Skip to main content
SpringerLink
Log in

Efficient synthesis of d-phenyllactic acid by a whole-cell biocatalyst co-expressing glucose dehydrogenase and a novel d-lactate dehydrogenase from Lactobacillus rossiae

  • Original Article
  • Published:
3 Biotech Aims and scope Submit manuscript

Abstract

d-Phenyllactic acid is a versatile natural organic acid, which is used as an antiseptic agent, monomer of the biodegradable material poly-phenyllactic acid and in the synthesis chiral intermediate of pharmaceuticals. In this report, the novel NADH-dependent d-lactate dehydrogenase LrLDH was identified by screening a shotgun genome of Lactobacillus rossiae. To improve cofactor regeneration, the Exiguobacterium sibiricum glucose dehydrogenase EsGDH was overexpressed together with LrLDH in E. coli BL21(DE3)-pCDFDuet-1-gdh-ldh. The total enzyme activity in the fermentation broth of E. coli BL 21(DE3)-pCDFDuet-1-gdh-ldh peaked at 2359.0 U l−1 when induced by 10 g l−1 lactose at 28 °C and 150 rpm for 14 h. The biocatalytic reduction of sodium phenylpyruvate to d-phenyllactic acid was successfully carried out using whole cells of the engineered E. coli. Under the optimized biocatalysis conditions, 50 g l−1 sodium phenylpyruvate was completely converted to d-phenyllactic acid with a space-time yield and enantiomeric excess of 262.8 g l−1 day−1 and > 99.5%, respectively. To our best knowledge, it is the highest productivity reported to date, with great potential for the mass production of d-phenyllactic acid.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Scheme 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Choudhury S, Baeg JO, Park NJ, Yadav RK (2014) A solar light-driven, eco-friendly protocol for highly enantioselective synthesis of chiral alcohols via photocatalytic/biocatalytic cascades. Green Chem 16(9):4389–4400

    Article  CAS  Google Scholar 

  • Cui YH, Wei P, Peng F, Zong MH, Lou WY (2018) Efficient biocatalytic stereoselective reduction of methyl acetoacetate catalyzed by whole cells of engineered E. coli. RSC Adv 8(18):9970–9978

    Article  CAS  Google Scholar 

  • Dallagnol AM, Catalan CAN, Mercado MI, Font de Valdez G, Rollan GC (2011) Effect of biosynthetic intermediates and citrate on the phenyllactic and hydroxyphenyllactic acids production by Lactobacillus plantarum CRL 778. J Appl Microbiol 111(6):1447–1455

    Article  CAS  PubMed  Google Scholar 

  • Dieuleveux V, Van Der Pyl D, Chataud J, Gueguen M (1998a) Purification and characterization of anti-Listeria compounds produced by Geotrichum candidum. Appl Environ Microbiol 64(2):800–803

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dieuleveux V, Lemarinier S, Guéguen M (1998b) Antimicrobial spectrum and target site of D-3-phenyllactic acid. Int J Food Microbiol 40(3):177–183

    Article  CAS  Google Scholar 

  • Ebdrup S, Ingrid P, Rasmussen HB, Heinz-Josef D, Anette FJ, Mortensen SB, Jan F, Lone P, Lars N, Per S (2003) Synthesis and biological and structural characterization of the dual-acting peroxisome proliferator-activated receptor alpha/gamma agonist ragaglitazar. J Med Chem 46(8):1306–1317

    Article  CAS  PubMed  Google Scholar 

  • Fujita T, Hieu Duc N, Ito T, Zhou S, Osada L, Tateyama S, Kaneko T, Takaya N (2013) Microbial monomers custom-synthesized to build true bio-derived aromatic polymers. Appl Microbiol Biotechnol 97(20):8887–8894

    Article  CAS  PubMed  Google Scholar 

  • He SB, Wang ZS, Zou Y, Chen SF, Xu XP (2014) Purification and characterization of a novel carbonyl reductase involved in oxidoreduction of aromatic beta-amino ketones/alcohols. Process Biochem 49(7):1107–1112

    Article  CAS  Google Scholar 

  • Jia BL, Pu ZJ, Tang K, Jia XM, Kim KH, Liu XL, Jeon CO (2018) Catalytic, computational, and evolutionary analysis of the d-lactate dehydrogenases responsible for d-lactic acid production in lactic acid bacteria. J Agric Food Chem 66(31):8371–8381

    Article  PubMed  Google Scholar 

  • Kawaguchi H, Uematsu K, Ogino C, Teramura H, Niimi-Nakamura S, Tsuge Y, Hasunuma T, Oinuma KI, Takaya N, Kondo A (2014) Simultaneous saccharification and fermentation of kraft pulp by recombinant Escherichia coli for phenyllactic acid production. Biochem Eng J 88(6):188–194

    Article  CAS  Google Scholar 

  • Kawaguchi H, Ogino C, Kondo A (2017) Microbial conversion of biomass into bio-based polymers. Bioresour Technol 245:1664–1673

    Article  CAS  PubMed  Google Scholar 

  • Lavermicocca P, Valerio FA (2003) Antifungal activity of phenyllactic acid against molds isolated from bakery products. Appl Environ Microbiol 69(1):634–640

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lavermicocca P, Valerio F, Evidente A, Lazzaroni S, Corsetti A, Gobbetti M (2000) Purification and characterization of novel antifungal compounds from the sourdough Lactobacillus plantarum strain 21B. Appl Environ Microbiol 66(9):4084–4090

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li L, Shin SY, Lee KW, Han NS (2014) Production of natural antimicrobial compound d-phenyllactic acid using Leuconostoc mesenteroides ATCC 8293 whole cells involving highly active d-lactate dehydrogenase. Lett Appl Microbiol 59(4):404–411

    Article  CAS  Google Scholar 

  • Mu WM, Yu SH, Zhu LJ, Zhang T, Jiang B (2012a) Recent research on 3-phenyllactic acid, a broad-spectrum antimicrobial compound. Appl Microbiol Biotechnol 95(5):1155–1163

    Article  CAS  PubMed  Google Scholar 

  • Mu WM, Yu SH, Zhu LJ, Jiang B, Zhang T (2012b) Production of 3-phenyllactic acid and 4-hydroxyphenyllactic acid by Pediococcus acidilactici DSM 20284 fermentation. Eur Food Res Technol 235(3):581–585

    Article  CAS  Google Scholar 

  • Ni Y, Xu JH (2012) Biocatalytic ketone reduction: a green and efficient access to enantiopure alcohols. Biotechnol Adv 30(6):1279–1288

    Article  CAS  PubMed  Google Scholar 

  • Schnürer J, Magnusson J (2005) Antifungal lactic acid bacteria as biopreservatives. Trends Food Sci Technol 16(1):70–78

    Article  Google Scholar 

  • Tang YP, Zhang GM, Wang Z, Liu D, Zhang LL, Zhou YF, Huang J, Yu FM, Yang ZS, Ding GF (2018) Efficient synthesis of a (S)-fluoxetine intermediate using carbonyl reductase coupled with glucose dehydrogenase. Bioresour Technol 250:457–463

    Article  CAS  PubMed  Google Scholar 

  • Urban FJ, Moore BS (1992) Synthesis of optically-active 2-benzyldihydrobenzopyrans for the hypoglycemic agent englitazone. J Heterocycl Chem 29(2):431–438

    Article  CAS  Google Scholar 

  • Valerio F, Lavermicocca P, Pascale M, Visconti A (2004) Production of phenyllactic acid by lactic acid bacteria: an approach to the selection of strains contributing to food quality and preservation. FEMS Microbiol Lett 233(2):289–295

    Article  CAS  PubMed  Google Scholar 

  • Wang JP, Yoo JS, Lee JH, Lee JH (2009) Effects of phenyllactic acid on production performance, egg quality parameters, and blood characteristics in laying hens. J Appl Poult Res 18(2):203–209

    Article  CAS  Google Scholar 

  • Wang JP, Lee JH, Yoo JS, Cho JH, Kim HJ, Kim IH (2010) Effects of phenyllactic acid on growth performance, intestinal microbiota, relative organ weight, blood characteristics, and meat quality of broiler chicks. Poultry Sci 89(7):1549–1555

    Article  CAS  Google Scholar 

  • Wang M, Zhu LF, Xu XL, Wang LM, Yin RC, Yu B (2016) Efficient production of enantiomerically pure d-phenyllactate from phenylpyruvate by structure-guided design of an engineered d-lactate dehydrogenase. Appl Microbiol Biot 100(17):7471–7478

    Article  CAS  Google Scholar 

  • Wang YJ, Shen W, Luo X, Liu ZQ, Zheng YG (2017a) Enhanced diastereoselective synthesis of t-Butyl 6-cyano-(3R,5R)-dihydroxyhexanoate by using aldo-keto reductase and glucose dehydrogenase co-producing engineered Escherichia coli. Biotechnol Prog 33(5):1235–1242

    Article  PubMed  Google Scholar 

  • Wang YJ, Ying BB, Chen M, Shen W, Liu ZQ, Zheng YG (2017b) An NADPH-dependent Lactobacillus composti short-chain dehydrogenase/reductase: characterization and application to (R)-1-phenylethanol synthesis. World J Microbiol Biotechnol 33(7):144

    Article  PubMed  Google Scholar 

  • Wang YJ, Ying BB, Shen W, Zheng RC, Zheng YG (2017c) Rational design of Kluyveromyces marxianus ZJB14056 aldo–keto reductase KmAKR to enhance diastereoselectivity and activity. Enzyme Microb Technol 107:32–40

    Article  CAS  PubMed  Google Scholar 

  • Wang FT, Wu HH, Jin PP, Sun ZL, Liu F, Du LH, Wang DY, Xu WM (2018) Antimicrobial activity of phenyllactic acid against Enterococcus faecalis and its effect on cell membrane. Foodborne Pathog Dis 15(10):645–652

    Article  CAS  PubMed  Google Scholar 

  • Weckwerth W, Miyamoto K, Iinuma K, Krause M, Glinski M, Storm T, Bonse G, Kleinkauf H, Zocher R (2000) Biosynthesis of PF1022A and related cyclooctadepsipeptides. J Biol Chem 275(23):17909–17915

    Article  CAS  PubMed  Google Scholar 

  • Wichmann R, Vasic-Racki D (2005) Cofactor regeneration at the lab scale. Adv Biochem Eng Biotechnol 92:225–260

    CAS  PubMed  Google Scholar 

  • Xu ZN, Jing KJ, Liu Y, Cen PL (2007) High-level expression of recombinant glucose dehydrogenase and its application in NADPH regeneration. J Ind Microbiol Biotechnol 34(1):83–90

    Article  CAS  PubMed  Google Scholar 

  • Xu GC, Zhang LL, Ni Y (2016) Enzymatic preparation of D-phenyllactic acid at high space-time yield with a novel phenylpyruvate reductase identified from Lactobacillus sp CGMCC 9967. J Biotechnol 222:29–37

    Article  CAS  PubMed  Google Scholar 

  • Yu SH, Zhou C, Zhang T, Jiang B, Mu WM (2015) Short communication: 3-Phenyllactic acid production in milk by Pediococcus pentosaceus SK25 during laboratory fermentation process. J Dairy Sci 98(2):813–817

    Article  CAS  Google Scholar 

  • Yu H, Qiu S, Chen F, Cheng YN, Wang YJ, Zheng YG (2019) Improving the catalytic efficiency of aldo-keto reductase KmAKR towards t-butyl 6-cyano-(3R,5R)-dihydroxyhexanoate via semi-rational design. Bioorg Chem 90:103018

    Article  CAS  PubMed  Google Scholar 

  • Zhang XQ, Zhang SL, Shi Y, Shen FD, Wang HK (2014) A new high phenyl lactic acid-yielding Lactobacillus plantarum IMAU10124 and a comparative analysis of lactate dehydrogenase gene. FEMS Microbiol Lett 356(1):89–96

    Article  CAS  PubMed  Google Scholar 

  • Zheng ZJ, Ma CQ, Gao C, Li FS, Qin JY, Zhang HW, Wang K, Xu P (2011) Efficient conversion of phenylpyruvic acid to phenyllactic acid by using whole cells of Bacillus coagulans SDM. PLoS One 6(4):e19030

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zheng ZJ, Sheng BB, Gao C, Zhang HW, Qin T, Ma CQ, Xu P (2013) Highly stereoselective biosynthesis of (R)-alpha-hydroxy carboxylic acids through rationally re-designed mutation of d-lactate dehydrogenase. Sci Rep 3:3401

    Article  PubMed  PubMed Central  Google Scholar 

  • Zhou XT, Zhang RZ, Xu Y, Liang HB, Jiang JW, Xiao R (2015) Coupled (R)-carbonyl reductase and glucose dehydrogenase catalyzes (R)-1-phenyl-1,2-ethanediol biosynthesis with excellent stereochemical selectivity. Process Biochem 50(11):1807–1813

    Article  CAS  Google Scholar 

  • Zhu YB, Hu FG, Zhu YY, Wang LM, Qi B (2015) Enhancement of phenyllactic acid biosynthesis by recognition site replacement of D-lactate dehydrogenase from Lactobacillus pentosus. Biotechnol Lett 37(6):1233–1241

    Article  CAS  PubMed  Google Scholar 

  • Zhu YB, Jiang ZY, Chen JH, Xu JM, Wang LM, Qi B (2017) Fusion of d-lactate dehydrogenase and formate dehydrogenase for increasing production of (R)-3-phenyllactic acid in recombinant Escherichia coli BL21 (DE3). J Biobased Mater Bioenergy 11(4):372–378

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was financially supported by the Natural Science Foundation for Young Scholars of Zhejiang Province (LQ19B060003) and the Scientific Research Foundation of Taizhou University (2017PY035).

Author information

Authors and Affiliations

  1. Institute of Biomass Resources, Taizhou University, Taizhou, 318000, Zhejiang, People’s Republic of China

    Xi Luo, Yingying Zhang, Longfei Yin, Weilong Zheng & Yongqian Fu

Authors
  1. Xi Luo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yingying Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Longfei Yin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Weilong Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yongqian Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yongqian Fu.

Ethics declarations

Conflict of interest

On behalf of all the authors, the corresponding author states that there is no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Luo, X., Zhang, Y., Yin, L. et al. Efficient synthesis of d-phenyllactic acid by a whole-cell biocatalyst co-expressing glucose dehydrogenase and a novel d-lactate dehydrogenase from Lactobacillus rossiae. 3 Biotech 10, 14 (2020). https://doi.org/10.1007/s13205-019-2003-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s13205-019-2003-2

Keywords

Search

Navigation