利用原生质体再生技术选育内切型菊粉酶高产菌株

The strain screen for high producing endo-inulinase breeding by protoplast preparation technique

利用原生质体制备与再生技术选育内切型菊粉酶高产菌株,以I200913作为出发菌株(酶活为0.413 u/m L),研究了在不同酶配比酶解液、不同酶解温度、不同酶解时间的条件下,对原生质体制备、再生以及内切型菊粉酶酶活的影响。研究结果表明,在酶配比为:纤维素酶0.7%+蜗牛酶0.2%+溶菌酶0.1%=1%,酶解温度为30℃,酶解时间为2.5h条件下,原生质体形成率最高,为22.32%。在酶配比为:纤维素酶0.7%+蜗牛酶0.2%+溶菌酶0.1%=1%,酶解温度为30℃,酶解时间为2.5h条件下,原生质体再生率最高,为16.02%。在酶配比为:纤维素酶0.7%+蜗牛酶0.2%+溶菌酶0.1%=1%,酶解温度为30℃,酶解时间为2.5h条件下,原生质体再生菌株内切型菊粉酶酶活最高为0.598 u/m L,比出发菌株提高了28%。

The protoplast preparation and regeneration technique was used to breed the strains with high producing yield of endo-inulinase. The strain 1200913 （enzyme activity, 0.413 u/mL） was used as original strain. The effects of different ratio of enzymatic hydrolysate, hydrolysis temperature and hydrolysis time on the protoplast preparation and regeneration as well as enzyme activity of endo-inulinase were studied. The results showed that the highest rate of protoplast formation of 22.32% was obtained at ： 1% enzymes, 0.7% cellulose, 0.2% glusulase and 0.1% lysozyme, 30℃ for 2.5 hours. The highest rate of protoplast regeneration of 16.02% was obtained at ： 1% enzymes, 0.7% cellulose, 0.2% glusulase and 0.1% lysozyme, 30 ℃ for 2.5 hours. The highest endo-inulinase activity producing by protoplast preparation was 0.598 u/mL, and 28% higher than the original strain, and it was obtained at. 1% enzymes, 0.7% cellulose, 0.2% glusulase and 0.1% lysozyme, 30 ℃ for 2.5 hours.