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Enzyme Microb Technol. 2013 Dec 10;53(6-7):365-72. doi: 10.1016/j.enzmictec.2013.08.001. Epub 2013 Aug 16.

Significant improvement of thermal stability of glucose 1-dehydrogenase by introducing disulfide bonds at the tetramer interface.

Author information

1
Institute of Microbiology, College of Life Science, Zhejiang University, Hangzhou 310058, China. Electronic address: htding@outlook.com.

Abstract

Rational design was applied to glucose 1-dehydrogenase (LsGDH) from Lysinibacillus sphaericus G10 to improve its thermal stability by introduction of disulfide bridges between subunits. One out of the eleven mutants, designated as DS255, displayed significantly enhanced thermal stability with considerable soluble expression and high specific activity. It was extremely stable at pH ranging from 4.5 to 10.5, as it retained nearly 100% activity after incubating at different buffers for 1h. Mutant DS255 also exhibited high thermostability, having a half-life of 9900min at 50°C, which was 1868-fold as that of its wild type. Moreover, both of the increased free energy of denaturation and decreased entropy of denaturation of DS255 suggested that the enzyme structure was stabilized by the engineered disulfide bonds. On account of its robust stability, mutant DS255 would be a competitive candidate in practical applications of chiral chemicals synthesis, biofuel cells and glucose biosensors.

KEYWORDS:

BmGDH-IWG3; Disulfide bond; GDH; GDH from Bacillus megaterium IWG3; GDH from Lysinibacillus sphaericus G10; Glucose 1-dehydrogenase; LsGDH; NAD; NAD(P)-dependent glucose 1-dehydrogenase; NADP; Protein engineering; Protein stability; nicotinamide adenine dinucleotide; nicotinamide adenine dinucleotide phosphate

PMID:
24315638
DOI:
10.1016/j.enzmictec.2013.08.001
[Indexed for MEDLINE]
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