链霉菌HMJ-7抑菌物质发酵优化及纯化

Optimizing the Fermentation Conditions and the Purifying the Antibacterial Substance Produced by Streptomyces HMJ-7

为提高HMJ-7菌株的抑菌能力并明确其抑菌物质,对前期筛选得到的具有显著抑菌活性的灰红链霉菌HMJ-7菌株的发酵条件进行了优化,结果表明,灰红链霉菌HMJ-7产生抑菌活性物质的最佳发酵条件为:温度37℃、起始pH7、接种量1%、转速180 r/min和发酵时间为4 d。经发酵条件优化,抑菌活性明显得到提高,抑菌圈直径能达到25 mm以上。利用硫酸铵分级沉淀法、Sephadex G-75分子筛、半制备型高效液相色谱(HPLC)和"724"阳离子交换树脂对其产生的抑菌活性物质进行了初步分离纯化。使用硫酸铵分级沉淀法、Sephadex G-75分子筛和半制备型高效液相色谱(HPLC)都获得了单一的活性组分,而利用"724"阳离子交换树脂未能活性组分分离开。说明Sephadex G-75分子筛和半制备型高效液相色谱(HPLC)适于分离纯化灰红链霉菌HMJ-7产生的抑菌物质。这些结果为HMJ-7产生的抑菌物质的结构鉴定及新型抗生素的开发提供了基础资料。

A Streptomyce named HMJ-7 with high antibacterial activation was screened by our lab, it was identified as Streptomyces griseoruber. For enhancing the antibacterial substance activation, it＇s fermentation conditions were optimized. The results showed that the optimal conditions were temperature 37 ℃, initial pH 7, rotation speed 180 r/min, and fermentation time 4 days. To separate and purify the antibacterial substance, the method of ammonium sulfate precipitation, “724” cation exchange resin adsorption, sephadex G-75 molecular sieve chromatography and HPLC were used. The results showed that it can be separated by ammonium sulfate precipitation, sephadex G-75 molecular sieve chromatography and HPLC, and can＇t be separated by “724”cation exchange resin adsorption. These maybe have some means for the next step Structure identification and new antibiotics development.