Abstract
In order to achieve efficient homo L-lactic acid fermentation from xylose, we first carried out addition of xylose assimilation ability to Lactococcus lactis IL 1403 by introducing a plasmid carrying the xylRAB genes from L. lactis IO-1 (pXylRAB). Then modification of xylose assimilation pathway was carried out. L. lactis has two pathways for xylose assimilation called the phosphoketolase pathway (PK pathway) that produces both lactic acid and acetic acid and the pentose phosphate pathway (PP pathway) that produces only lactic acid as a final product. Thus a mutant strain that disrupted its phosphokeolase gene (ptk) was constructed. The Δptk mutant harboring pXylRAB lacked the PK pathway and produced predominantly lactic acid from xylose via the PP pathway, although its fermentation rate slightly decreased. Further introduction of the transketolase gene (tkt) to disrupted ptk locus led restoration of fermentation rate and this was attributed to enhancement of the PP pathway. As a result, ptk::tkt strain harboring pXylRAB produced 50.1 g/l of L-lactic acid from xylose with a high optical purity of 99.6% and a high yield of 1.58 (moles per mole xylose consumed) that is close to theoretical value of 1.67 from xylose.
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Acknowledgments
We are grateful to Dr. Emmanuelle Maguin for supplying the E. coli VE7108 and VE6838 (VE7108 containing pG+host9 plasmid) strains and to Meiji Dairies Corporation for supplying the pSECE1 plasmid. We are grateful to Dr. Eiichi Satoh for supplying the L. lactis IL1403 wild type strain. This work was supported in part by a Special Coordination Funds for Promoting Science and Technology, Creation of Innovation Centers for Advanced Interdisciplinary Research Areas (Innovative Bioproduction Kobe), MEXT, Japan.
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Shinkawa, S., Okano, K., Yoshida, S. et al. Improved homo l-lactic acid fermentation from xylose by abolishment of the phosphoketolase pathway and enhancement of the pentose phosphate pathway in genetically modified xylose-assimilating Lactococcus lactis . Appl Microbiol Biotechnol 91, 1537–1544 (2011). https://doi.org/10.1007/s00253-011-3342-z
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DOI: https://doi.org/10.1007/s00253-011-3342-z