Transfer and expression of a Streptomyces cholesterol oxidase gene in Streptococcus thermophilus.
The recombinant plasmid pNCO937 (8.1 kbp) containing a Streptomyces sp. cholesterol oxidase gene was introduced into Streptococcus thermophilus by electrotransformation. Transformation frequency was 7.2 x 10(5) colony forming units/micrograms of DNA. The presence of the cholesterol oxidase gene in S. thermophilus was confirmed with Southern blot analysis using a biotinylated probe. Thin-layer chromatographic analysis showed the expression of the Streptomyces cholesterol oxidase gene resulting in the oxidation of cholesterol to 4-cholesten-3-one. S. thermophilus may be a suitable host for the expression of other genes regulating prokaryotic cholesterol metabolism.
Somkuti
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U.S. Department of Agriculture, ARS, Eastern Regional Research Center, Philadelphia, Pennsylvania 19118.
Solaiman
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Johnson
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Steinberg
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D000818:Animals; D001709:Biotechnology; D002789:Cholesterol Oxidase / Q000096:biosynthesis / Q000235:genetics*; D002791:Cholesterol, Dietary / Q000032:analysis; D003001:Cloning, Molecular; D005524:Food Technology; D015870:Gene Expression; D005798:Genes, Bacterial; D008892:Milk / Q000737:chemistry; D010957:Plasmids; D013291:Streptococcus / Q000201:enzymology* / Q000235:genetics; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics; D014162:Transfection
D002791:Cholesterol, Dietary; D002789:Cholesterol Oxidase