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High-yield secretion of multiple client proteins in Aspergillus

Paper ID Volume ID Publish Year Pages File Format Full-Text
17252 42654 2012 7 PDF Available
Title
High-yield secretion of multiple client proteins in Aspergillus
Abstract

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.

► High yield client protein secretion in Aspergillus. ► Stabilization of client protein secretion. ► Compound high yield client protein secretion from a single host. ► The use of recyclable markers enables high-yield secretion of multiple proteins from a single host.

Keywords
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
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 51, Issue 2, 15 July 2012, Pages 100–106
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us