Expression of 2-deoxy-scyllo-inosose synthase (kanA) from kanamycin gene cluster in Streptomyces lividans.
An actinomycetes expression vector (pIBR25) was constructed and applied to express a gene from the kanamycin biosynthetic gene cluster encoding 2-deoxy-scyllo-inosose synthase (kanA) in Streptomyces lividans TK24. The expression of kanA in pIBR25 transformants reached a maximum after 72 h of culture. The plasmid pIBR25 showed better expression than pSET152, and resulted in the formation of insoluble KanA when it was expressed in Escherichia coli. This strategy thus provides a valuable tool for expressing aminoglycoside-aminocyclitols (AmAcs) biosynthetic genes in Streptomyces spp.
Thuy
Mai Le
ML
Institute of Biomolecule Reconstruction, Sun Moon University, #100, Kalsan-ri, Tangjeong-myeon, Asan-si, Chungnam, 336-708, Korea.
Kharel
Madan Kumar
MK
Lamichhane
Rajan
R
Lee
Hei Chan
HC
Suh
Joo-Won
JW
Liou
Kwangkyoung
K
Sohng
Jae Kyung
JK
D015870:Gene Expression; D005822:Genetic Vectors; D007612:Kanamycin / Q000096:biosynthesis; D008190:Lyases / Q000096:biosynthesis* / Q000235:genetics*; D048170:Streptomyces lividans / Q000201:enzymology* / Q000235:genetics*
D007612:Kanamycin; D008190:Lyases; C085631:2-deoxy-scyllo-inosose synthase