해양에서 분리된 방선균을 이용한 항생제 내성균의 항균활성에 관한 연구

Abstract

43 strains of marine actinomycetes were isolated from marine soil sources in Jeju island. They were characterized by morphological and physio-biochemical properties, and the API 50CHB test. They were further confirm by molecular methods including partial sequencing of 16S rRNA. The phylogenic relationship and taxonomic position of the domain bacteria was investigated by performing a comparative sequence analysis of PCR-amplified 16S ribosomal RNA A neighbor-joining tree of partial 16S rRNA sequences divided in to 3 isolated HK-A13, HK-A43, HK-A44 of bacterial strains. major are, Gram positive bacteria/ Actinobacteria / Actinomycetales(order)/ Streptomycineae (suborder)/ Streptomycetaceae(family)/ Streptomyces(100%). All the 43 strains of Actinomtcetes, were grew in the various range of NaCl, pH and temperature. As the result, three strains of HK-A13, HK-A43, HK-A44 were for most similar to Streptomyces were named as Streptomyces sp. HK-A13 and Streptomyces sp. HK-A43, Streptomyces sp. HK-A44. HK-A13, HK-A43 and HK-A44 strains were showed that best growth on temperature between 20℃~40℃, and pH between 6~7. Streptomyces sp. HK-A13. showed 1% sucrose for carbon source and 1% ammonium nitrate for nitrogen source, and Streptomyces sp. HK-A43 showed 1% sucrose for carbon source and 1% yeast extract for nitrogen source, and Streptomyces sp. HK-A44 showed 1% lactose for carbon source and ammonium chloride or malt extract for nitrogen source indicating that it has the highest growth. As the Minimum InhInhtion Concysiration (MIC) result, methanol extraction from culture liquid of Streptomyces sp. HK-A13 was verified sp. Streptomyces sp. HK methanol extracts very high antimicrobial activation depending on density with Streptococcus parauberis SP-001, 003, 015, 044, 048, 060, 063, 065, 066, Proteus rettgeri CCARM 0263, Bacillus anthrasis CCARM 0217, SStreptococcus faecium CCARM 0210, Klebsiella pneumoniae CCARM 10258, Pseudomonas aeruginosa CCARM 0225 at 50㎎/㎖, Micrococcus luteus CCARM 0022, Staphylococcus aureus CCARM 0201, Enterobacter cloacae CCARM 0253 at 25㎎/㎖, Burkholderia cepacia CCARM 0226 at 6.3㎎/㎖. Methanol extraction of Streptomyces sp. HK-A43 had no sensitivity against human and fish diseases antibiotic-resistance bacteria other than Streptococcus parauberis SP-066, Micrococcus luteus CCARM 0022, Burkholderia cepacia CCARM 0226, Streptococcus faecium CCARM 0210, Enterobactrer cloacae CCARM 0253 causing human pathogen and fish diseases antibiotic-resistance bacteria. Methanol extraction of Streptomyces sp. HK-A44 in showed high sensitivity against Streptococcus parauberis SP-015, 044, 066, Micrococcus luteus CCARM 0022, Burkholderia cepacia CCARM 0226, Streptococcus faecium CCARM 0210 at 50 ㎎/㎖ and vibriosis causing fish diseases antibiotic-resistance bacteria Streptococcus parauberis SP-003, 063, 065 at 25㎎/㎖. The antioxidation activation of (DPPH radical scavenging activation) for each density of methanol extraction from isolated strains broth, was 1.4% HK-A13 at 0.05㎎/㎖, 14% for 0.25㎎/㎖, 25% for 0.5㎎/㎖ and 93% for 0.5㎎/㎖. (HK-A44 showed the RSA was 20%). Although there is no meaningful difference for Streptomyces broth methanol extract HK-A43. The antioxidation activities of 23% for addition of Streptomyces Broth methanol Extract HK-A13 way 0.25㎎/㎖ and 12 % 2.5㎎/㎖, and 83% 80% at 2.5㎎/㎖ of density. The result of Alkyl radical scavenging activity all tested bacterial showed signiticantly increased antioxidation activity at 0.25㎎/㎖. Extraction of HK-A13 atrain at 2.5㎎/㎖ showed the highest radical scavenging activation of 86%.