کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8326843 | 1540196 | 2018 | 35 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Purification, characterization and gene identification of a membrane-bound glucose dehydrogenase from 2-keto-d-gluconic acid industrial producing strain Pseudomonas plecoglossicida JUIM01
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
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چکیده انگلیسی
The membrane-bound glucose dehydrogenase (mGDH) is a rate-limiting enzyme for the industrial production of 2-keto-d-gluconic acid (2KGA) from glucose. In this study, mGDH was firstly purified from a 2KGA industrial producing strain Pseudomonas plecoglossicida JUIM01. The purified mGDH exhibited a specific activity of 16.85â¯U/mg and was identified as monomeric membrane-bound PQQ-dependent dehydrogenase with a molecular mass of ~87â¯kDa. The Km and Vmax value of d-glucose were 0.042â¯mM and 14.620â¯Î¼M/min, and the optimal pH and temperature were of 6.0 and 35â¯Â°C with favorable acid resistance and poor heat tolerance. Ca2+/Mg2+ showed a significantly positive effect on mGDH activity with 20% increase, whereas EDTA/EGTA had a negative influence, and Ca2+ was essential for enzyme activity. Furthermore, a 2412â¯bp-length gcd was amplified by genome walking technique and heterologously expressed in Escherichia coli. Bioinformatics analysis and heterologous expression further confirmed it as a mGDH encoding gene. mGDH contained binding sites of Ca2+, cofactor PQQ and polypeptide binding sites concluded from alignment results of mGDHs from different genera. This study would lay the foundation for improving 2KGA productivity through further strain modification.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: International Journal of Biological Macromolecules - Volume 118, Part A, 15 October 2018, Pages 534-541
Journal: International Journal of Biological Macromolecules - Volume 118, Part A, 15 October 2018, Pages 534-541
نویسندگان
Da-Ming Wang, Lei Sun, Wen-Jing Sun, Feng-Jie Cui, Jin-Song Gong, Xiao-Mei Zhang, Jin-Song Shi, Zheng-Hong Xu,