Nucleic acids encoding stable mutants of D-N-.alpha.-carbamoylas

Details Nucleic acids encoding stable mutants of D-N-.alpha.-carbamoylas Nucleic acids encoding stable mutants of D-N-.alpha.-carbamoylas

1997-07-25

1998-09-15

Carlson, Karen Cochrane

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

435183, 435325, 4353201, 536 232, C12P 2106, C12N 900, C07H 1700

Patent

active

058077103

ABSTRACT:
The present invention relates to mutants of D-N-.alpha.-carbamylase wherein at least one of the cysteines in position 243, 250 and 279 of the amino acid sequence of the wild type enzyme is substituted with a different residue selected from natural amino acids; a recombinant plasmid comprising a nucleotidic sequence which encodes for at least one of the mutants of D-N-.alpha.-carbamylase, host microorganisms transformed with said plasmid and a process for the preparation of these mutants by the culture of said microorganisms. The mutants of D-N-.alpha.-carbamoylase have a higher enzymatic stability than that of the wild type enzyme and are particularly useful in the preparation of D-.alpha.-amino acids providing an improvement in the production yields.

REFERENCES:
Alberto Buson et al, "Identification, Sequencing and Mutagenesis of the Gene for a D-Carbamoylase from Agrobacterium Radiobacter", FEMS Microbiology Letters, 145 (1996), pp. 55-62.
Jun Ogawa et al, "Thermostable N-Carbamoyl-D-Amino Acid Amidohydrolase: Screening, Purification and Characterization" Journal of Biotechnology 38 (1994), pp. 11-19.
Renata Grifantini et al, "Topological Mapping of the Cysteine Residues of N-Carbamyl-D-Amino-Acid Amidohydrolase and Their Role in Enzymatic Activity", Journal of Biotechnology 271, pp. 9326-9331.

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