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Information on EC 3.2.1.2 - beta-amylase and Organism(s) Bacillus cereus and UniProt Accession P36924
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3.2.1.2 beta-amylaseIUBMB Comments
Acts on starch, glycogen and related polysaccharides and oligosaccharides producing beta-maltose by an inversion. The term 'beta'' relates to the initial anomeric configuration of the free sugar group released and not to the configuration of the linkage hydrolysed.
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Word Map
- 3.2.1.2
- starch
- alpha-amylase
- maltose
- barley
- potato
- soybean
-
sweet
- amylopectin
-
amylolytic
- glucoamylase
- endosperm
- hordeum
- amylases
- pullulanase
-
debranching
- dextrin
- nutrition
- maltotetraose
- thermosulfurogenes
- isoamylase
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beta-limit
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starch-degrading
- amyloglucosidase
- polymyxa
- maltotriose
- maltopentaose
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granule-bound
- alpha-cyclodextrin
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extrachloroplastic
- industry
-
sporamin
- molecular biology
- brewing
The taxonomic range for the selected organisms is: Bacillus cereus
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
Reaction Schemes
Synonyms
beta-amylase, arath, spoii, tr-bamy, beta amylase, ct-bmy, bam-2, beta-amylase 1, bam-8, glycogenase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
(1-4)-alpha-D-glucan maltohydrolase
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1,4-alpha-D-glucan maltohydrolase
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amylase, beta-
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beta amylase
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glycogenase
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saccharogen amylase
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saccharogenamylase
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additional information
additional information
member of family 14 of the sequence-based classification of glycoside hydrolases
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of (1->4)-alpha-D-glucosidic linkages in polysaccharides so as to remove successive maltose units from the non-reducing ends of the chains
Hydrolysis of (1->4)-alpha-D-glucosidic linkages in polysaccharides so as to remove successive maltose units from the non-reducing ends of the chains
catalytic mechanism, reaction mechanism
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Hydrolysis of (1->4)-alpha-D-glucosidic linkages in polysaccharides so as to remove successive maltose units from the non-reducing ends of the chains
mechanism
Hydrolysis of (1->4)-alpha-D-glucosidic linkages in polysaccharides so as to remove successive maltose units from the non-reducing ends of the chains
catalytic mechanism
Hydrolysis of (1->4)-alpha-D-glucosidic linkages in polysaccharides so as to remove successive maltose units from the non-reducing ends of the chains
Glu367 is important in catalysis, the plant enzyme shows a reaction mechanism different from the bacterial enzyme, overview
PATHWAY SOURCE
PATHWAYS
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-, -
SYSTEMATIC NAME
IUBMB Comments
4-alpha-D-glucan maltohydrolase
Acts on starch, glycogen and related polysaccharides and oligosaccharides producing beta-maltose by an inversion. The term 'beta'' relates to the initial anomeric configuration of the free sugar group released and not to the configuration of the linkage hydrolysed.
CAS REGISTRY NUMBER
COMMENTARY
9000-91-3
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
maltopentaose + H2O
?
beta-amylase is an exo-enzyme that catalyzes the hydrolysis of the alpha-1,4-glucosidic linkage of the substrate liberating beta-maltose from the non-reducing end, Glu-172 and Glu-367 are catalytic residues, binding mode of substrate, substrate recognition mechanism, enzyme structure
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-
?
maltopentaose + H2O
?
binding mode of substrate in the active site
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?
maltopentaose + H2O
?
hydrolyzes the alpha-1,4-glucosidic linkage liberating beta-maltose from the non-reducing end of substrate, good substrate, mode of binding in the active site, catalytic mechanism, enzyme/domain structure
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-
?
starch + H2O
?
beta-amylase is an exo-enzyme that catalyzes the hydrolysis of the alpha-1,4-glucosidic linkage of the substrate liberating beta-maltose from the non-reducing end, Glu-172 and Glu-367 are catalytic residues, substrate recognition mechanism, enzyme structure
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-
?
starch + H2O
?
catalyzes the hydrolysis of alpha-1,4-glucosidic linkages of soluble starch, and liberates beta-anomeric maltose from the nonreducing ends, exo-acting enzyme, composed of two functional domains, a catalytic domain: domains A and B, and starch-binding domain: domain C, beta-amylase has three carbohydrate-binding sites aside from the active site: two in domain B named Site2 and Site3, one in domain C named Site1, roles of these sites in the catalytic reaction and raw starch-binding, beta-amylase hardly hydrolyzes raw starch from wheat, corn, potato or sweet potato, but binds to it strongly
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?
starch + H2O
?
hydrolyzes the alpha-1,4-glucosidic linkage liberating beta-maltose from the non-reducing end of substrate, enzyme/domain structure, starch binding site in domain C, catalytic mechanism
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?
maltopentaose + H2O
?
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beta-amylase is an inverting enzyme that hydrolyzes the alpha-1,4-glucosidic linkage of the substrate liberating beta-maltose from the non-reducing end, catalytic mechanism, Glu-172 acts as general acid, Glu-367 acts as general base
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?
starch + H2O
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beta-amylase hydrolyzes alpha-1,4-linkage, raw starch granules from potato, wheat, rice and corn, with the granules from rice being the best substrate, beta-amylase attacks very slowly on the starch granules, hydrolyzes corn granules efficiently at 45°C
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?
starch + H2O
?
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beta-amylase is an inverting enzyme that hydrolyzes the alpha-1,4-glucosidic linkage of the substrate liberating beta-maltose from the non-reducing end, catalytic mechanism, Glu-172 acts as general acid, Glu-367 acts as general base
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?
additional information
?
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no hydrolysis of alpha-1,6-glucosidic linkages
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?
additional information
?
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no hydrolysis of alpha-1,6-glucosidic linkages
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?
additional information
?
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beta-amylase does not catalyze a transglycosylation reaction
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?
additional information
?
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beta-amylase does not catalyze a transglycosylation reaction
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?
additional information
?
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analysis of effect of hydroxypropylation and beta-amylase treatment on complexation of debranched starch with naringenin using potato starch and Hylon VII. An increase in hydroxypropylation level improves recovery of soluble complexes, while total recovery remains unchanged. The beta-amylase treatment further increases soluble complex recovery. For the same treatment, the naringenin content is greater in Hylon VII complexes than in potato starch complexes
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?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2,3-epoxypropyl-alpha-D-glucopyranoside
affinity-labeling reagent, mode of binding, covalently bound to the catalytic residue Glu-172, inactivation mechanism
3,4-epoxybutyl-alpha-D-glucopyranoside
affinity-labeling reagent, mode of binding, covalently bound to the catalytic residue Glu-172
maltitol
behaves as a mixed-type or competitive inhibitor depending on the chain length of the substrate, inhibition mechanism, binds to Site2 in domain B and forms an abortive ESI complex when amylose is used as substrate
O-alpha-D-glucopyranosyl(1-4)O-alpha-D-glucopyranosyl(1-4)D-xylopyranose
mode of binding in the active site cleft
O-alpha-D-xylopyranosyl(1-4)O-alpha-D-glucopyranosyl(1-4)O-alpha-D-glucopyranoside
mixed-type inhibition, two molecules bind to enzyme
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.6
amylose
pH 7, 25°C, DPn: 16, S235A/Y249A/W449F/W495F quadruple mutant
additional information
additional information
binding parameters of wild-type and mutant enzymes to raw corn starch
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additional information
additional information
-
binding parameters of wild-type and mutant enzymes to raw corn starch
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additional information
additional information
kinetics, recombinant wild-type andmutant enzymes
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additional information
additional information
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kinetics, recombinant wild-type andmutant enzymes
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additional information
additional information
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Km for soluble starch is 0.4%
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.8
cyclohexaamylose
pH 7, 25°C, wild-type enzyme, amylose as substrate
0.45
O-alpha-D-xylopyranosyl(1-4)O-alpha-D-glucopyranosyl(1-4)O-alpha-D-glucopyranoside
pH 7, 25°C, wild-type enzyme, amylose as substrate
5.1
maltitol
pH 7, 25°C, S235A/Y249A double mutant, amylose as substrate
5.7
maltitol
pH 7, 25°C, S235A/Y249A/W449F/W495F quadruple mutant, amylose as substrate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
2182
purified commercial preparation, pH and temperature not specified in the publication
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
comparison of pH-dependency/pH-profiles of recombinant wild-type and mutant enzymes
additional information
-
comparison of pH-dependency/pH-profiles of recombinant wild-type and mutant enzymes
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4.5 - 8.5
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pH 4.5: about 40% of maximal activity, pH 8.5: about 60% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
28 - 61
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28°C: about 40% of maximal activity, 61°C: about 30% of maximal activity
45
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beta-amylase hydrolyzes corn starch granules efficiently at 45°C, 60% more active than soybean beta-amylase at 45°C
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
enzyme is composed of two functional domains, a catalytic domain: domains A and B, and starch-binding domain: domain C, beta-amylase has three carbohydrate-binding sites aside from the active site: two in domain B named Site2 and Site3, one in domain C named Site1, roles of these sites in the catalytic reaction and raw starch-binding
additional information
-
enzyme is composed of two functional domains, a catalytic domain: domains A and B, and starch-binding domain: domain C, beta-amylase has three carbohydrate-binding sites aside from the active site: two in domain B named Site2 and Site3, one in domain C named Site1, roles of these sites in the catalytic reaction and raw starch-binding
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
beta-amylase complexed with the inhibitors glucose, maltose and O-alpha-D-glucopyranosyl(1-4)O-alpha-D-glucopyranosyl(1-4)D-xylopyranose and the affinity-labeling reagents 2,3-epoxypropyl-alpha-D-glucopyranoside and 3,4-epoxybutyl-alpha-D-glucopyranoside, X-ray analysis
purified recombinant wild-type and mutant T47M/Y164E/T328N, hanging drop vapour diffusion method, 18°C, 0.005 ml of 15 mg/ml protein in 0.05 M sodium acetate is mixed with 0.005 ml mother liquor containing 15% PEG 6000, 5% saturated ammonium sulfate, 0.1 M phosphate, pH 6.5, crystallization of mutants Y164E and Y164F in the same way except for usage of 0.1 M sodium acetate buffer, pH 4.6, instead of phosphate buffer, X-ray diffraction structure determination and analysis at 1.72-1.95 A resolution, active site structure modelling
X-ray crystal structures of wild-type enzyme complexed with maltose and of E172A catalytic site mutant complexed with maltopentaose
crystal structure determined by the multiple isomorphous replacement method
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
S235A
binding parameters to raw corn starch, kinetic parameters for the hydrolysis of amylose, 88% of wild-type activity with soluble starch as substrate
S235A/Y249A
double mutant, binding parameters to raw corn starch, kinetic parameters for the hydrolysis of amylose, 63% of wild-type activity with soluble starch as substrate
S235A/Y249A/W449F/W495F
quadruple mutant, kinetic parameters for the hydrolysis of amylose, 51% of wild-type activity with soluble starch as substrate
T47M/Y164E/T328N
site-directed mutagenesis, the mutation of Y164 leads to disruption of the hydrogen bonding around the catalytic site, the mutant enzyme shows a shifted pH optimum and a 88% decreased kcat compared to the wild-type enzyme
W449F/W495F
double mutant, binding parameters to raw corn starch, kinetic parameters for the hydrolysis of amylose, 61% of wild-type activity with soluble starch as substrate
Y164E
site-directed mutagenesis, the mutation of Y164 leads to disruption of the hydrogen bonding around the catalytic site, the mutant enzyme shows a shifted pH optimum and a 59% decreased kcat compared to the wild-type enzyme
Y164F
site-directed mutagenesis, the mutation of Y164 leads to disruption of the hydrogen bonding around the catalytic site, the mutant enzyme shows a shifted pH optimum and a 64% decreased kcat compared to the wild-type enzyme
Y164H
site-directed mutagenesis, the mutation of Y164 leads to disruption of the hydrogen bonding around the catalytic site, the mutant enzyme shows a shifted pH optimum and a 97% decreased kcat compared to the wild-type enzyme
Y164Q
site-directed mutagenesis, the mutation of Y164 leads to disruption of the hydrogen bonding around the catalytic site, the mutant enzyme shows a shifted pH optimum and a 83% decreased kcat compared to the wild-type enzyme
Y249A
binding parameters to raw corn starch, kinetic parameters for the hydrolysis of amylose, 80% of wild-type activity with soluble starch as substrate
E172A
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catalytic site mutant, no hydrolytic activity, no rescue of activity by 2 M azide
E172A/E367A
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catalytic site double mutant, no hydrolytic activity, no rescue of activity by 2 M azide
E367A
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catalytic site mutant, no hydrolytic activity in the absence of azide, in the presence of 2 M azide the mutant enzyme hydrolyzes maltopentaose at pH 7 and 25°C producing maltose, mechanism
additional information
additional information
mutation of two of the three carbohydrate-binding sites aside from the active site: Site2 in domain B and Site1 in domain C
additional information
-
mutation of two of the three carbohydrate-binding sites aside from the active site: Site2 in domain B and Site1 in domain C
additional information
engineering of the enzyme's pH optimum, conversion of the pH optimum from the bacterial type with pH 6.7 to the higher-plant type with pH 5.4, from soybean enzyme, overview
additional information
-
engineering of the enzyme's pH optimum, conversion of the pH optimum from the bacterial type with pH 6.7 to the higher-plant type with pH 5.4, from soybean enzyme, overview
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant wild-type and mutant enzymes from Escherichia coli strain XL1-Blue
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression of wild-type and mutant enzymes in Escherichia coli strain XL1-Blue
wild-type and mutant enzymes, expression in Escherichia coli BL21(DE3)
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Takasaki, Y.
Purification and enzymatic properties of beta-amylase and pullulanase from Bacillus cereus var. mycoides
Agric. Biol. Chem.
40
1523-1530
1976
Bacillus cereus
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Nanmori, T.; Numata, Y.; Shinke, R.
Isolation and characterization of a Bacillus cereus mutant strain hyperproductive of exo-beta-amylase
Appl. Environ. Microbiol.
53
768-771
1987
Bacillus cereus
Nanmori, T.; Shinke, R.; Aoki, K.; Nishira, H.
Purification and characterization of beta-amylase from Bacillus cereus BQ10-S1 Spo II
Agric. Biol. Chem.
47
941-947
1983
Bacillus cereus
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Yamaguchi, T.; Matsumoto, Y.; Shirakawa, M.; Kibe, M.; Hibino, T.; Kozaki, S.; Takasaki, Y.; Nitta, Y.
Cloning, sequencing, and expression of a beta-amylase gene from Bacillus cereus var. mycoides and characterization of its products
Biosci. Biotechnol. Biochem.
60
1255-1259
1996
Bacillus cereus
Oyama, T.; Kusunoki, M.; Kishimoto, Y.; Takasaki, Y.; Nitta, Y.
Crystal structure of beta-amylase from Bacillus cereus var. Mycoides at 2.2 A resolution
J. Biochem.
125
1120-1130
1999
Bacillus cereus
Miyake, H.; Kurisu, G.; Kusunoki, M.; Nishimura, S.; Kitamura, S.; Nitta, Y.
Crystal structure of a catalytic site mutant of beta-amylase from Bacillus cereus var. mycoides cocrystallized with maltopentaose
Biochemistry
42
5574-5581
2003
Bacillus cereus (P36924), Bacillus cereus
Miyake, H.; Otsuka, C.; Nishimura, S.; Nitta, Y.
Catalytic mechanism of beta-amylase from Bacillus cereus var. mycoides: chemical rescue of hydrolytic activity for a catalytic site mutant (Glu367-->Ala) by azide
J. Biochem.
131
587-591
2002
Bacillus cereus
Oyama, T.; Miyake, H.; Kusunoki, M.; Nitta, Y.
Crystal structures of beta-amylase from Bacillus cereus var. mycoides in complexes with substrate analogs and affinity-labeling reagents
J. Biochem.
133
467-474
2003
Bacillus cereus (P36924), Bacillus cereus
Ye, Z.; Miyake, H.; Tatsumi, M.; Nishimura, S.; Nitta, Y.
Two additional carbohydrate-binding sites of beta-amylase from Bacillus cereus var. mycoides are involved in hydrolysis and raw starch-binding
J. Biochem.
135
355-363
2004
Bacillus cereus (P36924), Bacillus cereus
Sarikaya, E.; Higasa, T.; Adachi, M.; Mikami, B.
Comparison of degradation abilities of alpha- and beta-amylases on raw starch granules
Process Biochem.
35
711-715
2000
Bacillus cereus, Glycine max
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Hirata, A.; Adachi, M.; Utsumi, S.; Mikami, B.
Engineering of the pH optimum of Bacillus cereus beta-amylase: conversion of the pH optimum from a bacterial type to a higher-plant type
Biochemistry
43
12523-12531
2004
Bacillus cereus (P36924), Bacillus cereus
Niziolek, S.
beta-Amylase production by some Bacillus cereus, bacillus megaterium and Bacillus polymyxy strains
Acta Microbiol. Pol.
46
357-362
1997
Bacillus cereus, Paenibacillus polymyxa, Priestia megaterium
Gonzalez, A.; Wang, Y.; Staroszczyk, H.; Brownmiller, C.; Lee, S.
Effect of hydroxypropylation and beta-amylase treatment on complexation of debranched starch with naringenin
Starch Staerke
70
1700263
2018
Bacillus cereus (P36924)
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