常压室温等离子体诱变技术选育番茄红素高效-食品与生物技术学报.docVIP

基于ARTP技术快速选育番茄红素高效合成突变株 罗玮(， 周雅， 巩尊洋， 杜瑶， 余晓斌 （江南大学生物工程学院 糖化学与生物技术教育部重点实验室江苏 无锡 214122摘要：为了提高番茄红素的生物合成水平，本研究利用常压室温等离子体注入技术对番茄红素生产菌三孢布拉霉负菌（Blakeslea trispora(-)）进行诱变选育。首先建立了番茄红素含量分析的标准曲线和等离子体诱变致死率曲线。通过构建的“马鞍形”致死率曲线确定了合适的诱变处理时间，经过数轮肉眼初筛和摇瓶复筛，获得了一株产量为0.85 ± 0.04 g/L的突变株ARTP-3，比出发菌株番茄红素的产量提高51.4%，且传代实验表明该菌株的番茄红素产量遗传稳定性较好。本研究不仅获得了一株具有较大经济价值的番茄红素高效合成突变株，建立的育种方法也可为其他工业微生物特别是丝状真菌的选育提供有益的参考。关键词：等离子体番茄红素三孢布拉霉Rapid screening of high efficient lycopene-producing mutant by atmospheric and room temperature plasma mutagenesis LUO Wei*, ZHOU Ya, GONG Zun-Yang, DU Yao, YU Xiao-Bin (School of Biotechnology, Jiangnan University; Key Laboratory of Carbohydrate Chemistry Biotechnology, Ministry of Education, Wuxi, 214122, China) Abstract: In order to improve the level of lycopene biosynthesis, this study use atmospheric and room temperature pressure plasma to mutate Blakeslea trispora negative bacteria for lycopene efficient production. The standard curve of lycopene content analysis and the lethal rate curve were established. By constructing the saddle shaped lethal rate curve, the suitable mutation treatment time was determined. After several rounds of screening, a mutant strain ARTP-3 was obtained with the lycopene production of 0.85 ± 0.04 g/L, which was increased by 51.4% than that of the original strain. Furthermore, the subculture experiments showed that the ycopene production of ARTP-3 was much stable. In this study, we have not only obtained a high efficient lycopene producer with high economic value, but also can provide useful reference for the breeding of other industrial microorganisms, especially for of filamentous fungi. Keywords: Atmospheric and room temperature plasma (ARTP); Lycopene; Blakeslea trispora; High efficient synthetic mutant 番茄红素（lycopene）是一种重要的类胡萝卜素，具有极强的抗氧化和清除自由基能力，被广泛应用于食品、医药和化妆品领域[1]。目前番茄红素的生产方法主要包括天然提取法、化学合成法和微生物发酵法[2]。天然提取法获得的番茄红素产量不高，成本高且受制于原料来源；化学合成法存在安全隐患，而微生物发酵法具有工艺简单、生产效率高等优点，越来越受到关注[3]。三孢布拉霉（Blakeslea trispora）是一种丝状真菌，其番茄红素含量很高，是生产番茄