Alkaline active maltohexaose-forming α-amylase from Bacillus halodurans LBK 34
(2005) In Enzyme and Microbial Technology 36(1). p.139-146- Abstract
- The gene encoding Amy 34, a maltohexaose-forming α-amylase from Bacillus halodurans LBK 34 isolated from Lake Bogoria, Kenya, was cloned and sequenced. The mature peptide consists of 958 amino acids with a theoretical molecular weight of 107.2 kDa and pI 4.41, respectively. The gene was expressed in Escherichia coli and the recombinant enzyme purified to homogeneity by a combination of metal chelate affinity and size exclusion chromatography. The pure enzyme exhibited optimum activity at 60 °C and pH 10.5–11.5. The enzyme retained over 60% activity after incubation at 55 °C for 4 h and was most stable at pH 9.0. Complete inhibition of enzyme activity was observed in presence of 5 mM Cu2+, Fe2+, Fe3+, Mn2+ and 5 mM EDTA. The enzyme... (More)
- The gene encoding Amy 34, a maltohexaose-forming α-amylase from Bacillus halodurans LBK 34 isolated from Lake Bogoria, Kenya, was cloned and sequenced. The mature peptide consists of 958 amino acids with a theoretical molecular weight of 107.2 kDa and pI 4.41, respectively. The gene was expressed in Escherichia coli and the recombinant enzyme purified to homogeneity by a combination of metal chelate affinity and size exclusion chromatography. The pure enzyme exhibited optimum activity at 60 °C and pH 10.5–11.5. The enzyme retained over 60% activity after incubation at 55 °C for 4 h and was most stable at pH 9.0. Complete inhibition of enzyme activity was observed in presence of 5 mM Cu2+, Fe2+, Fe3+, Mn2+ and 5 mM EDTA. The enzyme displayed 80% of its original activity in presence of 1% (w/v) SDS and was stable in presence of up to 5 mM DTT. Maltohexaose (G6) was the main initial product of starch hydrolysis while other products formed were G4 > G2 > G5 > G3 and G1. The main end product of the enzyme's action on amylose, amylopectin and maltodextrin is maltotetraose. Amy 34 could not hydrolyse pullulan, α and β-cyclodextrin but could hydrolyse γ-cyclodextrin to produce glucose, maltose and maltotetraose. Maltotetraose was the smallest α-(1–4) linked maltooligosaccharide that could be hydrolysed by the enzyme. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/141629
- author
- Hashim, Suhaila LU ; Delgado, Osvaldo LU ; Martinez, Alejandra LU ; Hatti-Kaul, Rajni LU ; Mulaa, Francis J and Mattiasson, Bo LU
- organization
- publishing date
- 2005
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- B. halodurans, alkaliphile, maltohexaose, amylase
- in
- Enzyme and Microbial Technology
- volume
- 36
- issue
- 1
- pages
- 139 - 146
- publisher
- Elsevier
- external identifiers
-
- wos:000226195800018
- scopus:9944259026
- ISSN
- 0141-0229
- DOI
- 10.1016/j.enzmictec.2004.07.017
- language
- English
- LU publication?
- yes
- id
- 9c657bb0-0bda-4bbd-8f6a-bd98e9b5f3b7 (old id 141629)
- date added to LUP
- 2016-04-01 11:36:44
- date last changed
- 2022-01-26 07:37:30
@article{9c657bb0-0bda-4bbd-8f6a-bd98e9b5f3b7,
abstract = {{The gene encoding Amy 34, a maltohexaose-forming α-amylase from Bacillus halodurans LBK 34 isolated from Lake Bogoria, Kenya, was cloned and sequenced. The mature peptide consists of 958 amino acids with a theoretical molecular weight of 107.2 kDa and pI 4.41, respectively. The gene was expressed in Escherichia coli and the recombinant enzyme purified to homogeneity by a combination of metal chelate affinity and size exclusion chromatography. The pure enzyme exhibited optimum activity at 60 °C and pH 10.5–11.5. The enzyme retained over 60% activity after incubation at 55 °C for 4 h and was most stable at pH 9.0. Complete inhibition of enzyme activity was observed in presence of 5 mM Cu2+, Fe2+, Fe3+, Mn2+ and 5 mM EDTA. The enzyme displayed 80% of its original activity in presence of 1% (w/v) SDS and was stable in presence of up to 5 mM DTT. Maltohexaose (G6) was the main initial product of starch hydrolysis while other products formed were G4 > G2 > G5 > G3 and G1. The main end product of the enzyme's action on amylose, amylopectin and maltodextrin is maltotetraose. Amy 34 could not hydrolyse pullulan, α and β-cyclodextrin but could hydrolyse γ-cyclodextrin to produce glucose, maltose and maltotetraose. Maltotetraose was the smallest α-(1–4) linked maltooligosaccharide that could be hydrolysed by the enzyme.}},
author = {{Hashim, Suhaila and Delgado, Osvaldo and Martinez, Alejandra and Hatti-Kaul, Rajni and Mulaa, Francis J and Mattiasson, Bo}},
issn = {{0141-0229}},
keywords = {{B. halodurans; alkaliphile; maltohexaose; amylase}},
language = {{eng}},
number = {{1}},
pages = {{139--146}},
publisher = {{Elsevier}},
series = {{Enzyme and Microbial Technology}},
title = {{Alkaline active maltohexaose-forming α-amylase from Bacillus halodurans LBK 34}},
url = {{http://dx.doi.org/10.1016/j.enzmictec.2004.07.017}},
doi = {{10.1016/j.enzmictec.2004.07.017}},
volume = {{36}},
year = {{2005}},
}