CLONED DNA CONTAINING RAT L-GULONOLACTONE OXIDASE, FRAGMENT THEREOF, GENE RECOMBINANT VECTOR CONTAINING SAID CLONED DNA OR FRAGMENT INTEGRATED THEREIN, HOST CELL TRANSFORMED WITH SAID VECTOR AND PRODUCTION OF L-GULONOLACTONE OXIDASE OR SUBSTANCE ANALOGOUS THERETO USING SAID HOST CELL

摘要

PURPOSE:To mass-product L-gulonolactone oxidase useful in synthesis of L-ascorbic acid, by using a cloned DNA containing a base sequence coding an amino acid sequence of rat L-gulonolactone oxidase. CONSTITUTION:A rabbit is immunized with a high-purity L-gulonolactone oxidase derived from a rat to prepare an antiserum. A library expressing a cDNA of a lambdagt11 phage prepared from an mRNA derived from a rat liver is used to screen a clone of the L-gulonolactone oxidase by an enzyme immunoassay using the above-mentioned antiserum. The resultant phage DNA is then cleaved with a restriction enzyme EcoRI to afford the aimed cloned DNA of 2,100 nucleotides. A gene recombinant containing the above-mentioned cloned DNA or fragment thereof integrated therein is subsequently constructed to incorporate the afore-mentioned vector in a host cell and carry out transformation. The resultant tranformant cell is then cultivated to produce an L- gulonolactone oxidase.