Production of optically pure l(+)-lactic acid from waste plywood chips using an isolated thermotolerant Enterococcus faecalis SI at a pilot scale

  • Shuo-Fu Yuan
  • Teng-Chieh Hsu
  • Chun-An Wang
  • Ming-Feng Jang
  • Yang-Cheng Kuo
  • Hal S. AlperEmail author
  • Gia-Luen GuoEmail author
  • Wen-Song Hwang
Fermentation, Cell Culture and Bioengineering - Original Paper


Utilization of renewable and low-cost lignocellulosic wastes has received major focus in industrial lactic acid production. The use of high solid loadings in biomass pretreatment potentially offers advantages over low solid loadings including higher lactic acid concentration with decreased production and capital costs. In this study, an isolated Enterococcus faecalis SI with optimal temperature 42 °C was used to produce optically pure l-lactic acid (> 99%) from enzyme-saccharified hydrolysates of acid-impregnated steam explosion (AISE)-treated plywood chips. The l-lactic acid production increased by 10% at 5 L scale compared to the similar fermentation scheme reported by Wee et al. The fermentation with a high solid loading of 20% and 35% (w/v) AISE-pretreated plywood chips had been successfully scaled up to process development unit scale (100 L) and pilot scale (9 m3), respectively. This is the first report of pilot-scale lignocellulosic lactic acid fermentation by E. faecalis with high lactic acid titer (nearly 92 g L−1) and yield (0.97 kg kg−1). Therefore, large-scale l-lactic acid production by E. faecalis SI shows the potential application for industries.


Enterococcus faecalis l-Lactic acid Optical purity Plywood chips Pilot scale 



This study was supported by the National Energy Program-Phase II (Industry promotion platform for cellulosic ethanol with development of value-added biorefinery technology) from the government of Taiwan. The authors thank Mrs. Chun-Mei Huang for her technical support with analyzing the samples. The authors thank Dr. Wen-Hua Chen and her team for their assistance with the pretreatment tests.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10295_2018_2078_MOESM1_ESM.pdf (218 kb)
Supplementary material 1 (PDF 217 kb)


  1. 1.
    Agrawal R, Satlewal A, Kapoor M, Mondal S, Basu B (2017) Investigating the enzyme-lignin binding with surfactants for improved saccharification of pilot scale pretreated wheat straw. Bioresour Technol 224:411–418. CrossRefPubMedGoogle Scholar
  2. 2.
    Chafran LS, Campos JMC, Santos JS, Sales MJA, Dias SCL, Dias JA (2016) Synthesis of poly(lactic acid) by heterogeneous acid catalysis from d, l-lactic acid. J Polym Res 23:107. CrossRefGoogle Scholar
  3. 3.
    Chen W-H, Tsai C-C, Lin C-F, Tsai P-Y, Hwang W-S (2013) Pilot-scale study on the acid-catalyzed steam explosion of rice straw using a continuous pretreatment system. Bioresour Technol 128:297–304. CrossRefPubMedGoogle Scholar
  4. 4.
    Christopher LP, Kapatral V, Vaisvil B, Emel G, DeVeaux LC (2014) Draft genome sequence of a new homofermentative, lactic acid-producing Enterococcus faecalis isolate, CBRD01. Genome Announc 2:e00147–e00214. CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Eriksson T, Börjesson J, Tjerneld F (2002) Mechanism of surfactant effect in enzymatic hydrolysis of lignocellulose. Enzyme Microb Technol 31:353–364. CrossRefGoogle Scholar
  6. 6.
    Gupta A, Verma JP (2015) Sustainable bio-ethanol production from agro-residues: a review. Renew Sustain Energy Rev 41:550–567. CrossRefGoogle Scholar
  7. 7.
    Isikgor FH, Becer CR (2015) Lignocellulosic biomass: a sustainable platform for the production of bio-based chemicals and polymers. Polym Chem 6:4497–4559. CrossRefGoogle Scholar
  8. 8.
    Jin W, Chen L, Hu M, Sun D, Li A, Li Y, Hu Z, Zhou S, Tu Y, Xia T, Wang Y, Xie G, Li Y, Bai B, Peng L (2016) Tween-80 is effective for enhancing steam-exploded biomass enzymatic saccharification and ethanol production by specifically lessening cellulase absorption with lignin in common reed. Appl Energy 175:82–90. CrossRefGoogle Scholar
  9. 9.
    John R, Nampoothiri KM, Pandey A (2007) Fermentative production of lactic acid from biomass: an overview on process developments and future perspectives. Appl Microbiol Biotechnol 74:524–534. CrossRefPubMedGoogle Scholar
  10. 10.
    König H, Fröhlich J (2017) Lactic acid bacteria. In: König H, Unden G, Fröhlich J (eds) Biology of microorganisms on grapes, in must and in wine. Springer International Publishing, Cham, pp 3–41. CrossRefGoogle Scholar
  11. 11.
    Lin J, Zou Y, Cao K, Ma C, Chen Z (2016) The impact of heterologous catalase expression and superoxide dismutase overexpression on enhancing the oxidative resistance in Lactobacillus casei. J Ind Microbiol Biotechnol 43:703–711. CrossRefPubMedGoogle Scholar
  12. 12.
    Lu Y, Yang B, Gregg D, Saddler JN, Mansfield SD (2002) Cellulase adsorption and an evaluation of enzyme recycle during hydrolysis of steam-exploded softwood residues. Appl Biochem Biotechnol 98:641–654. CrossRefPubMedGoogle Scholar
  13. 13.
    Maas RW, Bakker R, Jansen MA, Visser D, de Jong E, Eggink G, Weusthuis R (2008) Lactic acid production from lime-treated wheat straw by Bacillus coagulans: neutralization of acid by fed-batch addition of alkaline substrate. Appl Microbiol Biotechnol 78:751–758. CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Nampoothiri KM, Nair NR, John RP (2010) An overview of the recent developments in polylactide (PLA) research. Bioresour Technol 101:8493–8501. CrossRefGoogle Scholar
  15. 15.
    Mazzoli R, Bosco F, Mizrahi I, Bayer EA, Pessione E (2014) Towards lactic acid bacteria-based biorefineries. Biotechnol Adv 32:1216–1236. CrossRefPubMedGoogle Scholar
  16. 16.
    Modenbach AA, Nokes SE (2012) The use of high-solids loadings in biomass pretreatment—a review. Biotechnol Bioeng 109:1430–1442. CrossRefPubMedGoogle Scholar
  17. 17.
    Odling-Smee L (2007) Biofuels bandwagon hits a rut. Nature 446:483. CrossRefPubMedGoogle Scholar
  18. 18.
    Ou MS, Ingram LO, Shanmugam KT (2011) l(+)-Lactic acid production from non-food carbohydrates by thermotolerant Bacillus coagulans. J Ind Microbiol Biotechnol 38:599–605. CrossRefPubMedGoogle Scholar
  19. 19.
    Ouyang J, Ma R, Zheng Z, Cai C, Zhang M, Jiang T (2013) Open fermentative production of l-lactic acid by Bacillus sp. strain NL01 using lignocellulosic hydrolyzates as low-cost raw material. Bioresour Technol 135:475–480. CrossRefPubMedGoogle Scholar
  20. 20.
    Palmqvist E, Hahn-Hägerdal B (2000) Fermentation of lignocellulosic hydrolysates. I: inhibition and detoxification. Bioresour Technol 74:17–24. CrossRefGoogle Scholar
  21. 21.
    Peng L, Wang L, Che C, Yang G, Yu B, Ma Y (2013) Bacillus sp. strain P38: an efficient producer of l-lactate from cellulosic hydrolysate, with high tolerance for 2-furfural. Bioresour Technol 149:169–176. CrossRefPubMedGoogle Scholar
  22. 22.
    Qing Q, Yang B, Wyman CE (2010) Impact of surfactants on pretreatment of corn stover. Bioresour Technol 101:5941–5951. CrossRefPubMedGoogle Scholar
  23. 23.
    Södergård A, Stolt M (2002) Properties of lactic acid based polymers and their correlation with composition. Prog Polym Sci 27:1123–1163. CrossRefGoogle Scholar
  24. 24.
    Sluiter A, Hames B, Ruiz R, Scarlata C, Sluiter J, Templeton D (2005) Determination of structural carbohydrates and lignin in biomass. Laboratory Analytical Procedure from the National Renewable Energy Laboratory Biomass Analysis Technology (NREL BAT) TeamGoogle Scholar
  25. 25.
    Subramanian MR, Talluri S, Christopher LP (2015) Production of lactic acid using a new homofermentative Enterococcus faecalis isolate. Microb Biotechnol 8:221–229. CrossRefPubMedGoogle Scholar
  26. 26.
    Wang Y, Tashiro Y, Sonomoto K (2015) Fermentative production of lactic acid from renewable materials: recent achievements, prospects, and limits. J Biosci Bioeng 119:10–18. CrossRefPubMedGoogle Scholar
  27. 27.
    Wee Y-J, Kim J-N, Yun J-S, Ryu H-W (2004) Utilization of sugar molasses for economical l(+)-lactic acid production by batch fermentation of Enterococcus faecalis. Enzyme Microb Technol 35:568–573. CrossRefGoogle Scholar
  28. 28.
    Wee Y-J, Yun J-S, Park D-H, Ryu H-W (2004) Biotechnological production of l(+)-lactic acid from wood hydrolyzate by batch fermentation of Enterococcus faecalis. Biotechnol Lett 26:71–74. CrossRefPubMedGoogle Scholar
  29. 29.
    Wee YJ, Yun JS, Kim D, Ryu HW (2006) Batch and repeated batch production of l(+)-lactic acid by Enterococcus faecalis RKY1 using wood hydrolyzate and corn steep liquor. J Ind Microbiol Biotechnol 33:431–435. CrossRefPubMedGoogle Scholar
  30. 30.
    Yuan SF, Guo GL, Hwang WS (2017) Ethanol production from dilute-acid steam exploded lignocellulosic feedstocks using an isolated multistress-tolerant Pichia kudriavzevii strain. Microb Biotechnol 10:1581–1590. CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Yun J-S, Wee Y-J, Ryu H-W (2003) Production of optically pure l(+)-lactic acid from various carbohydrates by batch fermentation of Enterococcus faecalis RKY1. Enzyme Microb Technol 33:416–423. CrossRefGoogle Scholar
  32. 32.
    Zabed H, Sahu JN, Suely A, Boyce AN, Faruq G (2017) Bioethanol production from renewable sources: current perspectives and technological progress. Renew Sustain Energy Rev 71:475–501. CrossRefGoogle Scholar
  33. 33.
    Zhao K, Qiao Q, Chu D, Gu H, Dao TH, Zhang J, Bao J (2013) Simultaneous saccharification and high titer lactic acid fermentation of corn stover using a newly isolated lactic acid bacterium Pediococcus acidilactici DQ2. Bioresour Technol 135:481–489. CrossRefPubMedGoogle Scholar

Copyright information

© Society for Industrial Microbiology and Biotechnology 2018

Authors and Affiliations

  • Shuo-Fu Yuan
    • 1
    • 2
  • Teng-Chieh Hsu
    • 1
  • Chun-An Wang
    • 1
  • Ming-Feng Jang
    • 1
  • Yang-Cheng Kuo
    • 1
  • Hal S. Alper
    • 2
    • 3
    Email author
  • Gia-Luen Guo
    • 1
    Email author
  • Wen-Song Hwang
    • 1
  1. 1.Chemistry DivisionInstitute of Nuclear Energy Research, Atomic Energy Committee, Executive YuanTaoyüanTaiwan, ROC
  2. 2.Institute for Cellular and Molecular BiologyThe University of Texas at AustinAustinUSA
  3. 3.McKetta Department of Chemical Engineering, The University of Texas at AustinAustinUSA

Personalised recommendations