Journal of Microbiology and Biotechnology

  • 제32권9호
  • /
  • Pages.1178-1185
  • /
  • 2022
  • /
  • 1017-7825(pISSN)
  • /
  • 1738-8872(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지
DOI QR코드

DOI QR Code

Development of a High-Titer Culture Medium for the Production of Cholesterol by Engineered Saccharomyces cerevisiae and Its Fed-Batch Cultivation Strategy

  • Wang, Ling-Xu (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology) ;
  • Zheng, Gao-Fan (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology) ;
  • Xin, Xiu-Juan (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology) ;
  • An, Fa-Liang (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology)
  • 투고 : 2021.06.10
  • 심사 : 2021.11.29
  • 발행 : 2022.09.28
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Steroids are a class of compounds with cyclopentane polyhydrophenanthrene as the parent nucleus, and they usually have unique biological and pharmacological activities. Most of the biosynthesis of steroids is completed by a series of enzymatic reactions starting from cholesterol. Synthetic biology can be used to synthesize cholesterol in engineered microorganisms, but the production of cholesterol is too low to further produce other high-value steroids from cholesterol as the raw material and precursor. In this work, combinational strategies were established to increase the production of cholesterol in engineered Saccharomyces cerevisiae RH6829. The basic medium for high cholesterol production was selected by screening 8 kinds of culture media. Single-factor optimization of the carbon and nitrogen sources of the culture medium, and the addition of calcium ions, zinc ions and citric acid, further increased the cholesterol production to 192.53 mg/l. In the 5-L bioreactor, through the establishment of strategies for glucose and citric acid feeding and dissolved oxygen regulation, the cholesterol production was further increased to 339.87 mg/l, which was 734% higher than that in the original medium. This is the highest titer of cholesterol produced by microorganisms currently reported. The fermentation program has also been conducted in a 50-L bioreactor to prove its stability and feasibility.

키워드

참고문헌

  1. Fernandes P, Cruz A, Angelova B, Pinheiro HM, Cabral JMS. 2003. Microbial conversion of steroid compounds: recent developments. Enzyme Microb. Technol. 32: 688-705. https://doi.org/10.1016/S0141-0229(03)00029-2
  2. Sultana N. 2018. Microbial biotransformation of bioactive and clinically useful steroids and some salient features of steroids and biotransformation. Steroids 136: 76-92. https://doi.org/10.1016/j.steroids.2018.01.007
  3. Chen Y, Tang Y-M, Yu S-L, Han Y-W, Kou J-P, Liu B-L, et al. 2015. Advances in the pharmacological activities and mechanisms of diosgenin. Chinese J. Nat. Med. 13: 578-587. https://doi.org/10.1016/S1875-5364(15)30053-4
  4. Bhatti HN, Khera RA. 2012. Biological transformations of steroidal compounds: a review. Steroids 77: 1267-1290. https://doi.org/10.1016/j.steroids.2012.07.018
  5. Giorgi V, Menendez P, Garcia-Carnelli C. 2019. Microbial transformation of cholesterol: reactions and practical aspects-an update. World J. Microbiol. Biotechnol. 35: 131. https://doi.org/10.1007/s11274-019-2708-8
  6. Parshikov IA, Sutherland JB. 2015. Biotransformation of steroids and flavonoids by cultures of Aspergillus niger. Appl. Biochem. Biotechnol. 176: 903-923. https://doi.org/10.1007/s12010-015-1619-x
  7. Thomas ST, VanderVen BC, Sherman DR, Russell DG, Sampson NS. 2011. Pathway profiling in Mycobacterium tuberculosis: elucidation of cholesterol-derived catabolite and enzymes that catalyze its metabolism. J. Biol. Chem. 286: 43668-43678. https://doi.org/10.1074/jbc.M111.313643
  8. Wang ZF, Huang YL, Rathman JF, Yang S-T. 2002. Lecithin-enhanced biotransformation of cholesterol to androsta-1,4-diene-3,17-dione and androsta-4-ene-3,17-dione. J. Chem. Technol. Biotechnol. 77: 1349-1357. https://doi.org/10.1002/jctb.728
  9. McLean KJ, Hans M, Munro AW. 2012. Cholesterol, an essential molecule: diverse roles involving cytochrome P450 enzymes. Biochem. Soc. Trans. 40: 587-593. https://doi.org/10.1042/BST20120077
  10. Hu Z, He B, Ma L, Sun Y, Niu Y, Zeng B. 2017. Recent advances in ergosterol biosynthesis and regulation mechanisms in S. cerevisiae. Indian J. Microbiol. 57: 270-277. https://doi.org/10.1007/s12088-017-0657-1
  11. Nagegowda DA, Gupta P. 2020. Advances in biosynthesis, regulation, and metabolic engineering of plant specialized terpenoids. Plant Sci. 294: 110457. https://doi.org/10.1016/j.plantsci.2020.110457
  12. Szczebara FM, Chandelier C, Villeret C, Masurel A, Bourot S, Duport C, et al. 2003. Total biosynthesis of hydrocortisone from a simple carbon source in yeast. Nat. Biotechnol. 21: 143-149. https://doi.org/10.1038/nbt775
  13. Li X, Wang Z, Zhang G, Yi L. 2019. Improving lycopene production in S. cerevisiae through optimizing pathway and chassis metabolism. Chem. Eng. Sci. 193: 364-369. https://doi.org/10.1016/j.ces.2018.09.030
  14. Hohmann HP, Lehmann M. 2012. Production of non-yeast sterols by yeast. US Patent. 20120231495.
  15. Lang C, Markus V.2011. Preparation of 7-dehydrocholesterol and/or the biosynthetic intermediates and/or secondary products thereof in transgenic organisms. US Patent. 12607017.
  16. Guo XJ, Xiao WH, Wang Y, Yao MD, Zeng BX, Liu H, et al. 2018. Metabolic engineering of S. cerevisiae for 7-dehydrocholesterol overproduction. Biotechnol. Biofuels 11: 192. https://doi.org/10.1186/s13068-018-1194-9
  17. Souza CM, Schwabe TM, Pichler H, Ploier B, Leitner E, Guan XL, et al. 2011. A stable yeast strain efficiently producing cholesterol instead of ergosterol is functional for tryptophan uptake, but not weak organic acid resistance. Metab. Eng. 13: 555-569. https://doi.org/10.1016/j.ymben.2011.06.006
  18. Cheng C, Zhang M, Xue C, Bai F, Zhao X. 2017. Development of stress tolerant S. cerevisiae strains by metabolic engineering: New aspects from cell flocculation and zinc supplementation. J. Biosci. Bioeng. 123: 141-146. https://doi.org/10.1016/j.jbiosc.2016.07.021
  19. Li S, Liu L, Chen J. 2015. Mitochondrial fusion and fission are involved in stress tolerance of Candida glabrata. Bioresour. Bioprocess. 2: 12. https://doi.org/10.1186/s40643-015-0041-0