[Effect of lat disruption on clavulanic acid production].
A 1.8kb fragment of lat was obtained from Streptomyces clavuligerus 27064, and replacement plasmid of pXAL1 and pXAL2 were constructed. PXAL1 and pXAL2 were used to disrupt the lat gene by bi-parental conjugation from E. coli to Streptomyces clavuligerus. A Am(r)Thio(S) transformant, named as XAL863, was obtained. The genome of Streptomyces clavuligerus 27064 and XAL863 was analyzed by southern blot technique, and the activity of lysine epsilon-aminotransferase in the two strains was also tested. Both results proved that the lat was disrupted in the XAL863. Streptomyces clavuligerus and XAL863 were cultured in the shaken flask respectively, and the production of clavulanic acid was analyzed by HPLC with the different incubation time interval, and the yield was approximately 1.8 times higher in the XAL863 at their highest production point.
Wang
Yong-Hu
YH
Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China. yonghw@263.net
Jing
Chen-Feng
CF
Tao
Mei-Feng
MF
Yang
Bo
B
Xu
An-Long
AL
D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D015139:Blotting, Southern; D002851:Chromatography, High Pressure Liquid; D019818:Clavulanic Acid / Q000096:biosynthesis*; D005285:Fermentation; D051306:L-Lysine 6-Transaminase / Q000235:genetics / Q000502:physiology*; D016133:Polymerase Chain Reaction; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D019818:Clavulanic Acid; D051306:L-Lysine 6-Transaminase