Cleavage of cellulose by a CBM33 protein
Version of Record online: 8 AUG 2011
Copyright © 2011 The Protein Society
Volume 20, Issue 9, pages 1479–1483, September 2011
How to Cite
Forsberg, Z., Vaaje-Kolstad, G., Westereng, B., Bunæs, A. C., Stenstrøm, Y., MacKenzie, A., Sørlie, M., Horn, S. J. and Eijsink, V. G.H. (2011), Cleavage of cellulose by a CBM33 protein. Protein Science, 20: 1479–1483. doi: 10.1002/pro.689
- Issue online: 17 AUG 2011
- Version of Record online: 8 AUG 2011
- Accepted manuscript online: 11 JUL 2011 04:19PM EST
- Manuscript Received: 1 JUL 2011
- Manuscript Accepted: 1 JUL 2011
- VISTA Program. Grant Number: 6505
- Norwegian Research Council. Grant Numbers: 190965/S60, 190877/S60, 196885/F20
Additional Supporting Information may be found in the online version of this article.
|PRO_689_sm_SuppinfoFig1.doc||869K||Supporting Information Figure 1. Analysis of an in-house generated aldonic acid standard (Glc4GlcA) generated by chemical oxidation of cellooligosaccharides. Note that the efficiency of the oxidation reaction is less than 100% and that some components partially co-elute in the subsequent purification step. Consequently, the Glc4GlcA sample also contains minor amounts of other compounds such as Glc5, Glc6 and Glc5GlnA. (A) Verification of the oxidized oligosaccharide by MALDI-TOF analysis. The mass spectrum shows the K and Na adducts of the lactone (Glc4GlcLA), aldonic acid (Glc4GlcA) and the native oligosaccharide (Glc5). Additionally, the K and Na adducts of the K and Na aldonic acid salts are observed. A distribution of ions representing the oxidized hexameric cellooligosaccharide (Glc5GlcLA/ Glc5GlcA/ Glc6) can also be observed. (B) HPAEC analysis. The picture shows an overlay of a chromatogram for the chemically oxidized oligosaccharide (dotted line) with a chromatogram for products generated by CelS2 action on Avicel (solid line). The chromatogram of the pentameric standard shows the same species as detected by MS analysis: the aldonic acid, the lactone and native form (Nb. Formally, the peak labeled Glc4GlcLA must be regarded as “unknown”; the peak was annotated as Glc4GlcLA by inference from the mass spectrum of panel A). The absence of the lactone form in the CelS2 product mix is due to the slow equilibrium between lactone and aldonic acid at the pH of the reaction (pH 6.5).|
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