Tamegai Hideyuki, Nango Eriko, Kuwahara Mieko, Yamamoto Hideki, Ota Yasumasa, Kuriki Hisako, Eguchi Tadashi, Kakinuma Katsumi
Department of Chemistry, Tokyo Institute of Technology, Japan.
J Antibiot (Tokyo). 2002 Aug;55(8):707-14. doi: 10.7164/antibiotics.55.707.
Using inverse PCR, two new genes (btrN and btrS) were identified upstream of the putative glycosyltransferase gene btrM in the butirosin-biosynthetic btr gene cluster of Bacillus circulans. The upstream gene btrS showed significant homology with stsC of Streptomyces griseus, which encodes L-glutamine:scyllo-inosose aminotransferase in the biosynthesis of streptomycin. The function of BtrS was further confirmed by heterologous expression in Escherichia coli and chemical identification of the conversion of 2-deoxy-scyllo-inosose into 2-deoxy-scyllo-inosamine. The identification of BtrS as L-glutamine:2-deoxy-scyllo-inosose aminotransferase is the first report of the aminotransferase gene responsible for 2-deoxystreptamine biosynthesis.
利用反向PCR技术,在环状芽孢杆菌的丁胺卡那霉素生物合成btr基因簇中,于假定的糖基转移酶基因btrM上游鉴定出两个新基因(btrN和btrS)。上游基因btrS与灰色链霉菌的stsC具有显著同源性,stsC在链霉素生物合成中编码L-谷氨酰胺:顺式-肌醇氨基转移酶。通过在大肠杆菌中的异源表达以及对2-脱氧-顺式-肌醇转化为2-脱氧-顺式-肌醇胺的化学鉴定,进一步证实了BtrS的功能。将BtrS鉴定为L-谷氨酰胺:2-脱氧-顺式-肌醇氨基转移酶,这是关于负责2-脱氧链霉胺生物合成的氨基转移酶基因的首次报道。