Functional characterisation of a recombinant xylanase from Pichia pastoris and effect of the enzyme on nutrient digestibility in weaned pigs

Br J Nutr. 2010 May;103(10):1507-13. doi: 10.1017/S0007114509993333. Epub 2010 Jan 12.

Abstract

The xyn2 gene of a filamentous mesophilic fungus, Trichoderma reesei, coding xylanase 2 (Xyn2) was previously expressed in Pichia pastoris. In the present study, the recombinant Xyn2 was prepared from a 15 litre fermenter, and subsequently characterised. It has been confirmed to have a molecular mass of 21 kDa, an optimal pH of 6.0 and an optimal temperature of 60 degrees C. When tested using oat-spelt xylan, it showed a Km and catalytic rate constant (kcat) of 1.1 mg/ml and 512.4/s, respectively. Analysis of the products from oat-spelt xylan degradation confirmed that the enzyme was an endoxylanase with xylotriose and xylobiose as the main degradation products. The unprocessed Xyn2 was supplemented to a xylan-containing diet to determine its influences on performance and nutrient digestibilities by weaned pigs. Results showed that the average body-weight gain increased 16.9 % when piglets received Xyn2 at a concentration of 500 U/kg diet. There also was a positive (0.05 < P < 0.10) effect on the digestibility values of crude protein, ash, Ca and acid-detergent fibre with Xyn2 supplementation. The potential benefits of Xyn2 in the nutrition of weaned pigs should make it an alternative applicant for industrial xylanase production.

Publication types

  • Randomized Controlled Trial
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animal Feed
  • Animal Nutritional Physiological Phenomena
  • Animals
  • Diet / veterinary
  • Digestion / physiology*
  • Female
  • Hydrogen-Ion Concentration
  • Hydrolysis
  • Male
  • Pichia / enzymology*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Substrate Specificity
  • Swine / growth & development
  • Swine / physiology*
  • Temperature
  • Weaning
  • Xylosidases / chemistry
  • Xylosidases / genetics
  • Xylosidases / metabolism*

Substances

  • Recombinant Proteins
  • Xylosidases