Conversion of soybean sterols into 3,17-diketosteroids using actinobacteria Mycobacterium neoaurum, Pimelobacter simplex, and Rhodococcus erythropolis

Андрюшина, В. А.; Rodina, N. V.; Stytsenko, T C; Luu, Duc Huy; Дружинина, А. В.; Ядерец, Вера Владимировна; Voishvillo, N. E.

doi:10.1134/s0003683811030021

Cited by 27 publications

(10 citation statements)

References 11 publications

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“…In a monophasic aqueous system, the product of AD can accumulate in culture liquid as large crystals for its low water‐solubility as the reaction proceeds . This agglomeration may also occur on the phytosterols solid particles or on the surface of Mycobacterium cells, thus restraining the bioconversion process.…”

Section: Resultsmentioning

confidence: 99%

Side-chain cleavage of phytosterols byMycobacteriumsp. MB 3683 in a biphasic ionic liquid/aqueous system

Yuan

Guan

Wang

et al. 2016

J. Chem. Technol. Biotechnol.

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BACKGROUND: Microbial transformation of phytosterols has been applied to produce androst-4-ene-3,17-dione (AD). However, low substrate solubility and product inhibition are major problems to be solved. Biphasic systems are efficient strategies, and ionic liquids (ILs), as one of the phases in biphasic systems, are promising alternatives to traditional organic solvents due to environmental constraints. RESULTS: In this work, nine water-immiscible ILs were used in phytosterols side-chain cleavage by Mycobacterium sp. MB 3683. The solubilities of phytosterols and AD in these ILs and distribution coefficients of AD between water and ILs were first measured.[PrMIM][PF 6 ] was determined to be a suitable candidate to establish a biphasic IL-aqueous system. Under optimal conditions, i.e. inoculum size of 12% with 48 h age, IL addition at 84 h, and phase ratio of 20:1 (v/v, aqueous/IL), AD production reached 2.23 g L −1 after 5 days of biotransformation with substrate concentration of 5 g L −1 in a scaled-up biphasic system of 200 mL. This bioconversion conducted in a biphasic IL-aqueous system was more efficient than that in a monophasic aqueous system. CONCLUSION: Owing to the environmental advantages of ILs and the above positive results, biphasic IL-aqueous systems have the potential to be applied in phytosterols side-chain cleavage. Selection of ILs for phytosterols biotransformationILs have been reported to have different effects on cellular membranes and biocompatibility was determined by the constitution of their anions and cations. Generally, ILs containing anion [PF 6 ] J Chem Technol Biotechnol 2016; 91: 2631-2637

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Section: Resultsmentioning

confidence: 99%

Side-chain cleavage of phytosterols byMycobacteriumsp. MB 3683 in a biphasic ionic liquid/aqueous system

Yuan

Guan

Wang

et al. 2016

J. Chem. Technol. Biotechnol.

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“…They are usually used to produce valuable chemicals by modifying their metabolic pathway . Many mycobacteria can use sterols as carbon and energy source and transform sterols into highly valuable sterol drug intermediates, such as 4‐androstene‐3,17‐dione (AD), androst‐1,4‐diene‐3,17‐dione (ADD), and 9α‐hydroxy‐AD . However, the poor water solubility of sterols limits the commercial use of most sterol‐transforming mycobacteria.…”

Section: Introductionmentioning

confidence: 99%

Bioconversion of cholesterol to 4‐cholesten‐3‐one by recombinant Bacillus subtilis expressing choM gene encoding cholesterol oxidase from Mycobacterium neoaurumJC‐12

Shao

Rao

Zhang

et al. 2014

J of Chemical Tech & Biotech

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BACKGROUND Bacillus subtilis 168 was used to express the cholesterol oxidase (EC 1.1.3.6) genes cloned from Mycobacterium neoaurum JC‐12 and the bioconversion of cholesterol to 4‐cholesten‐3‐one was realized. RESULTS Expression of cholesterol oxidase ChoM1 and ChoM2 from M. neoaurum in B. subtilis was confirmed by SDS‐PAGE and enzyme activity analysis. The activities of the recombinant ChoM1 and ChoM2 were 5.27 and 7.44 U mg−1, which were 5.2‐fold and 7.3‐fold, respectively, of the cholesterol oxidases activities in M. neoaurum JC‐12. Enzyme properties showed that the optimum pH and temperature for both ChoM1 and ChoM2 was 7.5 and 40 °C. Both enzymes were stimulated by Mg2+ and Mn2+ and showed higher activity towards the 3β‐hydroxy steroids with long alkyl chains at C17. Whole‐cells of the recombinant strains were used as catalysts, the recombinant strains B. subtilis 168/pMA5‐choM1 and B. subtilis 168/pMA5‐choM2 catalyzed the bioconversion of cholesterol to 4‐cholesten‐3‐one with a percentage conversion of 67% and 83% at 21 h. CONCLUSION The recombinant strains demonstrated efficient and stable enzyme activities and showed obvious pre‐eminence on the bioconversion of cholesterol into 4‐cholesten‐3‐one. As a safe microorganism, B. subtilis 168 would be a promising cell factory to produce 4‐cholesten‐3‐one in the pharmaceutical industry. © 2014 Society of Chemical Industry

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“…Utilizing natural sterols, such as cholesterol and phytosterols, as carbon and energy sources is a common physiological attribute of certain actinomycetes, such as mycobacteria and rhodococci 1 2 3 4 5 6 7 . The catabolism of sterols in pathogenic strains, such as M. tuberculosis , is highly concerned due to its close relevance to pathogenesis and persistence 1 2 .…”

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confidence: 99%

Unraveling and engineering the production of 23,24-bisnorcholenic steroids in sterol metabolism

Liu

Yao

et al. 2016

Sci Rep

134

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The catabolism of sterols in mycobacteria is highly important due to its close relevance in the pathogenesis of pathogenic strains and the biotechnological applications of nonpathogenic strains for steroid synthesis. However, some key metabolic steps remain unknown. In this study, the hsd4A gene from Mycobacterium neoaurum ATCC 25795 was investigated. The encoded protein, Hsd4A, was characterized as a dual-function enzyme, with both 17β-hydroxysteroid dehydrogenase and β-hydroxyacyl-CoA dehydrogenase activities in vitro. Using a kshAs-null strain of M. neoaurum ATCC 25795 (NwIB-XII) as a model, Hsd4A was further confirmed to exert dual-function in sterol catabolism in vivo. The deletion of hsd4A in NwIB-XII resulted in the production of 23,24-bisnorcholenic steroids (HBCs), indicating that hsd4A plays a key role in sterol side-chain degradation. Therefore, two competing pathways, the AD and HBC pathways, were proposed for the side-chain degradation. The proposed HBC pathway has great value in illustrating the production mechanism of HBCs in sterol catabolism and in developing HBCs producing strains for industrial application via metabolic engineering. Through the combined modification of hsd4A and other genes, three HBCs producing strains were constructed that resulted in promising productivities of 0.127, 0.109 and 0.074 g/l/h, respectively.

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Conversion of soybean sterols into 3,17-diketosteroids using actinobacteria Mycobacterium neoaurum, Pimelobacter simplex, and Rhodococcus erythropolis

Cited by 27 publications

References 11 publications

Side-chain cleavage of phytosterols byMycobacteriumsp. MB 3683 in a biphasic ionic liquid/aqueous system

Side-chain cleavage of phytosterols byMycobacteriumsp. MB 3683 in a biphasic ionic liquid/aqueous system

Bioconversion of cholesterol to 4‐cholesten‐3‐one by recombinant Bacillus subtilis expressing choM gene encoding cholesterol oxidase from Mycobacterium neoaurumJC‐12

Unraveling and engineering the production of 23,24-bisnorcholenic steroids in sterol metabolism

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