Overexpression of cytochrome p450 125 in Mycobacterium: a rational strategy in the promotion of phytosterol biotransformation
| Autor: | Zhihua Shang, Shuangping Xu, Xia Menglei, Yanbing Shen, Xingjuan An, Liqiu Su, Min Wang |
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| Rok vydání: | 2018 |
| Předmět: |
Proteomics
0301 basic medicine Bioengineering Applied Microbiology and Biotechnology Cofactor Mycobacterium Industrial Microbiology 03 medical and health sciences Cytochrome P-450 Enzyme System Biotransformation Tandem Mass Spectrometry Mycobacterium neoaurum Androstenediols biology Chemistry Phytosterol Androstenedione Phytosterols Cytochrome P450 Nontuberculous Mycobacteria biology.organism_classification Sterol Androstadienes 030104 developmental biology Biochemistry Steroid Hydroxylases biology.protein Fermentation Aryl Hydrocarbon Hydroxylases NAD+ kinase Oxidation-Reduction Metabolic Networks and Pathways Chromatography Liquid Biotechnology |
| Zdroj: | Journal of Industrial Microbiology and Biotechnology. 45:857-867 |
| ISSN: | 1476-5535 1367-5435 |
| Popis: | Androst-4-ene-3, 17-dione (AD) and androst-1, 4-diene-3, 17-dione (ADD) are generally produced by the biotransformation of phytosterols in Mycobacterium. The AD (D) production increases when the strain has high NAD+/NADH ratio. To enhance the AD (D) production in Mycobacterium neoaurum TCCC 11978 (MNR M3), a rational strategy was developed through overexpression of a gene involved in the phytosterol degradation pathway; NAD+ was generated as well. Proteomic analysis of MNR cultured with and without phytosterols showed that the steroid C27-monooxygenase (Cyp125-3), which performs sequential oxidations of the sterol side chain at the C27 position and has the oxidative cofactor of NAD+ generated, played an important role in the phytosterol biotransformation process of MNR M3. To improve the productivity of AD (D), the cyp125-3 gene was overexpressed in MNR M3. The specific activity of Cyp125-3 in the recombinant strain MNR M3C3 was improved by 22% than that in MNR M3. The NAD+/NADH ratio in MNR M3C3 was 131% higher than that in the parent strain. During phytosterol biotransformation, the conversion of sterols increased from 84 to 96%, and the yield of AD (D) by MNR M3C3 was increased by approximately 18% for 96 h fermentation. This rational strain modification strategy may also be applied to develop strains with important application values for efficient production of cofactor-dependent metabolites. |
| Databáze: | OpenAIRE |
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