Hyper-inducible expression system for streptomycetes.
Streptomycetes produce useful enzymes and a wide variety of secondary metabolites with potent biological activities (e.g., antibiotics, immunosuppressors, pesticides, etc.). Despite their importance in the pharmaceutical and agrochemical fields, there have been no reports for practical expression systems in streptomycetes. Here, we developed a "P(nitA)-NitR" system for regulatory gene expression in streptomycetes based on the expression mechanism of Rhodococcus rhodochrous J1 nitrilase, which is highly induced by an inexpensive and safe inducer, epsilon-caprolactam. Heterologous protein expression experiments demonstrated that the system allowed suppressed basal expression and hyper-inducible expression, yielding target protein levels of as high as approximately 40% of all soluble protein. Furthermore, the system functioned in important streptomycete strains. Thus, the P(nitA)-NitR system should be a powerful tool for improving the productivity of various useful products in streptomycetes.
Herai
Sachio
S
Institute of Applied Biochemistry and Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8572, Japan.
Hashimoto
Yoshiteru
Y
Higashibata
Hiroki
H
Maseda
Hideaki
H
Ikeda
Haruo
H
Omura
Satoshi
S
Kobayashi
Michihiko
M
D000619:Aminohydrolases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002209:Caprolactam / Q000031:analogs & derivatives / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D012240:Rhodococcus / Q000201:enzymology / Q000235:genetics; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
D001426:Bacterial Proteins; D002209:Caprolactam; D000619:Aminohydrolases; C021867:nitrilase