Biochemical studies on the microbial Δ1-dehydrogenation of cortisol by Pseudomonas fluorescens

Adham, Nehad Z.; El-Hady, Abeer Abd; Naim, Nadia

doi:10.1016/s0032-9592(02)00098-5

Cited by 23 publications

(8 citation statements)

References 16 publications

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“…The utilization of high amounts of the steroidal substrates is one of the important factors aVecting the economy of the transformation process [38]. In this study, increasing the concentration of ADD showed that the high concentration of the substrate (2, 4 and 5 mg ml ¡1 ) retarded the biotransformation process.…”

Section: Discussionmentioning

confidence: 57%

Studies on the microbial transformation of androst-1,4-dien-3,17-dione with Acremonium strictum

Faramarzi

Yazdi

Jahandar

et al. 2006

J IND MICROBIOL BIOTECHNOL

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The strain of Acremonium strictum PTCC 5282 was applied to investigate the biotransformation of androst-1,4-dien-3,17-dione (I; ADD). Microbial products obtained were purified by preparative TLC and the pure metabolites were characterized on the basis of their spectroscopic features (13C NMR, 1H NMR, FTIR, MS) and physical constants (melting points and optical rotations). The 15 alpha-Hydroxyandrost-1,4-dien-3,17-dione (II), 17 beta-hydroxyandrost-1,4-dien-3-one (III), androst-4-en-3,17-dione (IV; AD), 15 alpha-hydroxyandrost-4-en-3,17-dione (V), 15 alpha,17 beta-dihydroxyandrost-1,4-dien-3-one (VI) and testosterone (VII) were produced during this fermentation. Formation of the 15 alpha,17 beta-dihydroxy derivative of ADD is reported for the first time during steroid biotransformation. The bioconversion reactions observed were 1,2-hydrogenation, 15 alpha-hydroxylation and 17-ketone reduction. From the time course profile of this biotransformation, ketone reduction and 1,2-hydrogenation were observed from the first day of fermentation while 15 alpha-hydroxylation occurred from the third day. Optimum concentration of the substrate, which gave the maximum bioconversion efficiency, was 0.5 mg ml(-1) in one batch. The highest yield of the microbial products recorded in this work was achieved within the pH range 6.5-7.3 and at the temperature of 27 degrees C.

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Section: Discussionmentioning

confidence: 57%

Studies on the microbial transformation of androst-1,4-dien-3,17-dione with Acremonium strictum

Faramarzi

Yazdi

Jahandar

et al. 2006

J IND MICROBIOL BIOTECHNOL

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“…5). The results showed that KSDD activity was strongly stimulated by 1 mM K + , Na + and Ca 2+ , which had been revealed about other Δ 1 -dehydrogenation reaction [32]. However, in the presence of 1 mM Ag + , KSDD activity decreased to 28.05% compared to the control experiment.…”

Section: Characterization Of Ksddmentioning

confidence: 77%

“…Till now, no useful protectant has been reported. As reported that ions such as K + , Na + , Ca 2+ , Fe 2+ and Mg 2+ could stimulate the Δ 1 -dehydrogenation, we investigated the effect of mental ions on KSDD activity [32]. The results showed that KSDD activity was strongly stimulated by 1 mM K + , Na + and Ca 2+ .…”

Section: Discussionmentioning

confidence: 89%

Over-expression of Mycobacterium neoaurum 3-ketosteroid-Δ1-dehydrogenase in Corynebacterium crenatum for efficient bioconversion of 4-androstene-3,17-dione to androst-1,4-diene-3,17-dione

Zhang

Yang

et al. 2016

Electronic Journal of Biotechnology

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a b s t r a c tBackground: 3-Ketosteroid-Δ 1 -dehydrogenase (KSDD), a flavoprotein enzyme, catalyzes the bioconversion of 4-androstene-3,17-dione (AD) to androst-1,4-diene-3,17-dione (ADD). To date, there has been no report about characterization of KSDD from Mycobacterium neoaurum strains, which were usually employed to produce AD or ADD by fermentation. Results: In this work, Corynebacterium crenatum was chosen as a new host for heterologous expression of KSDD from M. neoaurum JC-12 after codon optimization of the KSDD gene. SDS-PAGE and western blotting results indicated that the recombinant C. crenatum harboring the optimized ksdd (ksdd II ) gene showed significantly improved ability to express KSDD. The expression level of KSDD was about 1.6-fold increased C. crenatum after codon optimization. After purification of the protein, we first characterized KSDD from M. neoaurum JC-12, and the results showed that the optimum temperature and pH for KSDD activity were 30°C and pH 7.0, respectively. The K m and V max values of purified KSDD were 8.91 μM and 6.43 mM/min. In this work, C. crenatum as a novel whole-cell catalyst was also employed and validated for bioconversion of AD to ADD. The highest transformation rate of AD to ADD by recombinant C. crenatum was about 83.87% after 10 h reaction time, which was more efficient than M. neoaurum JC-12 (only 3.56% at 10 h). Conclusions: In this work, basing on the codon optimization, overexpression, purification and characterization of KSDD, we constructed a novel system, the recombinant C. crenatum SYPA 5-5 expressing KSDD, to accumulate ADD from AD efficiently. This work provided new insights into strengthening sterol catabolism by overexpressing the key enzyme KSDD, for efficient ADD production.

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“…The lowest products yield (12.33 ± 0.58%) was obtained at substrate concentration of 50 mg/50 mL medium. This may be attributed to the toxic effect of the substrate on the activity of tested fungus as well as, the instantaneous accumulation of the substrate and its adsorption to the surface of mycelium decreased the substrate dissolution (Adham et al, 2003;Lu et al, 2006). According to Manosroi et al (2007) higher concentrations of the substrate that led to a decrease in the bioconversion activity, might be due to either the poor solubility of the substrate in the aqueous media or the limited activity of the enzyme.…”

Section: Effect Of Substrate Concentrationmentioning

confidence: 99%

Biotransformation of progesterone to hydroxysteroid derivatives by whole cells of Mucor racemosus

Mohamed¹,

El-Refai²,

Sallam³

et al. 2013

MJM

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Aims:The possibility of using Mucor racemosus cells in the biotransformation of progesterone to industrially important hydroxysteroid derivatives in one-step biotransformation process was investigated in this study Methodology and results: The fungal strain was inoculated into the transformation medium which supplemented with PR as a substrate (5-50 mg). The transformation products were separated and characterized on the bases of their GC/MS analysis as 11α-hydroxyprogesterone (11α-HP) as main product (I); 4-pregnen-18-al-11β,21-diol-3,20-dione (Aldosterone) (II) and 20-hydroxy-pregnan-18-oic acid (III) as minor products. The organism was tested for PR bioconversion at different transformation periods (6-96 h), as well as optimization of the basal medium through the addition of different concentrations of yeast extract and peptone (0.5 to 4 g/L) at various pH values (4-9). The optimal biotransformation conditions for maximum production of these PR derivatives were observed using 0.2 g/L of PR, 3 g/L of yeast extract and 3 g/L peptone after 48 h at pH value 5.5. Under these optimal conditions, cells total bioconversion efficiency reached about 96% of the original added PR. Conclusion, significance and impact of study: Under these optimum conditions, M. racemosus has the ability to biotransform PR to 11α-HP (I), Aldosterone (II) and 20-hydroxy-pregnan-18-oic acid (III) with total bioconversion efficiency of 96 ± 1.77%. These results may be of industrial importance because compounds II and III had not been previously recorded as biotransformation products of PR.

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Biochemical studies on the microbial Δ1-dehydrogenation of cortisol by Pseudomonas fluorescens

Cited by 23 publications

References 16 publications

Studies on the microbial transformation of androst-1,4-dien-3,17-dione with Acremonium strictum

Studies on the microbial transformation of androst-1,4-dien-3,17-dione with Acremonium strictum

Over-expression of Mycobacterium neoaurum 3-ketosteroid-Δ1-dehydrogenase in Corynebacterium crenatum for efficient bioconversion of 4-androstene-3,17-dione to androst-1,4-diene-3,17-dione

Biotransformation of progesterone to hydroxysteroid derivatives by whole cells of Mucor racemosus

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