单因素优化提高重组毕赤酵母β-呋喃果糖苷酶产量  

Production Improvement of β-fructofuranosidase in Pichia pastoris by Single-factor-at-a-time Optimization

作　　者：赵西浩 张慧杰 冯家勋[1,2] 秦秀林 

机构地区：[1]亚热带农业生物资源保护与利用国家重点实验室,南宁530004 [2]广西大学生命科学与技术学院,南宁530004

出　　处：《基因组学与应用生物学》2017年第9期3801-3806,共6页Genomics and Applied Biology

基　　金：国家自然科学基金项目(31300076);广西自然科学基金创新研究团队项目(2012GXNSFGA060005)和广西自然科学基金项目(2013GXNSFBA019096)共同资助

摘　　要：产生低聚果糖的β-呋喃果糖苷酶(β-fructofuranosidase,FFase)是一种具有广泛应用前景的工业酶。为了提高重组毕赤酵母生产FFase的产量,对重组毕赤酵母接种量、摇瓶装液量、甲醇添加量、山梨醇与甲醇共混4个因素进行单因素优化。研究结果表明,接种量为5%时酶活力达到8.35 U/m L,甲醇添加量为0.8%时酶活力达到6.89 U/m L,山梨醇添加量为1.5%时酶活力达到9.49 U/m L,装液量为20 m L时酶活力达到27.34 U/m L。在最优条件下诱导重组毕赤酵母,FFase酶活可达到33.46 U/m L,较优化前提高了4.86倍。该研究为进一步优化毕赤酵母FFase发酵条件和提高FFase产量奠定了一定基础。β-fructofuranosidase（FFase）,which can produce fructooligosaccharides（FOS）,is an industrial enzyme with wide application prospect.A single-factor-at-a-time optimization strategy was applied in the four significant critical conditions,including inoculum size,liquid volume in flask,methanol concentration and sorbitol concentration to improve the FFase production.Furthermore,the results of the single-factor-at-a-time experiments showed that the enzyme activity reached 8.35 U/m L when the inoculum size was 5%,the enzyme activity reached6.89 U/m L when the methanol concentration was 0.8%,the enzyme activity reached 9.49 U/m L when the sorbitol concentration was 1.5% and the enzyme activity reached 27.34 U/m L when the liquid volume in flask was 20 m L.Maximum yield of 33.46 U/m L was obtained under optimized conditions,which was 4.86 fold higher than that before optimization.This study laid a foundation for further optimization of the fermentation conditions of Pichia pastoris and FFase production.

关 键 词：毕赤酵母 发酵过程优化 Β-呋喃果糖苷酶 单因素优化 

分 类 号：TQ925[轻工技术与工程—发酵工程]