Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Genome Analysis and Optimization of Caproic Acid Production of Clostridium butyricum GD1-1 Isolated from the Pit Mud of Nongxiangxing Baijiu

  • Min Li (College of Bioengineering, Sichuan University of Science and Engineering) ;
  • Tao Li (College of Bioengineering, Sichuan University of Science and Engineering) ;
  • Jia Zheng (Wuliangye Yibin Co., Ltd.) ;
  • Zongwei Qiao (Wuliangye Yibin Co., Ltd.) ;
  • Kaizheng Zhang (College of Bioengineering, Sichuan University of Science and Engineering) ;
  • Huibo Luo (College of Bioengineering, Sichuan University of Science and Engineering) ;
  • Wei Zou (College of Bioengineering, Sichuan University of Science and Engineering)
  • Received : 2023.04.11
  • Accepted : 2023.06.07
  • Published : 2023.10.28
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Abstract

Caproic acid is a precursor substance for the synthesis of ethyl caproate, the main flavor substance of nongxiangxing baijiu liquor. In this study, Clostridium butyricum GD1-1, a strain with high caproic acid concentration (3.86 g/l), was isolated from the storage pit mud of nongxiangxing baijiu for sequencing and analysis. The strain's genome was 3,840,048 bp in length with 4,050 open reading frames. In addition, virulence factor annotation analysis showed C. butyricum GD1-1 to be safe at the genetic level. However, the annotation results using the Kyoto Encyclopedia of Genes and Genomes Automatic Annotation Server predicted a deficiency in the strain's synthesis of alanine, methionine, and biotin. These results were confirmed by essential nutrient factor validation experiments. Furthermore, the optimized medium conditions for caproic acid concentration by strain GD1-1 were (g/l): glucose 30, NaCl 5, yeast extract 10, peptone 10, beef paste 10, sodium acetate 11, L-cysteine 0.6, biotin 0.004, starch 2, and 2.0% ethanol. The optimized fermentation conditions for caproic acid production by C. butyricum GD1-1 on a single-factor basis were: 5% inoculum volume, 35℃, pH 7, and 90% loading volume. Under optimal conditions, the caproic acid concentration of strain GD1-1 reached 5.42 g/l, which was 1.40 times higher than the initial concentration. C. butyricum GD1-1 could be further used in caproic acid production, NXXB pit mud strengthening and maintenance, and artificial pit mud preparation.

Keywords

Acknowledgement

This work was supported by grants from the Cooperation Project of Wuliangye Group Co., Ltd., Sichuan University of Science & Engineering, China (CXY2019ZR011), and Sichuan University of Science & Engineering (Item No. 2020RC36). We would also like to thank International Science Editing (http://www.internationalscienceediting.com) for editing this manuscript.

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