High-yield secretion of multiple client proteins in Aspergillus

详细信息	查看全文 | 推荐本文 |

• 作者：Fernando Segato^a ; ² ; Andr&eacute ; R.L. Damá ; sio^a ; ¹ ; Thiago A. Gonç ; alves^c ; ² ; Rosymar C. de Lucas^a ; ¹ ; Fabio M. Squina^b ; Stephen R. Decker^d ; Rolf A. Prade^a ; ^{prade@okstate.edu}
• 关键词：5-FOA ; 5-fluorotic acid ; CAZy ; carbohydrate-active enZYmes Database (http ; //www.cazy.org/) ; CMC ; carboxymethylcellulose ; DNS ; dintirosalicilic acid ; HEPES ; 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid ; MM ; minimal Aspergillus medium ; PCR ; polymerase
• 刊名：Enzyme and Microbial Technology
• 出版年：2012
• 期刊代码：26_01410229
• 类别：ce
• 出版时间：15 July, 2012
• 卷：51
• 期：2
• 页码：100-106
• 文件大小：712 K

摘要

Production of pure and high-yield client proteins is an important technology that addresses the need for industrial applications of enzymes as well as scientific experiments in protein chemistry and crystallization. Fungi are utilized in industrial protein production because of their ability to secrete large quantities of proteins. In this study, we engineered a high-expression-secretion vector, pEXPYR that directs proteins towards the extracellular medium in two Aspergillii host strains, examine the effect of maltose-induced over-expression and protein secretion as well as time and pH-dependent protein stability in the medium. We describe five client proteins representing a core set of hemicellulose degrading enzymes that accumulated up to 50-100 mg/L of protein. Using a recyclable genetic marker that allows serial insertion of multiple genes, simultaneous hyper-secretion of three client proteins in a single host strain was accomplished.