【摘要】 为了提高辅酶Q10的产生菌米曲霉菌的基因组改组育种效率,以提高辅酶Q10的产量,本试验对米曲霉原生质体的制备条件进行了研究。通过对影响制备效果的条件,即米曲霉的菌丝菌龄、酶解温度、时间和原生质体稳定剂的浓度四个因素进行全面优化,并利用正交试验确定了原生质体制备的适宜条件:菌龄为12h、原生质体渗透压稳定剂NaCl浓度为0.85mol/L、酶解时间为3.5min、酶解温度为30℃时,米曲霉原生质体制备效果最好,所得原生质体为6.3×106个/mL,再生率达到38.06%。较高的原生质体生成量和再生率为该菌后序的基因组改组研究奠定了基础。更多还原

【Abstract】 In order to improve the breeding efficiency of Aspergillus oryzae by genome shuffling methods to produce more coenzyme Q10 products,the processing conditions to generate Aspergillus oryzae protoplasts are studied by optimum seeking method and orthogonal experiment in this paper.The results show that the optimal conditions for the protoplast formation and regeneration are as follows:the age of the mycelium of Aspergillus oryzae for protoplasts is 12 h,the hydrolysis reaction temperature is 30 ℃,the hydrolysis reaction time is 3.5min,the protoplast osmotic stabilizer NaCl concentration is 0.85mol/L,and under the optimal conditions,the protoplast formation amount and regeneration rate could respectively approach 6.3× 106 cells/mL and 38.06%.The high yield and high regeneration rate lay agood foundation for the genetic improvement of industrial coenzyme Q10-producing strain.更多还原