基于基因组改组的米曲霉沪酿3.042多亲株PEG介导融合育种  

Breeding of Aspergillus oryzae 3.042 by multiparental PEG-mediated protoplasts fusion based on Genome shuffling

作　　者：潘力[1] 梁燕嫦[1] 苗小康[1] 胡杰[1] 

机构地区：[1]华南理工大学生物科学与工程学院,广东广州510006

出　　处：《中国酿造》2008年第12期70-73,共4页China Brewing

基　　金：广东省科技攻关项目(2006B13001006)

摘　　要：利用经过分生孢子的原生质体紫外线-氯化锂、NTG复合诱变得到的米曲霉沪酿3.042的8株突变株作为候选株文库,采用基因组改组(genome shuffling),利用原生质体,进行多亲株双灭活PEG介导融合,优化融合条件。以酪蛋白平板初筛,以及固体发酵测定中性蛋白酶酶活复筛,通过2轮融合操作,筛选出6株高产中性蛋白酶的融合株。其中融合菌株FII-41酶活达到7412U/g(干基),比原始出发菌株4212U/g(干基)提高1.76倍,且遗传稳定。Mutant strains ofAspergillus oryzae 3.042 were obtained by compound mutagenesis of UV-LiCI and NTG chemomorphosis. Eight strains with improved neutral protease were selected and used as candidates of genome shuffling library. The eight strains were grouped： one group was inactivated by UV irradiation, another group was inactivated by heat. The technology of multiparental PEG was applied to mediate protoplasts fusion. The casein-degrading plate experiments and the protease actlveity of solid fermentation were used to screen high protease-producing mutants. Six stains with higher neutral protease were obtained after two cycles of confluent procedures. The activity of protease produced by mutant FII-41 reached 7412 U/g, which was 1.76 folds higher than that of the original strain. The protease activities of the mutants were verified to be genetically stable.

关 键 词：米曲霉沪酿3.042 中性蛋白酶 基因组改组 

分 类 号：Q813.2[生物学—生物工程]