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Greener L-lactic acid production through in situ extractive fermentation by an acid-tolerant Lactobacillus strain

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Abstract

Lactic acid (LA) fermentation requires a neutralizer for a physiologically acceptable range. However, a neutralizer generates a large amount of gypsum, an environmental pollutant. Furthermore, the downstream processing is complicated and expensive, comprising 50–70% of the total cost. We previously developed a Lactobacillus delbrueckii FM1, which can produce undissociated LA without neutralizer. Here, we improved FM1 by adaptive evolution at pH 4.5, which generated Adp FM1 showing an ~ 1.80-fold increase in LA production compared to FM1. The LA production via fed-batch fermentation yielded 36.2 g/L of LA, with a productivity of 0.500 g/L/h. However, cell viability was reduced due to the acidic pH and/or end-product inhibition. Therefore, an in situ LA recovery process using an extractive solvent was employed to maintain cell viability. Adp FM1 produced 49.2 g/L of LA via in situ LA-extractive fed-batch fermentation, which was ~ 1.4-fold higher than that without LA extraction. Adp FM1 provided a total LA productivity of 0.512 g/L/h in 96 h. Among the tested strains, Adp FM1 exhibited the highest H+-ATPase activity and a 415-fold increase in H+-ATPase gene expression compared to the parent strain. These results suggest that the in situ LA extractive fermentation process will ease downstream processing and prove to be a more economical and environmentally friendly option compared to the present fermentation. To our knowledge, this is the first report on the production of undissociated L-LA by Lactobacillus using an in situ recovery process, with high LA production levels and productivity.

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References

  • Abdel-Rahman MA, Tashiro Y, Zendo T, Shibata K, Sonomoto K (2011) Isolation and characterisation of lactic acid bacterium for effective fermentation of cellobiose into optically pure homo L-(+)-lactic acid. Appl Microbiol Biotechnol 89:1039–1049

    Article  PubMed  CAS  Google Scholar 

  • Abdel-Rahman MA, Tashiro Y, Sonomoto K (2013) Recent advances in lactic acid production by microbial fermentation processes. Biotechnol Adv 31:877–902

    Article  PubMed  CAS  Google Scholar 

  • Adsul MG, Varma AJ, Gokhale DV (2007) Lactic acid production from waste sugarcane bagasse derived cellulose. Green Chem 9:58–62

    Article  CAS  Google Scholar 

  • Akerberg C, Zacchi G (2000) An economic evaluation of the fermentative production of lactic acid from wheat flour. Bioresour Technol 75:119–126

    Article  CAS  Google Scholar 

  • Baek SH, Kwon EY, Kim SY, Hahn JS (2016) GSF2 deletion increases lactic acid production by alleviating glucose repression in Saccharomyces cerevisiae. Sci Rep 6:34812–34824

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Baek SH, Kwon EY, Bae SJ, Cho BR, Kim SY, Hahn JS (2017) Improvement of D-lactic acid production in Saccharomyces cerevisiae under acidic conditions by evolutionary and rational metabolic engineering. Biotechnol J 12:1700015–1700021

    Article  CAS  Google Scholar 

  • Benninga HA (1990) A history of lactic acid making. Kluyver Academic Publishers, Dordrecht

    Google Scholar 

  • Datta R, Henry M (2006) Lactic acid: recent advances in products, processes and technologies—a review. J Chem Technol Biotechnol 81:1119–1129

    Article  CAS  Google Scholar 

  • Datta R, Tsai S, Bonsignor P, Moon S, Frank J (1995) Technological and economic potential of poly(lactic acid) and lactic acid derivatives. FEMS Microbiol Rev 16:221–231

    Article  CAS  Google Scholar 

  • Davison BE, Llanos RL, Cancilla MR, Redman NC, Hillier AJ (1995) Current research on the genetics of lactic acid production in lactic acid bacteria. Int Dairy J 5:763–784

    Article  Google Scholar 

  • Fiske CH, Subbarow Y (1925) The colorimetric determination of phosphorous. J Biol Chem 66:375–389

    CAS  Google Scholar 

  • Gao MT, Shimamura T, Ishida N, Takahashi H (2011) pH-uncontrolled lactic acid fermentation with activated carbon as an adsorbent. Enz Microb Technol 48:526–530

    Article  CAS  Google Scholar 

  • Hongo M, Nomura Y, Iwahara M (1986) Novel method of lactic acid production by electrodialysis fermentation. Appl Environ Microbiol 52:314–319

    PubMed  PubMed Central  CAS  Google Scholar 

  • Joshi DS, Singhvi MS, Khire JM, Gokhale DV (2010) Strain improvement of Lactobacillus lactis for D-lactic acid production. Biotechnol Lett 32:517–520

    Article  PubMed  CAS  Google Scholar 

  • Juodeikiene G, Zadeike D, Bartkiene E, Klupsaite D (2016) Application of acid tolerant Pedioccocus strains for increasing the sustainability of lactic acid production from cheese whey. LWT Food Sci Technol 72:399–406

    Article  CAS  Google Scholar 

  • Kadam SR, Patil SS, Bastawde KB, Khire JM, Gokhale DV (2006) Strain improvement of Lactobacillus delbrueckii NCIM 2365 for lactic acid production. Process Biochem 41:120–126

    Article  CAS  Google Scholar 

  • Kawahata M, Masaki K, Fujii T, Iefuji H (2006) Yeast genes involved in response to lactic acid and acetic acid: acidic conditions caused by the organic acids in Saccharomyces cerevisiae cultures induce expression of intracellular metal metabolism genes regulated by Aft1p. FEMS Yeast Res 6:924–936

    Article  PubMed  CAS  Google Scholar 

  • Krzyzaniak A, Leeman M, Vossebeld F, Visser TJ, Schuur S (2013) Novel extractants for the recovery of fermentation derived lactic acid. Purif Technol 111:82–89

    Article  CAS  Google Scholar 

  • Marinova NA, Yankov DS (2009) Toxicity of some solvents and extractants towards Lactobacillus casei cells. Bulg Chem Commun 41:368–373

    CAS  Google Scholar 

  • Martak J, Sabolova E, Schlosser S, Rosenberg M, Kristofikova L (1997) Toxicity of organic solvents used in situ in fermentation of lactic acid by Rhizopus arrhizus. Biotechnol Tech 11:71–75

    Article  CAS  Google Scholar 

  • Matsumoto M, Nishimura M, Kobayashi H, Kondo K (2016) Extractive fermentation of lactic acid with Hiochi bacteria in a two-liquid phase system. Ferment Technol 5:1–6

    Article  CAS  Google Scholar 

  • Patnaik R, Louie S, Gavrilovic V, Perry K, Stemmer W, Ryan M, del Cardayre S (2002) Genome shuffling of Lactobacillus for improved acid tolerance. Nat Biotechnol 20:707–712

    Article  PubMed  CAS  Google Scholar 

  • Playne MJ, Smith BR (1983) Toxicity of organic extraction reagents to anaerobic bacteria. Biotechnol Bioeng 25:1251–1265

    Article  PubMed  CAS  Google Scholar 

  • Shobharani P, Halami PM (2014) Cellular fatty acid profile and H+-ATPase activity to assess acid tolerance of Bacillus sp. for potential probiotic functional attributes. Appl Microbiol Biotechnol 98:9045–9058

    Article  PubMed  CAS  Google Scholar 

  • Singhvi MS, Joshi DS, Adsul MG, Varma AJ, Gokhale DV (2010) D (−) lactic acid production from cellobiose and cellulose by Lactobacillus lactis mutant RM2-24. Green Chem 12:1106–1109

    Article  CAS  Google Scholar 

  • Singhvi MS, Gurjar GS, Gupta VS, Gokhale DV (2015) Biocatalyst development for lactic acid production at acidic pH using inter-generic protoplast fusion. RSC Adv 5:2024–2031

    Article  CAS  Google Scholar 

  • Singhvi MS, Zendo T, Iida H, Gokhale DV, Sonomoto K (2017) Stimulation of D-and L-lactate dehydrogenases transcriptional levels in presence of diammonium hydrogen phosphate resulting to enhanced lactic acid production by Lactobacillus strain. J Biosci Bioeng 124:674–679

    Article  PubMed  CAS  Google Scholar 

  • Sodergard A, Stolt M (2002) Properties of lactic acid based polymers and their correlation with composition. Prog Polym Sci 27:1123–1163

    Article  CAS  Google Scholar 

  • Suzuki T, Sakamoto T, Sugiyama M, Ishida N, Kambe H, Obata S, Kaneko Y, Takahashi, Harashima S (2013) Disruption of multiple genes whose deletion causes lactic-acid resistance improves lactic-acid resistance and productivity in Saccharomyces cerevisiae. J Biosci Bioeng 115:467–474

    Article  PubMed  CAS  Google Scholar 

  • Tik N, Bayraktar E, Mehmetoglu U (2001) In situ reactive extraction of lactic acid from fermentation media. J Chemical Technol Biotechnol 76:764–768

    Article  CAS  Google Scholar 

  • Tsuji F (2002) Autocatalytic hydrolysis of amorphous-made polylactides: effects of L-lactide content, tacticity, and enantiomeric polymer blending. Polymer 43:1789–1796

    Article  CAS  Google Scholar 

  • van Maris JA, Winkler AA, Porro D, van Dijken JP, Pronk JT (2004) Directed evolution of pyruvate decarboxylase-negative Saccharomyces cerevisiae, yielding a C2-independent, glucose-tolerant, and pyruvate-hyperproducing yeast. Appl Environ Microbiol 70:159–166

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Wang YH, Li Y, Pei XL, Yu L, Feng Y (2007) Genome-shuffling improved acid tolerance and L-lactic acid volumetric productivity in Lactobacillus rhamnosus. J Biotech 129:510–515

    Article  CAS  Google Scholar 

  • Wee YJ, Kim JN, Ryu HW (2006) Biotechnological production of lactic acid and its recent applications. Food Technol Biotechnol 44:163–172

    CAS  Google Scholar 

  • Yankov D, Molinier J, Albet J, Malmary G, Kyuchoukov G (2004) Lactic acid extraction from aqueous solutions with tri-n-octylamine dissolved in decanol and dodecane. Biochem Eng J 21:63–71

    Article  CAS  Google Scholar 

  • Yankov D, Molinier J, Kyuchoukov G, Albet J, Malmary G (2005) Improvement of the lactic acid extraction. Extraction from aqueous solutions and simulated fermentation broth by means of mixed extractant and TOA, partially loaded with HCl. Chem Biochem Eng Q 19:17–24

    CAS  Google Scholar 

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Acknowledgements

Mamata Singhvi was supported by a fellowship (no. P16100) from the Japan Society for the Promotion of Science (JSPS).

Funding

This work was partially supported by a Research Fellow JSPS grant (no. 16F16100) and a grant from JSPS KAKENHI (grant no. JP16F1610).

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Correspondence to Kenji Sonomoto.

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Singhvi, M., Zendo, T., Gokhale, D. et al. Greener L-lactic acid production through in situ extractive fermentation by an acid-tolerant Lactobacillus strain. Appl Microbiol Biotechnol 102, 6425–6435 (2018). https://doi.org/10.1007/s00253-018-9084-4

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