重组毕赤酵母发酵甘油制备α-葡聚糖酶的研究  

Study on the Fermentation Conditions for α-Glucanase Production by Recombinant Pichia pastoris Using Glycerol as Carbon Source

作　　者：吴兆鹏[1] 曾练强[1] 黄曾慰[1] 常国炜[1] 梁达奉[1,2] 李雨虹 

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]广西农垦糖业集团股份有限公司,广西糖业研发中心,广西南宁530002

出　　处：《广西科学》2014年第6期624-628,共5页Guangxi Sciences

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

摘　　要：【目的】考察甘油对组成型毕赤酵母（Pichiapastoris ）生长和α-葡聚糖酶表达量的影响。【方法】采用单因素实验法考察初始发酵培养基中甘油浓度、补料阶段甘油残留和通气量等对α-葡聚糖酶表达量的影响。【结果】发酵培养基中初始甘油浓度从50 g／L提高到150 g／L，菌体生长和α-葡聚糖酶表达量均未受到影响，继续提高到200 g／L时，α-葡聚糖酶表达量明显下降。补料过程甘油残留在0～5 g／L，菌体生长和α-葡聚糖酶表达量最佳，当甘油残留较多时，菌体生长和α-葡聚糖酶的表达量均受到影响。提高通气量有利于增加α-葡聚糖酶的表达量，发酵78 h为宜，在此条件下α-葡聚糖酶酶活力达1763 U。【结论】该工艺优化了以甘油作为碳源制备α-葡聚糖酶的发酵条件，提高了α-葡聚糖酶酶活力，为大规模生产α-葡聚糖酶奠定了基础。Objective]The aim of this paper is mainly to investigate the effects of glycerol on Pichia pastoris growth and constitutive expression ofα-glucanase.[Methods]The initial concen-tration of glycerol in fermentation medium,glycerin concentration in fed-batch,ventilation and other effects on constitutive expression ofα-glucanase were examined by means of single factor experiment.[Results]When the initial concentration of glycerol ranged from 50 g/L to 150 g/L,the growth andα-glucanase expression were not affected,while the growth andα-glucanase expression appeared to be markedly reduced when the glycerol increase to 200 g/L.Glycerol concentration between 0 to 5 g/L in fed-batch lead to the best growth andα-glucanase expres-sion.When glycerin concentration became higher,the growth of Pichia pastoris andα-glu-canase expression were restrained.Meanwhile,improving the ventilation was helpful to the ex-pression of α - glucanase, and suitable fermentation time was 78 h. Under this condition,α-glucanase activity reached about 1763 U.[Conclusion]The process optimized the fermentation conditions forα-glucanase prepara-tion with glycerol as carbon source,which great-ly improvedα-glucanase activity and laid a solid foundation forα-glucanase large-scale produc-tion.

关 键 词：毕赤酵母 α-葡聚糖酶 甘油 发酵条件 

分 类 号：TQ925[轻工技术与工程—发酵工程]