CAJ | 학술논문

目的从700多株海洋沉积物分离得到的细菌中筛选安莎类抗生素和6-脱氧己糖(6DOH)糖基化修饰的次级代谢产物产生菌.方法以3-氨基-5-羟基苯甲酸(AHBA)合酶基因和dTDP-葡萄糖-4,6-脱水酶基因为靶标,分别进行安莎类抗生素和6DOH糖基化修饰的次级代谢产物产生菌的分子筛选.对AHBA合酶及dTDP-葡萄糖-4,6-脱水酶基因阳性的菌株发酵液及提取物进行抗菌、抗肿瘤、抗病毒活性分析.采用利福霉素抗性及氢氧化钠显色初步鉴定安莎类化合物;采用α-萘酚硫酸显色初步鉴定6DOH糖基化修饰的化合物.利用16S rRNA序列对部分阳性菌株进行系统发育分析.结果共获得39株AHBA合酶基因阳性和10株dTDP-葡萄糖-4,6-脱水酶基因阳性菌株.阳性菌株中,78%具有不同程度的生物活性.化学初步鉴定结果表明:49%的AHBA合酶基因阳性菌株产生安莎类化合物:50%的dTDP-葡萄糖-4,6-脱水酶基因阳性菌株产生6DOH糖基化修饰的化合物.系统发育分析表明,大多数阳性菌株属于放线菌中的链霉菌属.结论基因探针筛选可作为一种合理、有效的方法从海洋细菌中发现活性代谢产物.
목적 종700다주해양침적물분리득도적세균중사선안사류항생소화6-탈양기당(6DOH)당기화수식적차급대사산물산생균.방법 이3-안기-5-간기분갑산(AHBA)합매기인화dTDP-포도당-4,6-탈수매기인위파표,분별진행안사류항생소화6DOH당기화수식적차급대사산물산생균적분자사선.대AHBA합매급dTDP-포도당-4,6-탈수매기인양성적균주발효액급제취물진행항균、항종류、항병독활성분석.채용리복매소항성급경양화납현색초보감정안사류화합물;채용α-내분류산현색초보감정6DOH당기화수식적화합물.이용16S rRNA서렬대부분양성균주진행계통발육분석.결과 공획득39주AHBA합매기인양성화10주dTDP-포도당-4,6-탈수매기인양성균주.양성균주중,78%구유불동정도적생물활성.화학초보감정결과표명:49%적AHBA합매기인양성균주산생안사류화합물:50%적dTDP-포도당-4,6-탈수매기인양성균주산생6DOH당기화수식적화합물.계통발육분석표명,대다수양성균주속우방선균중적련매균속.결론 기인탐침사선가작위일충합리、유효적방법종해양세균중발현활성대사산물.
Objective To screen ansamycin and 6-deoxyhexoses (6DOH) glycosylated secondary metabolites producers from more than 700 marine-sediment bacteria. Methods A gene-probe screening strategy was established targeting the 3-amino-5-hydroxybenzoic acid (AHBA) synthase and dTDP-glucose-4,6-dehydratase genes for discovering ansamycins and 6DOH glycosylated secondary metabolites, respectively. Bioactivities of the AHBA synthase and dTDP-glucose-4,6-dehydratase gene-positive strains were evaluated, including anti-bacterial, anti- tumoral and anti-viral activities. Rifamycin resistance profiles and color reactions with sodium hydroxyl or α naphthol were performed as preliminary identification of potential ansamycin or 6DOH glycosylated secondary metabolite producers. Taxonomic and phylogenetic analysis of some positive strains was conducted using partial 16S rRNA sequences. Results In total, 39 AHBA synthase gene-positive and 10 dTDP-glucose-4,6-dehydratase gene-positive strains were obtained. Of these positive strains, 78% showed varying degrees of biological activities. Preliminary chemical identification of metabolites showed that 49% of AHBA synthase gene-positive strains probably produced ansamycins while 50% of dTDP-glucose-4,6-dehydratase gene-positive strains might produce 6DOH glycosylated compounds. Taxonomic and phylogenetic analysis indicated that most of these positive strains belonged to Streptomyces. Conclusion The results suggested that gene-probe screening was a rational and effective strategy for discovering bioactive metabolites from marine bacteria.