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Kurzfassung

This technology provides D-Amino Acid Oxidase immobilized via surface display on yeast cells. It can be easily produced and used for the production of the Cephalosporin antibiotics precursor 7-ACA. No undesirable H2O2 is produced, and a single-pot conversion is possible saving valuable process time and production costs.

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Hintergrund

7-aminocephalosporanic acid (7-ACA) is a key precursor in the production of Cephalosporin antibiotics. In the traditional process D-Amino Acid Oxidase (DAAO) deaminates Cephalosporin C (Ceph C) under production of H2O2 and α-ketoadipyl-7-ACA (KA-7-ACA). H2O2 causes the degradation of KA-7-ACA into glutaryl 7-ACA (GL-7-ACA). GL-7-ACA Acylase (GA) transforms both compounds into 7-ACA. The main drawbacks of this process are that H2O2 causes undesirable byproducts and inactivates the enzymes, and a 2-3 step process is necessary.

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Lösung

 

 

 

 

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Vorteile

• Single-pot conversion of cephalosporin C to 7-aminocephalosporanic acid
• Instant H2O2 degradation by native yeast catalase (No formation of GL-7-ACA, No catalase or H2O2 addition necessary, No enzyme activity loss due to oxidation by H2O2
• Direct expression of enzyme in, and immediate immobilization on the surface during yeast fermentation
• Simple purification of SD cells by filtration of fermentation broth
• Stabilization of SD enzyme through immobilization
• Co-immobilization of Glutaryl 7-ACA Acylase possible

 

 

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Anwendungsbereiche

The D-Amino Acid Oxidase is immobilized on the surface of a yeast cell via surface display (SD). The modified yeast cells can be easily produced in large amounts and with high oxidase activity. Cells can be used for the conversion of Ceph C to 7-ACA. The yeast surface thereby catches the formed H2O2 and hydrolyzes it with its natural catalase. No H2O2 production can be detected in the process. The enzymes are stably immobilized on the cells and can easily be separated from the reaction mixture.

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Service

On behalf of University of Natural Resources and Life Sciences, Vienna (BOKU); TransMIT GmbH is looking for collaboration partners and license Agreements (exclusive/non-exclusive) worldwide.