Process optimization of fed-batch fermentation for pyruvic acid production with Candida glabrata

  • GUO Likun ,
  • ZENG Weizhu ,
  • ZHOU Jingwen
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  • 1(National Engineering Laboratory for Cereal Fermentation Technology (NELCF) (Jiangnan University), Wuxi 214122, China);
    2(School of Biotechnology, Jiangnan University, Wuxi 214122, China)

Received date: 2019-11-23

  Online published: 2020-05-19

Abstract

In order to improve the titer and yield of pyruvic acid, the fermentation conditions for pyruvic acid production by Candida glabrata mutants were systematically optimized. A strain C. glabrata 4H2 with better performance in accumulating pyruvic acid was firstly selected from seven screened strains. Then, various types and concentrations of nitrogen source were tested for seed cultivation in shake flasks. The optimum nitrogen source was determined to be soybean peptone and its optimal concentration was 10 g/L. As a result, the titer, yield and productivity of pyruvic acid of the optimized fermentation process reached (48.56±0.46) g/L, 0.46 g/g and 0.93 g/(L·h), respectively, after 52 h shake-flask cultivation at 30 ℃, which were increased by 25.0%, 43.8% and 52.5%, respectively. Further, the fermentation optimization was performed in a 15 L bioreactor. First, the optimum initial glucose concentration was determined to be 80 g/L. Then, when the concentration of residual glucose was lowered down to 55 g/L, 70 g/L glucose was supplemented into the bioreactor with a constant feeding approach. The final titer of pyruvic acid reached (86.63±0.29) g/L, which was 78.4% higher than that of shake-flask cultivation. The productivity and yield of pyruvic acid reached 1.07 g/(L·h) and 0.78 g/g, respectively. The results indicated that the fermentation optimization effectively improved pyruvic acid production in C. glabrata, which laid a foundation for the industrial production of pyruvic acid.

Cite this article

GUO Likun , ZENG Weizhu , ZHOU Jingwen . Process optimization of fed-batch fermentation for pyruvic acid production with Candida glabrata[J]. Food and Fermentation Industries, 2020 , 46(7) : 10 -16 . DOI: 10.13995/j.cnki.11-1802/ts.022872

References

[1] XU P, JIANHUA Q, CHAO G, et al. Biotechnological routes to pyruvate production [J]. Journal of Bioscience and Bioengineering, 2008, 105(3): 169-175.
[2] KERBER R C, FERNANDO M S. α-Oxocarboxylic acids [J]. Journal of Chemical Education, 2010, 87(10): 1 079-1 084.
[3] YANG M, XING J. Improvement of pyruvate production based on regulation of intracellular redox state in engineered Escherichia coli [J]. Biotechnology and Bioprocess Engineering, 2017, 22(4): 376-381.
[4] ARTHUR J L C, JAMES Z G, ALTON M. Synthesis and properties of the alpha-keto acids [J]. Chemical Reviews, 1983, 83(3): 321-358.
[5] SONG Y, LI J, SHIN H-D, et al. Biotechnological production of alpha-keto acids: Current status and perspectives [J]. Bioresource Technology, 2016, 219: 716-724.
[6] HOSSAIN G S, SHIN H-D, LI J, et al. Transporter engineering and enzyme evolution for pyruvate production from D/L-alanine with a whole-cell biocatalyst expressing L-amino acid deaminase from Proteus mirabilis [J]. RSC Advances, 2016, 6(86): 82 676-82 684.
[7] CHAO G, XIAOMAN X, CHUNHUI H, et al. Pyruvate producing biocatalyst with constitutive NAD-independent lactate dehydrogenases [J]. Process Biochemistry, 2010, 45(12): 1 912-1 915.
[8] EISENBERG A, SEIP J E, GAVAGAN J E, et al. Pyruvic acid production using methylotrophic yeast transformants as catalyst [J]. Journal of Molecular Catalysis B Enzymatic, 1997, 2(4): 223-232.
[9] KAWAI S, OHASHI K, YOSHIDA S, et al. Bacterial pyruvate production from alginate, a promising carbon source from marine brown macroalgae [J]. Journal of Bioscience and Bioengineering, 2014, 117(3): 269-274.
[10] MIYAZAKI M, SHIBUE M, OGINO K, et al. Enzymatic synthesis of pyruvic acid from acetaldehyde and carbon dioxide [J]. Chemical Communications, 2001, 33(18): 1 800-1 801.
[11] YANG S, CHEN X, XU N, et al. Urea enhances cell growth and pyruvate production in Torulopsis glabrata [J]. Biotechnology Progress, 2014, 30(1): 19-27.
[12] VAN MARIS A J A, GEERTMAN J M A, VERMEULEN A, et al. Directed evolution of pyruvate decarboxylase-negative Saccharomyces cerevisiae, yielding a C2-independent, glucose-tolerant, and pyruvate-hyperproducing yeast [J]. Applied and Environmental Microbiology, 2004, 70(1): 159-166.
[13] ZHU Y, EITEMAN M A, ALTMAN R, et al. High glycolytic flux improves pyruvate production by a metabolically engineered Escherichia coli strain [J]. Applied and Environmental Microbiology, 2008, 74(21): 6 649-6 655.
[14] JINGWEN Z, LUXI H, LIMING L, et al. Enhancement of pyruvate productivity by inducible expression of a F0F1-ATPase inhibitor INH1 in Torulopsis glabrata CCTCC M202019 [J]. Journal of Biotechnology, 2009, 144(2): 120-126.
[15] SHA X, JINGWEN Z, LIMING L, et al. Arginine: A novel compatible solute to protect Candida glabrata against hyperosmotic stress [J]. Process Biochemistry, 2011, 46(6): 1 230-1 235.
[16] LUO Z, ZENG W, DU G, et al. Enhanced pyruvate production in Candida glabrata by engineering ATP futile cycle system [J]. ACS Synthetic Biology, 2019, 8(4): 789-795.
[17] ZHOU J, LIU L, DU G, et al. Citrate protect the growth of Torulopsis glabrata CCTCC M202019 against acidic stress as additional ATP supplier [J]. Journal of Biotechnology, 2008, 136: S741.
[18] LUO Z, ZENG W, DU G, et al. A high-throughput screening procedure for enhancing pyruvate production in Candida glabrata by random mutagenesis [J]. Bioprocess and Biosystems Engineering, 2017, 40(5): 693-701.
[19] LUO Z, LIU S, DU G, et al. Enhanced pyruvate production in Candida glabrata by carrier engineering [J]. Biotechnology and Bioengineering, 2018, 115(2): 473.
[20] LI Y, CHEN J, LUN S Y, et al. Efficient pyruvate production by a multi-vitamin auxotroph of Torulopsis glabrata: key role and optimization of vitamin levels [J]. Applied Microbiology and Biotechnology, 2001, 55(6): 680-685.
[21] QIN Y, JOHNSON C H, LIU L, et al. Introduction of heterogeneous NADH reoxidation pathways into Torulopsis glabrata significantly increases pyruvate production efficiency [J]. Korean Journal of Chemical Engineering, 2011, 28(4): 1 078.
[22] LUO Z, LIU S, DU G, et al. Identification of a polysaccharide produced by the pyruvate overproducer Candida glabrata CCTCC M202019 [J]. Applied Microbiology and Biotechnology, 2017, 101(11): 4 447-4 458.
[23] CELI?SKA E. Debottlenecking the 1,3-propanediol pathway by metabolic engineering [J]. Biotechnology Advances, 2010, 28(4): 519-530.
[24] ENKLER L, RICHER D, MARCHAND A L, et al. Genome engineering in the yeast pathogen Candida glabrata using the CRISPR-Cas9 system [J]. Scientific Reports, 2016, 6: 35 766.
[25] CEN Y, FIORI A, DIJCK P V. Deletion of the DNA ligase IV gene in Candida glabrata significantly increases gene-targeting efficiency [J]. Eukaryotic Cell, 2015, 14(8): 783-791.
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