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.
Similar content being viewed by others
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
Abdel-Rahman MA, Tashiro Y, Sonomoto K (2013) Recent advances in lactic acid production by microbial fermentation processes. Biotechnol Adv 31:877–902
Adsul MG, Varma AJ, Gokhale DV (2007) Lactic acid production from waste sugarcane bagasse derived cellulose. Green Chem 9:58–62
Akerberg C, Zacchi G (2000) An economic evaluation of the fermentative production of lactic acid from wheat flour. Bioresour Technol 75:119–126
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
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
Benninga HA (1990) A history of lactic acid making. Kluyver Academic Publishers, Dordrecht
Datta R, Henry M (2006) Lactic acid: recent advances in products, processes and technologies—a review. J Chem Technol Biotechnol 81:1119–1129
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
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
Fiske CH, Subbarow Y (1925) The colorimetric determination of phosphorous. J Biol Chem 66:375–389
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
Hongo M, Nomura Y, Iwahara M (1986) Novel method of lactic acid production by electrodialysis fermentation. Appl Environ Microbiol 52:314–319
Joshi DS, Singhvi MS, Khire JM, Gokhale DV (2010) Strain improvement of Lactobacillus lactis for D-lactic acid production. Biotechnol Lett 32:517–520
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
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
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
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
Marinova NA, Yankov DS (2009) Toxicity of some solvents and extractants towards Lactobacillus casei cells. Bulg Chem Commun 41:368–373
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
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
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
Playne MJ, Smith BR (1983) Toxicity of organic extraction reagents to anaerobic bacteria. Biotechnol Bioeng 25:1251–1265
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
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
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
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
Sodergard A, Stolt M (2002) Properties of lactic acid based polymers and their correlation with composition. Prog Polym Sci 27:1123–1163
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
Tik N, Bayraktar E, Mehmetoglu U (2001) In situ reactive extraction of lactic acid from fermentation media. J Chemical Technol Biotechnol 76:764–768
Tsuji F (2002) Autocatalytic hydrolysis of amorphous-made polylactides: effects of L-lactide content, tacticity, and enantiomeric polymer blending. Polymer 43:1789–1796
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
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
Wee YJ, Kim JN, Ryu HW (2006) Biotechnological production of lactic acid and its recent applications. Food Technol Biotechnol 44:163–172
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
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
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).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval
This article does not contain any studies with human participants or animals performed by any of the authors.
Rights and permissions
About this article
Cite this article
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
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-018-9084-4