一株青藏高原来源的链霉菌菌株鉴定及其抑菌活性的初步分析  

Identification of A Streptomyces Strain Isolated from Qinghai-Tibet Plateau and Its Antibacterial Activity

作　　者：黄青兰[1] 袁文婧[1] 庄原[1] 罗清[1] 彭程[1] 叶波平[1] 

机构地区：[1]中国药科大学生命科学与技术学院,江苏南京210009

出　　处：《药物生物技术》2016年第4期308-312,共5页Pharmaceutical Biotechnology

摘　　要：根据菌株的形态和生理生化特征,结合16srRNA序列分析,将一株分离自中国青藏高原土壤的菌株167-2-28鉴定为链霉菌属放线菌。菌株167-2-28为革兰氏阳性菌,在高氏合成琼脂培养基上,基内菌丝呈红褐色,气生菌丝为灰粉色,产生浅褐色可溶性色素,孢子链为螺旋形,孢子呈椭圆形,大小约为（0.7~0.8）μm×（1.0~1.4）μm。菌株167-2-28的DNA G＋C含量为68.3 mol%,以MK-9（H6）为主要呼吸醌,其细胞中主要的脂肪酸为iso-C16：0（18.6%）、anteiso-C15：0（16.6%）、anteiso-C17：0（11.5%）和summed feature 3（C16：1ω7c/isoC15：0-2OH）（14.2%）。对16s rRNA序列的分析结果显示,该菌株与S.novaecaesareae NBRC 13368T（99.57%）、S.phaeoluteigriseus ISP 5182T（99.28%）和S.pseudovenezuelae NBRC 12904T（99.21%）等菌株亲缘关系较近,与菌株S.novaecaesareae ATCC 27452T的主要区别在于：菌株167-2-28在ISP3、ISP4、ISP5和ISP7培养基上可产生气生菌丝,并能产生H2S和利用柠檬酸。初步的活性分析结果表明：菌株167-2-28在高氏合成培养基中培养时,获得的乙酸乙酯提取浸膏对革兰氏阳性菌（金黄色葡萄球菌、枯草芽孢杆菌和藤黄微球菌）、耐甲氧西林金黄色葡萄球菌（MRSA）和耐甲氧西林表皮葡萄球菌（MRSE）的生长具有明显的抑制作用,并对黑曲霉有较弱的拮抗活性,其最低抑菌浓度（MIC）分别为32,32,16,50,100μg/m L和〉500μg/m L;而对革兰氏阴性菌（大肠埃希菌、铜绿假单胞菌、克雷伯氏菌）和真菌（白色念珠菌和禾谷丝核菌）的生长无抑制作用。Strain 167-2-28,which was isolated from soil of Qinghai-Tibet Plateau in west China,was identified as a possible member of the genus Streptomyces. It was Gram-staining-positive,brownish pigmented and could form red brown substrate mycelium and gray pink aerial mycelium on Gause＇s synthetic medium. Spiral spore chains were produced,and the spores were oval-shaped with 1. 0 ~1. 4 μm long and 0. 7-0. 8μm wide. The DNA G ＋ C content of strain 167-2-28 was about 68. 3 mol%. The predominant menaquinones were identified as MK-9（ H6）,and the major fatty acids of strain were iso-C16： 0（ 18. 6%）,anteiso-C15： 0（ 16. 6%）,anteiso-C17： 0（ 11. 5%） and summed feature 3（ C16： 1ω7c /iso-C15： 0-2OH）（ 14. 2%）. Phylogenetic analysis of the 16 S r DNA gene sequence showed that strain 167-2-28 was closely related to S. novaecaesareae ATCC 27452T（ 99. 57%）,S. phaeoluteigriseus ISP 5182T（ 99. 28%） and S. pseudovenezuelae NBRC 12904T（ 99. 21%）. However,unlike the reported strain of S. novaecaesareae ATCC27452 T,strain 167-2-28 could form aerial mycelium on ISP3,ISP4,ISP5 and ISP7 medium and produce H2 S and assimilate citrate.Further studies showed that the ethyl acetate extract from strain 167-2-28 cultured on Gause＇s synthetic medium inhibited the growth of Gram-positive bacteria and drug-resistance bacteria apparently with the minimum inhibitory concentrations（ MIC） value of 32,32,16,50 and 100 μg / m L against Staphylococcus aureus,Bacillus subtilis,Micrococcus luteus,Methicillin-resistant staphylococcus aureus（ MRSA） and Methicillin-resistant staphylococcus epidermidis（ MRSE）,respectively. In addition,the crude extract demonstrated weak activity against A. niger（ MIC〉 500 μg / m L）,while it could not suppress the growth of Candida albicans,Rhizoctonia cerealis and Gram-negative bacteria,such as Escherichia coli,Pseudomonas aeruginosa and Klebsiella aerogenes.

关 键 词：链霉菌属 青藏高原 分类 鉴定 抑菌活性 MRSA MRSE 

分 类 号：Q93[生物学—微生物学]