Applied Microbiology and Biotechnology

, Volume 85, Issue 3, pp 413–423 | Cite as

Biotechnological production of enantiomeric pure lactic acid from renewable resources: recent achievements, perspectives, and limits

  • Kenji Okano
  • Tsutomu Tanaka
  • Chiaki Ogino
  • Hideki Fukuda
  • Akihiko Kondo


Lactic acid (LA) is an important and versatile chemical that can be produced from renewable resources such as biomass. LA is used in the food, pharmaceutical, and polymers industries and is produced by microorganism fermentation; however, most microorganisms cannot directly utilize biomass such as starchy materials and cellulose. Here, we summarize LA production using several kinds of genetically modified microorganisms, such as LA bacteria, Escherichia coli, Corynebacterium glutamicum, and yeast. Using gene manipulation and metabolic engineering, the yield and optical purity of LA produced from biomass has been significantly improved. In this review, the drawbacks as well as improvements of LA production by fermentation is discussed.


Lactic acid fermentation Lactic acid bacteria Yeast Optically pure lactic acid 



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


  1. Adachi E, Torigoe M, Sugiyama M, Nikawa J, Shimizu K (1998) Modification of metabolic pathways of Saccharomyces cerevisiae by the expression of lactate dehydrogenase and deletion of pyruvate decarboxylase genes for the lactic acid fermentation at low pH value. J Ferment Bioeng 86:284–289CrossRefGoogle Scholar
  2. Adsul M, Khire J, Bastawde K, Gokhale D (2007) Production of lactic acid from cellobiose and cellotriose by Lactobacillus delbrueckii mutant Uc-3. Appl Environ Microbiol 73:5055–5057CrossRefGoogle Scholar
  3. Bianchi MM, Brambilla L, Protani F, Liu CL, Lievense J, Porro D (2001) Efficient homolactic fermentation by Kluyveromyces lactis strains defective in pyruvate utilization and transformed with the heterologous LDH gene. Appl Envion Microbiol 67:5621–5625CrossRefGoogle Scholar
  4. 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 21:793–798CrossRefGoogle Scholar
  5. 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 64:4720–4728Google Scholar
  6. Chang DE, Jung HC, Rhee JS, Pan JG (1999) Homofermentative production of D- or L-lactate in metabolically engineered Escherichia coli RR1. Appl Environ Microbiol 65:1384–1389Google Scholar
  7. Christensen CH, Rass-Hansen J, Marsden CC, Taarning E, Egeblad K (2008) The renewable chemicals industry. ChemSusChem 1:283–289CrossRefGoogle Scholar
  8. Dequin S, Barre P (1994) Mixed lactic acid-alcoholic fermentation by Saccharomyces cerevisiae expressing the Lactobacillus casei L(+)-LDH. Biotechnology 12:173–177CrossRefGoogle Scholar
  9. Dequin S, Baptista E, Barre P (1999) Acidification of grape musts by Saccharomyces cerevisiae wine yeast strains genetically engineered to produce lactic acid. Am J Enol Vitic 50:45–50Google Scholar
  10. Dodds DR, Gross RA (2007) Chemicals from biomass. Science 318:1250–1251CrossRefGoogle Scholar
  11. Gao MT, Shimamura T, Ishida N, Takahashi H (2009) Application of metabolically engineered Saccharomyces cerevisiae to extractive lactic acid fermentation. Biochem Eng J 44:251–255CrossRefGoogle Scholar
  12. Giraud E, Champailler A, Raimbault M (1994) Degradation of raw starch by a wild amylolytic strain of Lactobacillus plantarum. Appl Environ Microbiol 60:4319–4323Google Scholar
  13. Guyot JP, Calderon M, Morlon-Guyot J (2000) Effect of pH control on lactic acid fermentation of starch by Lactobacillus manihotivorans LMG 18010T. J Appl Microbiol 88:176–182CrossRefGoogle Scholar
  14. Helanto M, Kiviharju K, Leisola M, Nyyssölä A (2007) Metabolic engineering of Lactobacillus plantarum for production of L-ribulose. Appl Environ Microbiol 73:7083–7091CrossRefGoogle Scholar
  15. Hermann T (2003) Industrial production of amino acids by coryneform bacteria. J Biotechnol 104:155–172CrossRefGoogle Scholar
  16. Hofvendahl K, Hahn-Hägerdal B (1997) L-lactic acid production from whole wheat flour hydrolysate using strains of Lactobacilli and Lactococci. Enzyme Microb Technol 20:301–307CrossRefGoogle Scholar
  17. Hofvendahl K, Hahn-Hägerdal B (2000) Factors affecting the fermentative lactic acid production from renewable resources. Enzyme Microb Technol 26:87–107CrossRefGoogle Scholar
  18. Hohmann S (1991) Characterization of PDC6, a third structural gene for pyruvate decarboxylase in Saccharomyces cerevisiae. J Bacteriol 173:7963–7969Google Scholar
  19. Hohmann S, Cederberg H (1990) Autoregulation may control the expression of yeast pyruvate decarboxylase structural genes PDC1 and PDC5. Eur J Biochem 188:615–621CrossRefGoogle Scholar
  20. Ikada Y, Jamshidi K, Tsuji H, Hyon SH (1987) Stereocomplex formation between enantiomeric poly (lactides). Macromolecules 20:904–906CrossRefGoogle Scholar
  21. Ishida N, Saitoh S, Tokuhiro K, Nagamori E, Matsuyama T, Kitamoto K, Takahashi H (2005) Efficient production of L-lactic acid by metabolically engineered Sacchromyces cerevisiae with a genome-integrated L-lactate dehydrogenase gene. Appl Environ Microbiol 71:1964–1970CrossRefGoogle Scholar
  22. Ishida N, Saitoh S, Onishi T, Tokuhiro K, Nagamori E, Kitamoto K, Takahashi H (2006a) The effect of pyruvate decarboxylase gene knockout in Sacchromyces cerevisiae on L-lactic acid production. Biosci Biotechnol Biochem 70:1148–1153CrossRefGoogle Scholar
  23. Ishida N, Suzuki T, Tokuhiro K, Nagamori E, Onishi T, Saitoh S, Kitamoto K, Takahashi H (2006b) D-lactic acid production by metabolically engineered Sacchromyces cerevisiae. J Biosci Bioeng 101:172–177CrossRefGoogle Scholar
  24. Kalinowski J, Bathe B, Bartels D, Bischoff N, Bott M, Burkovski A, Dusch N, Eggeling L, Eikmanns BJ, Gaigalat L, Goesmann A, Hartmann M, Huthmacher K, Krämer R, Linke B, McHardy AC, Meyer F, Mockel B, Pfefferle W, Pühler A, Rey DA, Ruckert C, Rupp O, Sahm H, Wendisch VF, Wiegrabe I, Tauch A (2003) The complete Corynebacterium glutamicum ATCC 13032 genome sequence and its impact on the production of L-aspartate-derived amino acids and vitamins. J Biotechnol 104:5–25CrossRefGoogle Scholar
  25. Katahira S, Mizuike A, Fukuda H, Kondo A (2006) Ethanol fermentation from lignocellulosic hydrolysate by a recombinant xylose- and cellooligosaccharide-assimilating yeast strain. Appl Microbiol Biotechnol 72:1136–1143CrossRefGoogle Scholar
  26. Kawaguchi T, Enoki T, Tsurumaki S, Sumitani J, Ueda M, Ooi T, Arai M (1996) Cloning and sequencing of the cDNA encoding β-glucosidase 1 from Aspergillus aculeatus. Gene 173:287–288CrossRefGoogle Scholar
  27. Kawaguchi H, Vertès AA, Okino S, Inui M, Yukawa H (2006) Engineering of a xylose metabolic pathway in Corynebacterium glutamicum. Appl Environ Microbiol 72:3418–3428CrossRefGoogle Scholar
  28. Kawaguchi H, Sasaki M, Vertès AA, Inui M, Yukawa H (2008) Engineering of an L-arabinose metabolic pathway in Corynebacterium glutamicum. Appl Microbiol Biotechnol 77:1053–1062CrossRefGoogle Scholar
  29. Kotrba P, Inui M, Yukawa H (2001) The ptsI gene encoding enzyme I of the phosphotransferase system of Corynebacterium glutamicum. Biochem Biophys Res Commun 289:1307–1313CrossRefGoogle Scholar
  30. Kotrba P, Inui M, Yukawa H (2003) A single V317A or V317M substitution in Enzyme II of a newly identified β-glucoside phosphotransferase and utilization system of Corynebacterium glutamicum R extends its specificity towards cellobiose. Microbiology 149:1569–1580CrossRefGoogle Scholar
  31. Leskovac V, Trivić S, Pericin D (2002) The three zinc-containing alcohol dehydrogenases from baker’s yeast, Saccharomyces cerevisiae. FEMS Yeast Res 2:481–494Google Scholar
  32. Leuchtenberger W, Huthmacher K, Drauz K (2005) Biotechnological production of amino acids and derivatives: current status and prospects. Appl Microbiol Biotechnol 69:1–8CrossRefGoogle Scholar
  33. Litchfield JH (1996) Microbiological production of lactic acid. Adv Appl Microbiol 42:45–95CrossRefGoogle Scholar
  34. Matsui Y, Okada S, Uchimura T, Kondo A, Satoh E (2007) Determination and analysis of the starch binding domain of Streptococcus bovis 148 raw-starch-hydrolyzing α-amylase. J Appl Glycosci 54:217–222Google Scholar
  35. Narita J, Nakahara S, Fukuda H, Kondo A (2004) Efficient production of L-(+)-lactic acid from raw starch by Streptococcus bovis 148. J Biosci Bioeng 97:423–425Google Scholar
  36. Ohara H, Owaki M, Sonomoto K (2006) Xylooligosaccharide fermentation with Leuconostoc lactis. J Biosci Bioeng 101:415–420CrossRefGoogle Scholar
  37. 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 75:1007–1013CrossRefGoogle Scholar
  38. Okano K, Yoshida S, Tanaka T, Fukuda H, Kondo A (2009a) 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(15):5175–5178Google Scholar
  39. Okano K, Zhang Q, Shinkawa S, Yoshida S, Tanaka T, Fukuda H, Kondo A (2009b) Efficient production of optically pure D-lactic acid from raw corn starch by using genetically modified L-lactate dehydrogenase gene-deficient and α-amylase-secreting Lactobacillus plantarum strain. Appl Environ Microbiol 75:462–467CrossRefGoogle Scholar
  40. Okano K, Zhang Q, Yoshida S, Tanaka T, Ogino C, Fukuda H, Kondo A (2009c) D-Lactic acid production from cellooligosaccharides and β-glucan using L-LDH gene-deficient and endoglucanase-secreting Lactobacillus plantarum. Appl Microbiol Biotechnol (in press)Google Scholar
  41. Okino S, Inui M, Yukawa H (2005) Production of organic acids by Corynebacterium glutamicum under oxygen deprivation. Appl Microbiol Biotechnol 68:475–480CrossRefGoogle Scholar
  42. Okino S, Suda M, Fujikura K, Inui M, Yukawa H (2008) Production of D-lactic acid by Corynebacterium glutamicum under oxygen deprivation. Appl Microbiol Biotechnol 78:449–454CrossRefGoogle Scholar
  43. Porro D, Brambilla L, Ranzi BM, Martegani E, Alberghina L (1995) Development of metabolically engineered Saccharomyces cerevisiae cells for the production of lactic acid. Biotechnol Prog 11:294–298CrossRefGoogle Scholar
  44. Saitoh S, Mieno Y, Nagashima T, Kumagai C, Kitamoto K (1996) Breeding of a new type of baker’s yeast by δ-integration for overproduction of glucoamylase using a homothallic yeast. J Ferment Bioeng 81:98–103CrossRefGoogle Scholar
  45. Saitoh S, Ishida N, Onishi T, Tokuhiro K, Nagamori E, Kitamoto K, Takahashi H (2005) Genetically engineered wine yeast produces a high concentration of L-lactic acid of extremely high optical purity. Appl Environ Microbiol 71:2789–2792CrossRefGoogle Scholar
  46. Sasaki M, Jojima T, Inui M, Yukawa H (2008) Simultaneous utilization of D-cellobiose, D-glucose, and D-xylose by recombinant Corynebacterium glutamicum under oxygen-deprived conditions. Appl Microbiol Biotechnol 81:691–699CrossRefGoogle Scholar
  47. Satoh E, Niimura Y, Uchimura T, Kozaki M, Komagata K (1993) Molecular cloning and expression of two α-amylase genes from Streptococcus bovis 148 in Escherichia coli. Appl Environ Microbiol 59:3669–3673Google Scholar
  48. Serror P, Sasaki T, Ehrlich SD, Maguin E (2002) Electrotransformation of Lactobacillus delbrueckii subsp. bulgaricus and L. delbrueckii subsp. lactis with various plasmids. Appl Environ Microbiol 68:46–52CrossRefGoogle Scholar
  49. Skory CD (2003) Lactic acid production by Saccharomyces cerevisiae expressing a Rhizopus oryzae lactate dehydrogenase gene. J Ind Microbiol Biotechnol 30:22–27Google Scholar
  50. 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 60:160–167CrossRefGoogle Scholar
  51. Tarmy EM, Kaplan NO (1968) Kinetics of Escherichia coli B D-lactate dehydrogenase and evidence for pyruvate controlled change in conformation. J Biol Chem 243:2587–2596Google Scholar
  52. Tateno T, Fukuda H, Kondo A (2007a) Production of L-lysine from starch by Corynebacterium glutamicum displaying α-amylase on its cell surface. Appl Microbiol Biotechnol 74:1213–1220CrossRefGoogle Scholar
  53. Tateno T, Fukuda H, Kondo A (2007b) Direct production of L-lysine from raw corn starch by Corynebacterium glutamicum secreting Streptococcus bovis α-amylase using cspB promoter and signal sequence. Appl Microbiol Biotechnol 77:533–541CrossRefGoogle Scholar
  54. Tokuhiro K, Ishida N, Kondo A, Takahashi H (2008) Lactic fermentation of cellobiose by a yeast strain displaying β-glucosidase on the cell surface. Appl Microbiol Biotechnol 79:481–488CrossRefGoogle Scholar
  55. Tokuhiro K, Ishida N, Nagamori E, Saitoh S, Onishi T, Kondo A, Takahashi H (2009) Double mutation of the PDC1 and ADH1 genes improves lactate production in the yeast Saccharomyces cerevisiae expressing the bovine lactate dehydrogenase gene. Appl Microbiol Biotechnol 82:883–890CrossRefGoogle Scholar
  56. Wee YJ, Kim JN, Ryu HW (2006) Biotechnological production of lactic acid and its recent applications. Food Technol Biotechnol 44:163–172Google Scholar
  57. Wisselink HW, Toirkens MJ, del RF BM, Winkler AA, van Dijken JP, Pronk JT, van Maris AJA (2007) Engineering of Saccharomyces cerevisiae for efficient anaerobic alcoholic fermentation of L-arabinose. Appl Environ Microbiol 73:4881–4891CrossRefGoogle Scholar
  58. Yáñez R, Moldes AB, Alonso JL, Parajó JC (2003) Production of D(-)-lactic acid from cellulose by simultaneous saccharification and fermentation using Lactobacillus coryniformis subsp. torquens. Biotechnol Lett 25:1161–1164CrossRefGoogle Scholar
  59. Yoon HH (1997) Simultaneous saccharification and fermentation of cellulose for lactic acid production. Biotechnol Bioprocess Eng 2:101–104CrossRefGoogle Scholar
  60. Yukawa H, Omumasaba CA, Nonaka H, Kos P, Okai N, Suzuki N, Suda M, Tsuge Y, Watanabe J, Ikeda Y, Vertès AA, Inui M (2007) Comparative analysis of the Corynebacterium glutamicum group and complete genome sequence of strain R. Microbiology 153:1042–1058CrossRefGoogle Scholar
  61. Zhou S, Causey TB, Hasona A, Shanmugam KT, Ingram LO (2003a) Production of optically pure D-lactic acid in mineral salts medium by metabolically engineered Escherichia coli W3110. Appl Environ Microbiol 69:399–407CrossRefGoogle Scholar
  62. Zhou S, Shanmugam KT, Ingram LO (2003b) Functional replacement of the Escherichia coli D-(-)-lactate dehydrogenase gene (ldhA) with the L-(+)-lactate dehydrogenase gene (ldhL) from Pediococcus acidilactici. Appl Environ Microbiol 69:2237–2244CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Kenji Okano
    • 1
  • Tsutomu Tanaka
    • 2
  • Chiaki Ogino
    • 3
  • Hideki Fukuda
    • 2
  • Akihiko Kondo
    • 3
  1. 1.Department of Molecular Science and Material Engineering, Graduate School of Science and TechnologyKobe UniversityKobeJapan
  2. 2.Organization of Advanced Science and TechnologyKobe UniversityKobeJapan
  3. 3.Department of Chemical Science and Engineering, Graduate School of EngineeringKobe UniversityKobeJapan

Personalised recommendations