原生质体紫外诱变选育L-半胱氨酸高活力转化菌株  

Screening of High L-Cysteine Producing Strain Through Mutation of Protoplast Induced by UV Radiation

作　　者：王普[1] 蒋红军[1] 何军邀[1] 吴敏[1] 

机构地区：[1]浙江工业大学生物制药研究所

出　　处：《食品与发酵工业》2008年第3期1-4,共4页Food and Fermentation Industries

基　　金：浙江省科技厅重太项目资助(No.2006C13001)

摘　　要：对Pseudomonas sp．HJ14的原生质体制备和再生条件进行了研究，并对其原生质体紫外辐照诱变选育L-半胱氨酸高活力转化菌株。在37℃、2mg／mL溶菌酶作用下酶解60min，其原生质体的形成率和再生率分别为78．6％和28．6％。经过紫外辐照诱变Pseudomona sp．HJ-14原生质体，在含DL-2-氨基-△^2-噻唑啉-4-羧酸（DL-ATC）再生平板上筛选抗性菌株，获得1株酶活较高的正突变株B-3，该菌株具有良好的遗传稳定性和酶活稳定性。采用5L罐进行产酶培养，并在pH8．0、DL-ATC·3H2O浓度为10g／L、42℃酶促反应9h，L-半胱氨酸最高产量达4．63g／L，摩尔转化率为76．5％，较HJ-14提高了117．7％。Screening of a high L-Cysteine producing strain through protoplast mutation induced by UV radiation was studied. The results showed that the optimal conditions for the preparation of protoplast were enzymolysed with lysozyme 2mg/mL, at 37℃ for 60 minutes. The formation frequency and the regeneration frequency of the protoplasts were 78.6% and 28.6%, respectively. The protoplasts of Pseudomonas sp. HJ -14 were induced by UV radiation for 4 min, and the mutants were screened out on the regenerative plate containing DL-ATC 3-20g/L. A high-yield strain Pseudomonas sp. B-3 was obtained, which had a good stability of heredity and enzyme activation. In 5L bioreactor,4.63g/L of L-cysteine was produced by Pseudomonas sp. B-3 from 10 g/L DL-ATC · 3H2O at 42℃, pH 8.0 for 9h , with the molar yield of 76.5%, increased by 117.7% compared with HJ-14.

关 键 词：L-半胱氨酸 假单胞菌 原生质体 UV诱变 

分 类 号：Q933[生物学—微生物学]