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A carbohydrate binding module as a diversity-carrying scaffold

In English

Författare

Summary, in English

The growing field of biotechnology is in constant need of binding proteins with novel properties. Not just binding specificities and affinities but also structural stability and productivity are important characteristics for the purpose of large-scale applications. In order to find such molecules, libraries are created by diversifying naturally occurring binding proteins, which in those cases serve as scaffolds. In this study, we investigated the use of a thermostable carbohydrate binding module, CBM4-2, from a xylanase found in Rhodothermus marinus, as a diversity-carrying scaffold. A combinatorial library was created by introducing restricted variation at 12 positions in the carbohydrate binding site of the CBM4-2. Despite the small size of the library (1.6x10(6) clones), variants specific towards different carbohydrate polymers (birchwood xylan, Avicel and ivory nut mannan) as well as a glycoprotein (human IgG4) were successfully selected for, using the phage display method. Investigated clones showed a high productivity (on average 69 mg of purified protein/l shake flask culture) when produced in Escherichia coli and they were all stable molecules displaying a high melting transition temperature (75.7 +/- 5.3degreesC). All our results demonstrate that the CBM4-2 molecule is a suitable scaffold for creating variants useful in different biotechnological applications.

Publiceringsår

2004

Språk

Engelska

Sidor

213-221

Publikation/Tidskrift/Serie

Protein Engineering Design & Selection

Volym

17

Issue

3

Dokumenttyp

Artikel i tidskrift

Förlag

Oxford University Press

Ämne

  • Immunology in the medical area
  • Industrial Biotechnology

Status

Published

Projekt

  • Designed carbohydrate binding modules and molecular probes

ISBN/ISSN/Övrigt

  • ISSN: 1741-0126