Molecular cloning, characterization, and application of a novel thermostable α-glucosidase from the hyperthermophilic archaeon Pyrobaculum aerophilum strain IM2

被引：10
作者：
Jeon, Hyeyeon [1 ,2 ]
Lee, Hyewon [1 ,2 ]
Byun, Dahye [1 ,2 ]
Choi, Hyejeong [1 ,2 ]
Shim, Jae-Hoon [1 ,2 ]
机构：
[1] Hallym Univ, Dept Food Sci & Nutr, Chunchon 200702, Gangwon, South Korea
[2] Hallym Univ, Ctr Aging & Hlth Care, Chunchon 200702, Gangwon, South Korea
来源：
FOOD SCIENCE AND BIOTECHNOLOGY | 2015年 / 24卷 / 01期
基金：
新加坡国家研究基金会;
关键词：
alpha-glucosidase; hyperthermophilic enzyme; Pyrobaculum aerophilum str. IM2; saccharification; thermostability; ACID-SEQUENCE SIMILARITIES; SULFOLOBUS-SOLFATARICUS; PYROCOCCUS-FURIOSUS; MALTOGENIC AMYLASE; DIRECTED EVOLUTION; STARCH; GLUCOAMYLASE; PURIFICATION; ENZYMES; GENE;
D O I：
10.1007/s10068-015-0024-0
中图分类号：
TS2 [食品工业];
学科分类号：
0832 ;
摘要：
A novel hyperthermostable alpha-glucosidase from the hyperthermophilic archaeon Pyrobaculum aerophilum str. IM2 (PAE1968) was successfully expressed in Escherichia coli and characterized. Recombinant PAE1968 was purified using Ni-NTA affinity chromatography to reveal a glycosidase with a predicted molecular weight (Mw) of 76 kDa. PAE1968 liberated glucose from the non-reducing ends of oligosaccharides. Multiple sequence alignment confirmed that the enzyme belonged to glycoside hydrolase (GH) family 31, and a kinetic study showed that the enzyme had a substrate preference for maltose (G2), indicating a Type-II alpha-glucosidase. The optimum operating conditions for the enzyme were 90A degrees C and pH 6.0, while the enzyme retained at least 80% of maximal activity at pH 4.5-7.5. Transglycosylation and reversion reactions of PAE1968 were of a lower magnitude than for a commercial saccharification enzyme, indicating that this enzyme can be used for glucose production in the starch industry.
引用
收藏
页码：175 / 182
页数：8
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