Enhanced heterologous polyketide production in Streptomyces by exploiting plasmid co-integration.
A plasmid named pSMALL was discovered in a Streptomyces coelicolor strain that significantly enhanced the levels of production of 15-methyl-6-deoxyerythronolide B, a polyketide lactone normally produced in low amounts by engineered polyketide synthase (PKS) genes. It is a co-integrate between a conventional SCP2*-derived Streptomyces expression plasmid, pJRJ2, and SCP2@, a variant of the parental SCP2* plasmid. SCP2@ has a 45-bp deletion 35 bp upstream of the start codon of the repI gene in the replication region; and this correlated with an enhanced plasmid copy number and polyketide overproduction by its derivatives. This discovery was exploited to construct pBOOST, a high-copy-number cloning vector that can be used to achieve greatly elevated (at least 25-fold), stable metabolite production by PKS genes cloned in SCP2*-derived vectors by forming co-integrates with them.
Hu
Zhihao
Z
Kosan Biosciences, 3832 Bay Center Place, Hayward, CA 94545, USA. hu@kosan.com
Hopwood
David A
DA
Hutchinson
C Richard
CR
D001483:Base Sequence; D015964:Gene Expression Regulation, Bacterial; D007218:Industrial Microbiology / Q000379:methods; D018942:Macrolides / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics* / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism*
D018942:Macrolides; D009097:Multienzyme Complexes