduction of a disulfide bond into mutant DSB3, the mutant enzyme displays 15-21% increased specific activity against carboxymethylcellulose and beta-glucan, and increased thermostability compared to wild-type enzyme EGLII
-
S127C/A165C
-
site-directed mutagenesis, introduction of a disulfide bond into mutant DSB2, the mutant enzyme displays 15-21% increased specific activity against carboxymethylcellulose and beta-glucan, and increased thermostability compared to wild-type enzyme EGLII
-
Y171C/L201C
-
site-directed mutagenesis, introduction of a disulfide bond into mutant DSB3, the mutant enzyme displays 15-21% increased specific activity against carboxymethylcellulose and beta-glucan, and increased thermostability compared to wild-type enzyme EGLII
-
S127C/A165C
-
site-directed mutagenesis, introduction of a disulfide bond into mutant DSB2, the mutant enzyme displays 15-21% increased specific activity against carboxymethylcellulose and beta-glucan, and increased thermostability compared to wild-type enzyme EGLII
-
Y171C/L201C
-
site-directed mutagenesis, introduction of a disulfide bond into mutant DSB3, the mutant enzyme displays 15-21% increased specific activity against carboxymethylcellulose and beta-glucan, and increased thermostability compared to wild-type enzyme EGLII
-
S127C/A165C
-
site-directed mutagenesis, introduction of a disulfide bond into mutant DSB2, the mutant enzyme displays 15-21% increased specific activity against carboxymethylcellulose and beta-glucan, and increased thermostability compared to wild-type enzyme EGLII
-
Y171C/L201C
-
site-directed mutagenesis, introduction of a disulfide bond into mutant DSB3, the mutant enzyme displays 15-21% increased specific activity against carboxymethylcellulose and beta-glucan, and increased thermostability compared to wild-type enzyme EGLII
-
S127C/A165C
-
site-directed mutagenesis, introduction of a disulfide bond into mutant DSB2, the mutant enzyme displays 15-21% increased specific activity against carboxymethylcellulose and beta-glucan, and increased thermostability compared to wild-type enzyme EGLII
-
Y171C/L201C
-
site-directed mutagenesis, introduction of a disulfide bond into mutant DSB3, the mutant enzyme displays 15-21% increased specific activity against carboxymethylcellulose and beta-glucan, and increased thermostability compared to wild-type enzyme EGLII
-
D117A
-
activity with carboxymethyl cellulose is 0.03% of wild-type activity, activity with phosphoric acid-swollen cellulose is 0.02% of wild-type activity
D261A/R378K
-
Cel9A mutant, causes weaker binding to alpha-chitin than wild-type, mutation of residue near the catalytic center. Mutant has weak chitinase activity, but no soluble products are detected
D55A
activity with carboxymethyl cellulose is 0.2% of wild-type activity, activity with phosphoric acid-swollen cellulose is 1.2% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 1.6% of wild-type activity
D55A/D58A
activity with carboxymethyl cellulose is less than 0.1% of wild-type activity, activity with phosphoric acid-swollen cellulose is 0.13% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 0.5% of wild-type activity
D55N
activity with carboxymethyl cellulose is 0.3% of wild-type activity, activity with phosphoric acid-swollen cellulose is 1.6% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 2.2% of wild-type activity
D58A
activity with carboxymethyl cellulose is 0.4% of wild-type activity, activity with phosphoric acid-swollen cellulose is 1.8% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 3.5% of wild-type activity, mutant enzyme loses about 90% of the initial activity after 15 h at 65°C, compared to 10% loss of wild-type activity
D58N
activity with carboxymethyl cellulose is 0.45% of wild-type activity, activity with phosphoric acid-swollen cellulose is 2% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 2.7% of wild-type activity, mutant enzyme loses about 20% of the initial activity after 15 h at 65°C, compared to 10% loss of wild-type activity
DELTAT245-L251/R252K
activity with carboxymethyl cellulose, acid-swollen cellulose or bacterial microcrystalline cellulose from Acetobacter xylinum is nearly identical to wild-type activity
E424A
activity with carboxymethyl cellulose is 0.13% of wild-type activity, activity with phosphoric acid-swollen cellulose is 0.2% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 0.43% of wild-type activity, kcat/Km for 2,4-dinitrophenyl beta-D-cellobioside is 8.5fold higher than the wild-type value
E424G
activity with carboxymethyl cellulose is 0.3% of wild-type activity, activity with phosphoric acid-swollen cellulose is 1.1% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 2.5% of wild-type activity, kcat/Km for 2,4-dinitrophenyl beta-D-cellobioside is 123.8fold higher than the wild-type value
E424Q
activity with carboxymethyl cellulose is less than 0.1% of wild-type activity, activity with phosphoric acid-swollen cellulose is 0.15% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 1.1% of wild-type activity
G234S
-
Cel6B mutant, causes weaker binding to alpha-chitin than wild-type, mutation of residue near the catalytic center
R78A
-
activity with phosphoric acid-swollen cellulose is less than 1.3% of the wild-type activity, activity with carboxymethylcellulose is less than 0.9% of the wild-type activity, activity with bacterial microcrystalline cellulose is less than 18.7% of the wild-type activity
R78K
-
activity with phosphoric acid-swollen cellulose is 54% of the wild-type activity, activity with carboxymethylcellulose is 15% of the wild-type activity, activity with bacterial microcrystalline cellulose is 52% of the activity with wild-type enzyme
W329C
-
Cel6B mutant, causes weaker binding to alpha-chitin than wild-type, mutation of residue near the catalytic center
W332A
-
Cel6B mutant, causes weaker binding to alpha-chitin than wild-type, mutation of residue near the catalytic center
Y206F
Y206S
Y318A
activity with carboxymethyl cellulose is 5fold higher than wild-type activity, activity with phosphoric acid-swollen cellulose is 28% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 14.2% of wild-type activity, mutant enzyme loses about 30% of the initial activity after 15 h at 65°C, compared to 10% loss of wild-type activity
Y318F
activity with carboxymethyl cellulose is 6.7fold higher than wild-type activity, activity with phosphoric acid-swollen cellulose is 75% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 16.6% of wild-type activity
Y73F
-
activity with carboxymethyl cellulose is 8.4% of wild-type activity, activity with phosphoric acid-swollen cellulose is 5.7% of wild-type activity
Y73S
-
activity with carboxymethyl cellulose is 0.022% of wild-type activity, activity with phosphoric acid-swollen cellulose is 0.088% of wild-type activity
D55A
-
activity with carboxymethyl cellulose is 0.2% of wild-type activity, activity with phosphoric acid-swollen cellulose is 1.2% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 1.6% of wild-type activity
-
D55N
-
activity with carboxymethyl cellulose is 0.3% of wild-type activity, activity with phosphoric acid-swollen cellulose is 1.6% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 2.2% of wild-type activity
-
D58A
-
activity with carboxymethyl cellulose is 0.4% of wild-type activity, activity with phosphoric acid-swollen cellulose is 1.8% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 3.5% of wild-type activity, mutant enzyme loses about 90% of the initial activity after 15 h at 65°C, compared to 10% loss of wild-type activity
-
D58N
-
activity with carboxymethyl cellulose is 0.45% of wild-type activity, activity with phosphoric acid-swollen cellulose is 2% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 2.7% of wild-type activity, mutant enzyme loses about 20% of the initial activity after 15 h at 65°C, compared to 10% loss of wild-type activity
-
Y173F
Thermochaetoides thermophila
1.9fold increased the enzyme's specific activity. Mutation significantly improves the enzymes heat resistance at 80°C and 90°C
Y30F/Y173F
Thermochaetoides thermophila
mutant shows considerably higher stability at elevated temperatures but does not display the increased catalytic efficiency of its single mutant counterparts
A153V
mutant with increased activity on 1-ethyl-3-methylimidazolium acetate pretreated biomass. Mutant displays 130% of wild-type activity with carboxymethyl cellulose
E134C
site-directed mutagenesis, the catalytically inactive active-site mutant adopts a beta-jellyroll protein fold typical of the GH12-family enzymes, with two curved beta-sheets A and B and a central active-site cleft, crystal structure determination, overview
E225H/K207G
mutant based on homology modeling and rational design, display significantly improved activity and thermostability
E225H/K207G/D37V
mutant based on homology modeling and rational design, display significantly improved activity and thermostability
H138R
mutant with increased activity on 1-ethyl-3-methylimidazolium acetate pretreated biomass. Mutant displays 130% of wild-type activity with carboxymethyl cellulose
N236D
mutant with increased activity on 1-ethyl-3-methylimidazolium acetate pretreated biomass. Mutant displays 142% of wild-type activity with carboxymethyl cellulose
Y61G
about 170% of wild-type activity. Mutant also shows a wider range of working temperatures than does the wild type, along with retention of the hyperthermostability. The kcat and Km values of Y61G are both higher than those of the wild type. The higher endoglucanase activity is probably due to facile dissociation of the cleaved sugar moiety at the reducing end
Y66F
mutant with increased activity on 1-ethyl-3-methylimidazolium acetate pretreated biomass. Mutant displays 132% of wild-type activity with carboxymethyl cellulose
A153V
-
mutant with increased activity on 1-ethyl-3-methylimidazolium acetate pretreated biomass. Mutant displays 130% of wild-type activity with carboxymethyl cellulose
-
E225H/K207G
-
mutant based on homology modeling and rational design, display significantly improved activity and thermostability
-
E225H/K207G/D37V
-
mutant based on homology modeling and rational design, display significantly improved activity and thermostability
-
H138R
-
mutant with increased activity on 1-ethyl-3-methylimidazolium acetate pretreated biomass. Mutant displays 130% of wild-type activity with carboxymethyl cellulose
-
N236D
-
mutant with increased activity on 1-ethyl-3-methylimidazolium acetate pretreated biomass. Mutant displays 142% of wild-type activity with carboxymethyl cellulose
-
Y66F
-
mutant with increased activity on 1-ethyl-3-methylimidazolium acetate pretreated biomass. Mutant displays 132% of wild-type activity with carboxymethyl cellulose
-
G155C/G169C
introduction of disulfide bond for stabilization of the catalytic domain, melting temperature 54.9°C, decrease in activity with filter paper and avicel
G155C/N182C
introduction of disulfide bond for stabilization of the catalytic domain, melting temperature 56.5°C, decrease in activity with filter paper and avicel
G155C/N182C/N160C/G183C
introduction of disulfide bond for stabilization of the catalytic domain, melting temperature 56.9°C, decrease in activity with filter paper
G170C
Tm is 2.1°C higher than wild-type value, specific activity is 1.2fold higher than wild-type enzyme
G170C/P201C
Tm is 0.7°C higher than wild-type value, specific activity is 38% of wild-type value
G170C/P201C/V210C
Tm is 3.9°C higher than wild-type value, specific activity is 14% of wild-type value
G170C/V210C
specific activity is 1.4fold higher than wild-type value
G4C/E73C
introduction of disulfide bond for stabilization of the catalytic domain, melting temperature 56.9°C, decrease in activity with filter paper and avicel
G4C/F71C
introduction of disulfide bond for stabilization of the catalytic domain, melting temperature 56.9°C, decrease in activity with avicel
G4C/F71C/G155C/N182C
introduction of disulfide bond for stabilization of the catalytic domain, melting temperature 60.7°C, increase in activity with filter paper and avicel
G4C/F71C/G155C7N182C/N160C/G183C
introduction of disulfide bond for stabilization of the catalytic domain, melting temperature 60.6°C, decrease in activity with filter paper and avicel
G4C/F71C/N160C/G183C
introduction of disulfide bonds for stabilization of the catalytic domain, melting temperature 60.4°C, decrease in activity with filter paper and avicel
G4C/F71C/N160C/G183C/S168T
introduction of disulfide bond for stabilization of the catalytic domain, melting temperature 62.8°C, increase in activity with filter paper and avicel
G81C/V105C
introduction of disulfide bond for stabilization of the catalytic domain, melting temperature 55.9°C, decrease in activity with filter paper and avicel
K272F
mutation predicted to be thermostabilizing. Both high temperature and room temperature molecular dynamics simulations supported a stabilizing effect. Mutant exhibits higher thermostability compared with native EGI, but the specific activity of the mutant is lower
N160C/G183C
introduction of disulfide bond for stabilization of the catalytic domain, melting temperature 56.8°C, decrease in activity with filter paper and avicel
P201C
Q126F
mutation predicted to be thermostabilizing. Both high temperature and room temperature molecular dynamics simulations supported a stabilizing effect. Mutant exhibits higher thermostability compared with native EGI, but the specific activity of the mutant is lower
Q274V
mutation predicted to be thermostabilizing. Both high temperature and room temperature molecular dynamics simulations supported a stabilizing effect. Mutant exhibits higher thermostability compared with native EGI, but the specific activity of the mutant is lower
S213C/A296C
introduction of disulfide bond for stabilization of the catalytic domain, melting temperature 55.3°C, decrease in activity with filter paper and avicel
T57N/E53D/S79P/T80E/V101I/S133R/N155E/G189S/F191V/T233V/G239E/V265T/D271Y/G293A7S309W/S318P
thermostable mutant combining previously identified stabilizing mutations. Mutant has an optimal temperature 17°C higher than wild type and hydrolyzes 1.5 times as much cellulose over 60 h at its optimum temperature compared to the wild type enzyme at its optimal temperature
V210C
Y326C/G343C
introduction of disulfide bonds for stabilization of the catalytic domain, melting temperature 57.3°C, increase in activity with filter paper and decrease in activity with avicel
G155C/G169C
-
introduction of disulfide bond for stabilization of the catalytic domain, melting temperature 54.9°C, decrease in activity with filter paper and avicel
-
G155C/N182C
-
introduction of disulfide bond for stabilization of the catalytic domain, melting temperature 56.5°C, decrease in activity with filter paper and avicel
-
G4C/E73C
-
introduction of disulfide bond for stabilization of the catalytic domain, melting temperature 56.9°C, decrease in activity with filter paper and avicel
-
G4C/F71C
-
introduction of disulfide bond for stabilization of the catalytic domain, melting temperature 56.9°C, decrease in activity with avicel
-
G81C/V105C
-
introduction of disulfide bond for stabilization of the catalytic domain, melting temperature 55.9°C, decrease in activity with filter paper and avicel
-
T57N/E53D/S79P/T80E/V101I/S133R/N155E/G189S/F191V/T233V/G239E/V265T/D271Y/G293A7S309W/S318P
-
thermostable mutant combining previously identified stabilizing mutations. Mutant has an optimal temperature 17°C higher than wild type and hydrolyzes 1.5 times as much cellulose over 60 h at its optimum temperature compared to the wild type enzyme at its optimal temperature
-
D99A
106% of wild-type activity, temperature of the midpoint of the thermal denaturation transition is decreased by 1.9°C
Q40A/D99A
97% of wild-type activity, temperature of the midpoint of the thermal denaturation transition is decreased by 5.0°C
additional information
E289V
-
mutation identified by error-prone rolling circle amplification. Mutation results in a 7.93-fold increase in its enzymatic activity
E289V
-
mutation identified by error-prone rolling circle amplification. Mutation results in a 7.93-fold increase in its enzymatic activity
-
E201A
activity towards carboxymethyl cellulose is 0.01% of wild-type activity
E201A
site-directed mutagenesis, crystal structure determination with bound ligands
E342A
activity towards carboxymethyl cellulose is 0.08% of wild-type activity, no activity with p-nitrophenyl cellobiose
E342A
site-directed mutagenesis, crystal structure determination with bound ligands
W377A
activity towards carboxymethyl cellulose is 1.02% of wild-type activity
Y299F
activity towards carboxymethyl cellulose is 2.15% of wild-type activity. pH-optimum is 8.5, compared to 5.5-6 for wild-type enzyme
Y299F
site-directed mutagenesis, crystal structure determination with bound ligands, the mutant shows reduced activity compare to the wild-type enzyme, and a rare enzyme-substrate complex structure
Y206F
activity with carboxymethyl cellulose is 7% of wild-type activity, activity with phosphoric acid-swollen cellulose is 4.1% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 8.3% of wild-type activity, mutant enzyme loses about 20% of the initial activity after 15 h at 65°C, compared to 10% loss of wild-type activity
Y206F
-
Cel9A mutant, causes weaker binding to alpha-chitin than wild-type, mutation of residue near the catalytic center
Y206S
activity with carboxymethyl cellulose is 0.5% of wild-type activity, activity with phosphoric acid-swollen cellulose is 1.3% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 1.8% of wild-type activity
Y206S
-
Cel9A mutant, causes weaker binding to alpha-chitin than wild-type, mutation of residue near the catalytic center
P201C
Tm is 0.7°C higher than wild-type value, specific activity is 50% of wild-type value
P201C
Tm is 3.9°C higher than wild-type value, specific activity is 80% of wild-type value
V210C
Tm is 0.1°C higher than wild-type value, specific activity is 1.8fold of the wild-type value
V210C
Tm is identical to wild-type value, specific activity is 80% of wild-type value
additional information
-
construction of a His6-tagged truncated enzyme mutant CtLic26A-Cel5
additional information
-
replacement of carbohydrate-binding module either with a family 3 microcrystalline cellulose-directed carbohydrate-binding module from Clostridium josui scaffoldin, or a family 6 xylan-directed carbohydrate-binding module from Clostridium stercorarium xylanase 11A. Chimeric endoglucanases show enhanced activity that is affected by carbohydrate-binding module binding specificity. The chimeric enzymes can efficiently degrade milled lignocellulosic materials, such as corn hulls
additional information
construction of a C-terminally truncated enzyme, CtCel9QDELTAc
additional information
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construction of a C-terminally truncated enzyme, CtCel9QDELTAc
additional information
-
enzyme mutant CtGH5-F194A is fused with a beta-1,4-glucosidase, CtGH1 from Clostridium thermocellum to develop a chimeric enzyme. Improved structural integrity, thermostability and enhanced bifunctional enzyme activities of the chimeric mutant compared to the single point mutant
additional information
-
replacement of carbohydrate-binding module either with a family 3 microcrystalline cellulose-directed carbohydrate-binding module from Clostridium josui scaffoldin, or a family 6 xylan-directed carbohydrate-binding module from Clostridium stercorarium xylanase 11A. Chimeric endoglucanases show enhanced activity that is affected by carbohydrate-binding module binding specificity. The chimeric enzymes can efficiently degrade milled lignocellulosic materials, such as corn hulls
-
additional information
deletion of the Ig-like domain in the N-terminus of the catalytic domain increases the catalytic efficiency of the truncated enzyme up to 3fold without any significant changes in the Km of the enzyme and shifts pH and temperature optimum for activity from 6.5 to 7.5 and from 65 to 60°C, respectively
additional information
creation of a hybrid enzyme of GH5 endoglucanase AnCel5A from Aspergillus niger with elements of the mesophilic endoglucanase Cel5 from Stegonsporium opalus (SoCel5). The expressed hybrid enzyme exhibits increased enzyme activity relative to the values for the mesophilic parent enzyme. The mutant demonstrates improved catalytic efficiency on selected substrates. Method validation, overview
additional information
-
improvement of endoglucanase activity by utilizing error-prone rolling circle amplification, supplemented with 1.7 mM MnCl2. The procedure generates random mutations in the Bacillus amyloliquefaciens endoglucanase gene with a frequency of 10 mutations per kilobase
additional information
-
improvement of endoglucanase activity by utilizing error-prone rolling circle amplification, supplemented with 1.7 mM MnCl2. The procedure generates random mutations in the Bacillus amyloliquefaciens endoglucanase gene with a frequency of 10 mutations per kilobase
-
additional information
-
construction of chimeric enzymes between the Bacillus subtilis cellulase and an alkalophilic Bacillus cellulase
additional information
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C-terminus truncation mutant Egl330, truncation of the cellulose binding domain, a great improvement in thermal stability is observed in Egl330
additional information
-
variants (M44-11, S75 and S78) show 2.03fold to 2.68fold increased activities toward sodium carboxymethyl cellulose
additional information
generation of a set of chimeric proteins derived from the recombination of Geobacillus sp. CelA and Bacillus subtilis 168 Cel5A. The designed chimeras are assembled from 16 gene fragments of the two parents. Chimeric cellulase C10 shows significantly higher activity (22%-43%) and higher thermostability compared to the parental enzymes. A 310 helix is responsible for the improved thermostability. In the presence of ionic calcium and crown ether, the chimeric C10 retains 40% residual activity even after heat treatment at 90°C
additional information
cellulolytic activity of Paenibacillus sp. strain CAA11 is significantly enhanced by expressing a heterologous endoglucanase 168Cel5 from Bacillus subtilis under both aerobic and anaerobic conditions. The strain harboring the 168cel5 gene reveals 2fold bigger halo zone on Congo-red plate and 1.75fold more aerobic cellulose utilization in liquid medium compared with the negative control. Under anaerobic conditions, the recombinant strain expressing 168Cel5 consumes 1.83fold more cellulose (5.10 g/l) and produces 5fold more ethanol (0.65 g/l) along with 5fold more total acids (1.6 g/l) compared with the control, resulting in 2.73fold higher yields
additional information
-
cellulolytic activity of Paenibacillus sp. strain CAA11 is significantly enhanced by expressing a heterologous endoglucanase 168Cel5 from Bacillus subtilis under both aerobic and anaerobic conditions. The strain harboring the 168cel5 gene reveals 2fold bigger halo zone on Congo-red plate and 1.75fold more aerobic cellulose utilization in liquid medium compared with the negative control. Under anaerobic conditions, the recombinant strain expressing 168Cel5 consumes 1.83fold more cellulose (5.10 g/l) and produces 5fold more ethanol (0.65 g/l) along with 5fold more total acids (1.6 g/l) compared with the control, resulting in 2.73fold higher yields
additional information
-
generation of a set of chimeric proteins derived from the recombination of Geobacillus sp. CelA and Bacillus subtilis 168 Cel5A. The designed chimeras are assembled from 16 gene fragments of the two parents. Chimeric cellulase C10 shows significantly higher activity (22%-43%) and higher thermostability compared to the parental enzymes. A 310 helix is responsible for the improved thermostability. In the presence of ionic calcium and crown ether, the chimeric C10 retains 40% residual activity even after heat treatment at 90°C
-
additional information
-
cellulolytic activity of Paenibacillus sp. strain CAA11 is significantly enhanced by expressing a heterologous endoglucanase 168Cel5 from Bacillus subtilis under both aerobic and anaerobic conditions. The strain harboring the 168cel5 gene reveals 2fold bigger halo zone on Congo-red plate and 1.75fold more aerobic cellulose utilization in liquid medium compared with the negative control. Under anaerobic conditions, the recombinant strain expressing 168Cel5 consumes 1.83fold more cellulose (5.10 g/l) and produces 5fold more ethanol (0.65 g/l) along with 5fold more total acids (1.6 g/l) compared with the control, resulting in 2.73fold higher yields
-
additional information
-
variants (M44-11, S75 and S78) show 2.03fold to 2.68fold increased activities toward sodium carboxymethyl cellulose
-
additional information
-
C-terminus truncation mutant Egl330, truncation of the cellulose binding domain, a great improvement in thermal stability is observed in Egl330
-
additional information
construction of deletion mutants expressing solely the carboxyterminal domain containing the endoglucanase activity. Temperature optimum and stability of the deletion mutants are the same as wild-type
additional information
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construction of a chimeric xylanase/endoglucanase with an internal cellulase-binding domain by fusing the Bacillus subtilis xyn gene fragment to the 5'-end of the Cellulomonas fimi cenA. The hybrid protein behaves like the parental endoglucanase or xylanase when assayed on a number of soluble and insoluble substrates or xylans
additional information
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engineering of cellulase A from Cellulomonas fimi as a model to replace residues that potentially influence the pH-activity profile of the enzyme based on sequence alignments and analysis of the known three-dimensional structures of other CAZy family 6 glycoside hydrolases with the aim to lower its pH optimum, overview
additional information
EngHDELTACBM devoid of the carbohydrate binding module loses activity toward all substrates including carboxymethylcellulose
additional information
EngHDELTACBM devoid of the carbohydrate binding module loses activity toward all substrates including carboxymethylcellulose
additional information
EngHDELTACBM devoid of the carbohydrate binding module loses activity toward all substrates including carboxymethylcellulose
additional information
EngHDELTACBM devoid of the carbohydrate binding module loses activity toward all substrates including carboxymethylcellulose
additional information
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EngHDELTACBM devoid of the carbohydrate binding module loses activity toward all substrates including carboxymethylcellulose
additional information
EngMDELTSACBM, devoid of the carbohydrate binding module has activity toward CMC (0.003 U/mg), in contrast with no activity toward any other substrate
additional information
EngMDELTSACBM, devoid of the carbohydrate binding module has activity toward CMC (0.003 U/mg), in contrast with no activity toward any other substrate
additional information
EngMDELTSACBM, devoid of the carbohydrate binding module has activity toward CMC (0.003 U/mg), in contrast with no activity toward any other substrate
additional information
EngMDELTSACBM, devoid of the carbohydrate binding module has activity toward CMC (0.003 U/mg), in contrast with no activity toward any other substrate
additional information
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EngMDELTSACBM, devoid of the carbohydrate binding module has activity toward CMC (0.003 U/mg), in contrast with no activity toward any other substrate
additional information
EngYDELTSACBM, devoid of the carbohydrate binding module loses activity toward all substrates including carboxymethylcellulose
additional information
EngYDELTSACBM, devoid of the carbohydrate binding module loses activity toward all substrates including carboxymethylcellulose
additional information
EngYDELTSACBM, devoid of the carbohydrate binding module loses activity toward all substrates including carboxymethylcellulose
additional information
EngYDELTSACBM, devoid of the carbohydrate binding module loses activity toward all substrates including carboxymethylcellulose
additional information
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EngYDELTSACBM, devoid of the carbohydrate binding module loses activity toward all substrates including carboxymethylcellulose
additional information
C-terminal His-tagged form (tCfEG) shows hydrolytic activity on cellulosic substrates
additional information
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C-terminal His-tagged form (tCfEG) shows hydrolytic activity on cellulosic substrates
additional information
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fusion of the cellulose binding domain of cellobiohydrolase I from Trichoderma reesei to the C-terminus of Cryptococcus sp. carboxymethyl cellulase and expression in a recombinant expression system of Cryptococcus sp. S-2. The recombinant fusion enzymes display optimal pH similar to those of the native enzyme. Compared with Cryptococcus sp. carboxymethyl cellulase, the recombinant fusion enzymes have acquired an increased binding affinity to insoluble cellulose, and the cellulolytic activity toward insoluble cellulosic substrates, SIGMACELL and Avicel, is higher than that of native enzyme, confirming the presence of cellulose binding domains improve the binding and the cellulolytic activity of carboxymethyl cellulase on insoluble substrates
additional information
generation of a set of chimeric proteins derived from the recombination of Geobacillus sp. CelA and Bacillus subtilis 168 Cel5A. The designed chimeras are assembled from 16 gene fragments of the two parents. Chimeric cellulase C10 shows significantly higher activity (22%-43%) and higher thermostability compared to the parental enzymes. A 310 helix is responsible for the improved thermostability. In the presence of ionic calcium and crown ether, the chimeric C10 retains 40% residual activity even after heat treatment at 90°C
additional information
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generation of a set of chimeric proteins derived from the recombination of Geobacillus sp. CelA and Bacillus subtilis 168 Cel5A. The designed chimeras are assembled from 16 gene fragments of the two parents. Chimeric cellulase C10 shows significantly higher activity (22%-43%) and higher thermostability compared to the parental enzymes. A 310 helix is responsible for the improved thermostability. In the presence of ionic calcium and crown ether, the chimeric C10 retains 40% residual activity even after heat treatment at 90°C
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gene celF knockout prevents growth on cellulose although the mutant strain grows perfectly well on glucose, but does not affect other cellulase genes expressions, overview
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cellulolytic activity of Paenibacillus sp. strain CAA11 is significantly enhanced by expressing a heterologous endoglucanase 168Cel5 from Bacillus subtilis (DNA sequences encoding P43 promoter, signal peptide of Bacillus subtilis nprB, and mature Bacillus subtilis strain 168 Cel5 residues 30-499) under both aerobic and anaerobic conditions. The strain harboring the 168cel5 gene reveals 2fold bigger halo zone on Congo-red plate and 1.75fold more aerobic cellulose utilization in liquid medium compared with the negative control. Under anaerobic conditions, the recombinant strain expressing 168Cel5 consumes 1.83fold more cellulose (5.10 g/l) and produces 5fold more ethanol (0.65 g/l) along with 5fold more total acids (1.6 g/l) compared with the control, resulting in 2.73fold higher yields. Optimal growth of the engineered strain at 37°C, evaluation of optimal growth conditions, overview
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both the full-length enzyme and the catalytic domain have carboxymethylcellulase and filter paper hydrolase activity . The catalytic domain can also bind the cellulose substrate. The aromatic amino acids at the bottom of the barrel fold and those adjacent to the catalytic site significantly affect the cellulolytic activity and the cellulose binding affinity of the catalytic domain
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both the full-length enzyme and the catalytic domain have carboxymethylcellulase and filter paper hydrolase activity . The catalytic domain can also bind the cellulose substrate. The aromatic amino acids at the bottom of the barrel fold and those adjacent to the catalytic site significantly affect the cellulolytic activity and the cellulose binding affinity of the catalytic domain
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amino acids, E209 and E319, act as proton donor and nucleophile in the catalytic domain
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amino acids, E209 and E319, act as proton donor and nucleophile in the catalytic domain
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PopCel1 mRNA is accumulated in seven transgenic lines of Paraserianthes falcataria
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hybrid cellulase with the Thermomonospora fusca E2 cellulose-binding domain at its C terminus joined to the Prevotella ruminicola 40500 Da carboxymethylcellulase
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a truncated EGPf (EGPfDELTAN30) mutant lacking the proline and hydroxyl-residue rich region at the N-terminus is constructed
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a truncated EGPf (EGPfDELTAN30) mutant lacking the proline and hydroxyl-residue rich region at the N-terminus is constructed
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mutant enzyme lacking 5 residues at the C-terminus: hydrolytic activity towards carboxymethyl cellulose is 112% of wild-type activity, kcat/Km for p-nitrophenyl cellobiose is 1.2fold higher than wild-type value. Mutant enzyme lacking 5 residues at the C-terminus and 5 residues at the N-terminus: activity towards carboxymethyl cellulose is 111% of wild-type activity, kcat/Km for p-nitrophenyl cellobiose is 1.8fold higher than wild-type value. Thermostability is not significantly influenced
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mutant enzyme lacking 5 residues at the C-terminus: hydrolytic activity towards carboxymethyl cellulose is 112% of wild-type activity, kcat/Km for p-nitrophenyl cellobiose is 1.2fold higher than wild-type value. Mutant enzyme lacking 5 residues at the C-terminus and 5 residues at the N-terminus: activity towards carboxymethyl cellulose is 111% of wild-type activity, kcat/Km for p-nitrophenyl cellobiose is 1.8fold higher than wild-type value. Thermostability is not significantly influenced
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preparation of a fusion enzyme so that the thermostable chitin-binding domain of chitinase from Pyrococcus furiosus is joined to the C-terminus of EGPh and its variants. The fusion enzymes show stronger activities than the wild-type EGPh toward both carboxymethyl cellulose and crystalline cellulose (Avicel)
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preparation of a fusion enzyme so that the thermostable chitin-binding domain of chitinase from Pyrococcus furiosus is joined to the C-terminus of EGPh and its variants. The fusion enzymes show stronger activities than the wild-type EGPh toward both carboxymethyl cellulose and crystalline cellulose (Avicel)
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EGPhDELTAC5, truncated form of the enzyme shows similar enzymatic properties to the wild-type protein. EGPhDELTAC10, lacking ten residues at the C-terminus showed significantly decreased activity. Three truncated mutants (EGPhDELTAN5, EGPhDELTAC5 and EGPhDELTAN5C5) which show no change in enzymatic properties are prepared and screened for crystallization
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sequential deletion analyses from both N and C termini, removing 10 amino acids at a time, are carried out to determine whether a shorter enzyme with improved characteristics could becreated. Among the three C-terminal deletions, only the C10 mutant, which misses the last 10 amino acids, maintained activity. In contrast, all N-terminal deletion mutants (N10, N20, and N30) retains activity except N40. Detailed analysis of the aligned sequences reveals that the highly conserved sequence begins at L35, after the 34 N-terminal residues of EGPh. Therefore, a mutant with a deletion betweenN30 and N40 (N34) is prepared and expressed. This mutant retains enzymatic activity like N30, suggesting that the critical residues for enzyme activity start from L35. Each of the active N-terminal deletions is combined with the C10 deletion to establish the minimal sequence required for activity. When enzyme function is tested in the presence of CMC, only the N10C10 mutant exhibits activity, suggesting that the loss of activity may be due to the loss of thermostability. To test this, enzymatic activity assays are performed at 60°C. At this lower temperature, the N20C10, N30C10, and N34C10 mutants all exhibit carboxymethyl cellulase (CMCase) activity, but the N40, C20, and C30 mutants do not. Therefore, the shortest EGPh sequence maintaining hydrolytic activity isN34C10, representing an 11% reduction in amino acid residues. In addition to the decreased optimal temperature of C mutants,N and C combination mutants (except N10C10) are active at 60°C but not at 80°C. At 80°C, the wild type and the N10 mutant are stable, whereas N20, N30, and N34 show gradually decreasing activity. A longer deletion leads to a more severe decrease in activity, and theN34 mutant exhibits the shortest t1/2 of 8 h. Both C10 andN10C10 lose more than 50% activity in less than 2 h at 80°C, suggesting that the decreased activity of C-terminal deletion mutants is due to decreased thermostability
additional information
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sequential deletion analyses from both N and C termini, removing 10 amino acids at a time, are carried out to determine whether a shorter enzyme with improved characteristics could becreated. Among the three C-terminal deletions, only the C10 mutant, which misses the last 10 amino acids, maintained activity. In contrast, all N-terminal deletion mutants (N10, N20, and N30) retains activity except N40. Detailed analysis of the aligned sequences reveals that the highly conserved sequence begins at L35, after the 34 N-terminal residues of EGPh. Therefore, a mutant with a deletion betweenN30 and N40 (N34) is prepared and expressed. This mutant retains enzymatic activity like N30, suggesting that the critical residues for enzyme activity start from L35. Each of the active N-terminal deletions is combined with the C10 deletion to establish the minimal sequence required for activity. When enzyme function is tested in the presence of CMC, only the N10C10 mutant exhibits activity, suggesting that the loss of activity may be due to the loss of thermostability. To test this, enzymatic activity assays are performed at 60°C. At this lower temperature, the N20C10, N30C10, and N34C10 mutants all exhibit carboxymethyl cellulase (CMCase) activity, but the N40, C20, and C30 mutants do not. Therefore, the shortest EGPh sequence maintaining hydrolytic activity isN34C10, representing an 11% reduction in amino acid residues. In addition to the decreased optimal temperature of C mutants,N and C combination mutants (except N10C10) are active at 60°C but not at 80°C. At 80°C, the wild type and the N10 mutant are stable, whereas N20, N30, and N34 show gradually decreasing activity. A longer deletion leads to a more severe decrease in activity, and theN34 mutant exhibits the shortest t1/2 of 8 h. Both C10 andN10C10 lose more than 50% activity in less than 2 h at 80°C, suggesting that the decreased activity of C-terminal deletion mutants is due to decreased thermostability
additional information
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sequential deletion analyses from both N and C termini, removing 10 amino acids at a time, are carried out to determine whether a shorter enzyme with improved characteristics could becreated. Among the three C-terminal deletions, only the C10 mutant, which misses the last 10 amino acids, maintained activity. In contrast, all N-terminal deletion mutants (N10, N20, and N30) retains activity except N40. Detailed analysis of the aligned sequences reveals that the highly conserved sequence begins at L35, after the 34 N-terminal residues of EGPh. Therefore, a mutant with a deletion betweenN30 and N40 (N34) is prepared and expressed. This mutant retains enzymatic activity like N30, suggesting that the critical residues for enzyme activity start from L35. Each of the active N-terminal deletions is combined with the C10 deletion to establish the minimal sequence required for activity. When enzyme function is tested in the presence of CMC, only the N10C10 mutant exhibits activity, suggesting that the loss of activity may be due to the loss of thermostability. To test this, enzymatic activity assays are performed at 60°C. At this lower temperature, the N20C10, N30C10, and N34C10 mutants all exhibit carboxymethyl cellulase (CMCase) activity, but the N40, C20, and C30 mutants do not. Therefore, the shortest EGPh sequence maintaining hydrolytic activity isN34C10, representing an 11% reduction in amino acid residues. In addition to the decreased optimal temperature of C mutants,N and C combination mutants (except N10C10) are active at 60°C but not at 80°C. At 80°C, the wild type and the N10 mutant are stable, whereas N20, N30, and N34 show gradually decreasing activity. A longer deletion leads to a more severe decrease in activity, and theN34 mutant exhibits the shortest t1/2 of 8 h. Both C10 andN10C10 lose more than 50% activity in less than 2 h at 80°C, suggesting that the decreased activity of C-terminal deletion mutants is due to decreased thermostability
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post-translational modification of the N-terminal glutamine residue to diglutamate via glutaminyl cyclase enhances the stability of Cel7A and variants
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post-translational modification of the N-terminal glutamine residue to diglutamate via glutaminyl cyclase enhances the stability of Cel7A and variants
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creation of 10 hybrid enzymes of GH5 endoglucanase Egl5A from Talaromyces emersonii (TeEgl5A) with elements of the mesophilic endoglucanase Cel5 from Stegonsporium opalus (SoCel5). Five of the expressed hybrid enzymes exhibit enzyme activity. Two of these hybrids exhibit pronounced increases in the temperature optimum (10 and 20°C), the temperature T50 at which the protein loses 50% of its activity (15 and 19°C), and the melting temperature Tm (16.5 and 22.9°C) and extended half-lives (240fold and 650fold at 55°C) relative to the values for the mesophilic parent enzyme. The mutants demonstrate improved catalytic efficiency on selected substrates. Method validation, molecular dynamics simulations of both the SoCel5 and TeEgl5A parent enzymes, overview. Improved hydrophobic packing of the interface between alpha2 and alpha3 is the primary mechanism by which the hybrid enzymes increase their thermostability relative to that of the mesophilic parent SoCel5. Comparison of structures and molecular masses of the SoCel5-TeEgl5A hybrid enzymes. Mechanism of improved thermostability
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creation of 10 hybrid enzymes of GH5 endoglucanase Egl5A from Talaromyces emersonii (TeEgl5A) with elements of the mesophilic endoglucanase Cel5 from Stegonsporium opalus (SoCel5). Five of the expressed hybrid enzymes exhibit enzyme activity. Two of these hybrids exhibit pronounced increases in the temperature optimum (10 and 20°C), the temperature T50 at which the protein loses 50% of its activity (15 and 19°C), and the melting temperature Tm (16.5 and 22.9°C) and extended half-lives (240fold and 650fold at 55°C) relative to the values for the mesophilic parent enzyme. The mutants demonstrate improved catalytic efficiency on selected substrates. Method validation, molecular dynamics simulations of both the SoCel5 and TeEgl5A parent enzymes, overview. Improved hydrophobic packing of the interface between alpha2 and alpha3 is the primary mechanism by which the hybrid enzymes increase their thermostability relative to that of the mesophilic parent SoCel5. Comparison of structures and molecular masses of the SoCel5-TeEgl5A hybrid enzymes. Mechanism of improved thermostability
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symbiotic phenotype of an ANU843 celC2 knockout mutant derivative strain ANU843DELTAcelC2
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symbiotic phenotype of an ANU843 celC2 knockout mutant derivative strain ANU843DELTAcelC2
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symbiotic phenotype of an ANU843 celC2 knockout mutant derivative strain ANU843DELTAcelC2
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after bioinformatic analysis hybrid proteins based on beta-endoglucanase from Sulfolobus solfataricus P2 and beta-endoglucanase from Thermotoga maritima are constructed which should successfully combine the advantageous properties of both cellulases, i.e. recombinant expression in Escherichia coli, acidophily and thermophily. both hybrids are expressed insoluble in Escherichia coli, but one hybrid enzyme was successfully refolded from washed inclusion bodies
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targeted construction of chimeric enzymes of cellulase from Sulfolobus solfataricus P2 and cellulase CelA from Thermotoga maritima. The fusion protein SSO1949-CelA-SSO1949 consists of 193 amino acids SSO1949 followed by 76 amino acids CelA and 39 amino acids SSO1949. The catalytic center with the two catalytic glutamate residues is derived from SSO1949 whereas the reducing end of the substrate binding cleft comes from CelA. The fusion protein CelA-SSO1949-CelA consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA. Most of the active center is derived from SSO1949
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after bioinformatic analysis hybrid proteins based on beta-endoglucanase from Sulfolobus solfataricus P2 and beta-endoglucanase from Thermotoga maritima are constructed which should successfully combine the advantageous properties of both cellulases, i.e. recombinant expression in Escherichia coli, acidophily and thermophily. both hybrids are expressed insoluble in Escherichia coli, but one hybrid enzyme was successfully refolded from washed inclusion bodies
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additional information
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targeted construction of chimeric enzymes of cellulase from Sulfolobus solfataricus P2 and cellulase CelA from Thermotoga maritima. The fusion protein SSO1949-CelA-SSO1949 consists of 193 amino acids SSO1949 followed by 76 amino acids CelA and 39 amino acids SSO1949. The catalytic center with the two catalytic glutamate residues is derived from SSO1949 whereas the reducing end of the substrate binding cleft comes from CelA. The fusion protein CelA-SSO1949-CelA consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA. Most of the active center is derived from SSO1949
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post-transcriptional silencing of cel7 and cel9C1 with effects on nematode growth and development
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creation of 10 hybrid enzymes of GH5 endoglucanase Egl5A from Talaromyces emersonii (TeEgl5A) with elements of the mesophilic endoglucanase Cel5 from Stegonsporium opalus (SoCel5). Five of the expressed hybrid enzymes exhibit enzyme activity. Two of these hybrids exhibit pronounced increases in the temperature optimum (10 and 20°C), the temperature T50 at which the protein loses 50% of its activity (15 and 19°C), and the melting temperature Tm (16.5 and 22.9°C) and extended half-lives (240fold and 650fold at 55°C) relative to the values for the mesophilic parent enzyme. The mutants demonstrate improved catalytic efficiency on selected substrates. Method validation, molecular dynamics simulations of both the SoCel5 and TeEgl5A parent enzymes, overview. Improved hydrophobic packing of the interface between alpha2 and alpha3 is the primary mechanism by which the hybrid enzymes increase their thermostability relative to that of the mesophilic parent SoCel5. Comparison of structures and molecular masses of the SoCel5-TeEgl5A hybrid enzymes. Mechanism of improved thermostability
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creation of 10 hybrid enzymes of GH5 endoglucanase Egl5A from Talaromyces emersonii (TeEgl5A) with elements of the mesophilic endoglucanase Cel5 from Stegonsporium opalus (SoCel5). Five of the expressed hybrid enzymes exhibit enzyme activity. Two of these hybrids exhibit pronounced increases in the temperature optimum (10 and 20°C), the temperature T50 at which the protein loses 50% of its activity (15 and 19°C), and the melting temperature Tm (16.5 and 22.9°C) and extended half-lives (240fold and 650fold at 55°C) relative to the values for the mesophilic parent enzyme. The mutants demonstrate improved catalytic efficiency on selected substrates. Method validation, molecular dynamics simulations of both the SoCel5 and TeEgl5A parent enzymes, overview. Improved hydrophobic packing of the interface between alpha2 and alpha3 is the primary mechanism by which the hybrid enzymes increase their thermostability relative to that of the mesophilic parent SoCel5. Comparison of structures and molecular masses of the SoCel5-TeEgl5A hybrid enzymes. Mechanism of improved thermostability
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development of biocatalysts for enhanced hydrolysis of (hemi)cellulose into monosaccharides with random diversity by directed evolution of the gene coding for bacterial endo-beta-1,4-glucanase from Streptomyces sp. G12, improved catalysis of lignocellulose conversion, screening of a library of 10000 random mutants, detection of variants with higher activity than the wild-type enzyme, and structure-function relationships of the mutants, overview. Mutations T67N, D142E, T157I, and S218N are located in the catalytic module and mutations G251D, V259D, V242D, and D330E in the carbohydrate binding module (CBM)
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molecular dynamics simulations reveal that the introduced disulfide bond rigidified a global structure of DSB2 and DSB3 mutant variants, thus enhancing their thermostability
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molecular dynamics simulations reveal that the introduced disulfide bond rigidified a global structure of DSB2 and DSB3 mutant variants, thus enhancing their thermostability
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molecular dynamics simulations reveal that the introduced disulfide bond rigidified a global structure of DSB2 and DSB3 mutant variants, thus enhancing their thermostability
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molecular dynamics simulations reveal that the introduced disulfide bond rigidified a global structure of DSB2 and DSB3 mutant variants, thus enhancing their thermostability
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molecular dynamics simulations reveal that the introduced disulfide bond rigidified a global structure of DSB2 and DSB3 mutant variants, thus enhancing their thermostability
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additional information
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molecular dynamics simulations reveal that the introduced disulfide bond rigidified a global structure of DSB2 and DSB3 mutant variants, thus enhancing their thermostability
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molecular dynamics simulations reveal that the introduced disulfide bond rigidified a global structure of DSB2 and DSB3 mutant variants, thus enhancing their thermostability
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fusion enzyme (EG-M-Xyn) of endoglucanase (cellulase) from Teleogryllus emma and xylanase from Thermomyces lanuginosus
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the FnIII-like domain is deleted from Cel19A-90, reducing activity with bacterial microcrystalline cellulose to 43% of the wild-type
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the FnIII-like domain is deleted from Cel19A-90, reducing activity with bacterial microcrystalline cellulose to 43% of the wild-type
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the FnIII-like domain is deleted from Cel19A-90, reducing activity with bacterial microcrystalline cellulose to 43% of the wild-type
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exo-endo-1,4-beta-glucanase fusion protein
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targeted construction of chimeric enzymes of cellulase from Sulfolobus solfataricus P2 and cellulase CelA from Thermotoga maritima. The fusion protein SSO1949-CelA-SSO1949 consists of 193 amino acids SSO1949 followed by 76 amino acids CelA and 39 amino acids SSO1949. The catalytic center with the two catalytic glutamate residues is derived from SSO1949 whereas the reducing end of the substrate binding cleft comes from CelA. The fusion protein CelA-SSO1949-CelA consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA. Most of the active center is derived from SSO1949
additional information
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targeted construction of chimeric enzymes of cellulase from Sulfolobus solfataricus P2 and cellulase CelA from Thermotoga maritima. The fusion protein SSO1949-CelA-SSO1949 consists of 193 amino acids SSO1949 followed by 76 amino acids CelA and 39 amino acids SSO1949. The catalytic center with the two catalytic glutamate residues is derived from SSO1949 whereas the reducing end of the substrate binding cleft comes from CelA. The fusion protein CelA-SSO1949-CelA consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA. Most of the active center is derived from SSO1949
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engineering of mutants of isoform Cel5A with higher activity on 1-ethyl-3-methylimidazolium acetate pretreated biomass by screening of a random mutagenesis library. Twelve mutants with 2542% improvement in specific activity on carboxymethyl cellulose and up to 30% improvement on ionic-liquid pretreated switchgrass could be isolated and characterized. Mmost of the mutations in the improved variants are located distally to the active site on the protein surface and are not directly involved with substrate binding
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engineering of mutants of isoform Cel5A with higher activity on 1-ethyl-3-methylimidazolium acetate pretreated biomass by screening of a random mutagenesis library. Twelve mutants with 2542% improvement in specific activity on carboxymethyl cellulose and up to 30% improvement on ionic-liquid pretreated switchgrass could be isolated and characterized. Mmost of the mutations in the improved variants are located distally to the active site on the protein surface and are not directly involved with substrate binding
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engineering of mutants of isoform Cel5A with higher activity on 1-ethyl-3-methylimidazolium acetate pretreated biomass by screening of a random mutagenesis library. Twelve mutants with 2542% improvement in specific activity on carboxymethyl cellulose and up to 30% improvement on ionic-liquid pretreated switchgrass could be isolated and characterized. Mmost of the mutations in the improved variants are located distally to the active site on the protein surface and are not directly involved with substrate binding
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endoglucanase IV is reconstructed by fusing EGIV with an additional catalytic module (EGIVCM). The genes eg4 and eg4-cm are expressed in recombinant Pichia strains (Pichia pastoris EGIV1 and Pichia pastoris EGIV-CM1). The activities towards carboxymethyl cellulose of cultivation supernatant of Pichia pastoris EGIV1 and Pichia pastoris EGIV-CM1 are 2.4 U/ml and 4.3 U/ml, respectively. Modification of the EGIV structure with an additional catalytic module improves the specific activity about 4fold
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construction of disruption mutants of genes cbh1 and cbh2, encoding the 1,4-beta-D-glucan cellobiohydrolases, does not affect egl1 and egl3 expression, overview
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enzyme immobilization, e.g. by cross-linking with glutaraldehyde, enzyme activity and method evaluation
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usage of gene egl3 promoter for expression of gene bgl1, encoding a beta-glucosidase, in Trichoderma reesei, the mutant strain shows 4fold increased beta-glucosidase activity compared to the wild-type enzyme
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fusion of the cellulose binding domain of cellobiohydrolase I from Trichoderma reesei to the C-terminus of Cryptococcus sp. carboxymethyl cellulase and expression in a recombinant expression system of Cryptococcus sp. S-2. The recombinant fusion enzymes display optimal pH similar to those of the native enzyme. Compared with Cryptococcus sp. carboxymethyl cellulase, the recombinant fusion enzymes have acquired an increased binding affinity to insoluble cellulose, and the cellulolytic activity toward insoluble cellulosic substrates, SIGMACELL and Avicel, is higher than that of native enzyme, confirming the presence of cellulose binding domains improve the binding and the cellulolytic activity of carboxymethyl cellulase on insoluble substrates
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elimination of all of the glycosylation sites induces expression of the unfolded protein response target genes, and secretion of this CBH1 variant is severely compromised in a calnexin gene deletion strain. The thermal reactivity of CBH1 is significantly decreased by removal of either Asn45 or Asn384 glycosylation site during the catalyzed hydrolysis of soluble substrate. Combinatorial loss of these two N-linked glycans further exacerbates the temperature-dependent inactivation. Removal of N-glycosylation at Asn384 has a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270
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elimination of all of the glycosylation sites induces expression of the unfolded protein response target genes, and secretion of this CBH1 variant is severely compromised in a calnexin gene deletion strain. The thermal reactivity of CBH1 is significantly decreased by removal of either Asn45 or Asn384 glycosylation site during the catalyzed hydrolysis of soluble substrate. Combinatorial loss of these two N-linked glycans further exacerbates the temperature-dependent inactivation. Removal of N-glycosylation at Asn384 has a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270
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usage of gene egl3 promoter for expression of gene bgl1, encoding a beta-glucosidase, in Trichoderma reesei, the mutant strain shows 4fold increased beta-glucosidase activity compared to the wild-type enzyme
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a mutant lacking the immunoglobulin-like domain shows 1% of wild-type activity and a decrease in the temperature of the midpoint of the thermal denaturation transition by 6.3°C
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construction of mutant mgCel6ADELTACBM lacking the carbohydrate binding module 2 (CBM2)
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endoglucanase 1 (EG1) and its catalytic module (EG1-CM) exhibit very similar specific activities towards the soluble substrates carboxymethyl cellulose, lichenan and mannan, and insoluble H3PO4 acid-swollen cellulose, whereas the specific activities of EG1-CM towards the insoluble substrates alpha-cellulose, Avicel and filter paper are approximately 58%, 43% and 38%, respectively compared to EG1
additional information
the enzyme consists of an N-terminal signal peptide, two glycosyl hydrolase family 5 catalytic modules, two novel carbohydrate-binding modules, two linker sequences, and a C-terminal sequence with an unknown function. Removal of the carbohydrate-binding modules from rCel5A reduces the catalytic activities with various polysaccharides remarkably
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the enzyme consists of an N-terminal signal peptide, two glycosyl hydrolase family 5 catalytic modules, two novel carbohydrate-binding modules, two linker sequences, and a C-terminal sequence with an unknown function. Removal of the carbohydrate-binding modules from rCel5A reduces the catalytic activities with various polysaccharides remarkably
