Application of Immobilized Cholest-4-en-3-one Δ1-Dehydrogenase from Sterolibacterium Denitrificans for Dehydrogenation of Steroids

Mateusz Tataruch,Katarzyna Szymańska,Agnieszka M Wojtkiewicz,Patrycja Wójcik,Katarzyna Zaczyk,Maciej Szaleniec

doi:10.3390/catal10121460

Abstract

Cholest-4-en-3-one Δ1-dehydrogenase (AcmB) from Sterolibacterium denitrificans was successfully immobilized on 3-aminopropyltrimethoysilane functionalized mesoporous cellular foam (MCF) and Santa Barbara Amorphous (SBA-15) silica supports using adsorption or covalently with glutaraldehyde or divinyl sulfone linkers. The best catalyst, AcmB on MCF linked covalently with glutaraldehyde, retained the specific activity of the homogenous enzyme while exhibiting a substantial increase of the operational stability. The immobilized enzyme was used continuously in the fed-batch reactor for 27 days, catalyzing 1,2-dehydrogenation of androst-4-en-3-one to androst-1,4-dien-3-one with a final yield of 29.9 mM (8.56 g/L) and 99% conversion. The possibility of reuse of the immobilized catalyst was also demonstrated and resulted in a doubling of the product amount compared to that in the reference homogenous reactor. Finally, it was shown that molecular oxygen from the air can efficiently be used as an electron acceptor either reoxidizing directly the enzyme or the reduced 2,4-dichlorophenolindophenol (DCPIPH2).

Highlights

Steroids are an important group of naturally occurring or synthetic compounds belonging to nonsaponifiable lipids
0.91 mg mL−1 was immobilized on mesoporous cellular foam (MCF) and Santa Barbara Amorphous (SBA-15) silica carriers functionalized with 3-aminopropyltrimethoysilane (APTS), which provides
That way we proved that DCPIPH2 can be oxidized by molecular oxygen or H2 O2 without the help of the enzyme

Summary

Introduction

Steroids are an important group of naturally occurring or synthetic compounds belonging to nonsaponifiable lipids. Their structure is based on a cyclopenta[a]phenanthrene carbon skeleton which can be partially unsaturated and is usually substituted with a methyl group at C10 and C13 as well as with an alkyl group at C17 [1]. Steroid derivatives form one of the largest group of drugs currently on the market and their synthesis and modification are of utmost importance for the pharmaceutical industry. Due to their structural complexity and often multiple substituents, synthesis and modification of steroids were never entirely based on solely chemical methods.

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