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紫外诱变原生质体选育溶菌酶高产菌株的研究
冯学珍1, 郑媛2, 王跃军3, 孙谧4, 于建生5, 程江峰6
1.(1中国水产科学研究院黄海水产研究所,青岛 266071) (2青岛科技大学化工学院,266042) (3柳州医学高等专科学校药学教研室,545006);2.1中国水产科学研究院黄海水产研究所,青岛 266071;3.1中国水产科学研究院黄海水产研究所,青岛 266071;4.1中国水产科学研究院黄海水产研究所,青岛 266071;5.2青岛科技大学化工学院,266042;6.2青岛科技大学化工学院,266042
摘要:
以1株产海洋溶菌酶活性较高的菌株(S-12-86)为原始菌株,对其原生质体的制备、再生及紫外诱变育种进行了研究。实验所确定的原生质体的最佳制备条件为:菌株S-12-86培养18 h,溶菌酶浓度为1.0 mg/ml,在35 ℃下,酶解30 min,原生质体形成率为97.6%,再生率为23.6%。同时实验所确定的原生质体诱变的适宜条件为:30 W紫外灯下80 cm照射120 s。对大量再生突变株进行筛选,最终获得了1株遗传性能稳定的菌株R-J-101,其产酶活力达到1 808 U/mg,比原始菌株(1 290 U/mg)提高了40%。
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Breeding of a high lysozyme producingBacillusstrain by mutagenesis of protoplasts
Abstract:
The preparation, regeneration and UV mutagenesis of protoplasts of strain S-12-86, which had high lysozyme producing activity, were studied. The results showed that the optimal conditions of the preparation and regeneration of protoplast were as follows: incubation time of the strain S 12 86 was 18 h, concentrations of lysozyme was 1.0 mg/ml, and time of enzymolysis was 30 min at 35 ℃. The rates of formation and regeneration of protoplast under the optimal conditions were 97.6% and 23.6%, respectively. The optimal condition for mutagenesis of protoplasts was achieved when the protoplasts of strain S-12-86 were irradiated by a 30 w UV lamp at an 80 cm distance for 120 s. After being selected from a large amount of the regenerative mutants, a genetically stable mutant strain R-J-101, of which enzyme activity was 40% higher than that of the original strain S-12-86, was obtained under specific mutation conditions.
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