A Novel Thermostable β-Galactosidase from Geobacillus kaustophilus HTA42

doi:10.1007/s40242-014-4090-2

Chemical Research in Chinese Universities ›› 2014, Vol. 30 ›› Issue (5): 778-784.doi: 10.1007/s40242-014-4090-2

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A Novel Thermostable β-Galactosidase from Geobacillus kaustophilus HTA42

YU Shanshan¹, YIN Hongbing², WANG Xinying¹, FENG Li¹, XU Chunchun¹, LI Jing¹, HAN Hongxiang³, LIU Shuying^1,4

1. Jilin Ginseng Academy, Changchun 130117, P. R. China;
2. Affiliated Hospital, Changchun University of Chinese Medicine, Changchun 130117, P. R. China;
3. Jilin Agricultural University, Changchun 130118, P. R. China;
4. Changchun Center of Mass Spectrometry, Changchun Institute of Applied Chemistry, Chinese Academic of Science, Changchun 130022, P. R. China

Received:2014-03-17 Revised:2014-06-03 Online:2014-10-01 Published:2014-06-16
Contact: LIU Shuying E-mail:syliu19@ciac.ac.cn
Supported by:
Supported by the Scientific and Technological Poject of Jilin Province of China(Nos.20130101127JC, 20130303101YY).

Abstract

Abstract:

A novel thermostable β-galactosidase gene, designated as GkGal1A, from the thermophilic bacterium Geobacillus kaustophilus HTA426 was cloned and heterologously overexpressed in Escherichia coli(E. coli). Based on the sequence analysis, GkGal1A belongs to the glycosyl hydrolase family 1 that was the first β-galactosidase of bacterial origins expressed by us in this family. The apparent molecular weight of GkGal1A determined by sodium deodecyl sulfate-polyacrylamide gel electrophoresis is 52000. It exhibited the highest activity toward p-nitrophenyl-β-D-galactopyranoside at pH 7.8 and 70℃ and displayed high thermal stability. Divalent cations are prerequisite for the activity of GKGal1A, with the highest activity in the presence of Mn²⁺. Moreover, the three-dimensional structure of GkGal1A was modeled to speculate the structure of the catalytic residues and the reaction mechanism. The catalytic residues consisting of Glu166 and Glu355 were verified by site-directed mutagenesis.

Key words: Geobacillus kaustophilus HTA426, β-Galactosidase, Thermostability, Glycoside hydrolase

YU Shanshan, YIN Hongbing, WANG Xinying, FENG Li, XU Chunchun, LI Jing, HAN Hongxiang, LIU Shuying. A Novel Thermostable β-Galactosidase from Geobacillus kaustophilus HTA42[J]. Chemical Research in Chinese Universities, 2014, 30(5): 778-784.

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A Novel Thermostable β-Galactosidase from Geobacillus kaustophilus HTA42

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