{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,20]],"date-time":"2025-10-20T09:58:07Z","timestamp":1760954287997,"version":"build-2065373602"},"reference-count":30,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2001,10,15]],"date-time":"2001-10-15T00:00:00Z","timestamp":1003104000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotech &amp; Bioengineering"],"published-print":{"date-parts":[[2001,11,20]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>We have successfully expressed and observed secretion of the <jats:italic>Streptomyces clavuligerus<\/jats:italic> deacetoxycephalosporin C synthase (DAOCS) using the <jats:italic>Pichia pastoris<\/jats:italic> expression system. Two clones having multiple copies of the expression cassette were selected and used for protein\u2010expression analysis. SDS\u2010PAGE showed efficient expression and secretion of the bacterial recombinant DAOCS. The highest yield (120 \u03bcg\/mL) was obtained when expression was induced with 2% methanol. Free and immobilized protein were assayed for biological activity and found to expand penicillin N (its natural substrate) and penicillin G to deacetoxycephalosporin C (DAOC) and deacetoxycephalosporin G (DAOG), respectively. \u00a9 2001 John Wiley &amp; Sons, Inc. 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