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research article

Selection of catalytically active biotin ligase and trypsin mutants by phage display

Heinis, Christian  
•
Huber, Adrian
•
Demartis, Salvatore
Show more
2001
Protein Engineering

The efficiency of selection for catalysis by phage display library was studied using a ligase (the Escherichia coli biotin ligase BirA) and an endopeptidase (the rat trypsin His57->Ala mutant) as model enzymes. These enzymes could be displayed on phage as fusion proteins with calmodulin and the minor coat protein pIII. The display of functional enzyme and the efficiency of selection for catalysis were considerably improved by using phage vectors. In model selection expts., phage displaying BirA were consistently enriched (between 4- and 800-fold) per round of panning, relative to neg. controls. Phage displaying the trypsin His57->Ala mutant were also enriched (between 15- and 2000-fold), relative to neg. controls. A combinatorial phage display library of trypsin mutants was constructed to improve the catalytic properties of the trypsin His57->Ala mutant. Mutants with catalytic properties superior to those of trypsin His57->Ala mutant could not be isolated. [on SciFinder (R)]

  • Details
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Type
research article
DOI
10.1093/protein/14.12.1043
Author(s)
Heinis, Christian  
Huber, Adrian
Demartis, Salvatore
Bertschinger, Julian
Melkko, Samu
Lozzi, Luisa
Neri, Paolo
Neri, Dario
Date Issued

2001

Published in
Protein Engineering
Volume

14

Issue

12

Start page

1043

End page

1052

Subjects

Proteins Role: BUU (Biological use

•

unclassified)

•

BIOL (Biological study)

•

USES (Uses) (coat

•

fusion proteins with enzyme in phage display; selection of catalytically active biotin ligase and trypsin mutants by phage display); Calmodulins Role: BUU (Biological use

•

unclassified)

•

BIOL (Biological study)

•

USES (Uses) (fusion proteins with enzyme in phage display; selection of catalytically active biotin ligase and trypsin mutants by phage display); Fusion proteins Role: BUU (Biological use

•

unclassified)

•

BIOL (Biological study)

•

USES (Uses) (fusion with calmodulin and coat protein pIII; selection of catalytically active biotin ligase and trypsin mutants by phage display); Panning; Phage display; Phage display library; Protein engineering (selection of catalytically active biotin ligase and trypsin mutants by phage display)

•

phage display library biotin ligase trypsin

Note

CAN 136:305879

7-2

Enzymes

Inst. of Pharm. Sci.,Swiss Fed. Inst. of Technol.,Zurich,Switz.

Journal

written in English.

9002-07-7P (Trypsin); 39419-81-3P (Biotin ligase) Role: BPN (Biosynthetic preparation), BIOL (Biological study), PREP (Preparation) (selection of catalytically active biotin ligase and trypsin mutants by phage display)

Editorial or Peer reviewed

REVIEWED

Written at

OTHER

EPFL units
LPPT  
Available on Infoscience
November 17, 2008
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/31194
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