High titer production of tetracenomycins by heterologous expression of the pathway in a Streptomyces cinnamonensis industrial monensin producer strain.
Streptomyces cinnamonensis C730.1 and C730.7, are industrially mutagenized strains that produce moderate and high levels of the polyketide polyether antibiotic monensin A, respectively, in an oil-based fermentation medium. The possibility that these strains could be used for high titer production of a heterologous polyketide product was investigated by expression of the entire tetracenomycin (TCM) biosynthetic pathway using an integrative plasmid, pSET154. Expression in C730.1 led to stable production of approximately 0.44 g/l TCM C (the final biosynthetic product) and approximately 2.69 g/l TCM A2 (the penultimate biosynthetic product), and resulted in a 40% decrease in monensin production. Expression in the C730.7 led to higher levels of TCMs, approximately 0.6 g/l TCM C and approximately 4.35 g/l TCM A2, without any detectable decrease in the higher titer monensin production. Abrogation of monensin production in this strain through deletion of the corresponding biosynthetic genes did not lead to higher levels of TCM products. In the case of the C730.7 host, 85% of the TCM C and virtually all of the TCM A2 were intracellular, suggesting feedback inhibition leads to the accumulation of the final pathway intermediate. These observations contrast those made for the native producer Streptomyces glaucescens where the predominant product is TCM C and TCM titers are significantly lower levels (approximately 0.3 g/l), and demonstrate the potential utility of S. cinnamonensis strains as heterologous hosts for high level expression of a variety of polyketide synthase derived products.
Li
Chaoxuan
C
Department of Chemistry, Portland State University, Portland, OR 97207, United States.
Hazzard
Chris
C
Florova
Galina
G
Reynolds
Kevin A
KA
D024861:Genetic Enhancement / Q000379:methods*; D007218:Industrial Microbiology / Q000379:methods; D008985:Monensin / Q000378:metabolism*; D009279:Naphthacenes / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*; D013045:Species Specificity; D013302:Streptomyces / Q000145:classification / Q000502:physiology*
D009279:Naphthacenes; D011994:Recombinant Proteins; C080889:5838 DNI; D008985:Monensin