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Selection of a biotin protein ligase by phage display using a combination of in vitro selection and in vivo enzymatic activity

Paper ID Volume ID Publish Year Pages File Format Full-Text
22251 43264 2009 5 PDF Available
Title
Selection of a biotin protein ligase by phage display using a combination of in vitro selection and in vivo enzymatic activity
Abstract

We report the development of a phage display-based method for the direct selection of enzymatic activity by a combination of selection in vitro and an assay of enzymatic activity in vivo. We describe the selection of a biotin protein ligase (BPL) that specifically recognizes neurotensin as a substrate, as an example of the utility of our method. We constructed an enzyme library with a diversity of 8.8 × 106 by insertion of four randomized regions into a BPL backbone and used it for the selection of variant BPLs. We obtained an active BPL, which was identical to wild-type BPL, using the combination of selection in vitro and an assay of enzymatic activity in vivo. Our results indicate that our method is suitable for the selection of enzymes with specific functions of interest.

Keywords
ampr, ampicillin-resistance gene; BirA, gene for biotin protein ligase; BPL, biotin protein ligase; g3, gene III; g3p, pIII coat protein; g8p, pVIII coat protein; PBS, phosphate-buffered saline; RF, replicative formPhage display; Biotin protein ligase; Sc
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Selection of a biotin protein ligase by phage display using a combination of in vitro selection and in vivo enzymatic activity
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 107, Issue 3, March 2009, Pages 230–234
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