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Protoplast formation and regeneration from Streptomyces clavuligerus NRRL 3585 and clavulanic acid production

机译:克拉维链霉菌NRRL 3585原生质体的形成和再生以及克拉维酸的产生

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Protoplasts of the wild type Streptomyces clavuligerus NRRL 3585 (ATCC 27064) were formed from spores cultures obtained in the lag, exponential and stationary growth phases by using 0.5% glycine in the culture medium. The protoplasts were obtained by treatment of the cells with lysozyme (EC-3.2.1.17) 40,000 U (1mg/mL), in an osmotic solution for 90 min at 28oC. The frequency of regenerated protoplasts in the lag phase was 1.7x103 CFU/mL (28.97%), in the beginning of the exponential phase 0.4x102 CFU/mL (31.67%), in the exponential growth phase 2.5x103 CFU/mL (46.30%) and 1.0x10(5) CFU/mL in stationary phase (48.45%). Antibiotic production and activity of regenerated protoplasts were observed in all phases, except in the lag phase. The protoplast formation and regeneration techniques resulted in a new isolate strain of Streptomyces clavuligerus that produced approximately 2.5 fold more clavulanic acid.
机译:野生型克拉维链霉菌NRRL 3585(ATCC 27064)的原生质体是通过在培养基中使用0.5%甘氨酸,从在迟滞,指数和静止生长期获得的孢子培养物形成的。通过在28oC的渗透溶液中用40,000 U(1mg / mL)的溶菌酶(EC-3.2.1.17)处理细胞获得原生质体。滞后阶段的原生质体再生频率为1.7x103 CFU / mL(28.97%),在指数阶段开始时为0.4x102 CFU / mL(31.67%),在指数生长期为2.5x103 CFU / mL(46.30% )和1.0x10(5)CFU / mL的固定相(48.45%)。除滞后期外,所有阶段均观察到抗生素的产生和再生原生质体的活性。原生质体的形成和再生技术产生了一种新的分离株,即棒状链霉菌,产生了大约2.5倍的棒酸。

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