重组毕赤酵母发酵生产α-葡聚糖酶的放大研究  

Scale-up Fermentation of Recombinant Dextranase Expressed in Pichia Pastoris

作　　者：黄曾慰[1] 吴兆鹏[1] 常国炜[1] 黎志德[1] 张九花[1] 曾练强[1] 梁达奉[1,2] 

机构地区：[1]广州甘蔗糖业研究所广东省甘蔗改良与生物炼制重点实验室,广东广州510316 [2]广西糖业研发中心,广西南宁530002

出　　处：《甘蔗糖业》2016年第2期20-26,共7页Sugarcane and Canesugar

基　　金：现代农业产业技术体系建设专项资金(CARS-20-4-5);八桂学者建设工程专项经费

摘　　要：研究了毕赤酵母高密度发酵组成型表达α-葡聚糖酶,从6.8 L发酵罐逐级放大至5000 L发酵罐的放大工艺。采用恒定p H和控制甘油残留浓度和溶解氧相结合的调控策略,进行了多批次的发酵试验。结果表明:三磷酸甘油醛脱氢酶启动子驱动的α-葡聚糖酶的生成具有生长偶联特征,集中在对数生长期和稳定期。6.8 L发酵罐中上清酶活达1253 U/m L,中试放大至50 L发酵罐中产酶最高达1300 U/m L,500 L发酵罐中酶活为740 U/m L。5000 L规模发酵罐中产酶达到589 U/m L。建立了以甘油作为碳源,发酵调控简单的工艺,避免了使用甲醇带来的安全问题,适合放大生产。The high density fermentation of dextranase in the constitutive Pichia pastoris expression system scaled up from 6.8 to 5000 L in series was studied. By controlling residual concentration of glycerol and dissolved oxygen combined with p H-stat, multi batches of fermentation were performed. Results showed that the dextranase synthesis under GAP promoter is growth-associated and mainly in exponential phase and stationary phase. Activity of dextranase in 6.8 L, 50 L, 500 L and 5000 L fermenter reached 1253, 1300, 740 and 589 U/m L, respectively. A simple and convenient fermentation method using glycerol as carbon source has been developed, which obviates problems related with methanol safety concerns and is suitable for large scale production.

关 键 词：α-葡聚糖酶 毕赤酵母 组成型表达 发酵放大 

分 类 号：TS244[轻工技术与工程—制糖工程]