Cloning-independent expression and screening of enzymes using cell-free protein synthesis systems.

doi:10.1007/978-1-62703-782-2_6

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Article

Cloning-independent expression and screening of enzymes using cell-free protein synthesis systems.

Methods in molecular biology (Clifton, N.J.) - 1 Jan 2014

Kwon Yong-Chan, Song Jae-Kwang, Kim Dong-Myung

Abstract excerpt

We present a strategy for expression and screening of microbial enzymes without involving cloning procedures. Libraries of putative ω-transaminases (ω-TA) and mutated Candida antarctica lipase B (CalB) are PCR-amplified from bacterial colonies and directly expressed in an Escherichia coli-based cell-free protein synthesis system. The open nature of cell-free protein synthesis system also allows streamlined...

Topics

Cell-Free System
Enzyme Assays
Enzyme Stability
Escherichia coli
Fungal Proteins
Gene Library
Hydrolysis
Lipase
Mutation
Polymerase Chain Reaction
Protein Biosynthesis
Substrate Specificity
Temperature
Transaminases
Triglycerides

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Open in PubMed

Identifiers

PMID: 24395411
DOI: 10.1007/978-1-62703-782-2_6

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