Improvement of Natamycin Production by Cholesterol Oxidase Overexpression in Streptomyces gilvosporeus.
Natamycin is a widely used antifungal antibiotic. For natamycin biosynthesis, the gene pimE encodes cholesterol oxidase, which acts as a signalling protein. To confirm the positive effect of the gene pimE on natamycin biosynthesis, an additional copy of the gene pimE was inserted into the genome of Streptomyces gilvosporeus 712 under the control of the ermE* promoter (permE*) using intergeneric conjugation. Overexpression of the target protein engendered 72% and 81% increases in the natamycin production and cell productivity, respectively, compared with the control strain. Further improvement in the antibiotic production was achieved in a 1 L fermenter to 7.0 g/l, which was a 153% improvement after 120 h cultivation. Exconjugants highly expressing pimE and pimM were constructed to investigate the effects of both genes on the increase of natamycin production. However, the co-effect of pimE and pimM did not enhance the antibiotic production obviously, compared with the exconjugants highly expressing pimE only. These results suggest not only a new application of cholesterol oxidase but also a useful strategy to genetically engineer natamycin production.
Institute of Biopharmaceuticals of Shandong Province, Shandong Key Laboratory of Biological Drugs, Jinan 250101, P.R. China.|Shandong Academy of Pharmaceutical Science, Polysaccharide Drugs Engineering Laboratory of Shandong Province, Jinan 250101, P.R. China.
Miao
M
Wang
Institute of Biopharmaceuticals of Shandong Province, Shandong Key Laboratory of Biological Drugs, Jinan 250101, P.R. China.|Shandong Academy of Pharmaceutical Science, Polysaccharide Drugs Engineering Laboratory of Shandong Province, Jinan 250101, P.R. China.
Shaohua
S
Wang
Institute of Biopharmaceuticals of Shandong Province, Shandong Key Laboratory of Biological Drugs, Jinan 250101, P.R. China.|Shandong Academy of Pharmaceutical Science, Polysaccharide Drugs Engineering Laboratory of Shandong Province, Jinan 250101, P.R. China.
Gongli
G
Zong
Institute of Biopharmaceuticals of Shandong Province, Shandong Key Laboratory of Biological Drugs, Jinan 250101, P.R. China.|Shandong Academy of Pharmaceutical Science, Polysaccharide Drugs Engineering Laboratory of Shandong Province, Jinan 250101, P.R. China.
Zhongwen
Z
Hou
Institute of Biopharmaceuticals of Shandong Province, Shandong Key Laboratory of Biological Drugs, Jinan 250101, P.R. China.|Shandong Academy of Pharmaceutical Science, Polysaccharide Drugs Engineering Laboratory of Shandong Province, Jinan 250101, P.R. China.
Fei
F
Liu
Hormel Institute, University of Minnesota, Austin, MN55912, USA.
D Joshua
DJ
Liao
Institute of Biopharmaceuticals of Shandong Province, Shandong Key Laboratory of Biological Drugs, Jinan 250101, P.R. China.|Shandong Academy of Pharmaceutical Science, Polysaccharide Drugs Engineering Laboratory of Shandong Province, Jinan 250101, P.R. China.
Xiqiang
X
Zhu
D000900:Anti-Bacterial Agents / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D019149:Bioreactors; D002789:Cholesterol Oxidase / Q000235:genetics* / Q000378:metabolism*; D003470:Culture Media / Q000737:chemistry; D021901:DNA, Intergenic; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics; D005818:Genetic Engineering; D016680:Genome, Bacterial; D005810:Multigene Family; D010866:Natamycin / Q000096:biosynthesis* / Q000302:isolation & purification; D011401:Promoter Regions, Genetic; D013302:Streptomyces / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D003470:Culture Media; D021901:DNA, Intergenic; D010866:Natamycin; D002789:Cholesterol Oxidase
Natamycin; Streptomyces gilvosporeus; cholesterol oxidase; intergeneric conjugations