Biotechnology Letters

, Volume 40, Issue 4, pp 673–678 | Cite as

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

Original Research Paper

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.

Keywords

9α-Hydroxy-4-androstene-3,17-dione Mycobacteria Optimization Phytosterols Steroids Transport system 

Notes

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.

Supplementary material

10529_2018_2520_MOESM1_ESM.docx (44 kb)
Supplementary material 1 (DOCX 44 kb)

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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 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 EngineeringTianjin UniversityTianjinChina

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