柚苷酶高产菌株的选育及培养基优化

李耀; 陈晓龙; 李睿晓; 阚建全

doi:10.13386/j.issn1002-0306.2016.20.039

柚苷酶高产菌株的选育及培养基优化

Breeding and screening of a naringinase-producing strain and optimization of culture medium

摘要

摘要: 为了获得高产稳定的柚苷酶产生菌,以分离纯化出的36株黑曲霉菌为材料,利用常规筛选方法从中选出黑曲霉菌株M53,采用紫外线和Li Cl复合诱变,选育出一株柚苷酶高产突变株M53-7,并通过正交实验对该菌株液态发酵产酶的培养基组成进行优化。实验结果表明,正交优化后的培养基组成为蔗糖0.5%、豆饼粉3.0%、NH₄NO₃1.0%、K₂HPO₄0.1%,无机盐Ca Cl₂、Mg SO₄·7H₂O、Zn SO4在发酵培养基中的添加量分别为0.1%、0.5%、0.1%;采用最佳发酵培养基对突变菌株M53-7进行摇床发酵,其发酵液中柚苷酶活力可达到906.28 U/m L,且产酶性能稳定。

Abstract: In order to obtain a high yield and stable naringin enzyme producing bacteria,taking the 36 strains of Aspergillusnigeras the experimental material,a strain of naringinase production strain M53 was obtained by conventional screening methods. A mutational strain named M53- 7 was bred through multiple mutagenesis( Li Cl treatment and UV irradiation).The culture medium of M53-7 was optimized by orthogonal experiment. The results showed that the optimal parameters for the fermentation culture medium were as follows: sugar 0.5%,soybean meal 3.0%,NH_4NO_31.0%,K_2HPO_40.1%,Ca Cl_20.1%,Mg SO_4·7H_2O 0.5%,Zn SO_40.1%.Under the optimum conditions,the enzymatic activity of M53-7 was 906.28 U / m L.The research proved that the M53-7 was born with the stable performance at naringinase- producing,and it can be utilized in industrial production.