基于基因组重排技术的角蛋白酶高产菌株的筛选毕业论文设计.docVIP

本科生毕业论文 题 目 基于基因组重排技术的角蛋白酶 高产菌株的筛选 学 院 轻纺与食品学院 　 专 业 轻工生物技术 学生姓名 程 双 　 学 号 1043095006 年级 2010级 指导教师 吴 重 德 二Ο一 四 年 六 月 四 日  基于基因组重排技术的角蛋白酶高产菌株的筛选 专业：轻工生物技术 学生：程 双 指导老师：吴重德 摘 要 角蛋白酶以其特异性分解角蛋白的特性，在废弃羽毛的处理，清洁制革技术等领域中得到广泛应用。本论文以实验室保存的地衣芽孢杆菌(Bacillus licheniformis X-47)作为亲本菌株，利用基因组重排技术，筛选高产角蛋白酶的菌株。对基因组重组的条件进行了优化，得到Bacillus licheniformis X-4的最佳基因组重排条件为：菌龄8 h，溶菌酶酶浓度1.0 mg/mL，酶解时间30 min，酶解温度40，融合时间15 min，融合温度35，融合pH9.0，融合PEG浓度400 g/mL。通过筛选得到第一代融合子Y1-28和Y1-31，Y1-28角蛋白酶活力为240.9±7.2 U/mL，Y1-31角蛋白酶活力为257.5±17.5 U/mL，较原始菌株分别提高了68.4%和80.0%。以Y1-28和Y1-31为亲本，筛选的第二代融合子Y2-33角蛋白酶活力为318.2±31.2 U/mL，与较原始菌株相比提高了122.4%。 关键词：基因组重排技术 角蛋白酶 地衣芽孢杆菌 Screening of high keratinase-producing strains by genome shuffling Major: Biotechnology for Light Industry Student: Cheng Shuang Supervisor: Wu Chongde Abstract Keratinase, specialized in degradation of keratin, is used extensively in the treatment of waste feather and the technology of clean production in leather industry. This manuscript aimed to screen high keratinase-producing strain through genome shuffling by using Bacillus licheniformis X-47 as parental strain. The conditions for genome shuffling were optimized. the optimal cell age was 9 h, and the optimal lysozyme concentration, enzymatic time, and temperature were 1.0 mg/mL, 30 min, and 40℃, respectively. The fusion time, fusion temperature, pH, and PEF concentration were 15 min, 35℃, pH 9.0 and 400 g/mL, respectively. Under the optimal conditions, the first generation of fusants Y1-28 and Y1-31 were screened, and the keratinase activities were 240.9±7.2 U/mL and 257.5±17.5 U/mL, which increased 68.4%, and 80.0%, respectively, compared to the parental strain. The second generation of fusant Y2-33 was isolated by using Y1-28 and Y1-31 as part