Inhibition of steroid nucleus degradation in mycobacterial transformations

doi:10.1016/S0039-128X(68)80150-3

Steroids

Volume 11, Issue 3, March 1968, Pages 401-413

https://doi.org/10.1016/S0039-128X(68)80150-3 Get rights and content

Abstract

Screening a large number of Mycobacteria showed that most of them metabolize cholesterol (cholest-5-en-3β-ol) and cholest-4-en-3-one resp., and some of them accumulate androsta-1,4-diene-3,17-dione when the steroid nucleus break-down is inhibited. The 9α-hydroxylation which initiates the degradation of steroids was inhibited by chelating agents. The Mycobacterium phlei strain, studied extensively, formed Δ1,4-3-keto and rostadienes from 5α— and 5β-saturated, Δ1-, Δ4- and Δ5-double bond containing androstanes and 17β-side chain containing steroids reap.

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Cited by (36)

Application of microbial 3-ketosteroid Δ<sup>1</sup>-dehydrogenases in biotechnology
2021, Biotechnology Advances
Citation Excerpt :
Similarly, iron-chelating agents have been used to inhibit the enzyme in several different microorganisms, and to increase the yield of ADD (Wix et al., 1968). These chelating agents included 2,2’-dipyridyl, 2,2,2"-terpyridyl, 1,10-phenanthroline and derivatives, 8-hydroxyquinoline and derivatives, and cupferron, among others (Arima et al., 1968; Wix et al., 1968). However, they may be toxic and may adversely impact cell growth, and, therefore, they are generally introduced during the stationary phase of the microbial fermentation (Ahmad et al., 1992; Malaviya and Gomes, 2008).
3-Ketosteroid Δ¹-dehydrogenase catalyzes the 1(2)-dehydrogenation of 3-ketosteroid substrates using flavin adenine dinucleotide as a cofactor. The enzyme plays a crucial role in microbial steroid degradation, both under aerobic and anaerobic conditions, by initiating the opening of the steroid nucleus. Indeed, many microorganisms are known to possess one or more 3-ketosteroid Δ¹-dehydrogenases. In the pharmaceutical industry, 3-ketosteroid Δ¹-dehydrogenase activity is exploited to produce Δ¹-3-ketosteroids, a class of steroids that display various biological activities. Many of them are used as active pharmaceutical ingredients in drug products, or as key precursors to produce pharmaceutically important steroids. Since 3-ketosteroid Δ¹-dehydrogenase activity requires electron acceptors, among other considerations, Δ¹-3-ketosteroid production has been industrially implemented using whole-cell fermentation with growing or metabolically active resting cells, in which the electron acceptors are available, rather than using the isolated enzyme. In this review we discuss biotechnological applications of microbial 3-ketosteroid Δ¹-dehydrogenases, covering commonly used steroid-1(2)-dehydrogenating microorganisms, the bioprocess for preparing Δ¹-3-ketosteroids, genetic engineering of 3-ketosteroid Δ¹-dehydrogenases and related genes for constructing new, productive industrial strains, and microbial fermentation strategies for enhancing the product yield. Furthermore, we also highlight the recent development in the use of isolated 3-ketosteroid Δ¹-dehydrogenases combined with a FAD cofactor regeneration system. Finally, in a somewhat different context, we summarize the role of 3-ketosteroid Δ¹-dehydrogenase in cholesterol degradation by Mycobacterium tuberculosis and other mycobacteria. Because the enzyme is essential for the pathogenicity of these organisms, it may be a potential target for drug development to combat mycobacterial infections.
Androstenedione production by biotransformation of phytosterols
2008, Bioresource Technology
Citation Excerpt :
When 3-ketosteroid-1(2)-dehydrogenase alone is active, it inserts a double bond between the C1 and C2 position and ADD is formed; when 9α-hydroxylase alone is active, a hydroxyl group is inserted at the C9 position and 9α-hydroxy-4-androstene-3,17-dione is formed. When both enzymes are inactive, AD is the primary compound synthesized (Wix et al., 1967; Kieslich, 1985). In the presence of enzyme inhibitors, cholesterol, β-sitosterol, campesterol, stigmasterol and their mixtures can be effectively converted to C19, C22 or C27 steroids.
Androstenedione is a key intermediate of microbial steroid metabolism. It belongs to the 17-keto steroid family and is used as starting material for the preparation of different steroids. Androstenedione can be produced by microbial side chain cleavage of phytosterol, which is an alternative to multi-step chemical synthesis. In this review, various methods of biotransformation of sterols to androstenedione are surveyed. It begins with the history and current research status in this field. The existing methods of chemical and biochemical synthesis are examined. Various issues related to these methods and how researchers have addressed them is presented. Among these, the low solubility of sterols in aqueous systems is a critical problem since it limits the product yield. The main content of this review focuses on new methods of biotransformation that are being investigated. Recent biotechnological advances in this field are presented. The review ends with a note on future perspectives.
Steroid 17β-reduction by microorganisms - A review
2005, Process Biochemistry
17β-Reduction is one of key and widespread reactions of microbial steroid metabolism. It accompanies various steroid transformations by microorganisms and plays a role in the regulation of reducing equivalent pool and detoxication of exogenic steroids. The ability of microorganisms to reduce C₁₇-ketosteroids has been known for 60 years, but most progress in the biochemistry and biotechnology of the process has been recorded in the past two decades. The regulation of 17β-hydroxysteroid dehydrogenase activity is of importance for the selectivity of various steroid biotransformations. The potential application of microbial 17β-reduction and the reverse reaction – 17β-oxidation – in the pharmaceutical industry for the production of 17β-hydroxy and 17-oxosteroids is shown. Microbial side chain cleavage of sterols in combination with 17β-reduction represents the shortest way to obtain testosterone in a single biotechnological operation thus replacing multistep chemical and/or combined biotech-chemical syntheses. This review highlights progress in the understanding of the biochemical basis of 17β-reduction and the biotechnological applications of the process.
Physiological and biochemical improvement of the enzyme side-chain degradation of cholesterol by Fusarium solani
2005, Process Biochemistry
The conversion of cholesterol (C) to androstenedione (AD) and androstadienedione (ADD) was influenced significantly by the composition of the cultivation medium. A medium composed of (g/l): glucose, 50; peptone, 20; corn steep solid, 3; supported relatively high AD and ADD outputs. C convertibility was further accelerated when the cultivation medium was adjusted with phosphate buffer with the pH range 5.5–7.38. Supplementation of the medium with 8-hydroxyquinoline or methanol prevented the degradation of the C nucleus with concomitant formation of good AD and ADD yields.
Steroid transformations with Exophiala jeanselmei var. lecanii-corni and Ceratocystis paradoxa
1999, Steroids
The fungi Exophiala jeanselmei var. lecanii-corni [IMI (International Mycological Institute) 312989, UAMH (University of Alberta Microfungus Collection and Herbarium) 8783] and Ceratocystis paradoxa (IMI 374529, UAMH 8784) have been examined for their potential in steroid biotransformation. The study has determined that E. jeanselmei var. lecanii-corni effected overall anti-Markovnikov hydration on dehydroisoandrosterone, and side-chain degradation on a variety of pregnanes. Both ascomycetes were found to carry out redox reactions of alcohols and ketones as well as 1,4 reduction of α,β-unsaturated carbonyl systems.
Microbial transformation of β-sitosterol and stigmasterol into 26-oxygenated derivatives
1995, Steroids
The genetically modified Mycobacterium sp. BCS 396 strain has been used to transform sterols with stigmastane side chain in order to obtain 26-oxidized metabolites. β-Sitosterol (I) was transformed to 4-stigmasten-3-one (II), 26-hydroxy-4-stigmasten-3-one (III), and 3-oxo-4-stigmasten-26-oic acid (IV), while stigmasterol (V) was converted to 4,22-stigmastadien-3-one (VI), 6β-hydroxy-4,22-stigmastadien-3-one (VII), 26-hydroxy-4,22-stigmastadien-3-one (VIII), 3-oxo-4,22-stigmastadien-26-oic acid methyl ester (IX), and 3-oxo-1,4,22-stigmastatrien-26-oic acid methyl ester (X) with that strain. In both β-sitosterol and stigmasterol, 26-oxidation generates the R-configuration on C-25.

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⁺: Deceased in 1965.

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Inhibition of steroid nucleus degradation in mycobacterial transformations

Abstract

Biochim. Et Biophys. Acta

J. Biol. Chem.

Steroids

Steroids

Biochem. J.

Arch. Hyg.

Zentr. Bakteriol. Parazitenk., 2. ABT.

Antonie Van Leeuwenhoek. J. Microbiol. Serol.

J. Am. Chem. Soc.

Acta Microbiol. Acad. Sci. Hung.