响应面优化角质细胞生长因子自诱导发酵  

Optimization the Auto-induction Fermentation of Keratinocyte Growth Factor by Response Surface Design

作　　者：李希平 王娟 LI Xiping;WANG Juan(Hangzhou Xianghu High Tech Application Research Institute,Hangzhou 311200,China;Standard Sci-Tech Innovation(Qingdao)Pharmaceutical Technology Co.,Ltd.Qingdao 266114,China)

机构地区：[1]杭州市萧山区湘湖高新技术应用研究院,浙江杭州311200 [2]斯坦德科创医药科技(青岛)有限公司,山东青岛266114

出　　处：《药物生物技术》2024年第6期631-635,共5页Pharmaceutical Biotechnology

摘　　要：角质细胞生长因子(keratinocyte growth factor, KGF)具有重要的生物学作用,临床上已经用于治疗放化疗引起的急性口腔黏膜炎等疾病。通过基因工程菌发酵表达外源蛋白现今已得到广泛应用,但重组KGF蛋白对表达菌株存在毒性且表达量低等问题限制了其制备及应用。Studier FW在其文章[1]阐述了自诱导表达培养基,采用自诱导培养基制备毒性蛋白已有相关报道,但对自诱导培养基进一步优化的相关报道却很少。本文通过对自诱导培养基进行Plackett-Burman设计,最陡爬坡实验,Box-Behnken设计等实验,优化KGF在自诱导培养基中的表达。实验确认自诱导培养基中三个关键成分的量分别是yeast extract 11 g/L,MgSO_(4)·7H_(2)O 1.1 g/L,lactose 0.98 g/L。经优化KGF表达量提高约26.2%,可为类似蛋白的制备提供参考。The keratinocyte growth factor(KGF)plays a significant biological role and has been clinically used to treat conditions such as acute oral mucositis induced by radiotherapy or chemotherapy.The fermentation expression of exogenous proteins through genetic engineering in bacteria has been widely applied.However,limitations such as toxicity to the host strain and low expression levels of recombinant KGF protein have hindered its production and application.Studier FW discussed the use of auto-induction expression media in his article",which has been reported for the preparation of toxic proteins.However,there is limited literature on further optimization of auto-induction expression media.In this study,the authors optimized the expression of KGF in auto-induction expression media through experiments including Plackett-Burman design,steepest ascent experiments,and Box-Behnken design.The experiments confirmed that the optimal concentrations of three key components in the auto-induction expression media were as follows:yeast extract 11 g/L,MgSO_(4)·7H_(2)O 1.1 g/L,lactose 0.98 g/L.After optimization,the expression level of KGF increased by approximately 26.2%,which can serve as a reference for the preparation of similar proteins.

关 键 词：角质细胞生长因子 发酵 自诱导培养基 PLACKETT-BURMAN设计 最陡爬坡实验 BOX-BEHNKEN设计 

分 类 号：TQ920.6[轻工技术与工程—发酵工程]