PMID: 11342713May 9, 2001Paper

Do domain interactions of glycosyl hydrolases from Clostridium thermocellum contribute to protein thermostability?

Protein Engineering
Irina A KataevaLars G Ljungdahl

Abstract

Cellulolytic and hemicellulolytic enzymes usually have a domain composition. The mutual influence of a cellulose-binding domain and a catalytic domain was investigated with cellobiohydrolase CelK and xylanase XynZ from Clostridium thermocellum. CelK is composed of an N-terminal family IV cellulose-binding domain (CBDIV(CelK)), a family 9 glycosyl hydrolase domain (Gh9(CelK)) and a dockerin domain (DD). CelK without the DD, (CBDIV-Gh9)(CelK) and CBDIV(CelK) bound cellulose. The thermostability of (CBDIV-Gh9)(CelK) was significantly higher than that of CBDIV(CelK) and Gh9(CelK). The temperature optima of (CBDIV-Gh9)(CelK) and Gh9(CelK) were 65 and 45 degrees C, respectively. XynZ consists of an N-terminal feruloyl esterase domain (FAE(XynZ)), a linker (L), a family VI CBD (CBDVI(XynZ)), a DD and a xylanase domain. FAE(XynZ) and (FAE-L-CBDVI)(XynZ), used in the present study did not bind cellulose, but both were highly thermostable. Replacement of CBDVI(XynZ) with CBDIV(CelK) resulted in chimeras with feruloyl esterase activity and the ability to bind cellulose. CBDIV(CelK)-FAE(XynZ) bound cellulose with parameters similar to that of (CBDIV-Gh9)(CelK). (FAE-L)(XynZ)-CBDIV(CelK) and FAE(XynZ)-CBDIV(CelK) had lower relative affiniti...Continue Reading

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Citations

Sep 29, 2006·Extremophiles : Life Under Extreme Conditions·Gashaw MamoBo Mattiasson
Mar 3, 2007·Journal of Industrial Microbiology & Biotechnology·Sarath B GundllapalliRicardo R Cordero Otero
Jun 13, 2012·Proceedings of the National Academy of Sciences of the United States of America·Gilad GefenEdward A Bayer
Jul 7, 2009·Annual Review of Microbiology·Lars G Ljungdahl
Nov 9, 2012·Bioinformatics and Biology Insights·Santi NurbaitiRukman Hertadi
Aug 2, 2005·FEBS Letters·Irina A KataevaLars G Ljungdahl
Mar 15, 2006·Journal of Computational Chemistry·Li-Cheng WuBaw-Jhiune Liu
Apr 19, 2005·Biotechnology Letters·S B Gundllapalli MosesI S Pretorius
Mar 12, 2017·Extremophiles : Life Under Extreme Conditions·Debamitra ChakravortySanjukta Patra

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