Quinones as external electron acceptors in steroid dehydrogenation with entrapped cells in organic medium

• A series of quinone-based compounds were tested for their ability to act as external electron acceptors in the 1-dehydrogenation of-αmethyl-hydrocortisone-21-acetate, with polyurethane-entrapped Arthrobacter simplex cells in buffer-saturated n-decan-1-ol. This organic solvent was needed to solubilize the steroid substrate. In aqueous medium, the conversion with free cells virtually stopped after one hour, probably due to substrate limitation. All the tested quinones acted as external electron acceptors, increasing the bioconversion rate. The process kinetics were complex. However, when keeping the concentration of one of the substrates (steroid or quinone) constant and varying that of the other, Michaelis-Menten kinetics provided a reasonably good model for the initial reaction rates, and apparent kinetic constants were estimated. The most effective of the tested external electron acceptors were 2,6-dimethyl-p-benzoquinone and menadione. Mass transfer limitations seemed to appear after some hours of reaction, with low concentrations of the more efficient quinones, when the biocatalyst microenvironment was quinone- and possibly oxygen-depleted. Monosodium glutamate was included with the cells in the immobilisation foam, as an activity-stabilizing agent. It was observed that some of the quinones apparently formed complexes with this glutamate, thereby influencing the kinetics of the process. The catalytic half-life of the system depended on the quinone concentration and optimal values (60-80 h) were observed at 1 mM levels of 2,6-dimethyl-p-benzoquinone or menadione. Quinone toxicity, direct or through the formation of peroxides in the aerobic reoxidation process, may be at the origin of enzyme deactivation.

Ämnesord

TEKNIK OCH TEKNOLOGIER  -- Kemiteknik -- Kemiska processer (hsv//swe)

ENGINEERING AND TECHNOLOGY  -- Chemical Engineering -- Chemical Process Engineering (hsv//eng)

Nyckelord

Arthrobacter simplex

External electron acceptor

Organic medium

Polyurethane entrapment

Quinones

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