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Characterization of a GH Family 43 ß-Xylosidase Having a Novel Carbohydratebinding Module from Paenibacillus xylaniclasticus Strain TW1.
- Source :
- Journal of Applied Glycoscience; 2022, Vol. 69 Issue 3, p65-71, 7p
- Publication Year :
- 2022
-
Abstract
- Paenibacillus xylaniclasticus strain TW1, a gram-positive facultative anaerobic bacterium, was isolated as a xylanolytic microorganism from the wastes of a pineapple processing factory. A gene encoding one of its xylanolytic enzymes, a ß-xylosidase, was cloned and sequenced. Sequence analysis revealed that this ß-xylosidase, named PxXyl43A, was composed of a glycoside hydrolase (GH) family 43 subfamily 12 catalytic module and an unknown function module (UM). The full-length PxXyl43A (PxXyl43A) was heterologously expressed in Escherichia coli and purified. Recombinant PxXyl43A exhibited hydrolysis activity against both p-nitrophenyl-ß-D-xylopyranoside (pNPX) and p-nitrophenyl-a-L-arabinofuranoside at specific activities of 250 and 310 mU/mg, respectively. The optimal reaction pH and temperature for pNPX hydrolysis were 7.1 and 54 °C, respectively. At pH 7.0 and 54 °C, the Km and kcat for pNPX were 1.2 mM and 2.8 ± 0.15 s<superscript>-1</superscript>, respectively. It was also discovered that the recombinant unknown function module of PxXyl43A (PxXyl43A-UM) could bind to insoluble xylans like birchwood xylan and oat spelt xylan, whereas it did not bind to cellulosic substrates such as ball-milled cellulose, carboxymethyl cellulose or lichenan. The PxXyl43A-UM's binding constant value Ka for oat spelt xylan was 2.0 × 10<superscript>-5</superscript> M<superscript>-1</superscript>. These results suggest that PxXyl43A possesses a novel carbohydrate-binding module, named as CBM91, specific for xylan-containing polysaccharides. [ABSTRACT FROM AUTHOR]
- Subjects :
- XYLOSIDASES
ANAEROBIC bacteria
PAENIBACILLUS
ESCHERICHIA coli
POLYSACCHARIDES
Subjects
Details
- Language :
- English
- ISSN :
- 13447882
- Volume :
- 69
- Issue :
- 3
- Database :
- Complementary Index
- Journal :
- Journal of Applied Glycoscience
- Publication Type :
- Academic Journal
- Accession number :
- 159751066
- Full Text :
- https://doi.org/10.5458/jag.jag.JAG-2022_0001