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Homo-d-lactic acid production from mixed sugars using xylose-assimilating operon-integrated Lactobacillus plantarum

Abstract

In order to achieve efficient d-lactic acid fermentation from a mixture of xylose and glucose, the xylose-assimilating xylAB operon from Lactobacillus pentosus (PXylAB) was introduced into an l-lactate dehydrogenase gene (ldhL1)-deficient Lactobacillus plantarumldhL1-xpk1::tktxpk2) strain in which the phosphoketolase 1 gene (xpk1) was replaced with the transketolase gene (tkt) from Lactococcus lactis, and the phosphoketolase 2 (xpk2) gene was deleted. Two copies of xylAB introduced into the genome significantly improved the xylose fermentation ability, raising it to the same level as that of ΔldhL1-xpk1::tktxpk2 harboring a xylAB operon-expressing plasmid. Using the two-copy xylAB integrated strain, successful homo-d-lactic acid production was achieved from a mixture of 25 g/l xylose and 75 g/l glucose without carbon catabolite repression. After 36-h cultivation, 74.2 g/l of lactic acid was produced with a high yield (0.78 g per gram of consumed sugar) and an optical purity of d-lactic acid of 99.5%. Finally, we successfully demonstrated homo-d-lactic acid fermentation from a mixture of three kinds of sugar: glucose, xylose, and arabinose. This is the first report that describes homo-d-lactic acid fermentation from mixed sugars without carbon catabolite repression using the xylose-assimilating pathway integrated into lactic acid bacteria.

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References

  1. Bustos G, Moldes AB, Cruz JM, Domínguez JM (2005) Influence of the metabolism pathway on lactic acid production from hemicellulosic trimming vine shoots hydrolyzates using Lactobacillus pentosus. Biotechnol Prog 21:793–798

  2. Chaillou S, Bor YC, Batt CA, Postma PW, Pouwels PH (1998) Molecular cloning and functional expression in Lactobacillus plantarum 80 of xylT, encoding the d-xylose-H+ symporter of Lactobacillus brevis. Appl Environ Microbiol 64:4720–4728

  3. Cook GM, Janssen PH, Morgan HW (1993) Simultaneous uptake and utilization of glucose and xylose by Clostridium thermohydrosulfuricum. FEMS Microbiol Lett 109:55–61

  4. Deutscher J (2008) The mechanisms of carbon catabolite repression in bacteria. Curr Opin Microbiol 11:87–93

  5. Deutscher J, Francke C, Postma PW (2006) How phosphotransferase system-related protein phosphorylation regulates carbohydrate metabolism in bacteria. Microbiol Mol Biol Rev 70:939–1031

  6. Gorke B, Stulke J (2008) Carbon catabolite repression in bacteria: many ways to make the most out of nutrients. Nat Rev Microbiol 6:613–624

  7. Ho NW, Chen Z, Brainard AP (1998) Genetically engineered Saccharomyces yeast capable of effective cofermentation of glucose and xylose. Appl Environ Microbiol 64:1852–1859

  8. Hofvendahl K, Hahn-Haegerdal B (2000) Factors affecting the fermentative lactic acid production from renewable resources. Enz Microb Technol 26:87–107

  9. Ikada Y, Jamshidi K, Tsuji H, Hyon SH (1987) Stereocomplex formation between enantiomeric poly (lactides). Macromolecules 20:904–906

  10. Kastner JR, Jones WJ, Roberts RS (1998) Simultaneous utilization of glucose and d-xylose by Candida shehatae in a chemostat. J Ind Microbiol Biotechnol 20:339–343

  11. Kennes C, Veiga MC, Dubourguier HC, Touzel JP, Albagnac G, Naveau H, Nyns EJ (1991) Trophic relationships between Saccharomyces cerevisiae and Lactobacillus plantarum and their metabolism of glucose and citrate. Appl Environ Microbiol 57:1046–1051

  12. Kim JH, Shoemaker SP, Mills DA (2009) Relaxed control of sugar utilization in Lactobacillus brevis. Microbiology 155:1351–1359

  13. Kim JH, Block DE, Mills DA (2010) Simultaneous consumption of pentose and hexose sugars: an optimal microbial phenotype for efficient fermentation of lignocellulosic biomass. Appl Microbiol Biotechnol 88:1077–1085

  14. Lawford HG, Rousseau JD (2002) Performance testing of Zymomonas mobilis metabolically engineered for cofermentation of glucose, xylose, and arabinose. Appl Biochem Biotechnol 98:429–448

  15. Lokman BC, van Santen P, Verdoes JC, Krüse J, Leer RJ, Posno M, Pouwels PH (1991) Organization and characterization of three genes involved in d-xylose catabolism in Lactobacillus pentosus. Mol Gen Genet 230:161–169

  16. Maheshwari R, Balasubramanyam PV (1988) Simultaneous utilization of glucose and sucrose by thermophilic fungi. J Bacteriol 170:3274–3280

  17. Mahr K, Hillen W, Titgemeyer F (2000) Carbon catabolite repression in Lactobacillus pentosus: analysis of the ccpA region. Appl Environ Microbiol 66:277–283

  18. Marasco R, Muscariello L, Varcamonti M, De Felice M, Sacco M (1998) Expression of the bglH gene of Lactobacillus plantarum is controlled by carbon catabolite repression. J Bacteriol 180:3400–3404

  19. Morel F, Frot-Coutaz J, Aubel D, Portalier R, Atlan D (1999) Characterization of a prolidase from Lactobacillus delbrueckii subsp. bulgaricus CNRZ 397 with an unusual regulation of biosynthesis. Microbiology 145:437–446

  20. Narita J, Okano K, Kitao T, Ishida S, Sewaki T, Sung MH, Fukuda H, Kondo A (2006) Display of α-amylase on the surface of Lactobacillus casei cells by use of the PgsA anchor protein, and production of lactic acid from starch. Appl Environ Microbiol 72:269–275

  21. Ohara H, Owaki M, Sonomoto K (2007) Calculation of metabolic flow of xylose in Lactococcus lactis. J Biosci Bioeng 103:92–94

  22. Okano K, Kimura S, Narita J, Fukuda H, Kondo A (2007) Improvement in lactic acid production from starch using α-amylase-secreting Lactococcus lactis cells adapted to maltose or starch. Appl Microbiol Biotechnol 75:1007–1013

  23. Okano K, Zhang Q, Shinkawa S, Yoshida S, Tanaka T, Fukuda H, Kondo A (2009a) Efficient production of optically pure d-lactic acid from raw corn starch using genetically modified l-lactate dehydrogenase gene-deficient and α-amylase-secreting Lactobacillus plantarum. Appl Environ Microbiol 75:462–467

  24. Okano K, Yoshida S, Yamada R, Tanaka T, Ogino C, Fukuda H, Kondo A (2009b) Improved production of homo-d-lactic acid via xylose fermentation by introduction of xylose assimilation genes and redirection of the phosphoketolase pathway to the pentose phosphate pathway in l-lactate dehydrogenase gene-deficient Lactobacillus plantarum. Appl Environ Microbiol 75:7858–7861

  25. Okano K, Yoshida S, Tanaka T, Fukuda H, Kondo A (2009c) Homo d-lactic acid fermentation from arabinose by redirection of phosphoketolase pathway to pentose phosphate pathway in l-lactate dehydrogenase gene-deficient Lactobacillus plantarum. Appl Environ Microbiol 75:5175–5178

  26. Ramos A, Jordan KN, Cogan TM, Santos H (1994) 13 C nuclear magnetic resonance studies of citrate and glucose cometabolism by Lactococcus lactis. Appl Environ Microbiol 60:1739–1748

  27. Saier MH (1998) Multiple mechanisms controlling carbon metabolism in bacteria. Biotechnol Bioeng 58:170–174

  28. Schick J, Weber B, Klein JR, Henrich B (1999) PepR1, a CcpA-like transcription regulator of Lactobacillus delbrueckii subsp. lactis. Microbiology 145:3147–3154

  29. Stulke J, Hillen W (1999) Carbon catabolite repression in bacteria. Curr Opin Microbiol 2:195–201

  30. Tanaka K, Komiyama A, Sonomoto K, Ishizaki A, Hall SJ, Stanbury PF (2002) Two different pathways for d-xylose metabolism and the effect of xylose concentration on the yield coefficient of l-lactate in mixed-acid fermentation by the lactic acid bacterium Lactococcus lactis IO-1. Appl Microbiol Biotechnol 60:160–167

  31. Titgemeyer F, Hillen W (2002) Global control of sugar metabolism: a gram-positive solution. Antonie Van Leeuwenhoek 82:59–71

  32. Veyrat A, Monedero V, Perez-Martinez G (1994) Glucose transport by the phosphoenolpyruvate: mannose phosphotransferase system in Lactobacillus casei ATCC 393 and its role in carbon catabolite repression. Microbiology 140:1141–1149

  33. Viana R, Monedero V, Dossonnet V, Vadeboncoeur C, Perez-Martinez G, Deutscher J (2000) Enzyme I and HPr from Lactobacillus casei: their role in sugar transport, carbon catabolite repression and inducer exclusion. Mol Microbiol 36:570–584

  34. Yamada R, Tanaka T, Ogino C, Kondo A (2010) Gene copy number and polyploidy on products formation in yeast. Appl Microbiol Biotechnol 88:849–857

  35. Zhang M, Eddy C, Deanda K, Finkelstein M, Picataggio S (1995) Metabolic engineering of a pentose metabolism pathway in ethanologenic Zymomonas mobilis. Science 267:240–243

  36. Zuniga M, Champomier-Verges M, Zagorec M, Perez-Martinez G (1998) Structural and functional analysis of the gene cluster encoding the enzymes of the arginine deiminase pathway of Lactobacillus sake. J Bacteriol 18:4154–4159

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Acknowledgments

This work was supported by Special Coordination Funds for Promoting Science and Technology, Creation of Innovation Centers for Advanced Interdisciplinary Research Areas (Innovative Bioproduction Kobe), MEXT, Japan.

Conflicts of interest

The authors declare that they have no conflicts of interest.

Author information

Correspondence to Akihiko Kondo.

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Yoshida, S., Okano, K., Tanaka, T. et al. Homo-d-lactic acid production from mixed sugars using xylose-assimilating operon-integrated Lactobacillus plantarum . Appl Microbiol Biotechnol 92, 67–76 (2011). https://doi.org/10.1007/s00253-011-3356-6

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Keywords

  • Lactic acid
  • Co-fermentation
  • Xylose
  • Fermentation