Chen Chao-Hsien, Hu Hui-Yu, Cho Yen-Ching, Hsu Wen-Hwei
Department of Medical Laboratory Science, China Medical University, Taichung, 404, Taiwan.
Curr Microbiol. 2006 Aug;53(2):108-12. doi: 10.1007/s00284-005-0276-7. Epub 2006 Jun 26.
A simple method of using compactin for effective screening of microbial strains with high hydroxylation activity at the 6beta position of compactin was developed. Agar plates containing different carbon sources and 500 microg compactin mL(-1) were used to screen the microorganisms that can convert compactin to pravastatin. About 100 compactin-resistant strains were isolated from the Basal agar containing 7% (w/v) mannitol as a carbon source, in which two bacteria, Pseudomocardia autotrophica BCRC 12444 and Streptomyces griseolus BCRC 13677, capable of converting compactin to pravastatin with the yield of 20 and 32% (w/w), respectively, were found. High-performance liquid chromatography using C-18 column and two sequential mobile phases, 30% and 50% (v/v) acetonitrile, was also established to simultaneously determine the concentration of compactin and pravastatin in the culture broth. As such, about 2% of target microorganisms could be obtained from the screening program.
开发了一种使用康帕丁有效筛选在康帕丁6β位具有高羟基化活性的微生物菌株的简单方法。含有不同碳源和500微克/毫升康帕丁的琼脂平板用于筛选可将康帕丁转化为普伐他汀的微生物。从含有7%(w/v)甘露醇作为碳源的基础琼脂中分离出约100株抗康帕丁菌株,其中发现两株细菌,即自养假单胞菌BCRC 12444和灰色链霉菌BCRC 13677,能够将康帕丁转化为普伐他汀,产率分别为20%和32%(w/w)。还建立了使用C-18柱和两种连续流动相(30%和50%(v/v)乙腈)的高效液相色谱法,以同时测定培养液中康帕丁和普伐他汀的浓度。因此,从筛选程序中可获得约2%的目标微生物。