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Stabilization ofd-amino-acid oxidase fromTrigonopsis variabilis by manganese dioxide

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Abstract

Stabilization of immobilizedd-amino-acid oxidase was achieved as follows. YeastTrigonopsis variabilis producingd-amino-acid oxidase was used to deaminate cephalosporin C to glutaryl07-aminocephalosporanic acid. Permeabilized cells were co-immobilized with manganese dioxide by entrapment in (poly)acrylamide gel so that hydrogen peroxide, liberated in the reaction, could be partially deactivated and both the enzyme and the substrate could be stabilized. Activity of entrapped cells was determined by HPLC and enzyme flow microcalorimetry. The process was evaluated in terms of activity, immobilization yield, storage stability and oxo-product formation by immobilized preparations. The storage stability of immobilized biocatalysts with MnO2 was nearly doubled and production of 2-oxoadipyl-7-aminocephalosporanic acid was 2–3-fold higher than by entrapped cells without MnO2. Glutaryl-7-aminocephalosporanic acid can be easily obtained from the resulting oxo-product by a non-enzymic reactionvia externally added hydrogen peroxide.

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Abbreviations

d-AAO:

d-amino-acid oxidase (EC 1.4.3.3)

7-ACA:

7-aminocephalosporanic acid

CPC:

cephalosporin C

EFMC:

enzyme flow microcalorimetry

GA:

glutaraldehyde

Glu-7-ACA:

glutaryl-7-aminocephalosporanic acid

2-OA-7-ACA:

2-oxoadipyl-7-aminocephalosporanic acid

PAG:

(poly)acrylamide gel

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Authors and Affiliations

  1. Institute of Chemistry, Slovak Academy of Sciences, 842 38, Bratislava, Slovakia

    A. Vikartovská-Welwardová & P. Gemeiner

  2. Biotika Pharmaceutical Plant, 976 13, Slovenská L'upča, Slovakia

    E. Michalková & L. Welward

Authors
  1. A. Vikartovská-Welwardová
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  2. E. Michalková
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  3. P. Gemeiner
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  4. L. Welward
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Vikartovská-Welwardová, A., Michalková, E., Gemeiner, P. et al. Stabilization ofd-amino-acid oxidase fromTrigonopsis variabilis by manganese dioxide. Folia Microbiol 44, 380–384 (1999). https://doi.org/10.1007/BF02903709

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