Endo-xylogalacturonan hydrolase, a novel pectinolytic enzyme.

Abstract:

:We screened an Aspergillus tubingensis expression library constructed in the yeast Kluyveromyces lactis for xylogalacturonan-hydrolyzing activity in microwell plates by using a bicinchoninic acid assay. This assay detects reducing carbohydrate groups when they are released from a carbohydrate by enzymatic activity. Two K. lactis recombinants exhibiting xylogalacturonan-hydrolyzing activity were found among the 3,400 colonies tested. The cDNA insert of these recombinants encoded a 406-amino-acid protein, designated XghA, which was encoded by a single-copy gene, xghA. A multiple-sequence alignment revealed that XghA was similar to both polygalacturonases (PGs) and rhamnogalacturonases. A detailed examination of conserved regions in the sequences of these enzymes revealed that XghA resembled PGs more. High-performance liquid chromatography and matrix-assisted laser desorption ionization-time of flight mass spectrometry of the products of degradation of xylogalacturonan and saponified modified hairy regions of apple pectin by XghA demonstrated that this enzyme uses an endo type of mechanism. XghA activity appeared to be specific for a xylose-substituted galacturonic acid backbone.

journal_name

Appl Environ Microbiol

authors

van der Vlugt-Bergmans CJ,Meeuwsen PJ,Voragen AG,van Ooyen AJ

doi

10.1128/aem.66.1.36-41.2000

keywords:

subject

Has Abstract

pub_date

2000-01-01 00:00:00

pages

36-41

issue

1

eissn

0099-2240

issn

1098-5336

journal_volume

66

pub_type

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