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Glucokinase contributes to glucose phosphorylation in d-lactic acid production by Sporolactobacillus inulinus Y2-8

  • Genetics and Molecular Biology of Industrial Organisms
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Sporolactobacillus inulinus, a homofermentative lactic acid bacterium, is a species capable of efficient industrial d-lactic acid production from glucose. Glucose phosphorylation is the key step of glucose metabolism, and fine-tuned expression of which can improve d-lactic acid production. During growth on high-concentration glucose, a fast induction of high glucokinase (GLK) activity was observed, and paralleled the patterns of glucose consumption and d-lactic acid accumulation, while phosphoenolpyruvate phosphotransferase system (PTS) activity was completely repressed. The transmembrane proton gradient of 1.3–1.5 units was expected to generate a large proton motive force to the uptake of glucose. This suggests that the GLK pathway is the major route for glucose utilization, with the uptake of glucose through PTS-independent transport systems and phosphorylation of glucose by GLK in S. inulinus d-lactic acid production. The gene encoding GLK was cloned from S. inulinus and expressed in Escherichia coli. The amino acid sequence revealed significant similarity to GLK sequences from Bacillaceae. The recombinant GLK was purified and shown to be a homodimer with a subunit molecular mass of 34.5 kDa. Strikingly, it demonstrated an unusual broad substrate specificity, catalyzing phosphorylation of 2-deoxyglucose, mannitol, maltose, galactose and glucosamine, in addition to glucose. This report documented the key step concerning glucose phosphorylation of S. inulinus, which will help to understand the regulation of glucose metabolism and d-lactic acid production.

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Acknowledgments

We acknowledge the support of the National Basic Research Program of China (2011CB707400) and the National High Technology Research and Development Program of China (2011AA02A202). This work was also supported by the projects funded by PCSIRT, PAPD and CXZZ11_0358.

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Authors and Affiliations

  1. College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing, 211816, China

    Lu Zheng, Zhongzhong Bai & Bingfang He

  2. College of Pre-clinical Medicine, Nanjing University of Chinese Medicine, Nanjing, 210046, China

    Tingting Xu

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  1. Lu Zheng
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  2. Zhongzhong Bai
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  3. Tingting Xu
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  4. Bingfang He
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Correspondence to Bingfang He.

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Zheng, L., Bai, Z., Xu, T. et al. Glucokinase contributes to glucose phosphorylation in d-lactic acid production by Sporolactobacillus inulinus Y2-8. J Ind Microbiol Biotechnol 39, 1685–1692 (2012). https://doi.org/10.1007/s10295-012-1176-z

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  • DOI: https://doi.org/10.1007/s10295-012-1176-z

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