Comparative biochemical characterization of mammalian-derived CYP11A1s with cholesterol side-chain cleavage activities

Liu, Ruxin; Pan, Yunjun; Wang, Ning; Tang, Dandan; Urlacher, Vlada B.; Li, Shengying

doi:10.1016/j.jsbmb.2023.106268

Cited by 4 publications

(3 citation statements)

References 71 publications

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“…The lengthening of the side chain, in the case of phytosterol, led to an increase in the time of complete conversion of the substrate compared to cholesterol. The results correlate with experimental data, where it was shown, that CYP11A1 converted plant sterols (β-sitosterol, campesterol) to pregnenolone in vitro with less efficiency compared to cholesterol [ 30 , 31 ]. The presence of a methoxymethyl substituent at C3 in cholesterol and phytosterol reduced the rate of substrate transformation possibly because the MOM-derived sterols are even much more hydrophobic as compared with natural sterols.…”

Section: Discussionsupporting

confidence: 79%

Pregnenolone and progesterone production from natural sterols using recombinant strain of Mycolicibacterium smegmatis mc2 155 expressing mammalian steroidogenesis system

Karpov,

Strizhov,

Novikova

et al. 2024

Microb Cell Fact

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Background Pregnenolone and progesterone are the life-important steroid hormones regulating essential vital functions in mammals, and widely used in different fields of medicine. Microbiological production of these compounds from sterols is based on the use of recombinant strains expressing the enzyme system cholesterol hydroxylase/C20-C22 lyase (CH/L) of mammalian steroidogenesis. However, the efficiency of the known recombinant strains is still low. New recombinant strains and combination approaches are now needed to produce these steroid hormones. Results Based on Mycolicibacterium smegmatis, a recombinant strain was created that expresses the steroidogenesis system (CYP11A1, adrenodoxin reductase, adrenodoxin) of the bovine adrenal cortex. The recombinant strain transformed cholesterol and phytosterol to form progesterone among the metabolites. When 3-methoxymethyl ethers of sterols were applied as bioconversion substrates, the corresponding 3-ethers of pregnenolone and dehydroepiandrosterone (DHEA) were identified as major metabolites. Under optimized conditions, the recombinant strain produced 85.2 ± 4.7 mol % 3-methoxymethyl-pregnenolone within 48 h, while production of 3-substituted DHEA was not detected. After the 3-methoxymethyl function was deprotected by acid hydrolysis, crystalline pregnenolone was isolated in high purity (over 98%, w/w). The structures of steroids were confirmed using TLC, HPLC, MS and 1H- and 13C-NMR analyses. Conclusion The use of mycolicybacteria as a microbial platform for the expression of systems at the initial stage of mammalian steroidogenesis ensures the production of valuable steroid hormones—progesterone and pregnenolone from cholesterol. Selective production of pregnenolone from cholesterol is ensured by the use of 3-substituted cholesterol as a substrate and optimization of the conditions for its bioconversion. The results open the prospects for the generation of the new microbial biocatalysts capable of effectively producing value-added steroid hormones.

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Section: Discussionsupporting

confidence: 79%

Pregnenolone and progesterone production from natural sterols using recombinant strain of Mycolicibacterium smegmatis mc2 155 expressing mammalian steroidogenesis system

Karpov,

Strizhov,

Novikova

et al. 2024

Microb Cell Fact

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“…Furthermore, turmeric is a relatively expensive raw material, and recently, its prices have been constantly fluctuating in the world [165]. So far, turmeric has been used, among others, as a raw material for the production of steroids, thanks to the diosgenin extracted from it.…”

Section: Bioavailability Disadvantages and Hazards Of The Use Of Curc...mentioning

confidence: 99%

Turmeric and Curcumin—Health-Promoting Properties in Humans versus Dogs

Kępińska-Pacelik,

Biel

2023

IJMS

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The growing popularity of the use of nutraceuticals in the prevention and alleviation of symptoms of many diseases in humans and dogs means that they are increasingly the subject of research. A representative of the nutraceutical that deserves special attention is turmeric. Turmeric belongs to the family Zingiberaceae and is grown extensively in Asia. It is a plant used as a spice and food coloring, and it is also used in traditional medicine. The biologically active factors that give turmeric its unusual properties and color are curcuminoids. It is a group of substances that includes curcumin, de-methoxycurcumin, and bis-demethoxycurcumin. Curcumin is used as a yellow-orange food coloring. The most important pro-health effects observed after taking curcuminoids include anti-inflammatory, anticancer, and antioxidant effects. The aim of this study was to characterize turmeric and its main substance, curcumin, in terms of their properties, advantages, and disadvantages, based on literature data.

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“…Steroids represent a significant class of metabolites present in almost all organisms, exhibiting various biological activities, including antitumor, [1] anticancer, [2] antifungal, [3] antidiabetic, [4] and antiinflammatory, [5] and are considered the second largest class of pharmaceuticals, following with antibiotics. [6] 22(R)-hydroxycholesterol (22(R)-HCHO) is an oxidized derivative of cholesterol (CHO) that exists in plants and animals. It is a crucial precursor for the biosynthesis of numerous high-value steroidal saponins and steroid alkaloids, includ-ing diosgenin, [7] pregnenolone, [8] and cyclopamine.…”

Section: Introductionmentioning

confidence: 99%

Efficient production of 22(R)‐hydroxycholesterol via combination optimization of Saccharomyces cerevisiae

Pang,

Cheng,

Ban

et al. 2024

Biotechnology Journal

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Abstract22(R)‐hydroxycholesterol (22(R)‐HCHO) is a crucial precursor of steroids biosynthesis with various biological functions. However, the production of 22(R)‐HCHO is expensive and unsustainable due to chemical synthesis and extraction from plants or animals. This study aimed to construct a microbial cell factory to efficiently produce 22(R)‐HCHO through systems metabolic engineering. First, we tested 7‐dehydrocholesterol reductase (Dhcr7s) and cholesterol C22‐hydroxylases from different sources in Saccharomyces cerevisiae, and the titer of 22(R)‐HCHO reached 128.30 mg L−1 in the engineered strain expressing Dhcr7 from Columba livia (ClDhcr7) and cholesterol 22‐hydroxylase from Veratrum californicum (VcCyp90b27). Subsequently, the 22(R)‐HCHO titer was significantly increased to 427.78 mg L−1 by optimizing the critical genes involved in 22(R)‐HCHO biosynthesis. Furthermore, hybrid diploids were constructed to balance cell growth and 22(R)‐HCHO production and to improve stress tolerance. Finally, the engineered strain produced 2.03 g L−1 of 22(R)‐HCHO in a 5‐L fermenter, representing the highest 22(R)‐HCHO titer reported to date in engineered microbial cell factories. The results of this study provide a foundation for further applications of 22(R)‐HCHO in various industrially valuable steroids.

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Comparative biochemical characterization of mammalian-derived CYP11A1s with cholesterol side-chain cleavage activities

Cited by 4 publications

References 71 publications

Pregnenolone and progesterone production from natural sterols using recombinant strain of Mycolicibacterium smegmatis mc2 155 expressing mammalian steroidogenesis system

Pregnenolone and progesterone production from natural sterols using recombinant strain of Mycolicibacterium smegmatis mc2 155 expressing mammalian steroidogenesis system

Turmeric and Curcumin—Health-Promoting Properties in Humans versus Dogs

Efficient production of 22(R)‐hydroxycholesterol via combination optimization of Saccharomyces cerevisiae

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