Cao Zhisheng, Yoshida Ryuhei, Kinashi Haruyasu, Arakawa Kenji
Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Hiroshima, Japan.
J Antibiot (Tokyo). 2015 May;68(5):328-33. doi: 10.1038/ja.2014.160. Epub 2014 Dec 3.
In our effort to find the key intermediates of lankacidin biosynthesis in Streptomyces rochei, three UV-active compounds were isolated from mutant FS18, a gene disruptant of lkcA encoding a non-ribosomal peptide synthetase (NRPS)-polyketide synthase (PKS) hybrid enzyme. Their structures were elucidated on the basis of spectroscopic data of NMR and MS. Two compounds of a higher mobile spot on silica gel TLC (Rf=0.45 in CHCl3-MeOH=20:1) were determined to be an epimeric mixture of citreodiol and epi-citreodiol at the C-6 position in the ratio of 2:1. In contrast, the compound of a lower mobile spot (Rf=~0 in CHCl3-MeOH=20:1) was identical to a 28-membered polyene macrolide pentamycin. The yields of citreodiols and pentamycin in FS18 were 5- and 250-fold higher compared with the parent strain. Introduction of a second mutation of srrX, coding a biosynthetic gene of the signaling molecules SRBs, into mutant FS18 did not affect the production of three metabolites. Thus, their production was not regulated by the SRB signaling molecules in contrast to lankacidin or lankamycin.
在我们寻找罗氏链霉菌中兰卡杀菌素生物合成关键中间体的过程中,从突变体FS18中分离出了三种具有紫外活性的化合物,该突变体是lkcA基因的破坏菌株,lkcA编码一种非核糖体肽合成酶(NRPS)-聚酮合酶(PKS)杂交酶。基于核磁共振(NMR)和质谱(MS)的光谱数据阐明了它们的结构。硅胶薄层层析(TLC)上移动性较高的两个化合物(在氯仿-甲醇=20:1中的Rf = 0.45)被确定为柠檬二醇和表柠檬二醇在C-6位的差向异构体混合物,比例为2:1。相比之下,移动性较低的化合物(在氯仿-甲醇=20:1中的Rf =~0)与一种28元多烯大环内酯类化合物戊霉素相同。与亲本菌株相比,FS18中柠檬二醇和戊霉素的产量分别高出5倍和250倍。将编码信号分子SRBs生物合成基因的srrX的第二个突变引入突变体FS18中,并不影响这三种代谢产物的产生。因此,与兰卡杀菌素或兰卡霉素不同,它们的产生不受SRB信号分子的调控。