3.2.1.21: beta-glucosidase

H0HC94

contrary to uncompetitive inhibition of wild-type by glucose, mutant is competitively inhibited

H229S

H0HC94

contrary to uncompetitive inhibition of wild-type by glucose, mutant is competitively inhibited

L178E

2 entries

V176A

H0HC94

slight increase in specific activity

W127F

H0HC94

mutant shows enhanced thermostability

Agrobacterium tumefaciens 5A

C174V

contrary to uncompetitive inhibition of wild-type by glucose, mutant is competitively inhibited

H229S

Agrobacterium tumefaciens 5A

contrary to uncompetitive inhibition of wild-type by glucose, mutant is competitively inhibited

L178E

2 entries

W127F

Agrobacterium tumefaciens 5A

mutant shows enhanced thermostability

D463A

the variant exhibits kinetic constants similar to the wild type enzyme

D487A

the variant exhibits kinetic constants similar to the wild type enzyme

D496A

the variant exhibits kinetic constants similar to the wild type enzyme

D501A

the variant exhibits kinetic constants similar to the wild type enzyme

E490A

the variant exhibits greatly reduced kcat and kcat/Km values toward the natural substrate cellobiose (67000 and 61000fold, respectively)

W49A

site-directed mutagenesis, the mutation reduces the transglucosylation activity of the enzyme due to a large decrease in binding capacity for Glc at site +1 and an increase in binding ability at site-1, while the hydrolytic activity is only slightly decreased compared to the wild-type enzyme, the mutant shows altered kinetics, overview

W49D

W49G

W49N

D463A

the variant exhibits kinetic constants similar to the wild type enzyme

D487A

the variant exhibits kinetic constants similar to the wild type enzyme

D496A

the variant exhibits kinetic constants similar to the wild type enzyme

D501A

the variant exhibits kinetic constants similar to the wild type enzyme

E490A

the variant exhibits greatly reduced kcat and kcat/Km values toward the natural substrate cellobiose (67000 and 61000fold, respectively)

G294A

site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme

G294C

site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme

G294D

site-directed mutagenesis, the mutant shows similar activity as the wild-type enzyme

G294E

site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme

G294F

site-directed mutagenesis, the mutant shows 1.5fold higher activities for substrate recognition than the wild-type enzyme

G294H

site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme

G294I

site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme

G294K

site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme

G294L

site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme

G294M

site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme

G294N

site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme

G294P

site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme

G294Q

site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme

G294R

site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme

G294S

site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme

G294T

site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme

G294V

site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme

G294W

site-directed mutagenesis, the mutant shows 1.5fold higher activities for substrate recognition than the wild-type enzyme

G294Y

site-directed mutagenesis, the mutant shows 1.6fold higher activities for substrate recognition than the wild-type enzyme

E495L

Avena sativa

isoform AsGlu2. Mutation assembles the AsGlu2 into fibrillar homomultimers. The mutant AsGlu2 homomultimer is highly stable and has relatively faster electric mobility in native gel than the AsGlu1 homomultimer. Multimerization increases enzyme affinity to substrates

696468

F417S

significantly impaired catalytic activity with cellobiose and lactose as substrates, 1000fold decrease in apparent kcat of cellobiose, 47fold decrease in kcat of lactose. Mutant produces improved total galactooligosaccharides yields by transgalactosylation

F417Y

mutant retains the capacity to hydrolyze both cellobiose and lactose with the same relative substrate selectivity as the wild-type, but with about 10fold lower turnover numbers. Mutant produces improved total galactooligosaccharides yields by transgalactosylation

N222F

significantly impaired catalytic activity with cellobiose and lactose as substrates

N294T

significantly impaired catalytic activity with cellobiose and lactose as substrates

Y296F

significantly impaired catalytic activity with cellobiose and lactose as substrates. Mutant produces improved total galactooligosaccharides yields by transgalactosylation

F417S

F417Y

N222F

significantly impaired catalytic activity with cellobiose and lactose as substrates

N294T

significantly impaired catalytic activity with cellobiose and lactose as substrates

Y296F

significantly impaired catalytic activity with cellobiose and lactose as substrates. Mutant produces improved total galactooligosaccharides yields by transgalactosylation

D409H

naturally occurring mutation involved in Gaucher's disease development with reduced enzyme activity, obligated GD heterozygotes

E165P

crystallization data of covalent intermediate with substrate, in which glucose is bound to nucleophile E373

696002

F225S

0.65% of wild-type specific activity with p-nitrophenyl-beta-D-glucopyranoside

L444P

naturally occurring mutation involved in Gaucher's disease development with reduced enzyme activity, obligated GD heterozygotes

M172L

site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, and altered substrate specificity

N370S

naturally occurring mutation involved in Gaucher's disease development with reduced enzyme activity, obligated GD heterozygotes, and homozygote GD patients

N370S/D409H

naturally occurring mutation involved in Gaucher's disease development with reduced enzyme activity, homozygote GD patients

N370S/L444P

naturally occurring mutation involved in Gaucher's disease development with reduced enzyme activity, homozygote GD patients

Q307N

site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, and altered substrate specificity

V168Y

12% of wild-type specific activity with p-nitrophenyl-beta-D-glucopyranoside

W345A

site-directed mutagenesis, almost inactive mutant

W345M

site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, and altered substrate specificity

Y308A

0.3% of wild-type specific activity with p-nitrophenyl-beta-D-glucopyranoside

Y308F

6% of wild-type specific activity with p-nitrophenyl-beta-D-glucopyranoside

F304I

mutant enzyme shows no activity with 4-nitrophenyl beta-D-glucopyranoside

I278F

decreased activities towards 4-nitrophenyl beta-D-glucopyranoside, hydrolysis activity on sophorose, which is much higher than the activity on 4-nitrophenyl beta-D-glucopyranoside

I48W

decreased activities towards 4-nitrophenyl beta-D-glucopyranoside, hydrolysis activity on sophorose, which is much higher than the activity on 4-nitrophenyl beta-D-glucopyranoside

I48W/I278F/T484Y

decreased activities towards 4-nitrophenyl beta-D-glucopyranoside, hydrolysis activity on sophorose, which is much higher than the activity on 4-nitrophenyl beta-D-glucopyranoside

T484Y

decreased activities towards 4-nitrophenyl beta-D-glucopyranoside, hydrolysis activity on sophorose, which is much higher than the activity on 4-nitrophenyl beta-D-glucopyranoside

W69I

mutant enzyme shows no activity with 4-nitrophenyl beta-D-glucopyranoside

Y509T

introduction of mutation Y509T causes the increase of the Km value, but does not affect the kcat value

I278F

decreased activities towards 4-nitrophenyl beta-D-glucopyranoside, hydrolysis activity on sophorose, which is much higher than the activity on 4-nitrophenyl beta-D-glucopyranoside

I48W

decreased activities towards 4-nitrophenyl beta-D-glucopyranoside, hydrolysis activity on sophorose, which is much higher than the activity on 4-nitrophenyl beta-D-glucopyranoside

I48W/I278F/T484Y

decreased activities towards 4-nitrophenyl beta-D-glucopyranoside, hydrolysis activity on sophorose, which is much higher than the activity on 4-nitrophenyl beta-D-glucopyranoside

T484Y

decreased activities towards 4-nitrophenyl beta-D-glucopyranoside, hydrolysis activity on sophorose, which is much higher than the activity on 4-nitrophenyl beta-D-glucopyranoside

W69I

mutant enzyme shows no activity with 4-nitrophenyl beta-D-glucopyranoside

Y509T

Listeria innocua Clip11262

introduction of mutation Y509T causes the increase of the Km value, but does not affect the kcat value

D270A

Q92AS9

nucleophile mutant, crystallization data

760014

R572K

Listeria innocua Clip11262

Q92AS9

mutation in residue important for substrate recognition, 10fold increase in Km value for sophorose

760014

D270A

Listeria innocua Clip11262 ATCC BAA-680

nucleophile mutant, crystallization data

R572K

Listeria innocua Clip11262 ATCC BAA-680

mutation in residue important for substrate recognition, 10fold increase in Km value for sophorose

E176A

inactive under standard assay conditions, mutant can be rescued by (small nucleophiles) acetate and azide but not as mutant E176Q

E176D

inactive under standard assay conditions, mutant can not be rescued by any small nucleophile

E176Q

E176Q active site mutant can be effectively rescued by small nucleophiles, such as acetate, azide and ascorbate, for hydrolysis of aryl glycosides in a pH-independent manner above pH 5. Cellotriose, cellotetraose, cellopentaose, cellohexaose and laminaribiose are not hydrolyzed by the mutant and instead exhibit competitive inhibition

E386D

3000fold less active than wild-type

E386Q

60000fold less active than wild-type

I179V

decrease in catalytic efficiencies

L442R

catalytic efficiencies similar to wild-type

N190H

threefold increase in catalytic efficiency towards cellobiose

N245V

twofold increase in catalytic efficiency towards cellobiose

E172A

A1C3J9

site-directed mutagenesis, inactive catalytic residue mutant

681491

E173A

Q6QGY5

site-directed mutagenesis, inactive mutant

664360

E362A

Q6QGY5

site-directed mutagenesis, inactive mutant

664360

E370A

A1C3J9

site-directed mutagenesis, inactive catalytic residue mutant

681491

D229N

site-directed mutagenesis, mutation of the isozyme BGL1A residue from subsite +1 to the correspondent residue of isozyme BGL1B, the mutant shows decreased catalytic efficiency compared to the wild-type BGL1A

D229N/K253A

site-directed mutagenesis, mutation of the isozyme BGL1A residues from subsite +1 to the correspondent residues of isozyme BGL1B, the double mutant has a hydrolytic activity at neutral pH that is restored to that of the wild-type enzyme

H231D

K253A

M177L

site-directed mutagenesis, mutation of the isozyme BGL1A residue from subsite +1 to the correspondent residue of isozyme BGL1B, the mutant shows catalytic efficiency similar to the wild-type BGL1A

V173C

site-directed mutagenesis, mutation of the isozyme BGL1A residue from subsite +1 to the correspondent residue of isozyme BGL1B, the mutant shows catalytic efficiency similar to the wild-type BGL1A

E173N

mutant enzyme shows no activity

E389N

mutant enzyme shows no activity

E446N

beta-fucosidase activity is 26.49% compared to wild-type activity, beta-glucosidase activity is 21.37% compared to wild-type activity, beta-galactosidase activity is 15.7% compared to wild-type activity

N172A

beta-fucosidase activity is 18.38% compared to wild-type activity, beta-glucosidase activity is 17.65% compared to wild-type activity, beta-galactosidase activity is 13.4% compared to wild-type activity

E417S

mutant with reduced pH optimum, increased Km and turnover number for 2-nitrophenyl-beta-1,4-D-glucopyranoside

326270

F426Y

mutant with unchanged pH optimum, slight increased Km and slight reduced turnover number for 2-nitrophenyl-beta-1,4-D-glucopyranoside

326270

M424K

mutant with increased pH optimum, slight increased Km and reduced turnover number for 2-nitrophenyl-beta-1,4-D-galactopyranoside

326270

R170A/R220A/Y227F

mutant enzyme occurs as dimer

R170A/R220A/Y227F/R448E

mutant enzyme occurs as monomer, melting temperature is decreased form 110°C for the wild-type enzyme to 73-75°C for the mutant enzyme

R170A/R220A/Y227F/R449R

mutant enzyme occurs as monomer, melting temperature is decreased form 110°C for the wild-type enzyme to 73-75°C for the mutant enzyme

R170A/R220A/Y227F/R459G

mutant enzyme occurs as monomer, melting temperature is decreased form 110°C for the wild-type enzyme to 73-75°C for the mutant enzyme

E191A

site-directed mutagenesis, inactive mutant

E407A

site-directed mutagenesis, inactive mutant

F198A

F471Y

site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme

G464F

site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme

G464S

site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme

G464S/S465L

site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme

S465L

site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme

Y378A

site-directed mutagenesis, the mutant shows 300-380% increased catalytic eficiency with substrates 2-O-beta-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one and 2-O-beta-D-glucopyranosyl-4-hydroxy-7-demethoxy-1,4-benzoxazin-3-one, respectively

Y378F

site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme

A375P

Km (2-nitrophenyl beta-D-glucopyranoside) decreased compared to wild-type, kcat (2-nitrophenyl beta-D-glucopyranoside) decreased. Relative efficacy toward (2-nitrophenyl beta-D-glucopyranoside) similar to wild-type. Km (dhurrin) similar to wild-type, kcat drastically decreased

E189D

substrate binding structure analysis from crystal structure

665542

F469Y

Km (2-nitrophenyl beta-D-glucopyranoside) increased compared to wild-type, kcat (2-nitrophenyl beta-D-glucopyranoside) decreased. Relative efficacy toward (2-nitrophenyl beta-D-glucopyranoside) significantly decreased by factor 2.5. Km (dhurrin) similar to wild-type, kcat drastically decreased

V196F/L203F

E190Q

mutant construcuted for analysis of the relative contribution of the residue to specificity and catalysis. E190 favors the binding of the initial portion of alkyl-type aglycones

E194A

mutant construcuted for analysis of the relative contribution of the residue to specificity and catalysis. E190 favors the binding of the initial portion of alkyl-type aglycones

E451A

the double mutant shows decreased activity compared to the wild type enzyme

E451S

the double mutant shows decreased activity compared to the wild type enzyme

K201A

mutant construcuted for analysis of the relative contribution of the residue to specificity and catalysis. K201 is relevant for binding of D-glucose units

K201F

mutant construcuted for analysis of the relative contribution of the residue to specificity and catalysis. K201 is relevant for binding of D-glucose units

M453A

mutant construcuted for analysis of the relative contribution of the residue to specificity and catalysis. M453 favors the binding of the second D-glucose unit of oligosaccharidic aglycones

Q39E

the double mutant shows decreased activity compared to the wild type enzyme

Q39E/E451A

the double mutant shows decreased activity compared to the wild type enzyme

Q39E/E451S

the double mutant shows decreased activity compared to the wild type enzyme

Q39N

the double mutant shows decreased activity compared to the wild type enzyme

Q39N/E451S

the double mutant shows decreased activity compared to the wild type enzyme

E178A

3500fold reduction in catalytic efficiency, E178 ist the general acid/base catalyst of Bgl3

E178Q

250fold reduction in catalytic efficiency

E383A

100000fold reduction in catalytic efficiency

E383Q

1000000fold reduction in catalytic efficiency, E383 is the catalytic nucleophile of Bgl3

A433V

random mutagenesis, identification of the thermostable mutant, kinetics in comparison to the wild-type enzyme

L444F

random mutagenesis, identification of the thermostable mutant, kinetics in comparison to the wild-type enzyme

N178I

random mutagenesis, identification of the thermostable mutant, substrate-saturation study, binding of a second substrate molecule increases the turnover with D-cellobiose

N317Y

random mutagenesis, identification of the thermostable mutant, kinetics in comparison to the wild-type enzyme

N317Y/L444F

random mutagenesis, identification of the thermostable mutant, substrate-saturation study, binding of a second substrate molecule increases the turnover with D-cellobiose

N317Y/L444F/A422V

random mutagenesis, identification of the thermostable mutant, substrate-saturation study, binding of a second substrate molecule increases the turnover with D-cellobiose

N439S

random mutagenesis, identification of the thermostable mutant, kinetics in comparison to the wild-type enzyme

S319C

random mutagenesis, identification of the thermostable mutant, the mutant shows highly increased thermotolerance and activity with D-cellobiose compared to the wild-type enzyme

D206N

catalytically active mutant enzyme, similar temperature optimum like wild-type enzyme. The high-catalytic turn-over rate by D206N for beta-glucosidase activity makes it a useful enzyme in cellulose degradation at high temperatures

D206Q

mutant enzyme shows less than 10% hydrolytic activity than the wild-type toward 4-nitrophenyl glycosides

E207S

no hydrolytic activity

E399S

no hydrolytic activity

Q77R

mutant enzyme shows less than 10% hydrolytic activity than the wild-type toward 4-nitrophenyl glycosides

N62Y/Q63K/L231F/V303A/A412V

Thermosphaera aggregans

Q9YGA8

mutations observed in all the clones isolated

735665

N62Y/Q63K/L231F/V303A/E386G/A412V

Thermosphaera aggregans

Q9YGA8

additional mutation in nucleophile residue E386, mutation completely abolishes activity. The addition of 2 M sodium formate as an external nucleophile leads to the recovery of 4.60% activity with accumulation of oligosaccharides. At pH 3.0 and low concentrations of sodium formate buffer, the hyperthermophilic glycosynthase shows kcat values similar to those of the wild-type and 17fold higher than those observed at the usual reactivation conditions in 2 M sodium formate at pH 6.5

735665

D242G

mutant loses 96% of wild-type activity, activity is partially restored after addition of NaN3

D58A

dramatic decrease in kcat compared to wild-type, activity is highly recovered after addition of NaN3

E349G

crystallization data

E458G

dramatic decrease in kcat compared to wild-type, activity is partially restored after addition of NaN3

F224I

mutation the aglycone binding region, flavonoid glucoside hydrolysis is not affected

F310E

mutation the aglycone binding region, flavonoid glucoside hydrolysis is not affected

F310L

mutation the aglycone binding region, flavonoid glucoside hydrolysis is not affected

N220F

mutation the aglycone binding region

N220S

mutation the aglycone binding region, increases the catalytic efficiency on both quercetin 3-glucoside and 4-nitrophenyl-beta-D-glucoside

N221F

mutation the aglycone binding region

N221S

mutation the aglycone binding region, increases the catalytic efficiency on both quercetin 3-glucoside and 4-nitrophenyl-beta-D-glucoside

W243F

Km (4-nitrophenyl beta-D-glucopyranoside) close to wild-type, kcat decreased to 34% compared to wild-type

W322A

mutation the aglycone binding region, residue is important for substrate accomodation. Mutation W322A results in a large reduction of hydrolytic activity on 3-glucosylated flavonoids

E349G

crystallization data

F224I

mutation the aglycone binding region, flavonoid glucoside hydrolysis is not affected

F310L

mutation the aglycone binding region, flavonoid glucoside hydrolysis is not affected

N220S

mutation the aglycone binding region, increases the catalytic efficiency on both quercetin 3-glucoside and 4-nitrophenyl-beta-D-glucoside

W322A

mutation the aglycone binding region, residue is important for substrate accomodation. Mutation W322A results in a large reduction of hydrolytic activity on 3-glucosylated flavonoids

P172L/L167W

Trichoderma reesei

O93785

the enzyme has mutations on the left wall of the substrate binding site and shows decreased sensitivity of the catalytic activity to pH and, thus, the pH profile for optimal activity is expanded from pH 6.0 to a pH range from pH 5.0 through pH 7.0

750243

E191A

Triticum aestivum

Q1XH04, Q1XH05, Q1XIR9

site-directed mutagenesis, inactive TaGlu1a mutant

E407A

Triticum aestivum

Q1XH04, Q1XH05, Q1XIR9

site-directed mutagenesis, inactive TaGlu1a mutant

F198A

Triticum aestivum

Q1XH04, Q1XH05, Q1XIR9

site-directed mutagenesis, the TaGlu1a mutant shows reduced activity with substrates 2-O-beta-D-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one and 2-O-beta-D-glucopyranosyl-4-hydroxy-7-demethoxy-1,4-benzoxazin-3-one with highly incresed Km values, respectively

D772A

uncultured bacterium

Q3HXC2

the mutant shows less than 1/10000 activity of the wild type enzyme

702747

N223Q

uncultured microorganism

A0A1E1FFN6

mutation results in reduced glucose tolerance and transglycosylation activity, and drastically changed substrate specificity

744881

N223Y

uncultured microorganism

A0A1E1FFN6

mutation results in reduced glucose tolerance and transglycosylation activity, and drastically changed substrate specificity

744881

D261N

the D261N substitution leads to a drastic decrease in relative efficiency of ZmGlu1 on all substrates tested, but little effect on the Km

E191D

substrate binding structure analysis from crystal structure

665542

E191D/F198V

inactive mutant, F198 substitution causes a rearrangement of residues F205, F466, and E464 involved in aglycone binding

665542

F193A

increase in affinity for a small polar aglycone, deep decrease in kcat value. 7.8% of wild-type activity for substrate 4-nitrophenyl-beta-D-glucoside, 3% for substrate 4-methylumbelliferyl-beta-D-glucoside, respectively

F193A/F200K/W373K/F461L

almost complete loss of catalytic activity, extensive alterations in strucutre

F198V

mutant shows drastic effect on catalytic efficacy, kcat is reduced by 90% for hydrolysis of 4-methylumbelliferyl beta-D-glucopyranoside compared to wild-type. Mutant is unable to hydrolyze 4-nitrophenyl beta-D-glucopyranoside. Only negligible activity toward substrate 2-nitrophenyl beta-D-glucopyranoside compared to wild-type

F198V/F205L/P377A

mutant shows a drastic reduced relative efficacy for all test substrated due to a decreased turnover

F200K

1.1% of wild-type activity for substrate 4-nitrophenyl-beta-D-glucoside, 0.3% for substrate 4-methylumbelliferyl-beta-D-glucoside, respectively

F205L

Mutant shows similar relative efficacy compared to wild-type toward all substrates tested. Mutant shows no significant effect on hydrolysis of 4-nitrophenyl beta-D-glucopyranoside other than doubling the kcat value compared to wild-type

F461L

120% of wild-type activity for substrate 4-nitrophenyl-beta-D-glucoside, 119% for substrate 4-methylumbelliferyl-beta-D-glucoside, respectively

F466S

mutant exhibits a marked increase in relative efficiency in comparison to wild-type ZmGlu1 on all substrates tested (from 28 to 123%), because of increased substrate turnover. Mutant has little effect on dhurrin hydrolysis compared to wild-type (3% of relative efficacy compared to Sorghum Dhr1)

F466S/A467S

the A467S substitution in ZmGlu1 has no detectable effect on substrate specificity and catalytic efficiency in the case of 4-nitrophenyl beta-D-glucopyranoside or 2-nitrophenyl beta-D-glucopyranoside hydrolysis, and only a lower Km is obtained for 4-methylumbelliferyl beta-D-glucuronide compared to mutant F466S. Mutant has little effect on dhurrin hydrolysis compared to wild-type (3% of relative efficacy compared to Sorghum Dhr1)

K81E

mutant not able to bind chimeric lectin beta-glucosidase aggregating factor

696319

M263F

mutant shows reduced relative efficacy by 40 to 50% on 4-nitrophenyl beta-D-glucopyranoside and 4-methylumbelliferyl beta-D-glucopyranoside whereas it shows an increase of about 60% relative efficacy on 2-nitrophenyl beta-D-glucopyranoside compared to wild-type

N481E

mutant with lowered affinity to cimeric lectin beta-glucosidase aggregating factor

696319

P377A

P377A mutation leads to changes (decreases) in both kcat and Km for all substrated with no decrease in the overall relative efficiency

T82Y

mutant not able to bind chimeric lectin beta-glucosidase aggregating factor

696319

W373K

large decrease in kcat value. 0.5% of wild-type activity for substrate 4-nitrophenyl-beta-D-glucoside, 0.04% for substrate 4-methylumbelliferyl-beta-D-glucoside, respectively

Y473F

kcat is increased by 110% to 450% depending on the substrate tested. Enhanced turnover is compensated by a slight increase in Km of this mutant giving a 40% to 250% (depending on the substrate tested) better relative efficacy over the wild-type enzyme. Mutation leads to a significant effect towards dhurrin hydrolysis (10% relative efficacy as compared to Sorghum Dhr1)