产番茄红素红酵母基因组重排实验条件的优化
详细信息Optimizing process of genome shuffling in Rhodotorula sp. producing lycopene
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摘要: 目的:探索基因组重排技术应用于产番茄红素红酵母育种中关键步骤的最优条件。方法:本实验以3株产生番茄红素的红酵母cp-2-11、cp-2-6、cp-28为实验菌株,研究菌体培养时间、酶解浓度、酶解时间、灭活条件以及融合时间对基因组重排的影响。结果:当菌体培养12h,酶解浓度为2%,酶解70min时,原生质体制备与再生率最佳;15W紫外灯,照射距离为35cm,磁力搅拌50r/min的条件下照射18min,60℃下灭活35min作为双亲灭活条件;35%聚乙二醇4000（PEG-4000）作为融合剂,处理23min为最佳融合时间。结论:通过基因组重排最佳条件的确定,后续重排实验的效率得到很大的提升,最终筛选出了高产的目的菌株,产量较融合亲本提高了122%。Abstract: Objective:Exploring the optimum conditions of the genome shuffling which applied to the key steps of Rhodotorula. Method:With Rhodotorula cp-2-11、cp-2-6、cp-28 as the tested material, we explored the effects of several factors including cell age, enzyme concentration, hydrolysis time, inactivation conditions, fusion time on genome shuffling. Results:The protoplast preparation rate and regeneration rate was supreme when the cell age was 12 hours, the enzyme concentration was 2% and the hydrolysis time was 70 minute.15W UV light, the irradiation distance was 35cm irradiates 18 minute and 60℃ water bath inactive 35 minutes were used as the conditions of Inactivation, fused 23 minute in 35% PEG as the fusion condition. Conclusions: Through determining the optimum conditions of genome shuffling, the follow-up work was improved effectively. After several rounds of genome shuffling, the high yield strains were breed, which increased by 122% than original strains.
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Keywords:
- genome shuffling;Rhodotorula;lycopene; /
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[1] Zhang Y, Perry K, Vinci V, et al.Genome shuffling leads to rapid phenotypic improvement in bacteria[J].Nature, 2002, 415:644-646.
[2] Stephanopoulos G.Metabolic engineering by genome shuffling[J].Nature Biotechnology, 2002, 20:666-668.
[3] 赵靖.基因组重排技术原理及应用研究[J].广州化工, 2012, 40 (6) :29-30. [4] Nguyen M, L Schwartz S J.Lycopene:chemieal and biological properties[J].Food Technology, 1999, 53 (2) :38-45.
[5] 李灿明, 黄时海, 张云开, 等.灭活原生质体融合技术提高豆豉纤溶酶菌产酶量[J].食品工业科技, 2009 (8) :140-142. [6] 肖春玲.樱桃番茄色素的抗氧化活性研究[J].食品工业科技, 2011, 32 (4) :117-120. [7] 邓建军.番茄红素功能活性研究进展[J].中国食品工业, 2012 (4) :50-51. [8] 解瑞宁, 沈新南.番茄红素生物学作用及与慢性病的关系[J].中国公共卫生, 2005, 21 (1) :112-113. [9] 陈学荣.粘红酵母生产类胡萝素b的发酵条件研究[J].食品工业科技, 2010 (6) :223 [10] 詹萍, 苏龙, 周乃东.红酵母原生质体制备和再生条件研究[J].安徽农业科学, 2010, 38 (3) :1154-1155, 1172. [11] 王海兵, 吴晓英, 刘世龙, 等.红酵母番茄红素提取工艺优化[J].食品科学, 2011 (32) :45-48. [12] 顾蕾, 陆玲, 袁生.红酵母原生质体制备及其紫外诱变育种的研究[J].食品工业科技, 2004 (4) :60-62, 65. [13] 廖宇静, 张利平, 谢响明, 等.微生物遗传育种学[M].第一版.北京:气象出版社, 2010, 8:570-575. [14] 高艳, 谭兴和, 周红丽, 等.南阳酵母2-577原生质体制备和再生条件的研究[J].食品科技, 2008 (8) :5-8. [15] 张宁, 虞龙, 沈以凌.氦离子注入番茄红素产生菌诱变选育的研究[J].辐射研究与辐射工艺学报, 2008, 26 (5) :286-288.
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