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Engineering phytosterol transport system in Mycobacterium sp. strain MS136 enhances production of 9α-hydroxy-4-androstene-3,17-dione

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Abstract

Objectives

To enhance the yield of 9α-hydroxy-4-androstene-3,17-dione (9-OHAD) from phytosterols, a phytosterol transport system was constructed in Mycobacterium sp. strain MS136.

Results

9-OHAD can be produced via the controlled degradation of phytosterols by mycobacteria. This involves an active transport process that requires trans-membrane proteins and ATP. A phytosterol transport system from Mycobacterium tuberculosis H37Rv was constructed in Mycobacterium sp. strain MS136 by co-expression of an energy-related gene, mceG, and two integrated membrane protein genes, yrbE4A and yrbE4B. The resultant of the Mycobacterium sp. strain MS136-GAB gave 5.7 g 9-OHAD l−1, which was a 20% increase over 4.7 g l−1 by the wild-type strain. The yield of 9-OHAD was increased to 6.0 g l−1 by optimization of fermentation conditions, when 13 g phytosterols l−1 were fermented for 84 h in 30 ml biotransformation medium in shake flasks.

Conclusions

Phytosterol transport system plays an active role in the uptake and transport of sterols, cloning of the system improved the mass transfer of phytosterols and increased the production of 9-OHAD.

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References

  • Donova MV, Egorova OV (2012) Microbial steroid transformations: current state and prospects. Appl Microbiol Biotechnol 94:1423–1447

    Article  CAS  PubMed  Google Scholar 

  • Fernandes P, Cruz A, Angelova B, Pinheiro HM, Cabral JMS (2003) Microbial conversion of steroid compounds: recent developments. Enzym Microbial Technol 32:688–705

    Article  CAS  Google Scholar 

  • Gao XQ (2016) Enhanced steroid metabolites production by resting cell phytosterol bioconversion. Chem Biochem Eng Q 29:567–573

    Article  Google Scholar 

  • Geize RVD, Yam K, Heuser T, Wilbrink MH, Hara H et al (2007) A gene cluster encoding cholesterol catabolism in a soil actinomycete provides insight into Mycobacterium tuberculosis survival in macrophages. Proc Natl Acad Sci USA 104:1947–1952

    Article  PubMed  PubMed Central  Google Scholar 

  • Joshi SM, Pandey AK, Capite N, Fortune SM, Rubin EJ, Sassetti CM (2006) Characterization of mycobacterial virulence genes through genetic interaction mapping. Proc Natl Acad Sci USA 103:11760–11765

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Klepp LI, Forrellad MA, Osella AV, Blanco FC et al (2012) Impact of the deletion of the six mce operons in Mycobacterium smegmatis. Microbes Infect 14:590–599

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Knol J, Bodewits K, Hessels GI, Dijkhuizen L, Geize RVD (2008) 3-Keto-5alpha-steroid delta(1)-dehydrogenase from Rhodococcus erythropolis SQ1 and its orthologue in Mycobacterium tuberculosis H37Rv are highly specific enzymes that function in cholesterol catabolism. Biochem J 410:339–346

    Article  CAS  PubMed  Google Scholar 

  • Mohn WW, Geize RVD, Stewart GR, Okamoto S, Liu J, Dijkhuizen L, Eltis LD (2008) The actinobacterial mce4 locus encodes a steroid transporter. J Biol Chem 283:35368–35374

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pandey AK, Sassetti CM (2008) Mycobacterial persistence requires the utilization of host cholesterol. Proc Natl Acad Sci USA 105:4376–4380

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang Z, Zhao F, Chen D, Li D (2006) Biotransformation of phytosterol to produce androsta-diene-dione by resting cells of Mycobacterium in cloud point system. Process Biochem 41:557–561

    Article  CAS  Google Scholar 

  • Wei W, Wang FQ, Fan SY, Wei DZ (2010) Inactivation and augmentation of the primary 3-ketosteroid-Δ1-dehydrogenase in Mycobacterium neoaurum NwIB-01: biotransformation of soybean phytosterols to 4-androstene-3,17-dione or 1,4-androstadiene-3,17-dione. Appl Environ Microbiol 76:4578–4582

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wilbrink MH, Petrsma M, Dijkhuizen L, Geize RVD (2011) FadD19 of Rhodococcus rhodochrous DSM43269, a steroid-coenzyme a ligase essential for degradation of C-24 branched sterol side chains. Appl Environ Microbiol 77:4455–4464

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

The authors acknowledge the financial support by a startup funding from Tianjin University.

Supporting information

Supplementary Table 1—Strains, primers and plasmids used in this study.

Supplementary Fig. 1—HPLC analysis of 9-OHAD produced by engineered strains.

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Authors and Affiliations

  1. Key Laboratory of Systems Bioengineering, Ministry of Education, School of Chemical Engineering and Technology, and SynBio Research Platform, Collaborative Innovation Centre of Chemical Science and Engineering, Tianjin University, Tianjin, 300072, China

    Kun He, Hong Sun & Hao Song

Authors
  1. Kun He
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  2. Hong Sun
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  3. Hao Song
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Corresponding author

Correspondence to Hao Song.

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He, K., Sun, H. & Song, H. Engineering phytosterol transport system in Mycobacterium sp. strain MS136 enhances production of 9α-hydroxy-4-androstene-3,17-dione. Biotechnol Lett 40, 673–678 (2018). https://doi.org/10.1007/s10529-018-2520-9

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  • DOI: https://doi.org/10.1007/s10529-018-2520-9

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