US 6,187,574 B1 · Process for producing the enzyme D-amino acid oxidase of Rhodotorula gracilis in host cells

Abstract

Process for producing the enzyme D-amino acid oxydase of Rhodotorula gracilis in host cells. The process for the expression of the enzyme comprises isolating the complementary DNA corresponding to the messager RNA of the gene which codes for the D-amino acid oxydase of any strain of Rhodotorula gracillis producing said enzyme; fusing the fragment of DNA which codes for D-amino acid oxydase of Rhodotorula gracillis with a DNA sequence which contains a site of union to the ribosome and a high efficiency promoter sequence for the express of genes in host cells; inserting the DNA fragment into a plasmid appropriate for the host cell; cultivating the host cells transformed with said plasmid; and collecting the enzyme.

Claims12

Dependent claims

A process comprising producing a Rhodotorula gracilis D-amino acid oxidase enzyme by the steps of:

(a) transforming a prokaryotic host cell with a recombinant DNA molecule comprising
(i) a DNA sequence encoding said enzyme, wherein said DNA sequence comprises SEQ ID NO:8;
(ii) a ribosome binding site and
(iii) a promoter operatively coupled to said DNA sequence to direct expression of the enzyme in said host cell, wherein the prokaryotic host cell is capable of stable production of said enzyme and is devoid of any other enzyme that would degrade glutaryl-7-aminocephalosporanic acid or hydrogen peroxide;
(b) growing the transformed host cell in a culture medium under conditions that cause the host cell to produce said enzyme;
and
(c) recovering the enzyme from the culture medium.

4 more elements

Dependencies are inferred from the claim text.

Process for producing the enzyme D-amino acid oxidase of Rhodotorula gracilis in host cells

Life of this patent

Process for producing the enzyme D-amino acid oxidase of Rhodotorula gracilis in host cells

Life of this patent

Abstract

Claims12