嗜水性產氣單胞桿菌CKH-29蛋白質分泌突變株及性質

Characteristics of Extracellular Protein Secretion Mutants of Aeromonas hydrophila CKH-29

陳素漪(Su-Yi Chen) ； 賴怡岑(Yi-Tsen Lai) ； 劉俊民(Chung-Ming Liou)

《中華生質能源學會會誌》 26卷1&2期 (2007/06) Pp. 67-77

https://doi.org/10.6385/JBESC.200706.0067

摘要

嗜水性產氣單胞桿菌為人類與動物常見之致病菌，其致病因子中含多種外泌性蛋白質，因此蛋白質之胞外分泌機制及其調控可能為其致病性之重要因子。為明瞭蛋白質分泌機制對本菌之影響及其與環境因子之關係，本研究中以臨床株為野生型菌株誘導菌株突變，由其中篩選無法測得外泌蛋白酶活性之突變株Mut1～Mut6。測定各菌株其他外泌性酵素活性，及與蛋白酶外泌機制相關之抗生素耐受性，以分析其突變性質，顯示突變株Mut1〜Mut3 為蛋白質分泌系統突變株，突變株Mut4～Mut6 為細胞構造突變株。實驗又顯示蛋白質分泌系統之變異會影響菌株之胞內及胞外蛋白質之種類及分布，其生長比野生型菌株為差。但突變株生長之差異在常溫及高溫下相似，顯示蛋白質分泌系統之變異與菌株在高溫下生長之轉變無關，因此推測蛋白質分泌系統之調控可能與溫度無直接關係。

關鍵字

嗜水性產氣單胞桿菌 ； 蛋白質分泌性 ； 突變株 ； 致病性

並列摘要

Aeromonas hydrophila is a Gram negative, facultative anaerobic bacterium occurring widely in aquatic and soil environments. This bacterium has been regarded as a pathologenic bacterium, can cause diarrheal, intestinal and several human and animal diseases. The pathogenetic mechanisms of this bacterium are not clear, but many studies have revealed that the secretive proteins, including haemolysin, lipase, protease and enterotoxin, are critical virulent factors, may be the crucial roles in its pathogenicity. Due to the possible relativeness of secretion system with pathogenicity, studying on the secretion system of A. hydrophila may help to acquaint the mechanism o f it. In this study, clinical strain A. hydrophila CKH-29 was carried to irradiation with UV to induce mutagenesis, screening of secretion mutants after irradiation and dark cultivation acquired secretion mutants Mut1~Mut6, which were deficient in extracellular enzyme activities o f amylase and lipase in addition to proteinase. The antibiotics ampicillin and tetracyclin-resistance test showed that Mut1~Mut3 are the secretion system mutants and Mut4~Mut6 are the cellular structures mutants related o f protein secretion. SDS-PAGE of extracellular, periplasmic and intracellular fraction proteins in Mut1Mut3 and Mut4~Mut6 showed alteration o f pattern and quantity, the results appealed that mutation in secretion system affected in the protein synthesis. Compare CKH-29, Mut1~Mut3 and Mut4~Mut6 growing at 30 and 37℃, temperature 37℃ showed decline o f log phase period in all bacteria but mutation of secretion system have no effect on the temperature tolerance.

並列關鍵字

Aeromonas hydrophila ； protein secretion ； mutant ； pathogenicity