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Gewählte Publikation:

Klug-Santner, BG; Schnitzhofer, W; Vrsansk, M; Weber, J; Agrawal, PB; Nierstrasz, VA; Guebitz, GM.
(2006): Purification and characterization of a new bioscouring pectate lyase from Bacillus pumilus BK2.
J Biotechnol. 2006; 121(3):390-401 FullText FullText_BOKU

Abstract:
An alkalophilic bacterium was isolated based on the potential of extra-cellular enzymes for bioscouring. The bacterium was identified as a new strain of Bacillus pumilus BK2 producing an extra-cellular endo-pectate lyase PL (EC 4.2.2.2). PL was purified to homogeneity in three steps and has a molecular mass of 37.3 +/- 4.8 kDa as determined by SDS-PAGE and an isoelectric point of pH 8.5. Peptide mass mapping by nano-LC-MS of PL revealed 15% homology with a pectate lyase from Bacillus sp. The pectate lyase exhibited optimum activity at pH 8.5 and around 70 degrees C in Tris/HCl buffer. It showed a half-life at 30 degrees C of more than 75 h. Stability decreased with increasing temperature, extremely over 60 degrees C. The enzyme did not require Ca2+, ions for activity, and was strongly inhibited by EDTA and Co2+. PL was active on polygalacturonic acid and esterified pectin, but the affinity showed a maximum for intermediate esterified pectins and decreased over a value of 50% of esterification. The best substrate was 29.5% methylated pectin. PL cleaved polygalacturonic acid via a beta-elimination mechanism as shown by NMR analysis. PL released unsaturated tetragalacturonic acid from citrus pectin and polygalacturome acid, but did not show any side activities on other hemicelluloses. On polygalacturonic acid PL showed a K-m of 0.24 g l(-1) and a v(max) of 0.72 g l(-1) min(-1). The applicability of pectate lyase for the bioscouring process was tested on a cotton fabric. Removal of up to 80% of pectin was proven by means of ruthenium red dyeing and HPAEC (65%). Structural contact angle measurements clearly indicated the increased hydrophilicity of enzyme treated fabrics. (c) 2005 Elsevier B.V. All rights reserved.
Autor*innen der BOKU Wien:
Gübitz Georg
BOKU Gendermonitor:

Find related publications in this database (using NML MeSH Indexing)
Bacillus - classification
Bacterial Proteins - analysis
Chromatography, Ion Exchange -
Cobalt - pharmacology
Edetic Acid - pharmacology
Electrophoresis, Polyacrylamide Gel -
Enzyme Stability -
Half-Life -
Hydrogen-Ion Concentration -
Hydrolysis -
Kinetics -
Molecular Weight -
Nuclear Magnetic Resonance, Biomolecular -
Peptide Mapping -
Polysaccharide-Lyases - analysis
Substrate Specificity -
Temperature -
Viscosity -

Find related publications in this database (Keywords)
bioscouring
pectate lyase
Bacillus pumilus


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