[Cloning and expression of alpha-hydroxy-gamma-aminobutyl acylase gene of Bacillus circulans NRRL-B3312].
With shot-gun cloning strategy, we used pUB110 plasmid as a vactor to clone DNA fragment of Bacillus circulans NRRL-B3312, which is butirosin producer, into Bacllus subtilis 168. Among the transformants, the results of TLC, bioautography and FAB mass, spectrum analysis for the bioconversion product of No. 733 transformant showed that this transformant could transform kanamycin into amikacin. According to these results, the HABA acylase gene locates on the insert fragment of pUBC733 plasmid harbouring No. 733 transformant. We can confirm that the HABA acylase gene was cloned and expressed in B. subtilis 168. Molecular weight of pUBC733 is 7.3kb. Southern hybridization demonstrated that the 2.8 kb inserted fragment of this plasmid originated from B. circulans NRRL-B3312. The restriction map of pUBC733 plasmid was constructed.
Li
Y
Y
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing.
Liu
B
B
Li
X
X
Yu
H
H
D000217:Acyltransferases / Q000235:genetics*; D000583:Amikacin / Q000378:metabolism*; D001407:Bacillus / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular; D015870:Gene Expression; D005798:Genes, Bacterial; D007612:Kanamycin / Q000378:metabolism*; D010957:Plasmids
D007612:Kanamycin; D000583:Amikacin; D000217:Acyltransferases; C081850:kanamycin - alpha-hydroxy-gamma-aminobutyl acylase