Transformation of 11-trifluoroacetate 6α-methylhydrocortisone (11-TFA MHC) by cells of the actinobacteria Arthrobacter (Nocardioides) was carried out in the presence of α-cyclodextrin (α-CD). The composition and dynamics of the accumulation of the transformation products in the culture medium at various pHs and the ratio of the α-CD/substrate were studied. It was shown that the addition of α-CD to the transformation medium at pH 7 promotes an increase in the rate of 1,2-dehydrogenation with the formation of 6 α-methylprednisolone 11-trifluoroacetate (11-TFA MPL). At pH 7, the primary process is the hydrolysis of the 11β-trifluoroacetyloxy group. In this case, the participation of α-CD in these processes as an acceptor of the trifluoroacetyl ion is not excluded.

An efficient procedure was proposed for the preparation of pregna-4-en-17 alpha,21-dio1-3,20-dioneby using nitrile method to result in high yield up to 56.0-57.0 % from androst-4-en-3,20-dione, a key intermediate for producing corticoids.

A novel Aspergillus nidulans lac№4 (argB–) strain, a producer of recombinant laccase A from basidiomycete Trametes hirsuta 072, was obtained. This strain was used for the biocatalytic transformation of progesterone (PG). The major biotransformation products are 11α-hydroxy-PG, 11α-acetoxy-PG and 6β, 11α-dihydroxy-PG. Macronet MN-200 sorbent was used to study the adsorption kinetics of PG, and the main biotransformation products from the transformation medium. The solid-phase extraction of steroids without preliminary removal of the mycelium was shown to be efficient.

In this study, we developed a method for the production of androsta-9(11)-diene-3,17-dione (Δ9(11)-AD), which is a combination of phytosterol side-chain microbial oxidation with simultaneous 9α‑hydroxylation and subsequent chemical regio-selective dehydration of 9α-hydroxy-3,17-diketo-intermediate without isolation and purification. Phytosterol was converted into 9α-hydroxyandrost-4-ene-3,17-dione (9-OH-AD) with the use of the wild-type Mycobacterium sp. VKM Ac-1817D strain. The product was extracted from the culture medium with the use of an organic solvent and dehydrated in the extract with mineral acid. The resulting Δ9(11)-AD was purified with the selective crystallization method. Minor products were isolated and identified. It has been shown that this strain is capable of sterol transformation with the formation of methyl ester of 9α-hydroxypregn-4-ene-3-one-20-carboxylic acid. Our approach makes it possible to simplify the flow chart for production of the target compound. It not only eliminates 9-OH-AD loss but also minimizes the amount of production waste.

pCXSN-Ch140mb, almost complete attenuation of the GFP fluorescence was observed. At the same time, qPCR analysis confirmed that there were no changes in GFP expression in all samples, indicating that the amount GFP was only regulated post-translationally. Thus, we have created the system which allows us to perform the selective degradation of plant proteins via the UPS. Supported by RFBR #15-04-09365.

The ability of the ascomycete Aspergillus niger N402 to transform exogenous progesterone was investigated. We found that this strain has steroid-hydroxylating activity and can introduce a hydroxyl group into the progesterone molecule mainly at positions C11(α) and C21 with predominant formation of 21-hydroxyprogesterone (deoxycortone). In addition, formation of 6β,11α-dihydroxyprogesterone was also observed. Studying the effects of the growth medium composition and temperature on progesterone conversion by A. niger N402 showed that the most intense accumulation of 21-hydroxyprogesterone occurred in minimal synthetic medium at 28°C. Increasing the cultivation temperature to 37°C resulted in almost complete inhibition of the hydroxylase activity in the minimal medium. In the complete medium, a similar increase in temperature inhibited 11α-hydroxylase activity and completely suppressed 6β-hydroxylase activity, but it produced no effect on 21-hydroxylating activity.

An effective method of dehydrating 9_-hydroxyandrost-4-ene-3,17-dione by mineral acids in organic solvents, producing essentially quantitative formation of androsta-4,9(11)-diene-3,17-dione is proposed. Quantitative isomerization of the side product androsta-4,8(9)-diene-3,17-dione to target product was shown to be pssible.

The newly synthesized (alpha/beta)-diastereomers of 6-(N-methyl-N-phenyl)aminomethylandrost-4-ene-3,17-dione (5) and 6-(N-methyl-N-phenyl)aminomethylandrost-4-en-17 beta-ol-3-one (6) were firstly investigated as substrates for the whole cells of Nocardioides simplex VKM Ac-2033D in comparison with their unsubstituted analogs, - androst-4-ene-3,17-dione (1) and androst-4-en-17 beta-ol-3-one (2).1(2)-Dehydroderivatives were identified as the major bioconversion products from all the substrates tested. When using the mixtures of (alpha/beta)-stereoisomers of 5 and 6 as the substrates, only beta-stereoisoiners of the corresponding 1,4-diene-steroids were formed. Along with 1(2) -dehydrogenation, N. simplex VKM Ac -2033D promoted oxidation of the hydroxyl group at C-17 position of 6: both 6(alpha) and 6(beta) were transformed to the corresponding 17-keto derivatives. No steroid core destruction was observed during the conversion of the 6-substituted androstanes 5 and 6, while it was significant when 1 or 2 was used as the substrate.The results suggested high potentials of N. simplex VKM Ac-2033D for the generation of novel 1(2)dehydroanalogs. (C) 2016 Elsevier Inc. All rights reserved.

9α-Hydroxyandrost-4-en-3,17-dione was prepared from a mixture of phytosterols using microbiological methods to degrade soybean sterols to androstenedione followed by its 9α-hydroxylation. Phytosterols could be extracted from deodorizer distillate sludge produced during production of soybean oil with low sterol content (5.2 %) with a level of extraction of greater than 96 % and a total sterol content of at least 90 %. The 9α-hydroxyandrost-4-en-3,17-dione yield from the phytosterol mixture was 38 %.

9α-Hydroxyandrost-4-ene-3,17-dione has been obtained from a phytosterol mixture by using of microbiological methods of sterol side chain degradation with the formation of androstenedione, followed by 9α-hydroxylation. It is shown, that phytosterols can be isolated from deodorizer sludge of soybean oil production with the low content of sterols (5.2%) at a yield of more than 96% and a total sterol content no less than 90%. The yield of 9α-hydroxyandrost-4-ene-3,17-dione from the phytosterol mixture was amounted to 38 %.

An efficient procedure was proposed for the synthesis of 3β-acetoxy-17α-hydroperoxy-16α-methylpregn-5-en-20-one. Optimal conditions were found for the combined process including 1,4-addition of methylmagnesium bromide at the Δ 16 -20-oxo fragment of dehydropregnenolone acetate and autooxidation of resulting bromomagnesium 3β-acetoxy-16α-methylpregna-5,17(20)-dien-20-olate. The subsequent reduction of the 17α-hydroperoxy group and hydrolysis of the 3β-acetoxy group afforded 17α-hydroxy-16α-methyl-substituted dehydropregnenolone acetate and its 3-hydroxy analog in high yield.

Two methods of C 6 -alkylation of hydrocortisone, C 6 -formylation and direct methylenation, have been studied. It is shown that both methods are effective and can be of practical interest because they can produce 6α-methylhydrocortisone from hydrocortisone with an overall yield of 50–55%. A short and convenient pathway has been found for producing Δ 9(11) -6α-methylcorticosteroids used as intermediates in the synthesis of highly active 6α-methyl-9α-halo-substituted hydrocortisone derivatives.

Продукт микробиологического расщепления боковой цепи стеринов – андроста-1,4-диен-3,17-дион – токсичен для бактерий, в частности актинобактерий родов Mycobacterium и Аrthrobacter. Осуществлено превращение стеринов в андроста-1,4-диен-3,17-дион с помощью культуры мутантного штамма M. neoaurum ВКПМ Ас-1656 с высоким выходом при устранении контакта клеток бактерии с продуктом с помощью сорбента. При этом расщепление более разветвленной боковой цепи фитостеринов с помощью M. neoaurum, в отличие от боковой цепи холестерина, протекало с высокой скоростью только в условиях турбулентного перемешивания культуральной жидкости, содержащей СаСО3, благодаря чему in situ интенсифицировалось образование гидрокарбонатного иона.