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Bidirectional Cell-Surface Anchoring Function of C-Terminal Repeat Region of Peptidoglycan Hydrolase of Lactococcus lactis IL1403

Paper ID Volume ID Publish Year Pages File Format Full-Text
21934 43244 2008 6 PDF Available
Title
Bidirectional Cell-Surface Anchoring Function of C-Terminal Repeat Region of Peptidoglycan Hydrolase of Lactococcus lactis IL1403
Abstract

With the aim of constructing an efficient protein display system for lactic acid bacteria (LABs), the effect of fusion direction on the cell-surface binding activity of the C-terminal region of the peptidoglycan hydrolase (CPH) of Lactococcus lactis IL1403 was studied. CPH fused to the α-amylase (AMY) of Streptococcus bovis 148 either at its C-terminus (CPH-AMY) or at its N-terminus (AMY-CPH) was expressed intracellularly in Escherichia coli. This domain was able to direct binding of AMY to the surface of L. lactis ATCC 19435 in both constructs. However, the number of bound molecules per cell and the specific activity for starch digestion in the case of CPH-AMY were 3 and 14 times greater than those in the case of AMY-CPH, respectively. Of the LABs tested, L. lactis ATCC 19435 showed the highest binding capability for CPH-AMY, up to 6×104 molecules per cell, with a dissociation rate constant of 5.00×10−5 s−1. The binding of CPH-AMY to the surface of Lactobacillus delbrueckii ATCC 9649 cells was very stable with a dissociation rate constant of 6.96×10−6 s−1. The production of CPH-AMY in the soluble form increased 3-fold as a result of coexpression with a molecular chaperone, trigger factor. The results of this study suggest the usefulness of CPH as a bidirectional anchor protein for the production of cell-surface adhesive enzymes in E. coli. Furthermore, the importance of the fusion direction of CPH in determining cell-surface binding and enzymatic activities was shown.

Keywords
Lactococcus lactis IL1403; Lactobacillus delbrueckii; surface display; peptidoglycan hydrolase; α-amylase; dissociation rate constant; Escherichia coli
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Bidirectional Cell-Surface Anchoring Function of C-Terminal Repeat Region of Peptidoglycan Hydrolase of Lactococcus lactis IL1403
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 105, Issue 2, February 2008, Pages 116–121
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