| 摘要 |
PURPOSE: To obtain the subject enzyme useful for treatment, etc., of pancreatic insufficiency, etc., in high yield by expressing a plasmid containing a bile acid salt-activated lipase gene or a gene prepared by connecting a gene fragment partially deficient therein to a secretory signal gene in a yeast of the genus Pichia. CONSTITUTION: A gene coding for a bile acid salt-activated lipase or a gene prepared by connecting a gene fragment deficient in a part or all of a gene coding for an amino acid sequence of a repetitive structure of 16 times in a peptide comprising 11 amino acid residues present in a carboxyl terminal region of a gene sequence as a unit to a gene coding for a secretion signal and taken up by an EcoRI site of a plasmid pHIL301. The resultant recombinant plasmid is transduced into a Pichia pastoris GS115 strain to transform the Pichia pastoris. The transformant is then proliferated under conditions of 30-50% dissolved oxygen in saturation with air at 30-37 deg.C temperature and pH2.5-6 and the gene is expressed in a culture medium containing methanol and a protein under conditions of <=20% dissolved oxygen in saturation with air at 15-30 deg.C. Thereby, the lipase is obtained. |
