Biotechnological Transformation of Hydrocortisone into 16α-Hydroxyprednisolone by Coupling Arthrobacter simplex and Streptomyces roseochromogenes.

Odile Francesca Restaino,Addolorata Perna,Maria Giovanna Borzacchiello,Chiara Schiraldi,Marcella Cammarota,Antonio Fiorentino,Simona Barbuto Ferraiuolo

doi:10.3390/molecules25214912

Odile Francesca Restaino, Addolorata Perna + Show 5 more

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https://doi.org/10.3390/molecules25214912

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Abstract

16α-Hydroxyprednisolone, an anti-inflammatory drug, could be potentially obtained from hydrocortisone bioconversion by combining a 1,2-dehydrogenation reaction performed by Arthrobacter simplex ATCC31652 with a 16α-hydroxylation reaction by Streptomyces roseochromogenes ATCC13400. In this study we tested, for the first time, potential approaches to couple the two reactions using similar pH and temperature conditions for hydrocortisone bioconversion by the two strains. The A. simplex capability to 1,2-dehydrogenate the 16α-hydroxyhydrocortisone, the product of S. roseochromogenes transformation of hydrocortisone, and vice versa the capability of S. roseochromogenes to 16α-hydroxylate the prednisolone were assessed. Bioconversions were studied in shake flasks and strain morphology changes were observed by SEM. Whole cell experiments were set up to perform the two reactions in a sequential mode in alternate order or contemporarily at diverse temperature conditions. A. simplex catalyzed either the dehydrogenation of hydrocortisone into prednisolone efficiently or of 16α-hydroxyhydrocortisone into 16α-hydroxyprednisolone in 24 h (up to 93.9%). Surprisingly S. roseochromogenes partially converted prednisolone back to hydrocortisone. A 68.8% maximum of 16α-hydroxyprednisolone was obtained in 120-h bioconversion by coupling whole cells of the two strains at pH 6.0 and 26 °C. High bioconversion of hydrocortisone into 16α-hydroxyprednisolone was obtained for the first time by coupling A. simplex and S. roseochromogenes.

Highlights

Microbial transformations are widely used to obtain steroidal drugs with high regio- and stereo-specificity [1,2,3,4,5]
The transformation of cortisone acetate into prednisone acetate by Arthrobacter simplex whole or immobilized cells has been widely described [11,12,13]. 16α-hydroxyprednisolone (16α-OH-PD) or desfluorotriamcinolone is a highly effective anti-inflammatory drug that could potentially be obtained through the biotechnological bioconversion (BC) of hydrocortisone (HC) by coupling a 1,2 -dehydrogenation reaction with a 16α-hydroxylation [14]
A. simplex physiological studies were performed in shake flasks on two different media at two diverse pH and temperature values in order to test the effect of various conditions on both the microbial diverse pH and temperature values in order to test the effect of various conditions on both the growth and the bioconversion of HC into prednisolone (Figure 2)

Summary

Introduction

Microbial transformations are widely used to obtain steroidal drugs with high regio- and stereo-specificity [1,2,3,4,5]. Molecules 2020, 25, 4912 including estrane, pregnane and androstane at specific positions of the rings with high selectivity [6,7] Both hydroxylation and dehydrogenation reactions performed by whole microbial cells or their enzymes, as biocatalysts, have been widely used to modify steroidal drugs to obtain higher pharmaceutical efficacy [7]. Streptomyces species (e.g., Streptomyces argenteolus and Streptomyces roseochromogenes) are able to convert different substrates such as testosterone, 1-dehydrotestolactone, pregnenolone, progesterone and dehydroepiandrosterone into their 16α-hydroxylated derivates, with high specificity thanks to a cytochrome P450 multi enzymatic complex These species could be employed to obtain new efficient anti-inflammatory agents with higher glucocorticoid activities [3,8].

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