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

The potential benefits of Xyn2 in the nutrition of weaned pigs should make it an alternative for industrial xylanase production.
10 February 2011
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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 60ºC. 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.

Twenty-four weaned pigs (Duroc x Landrace x Yorkshire) with an average initial body weight of 10.2 ± 0.35 kg were selected and randomly allotted to four dietary treatments (n=6) with equal numbers of males and females in each group. Treatments consisted of: (1) control without Xyn2 supplementation; (2) Xyn2 added at a concentration of 500 U/kg diet (LD); (3) Xyn2 added at a concentration of 1000 U/kg diet (MD); (4) Xyn2 added at a concentration of 1500 U/kg diet (HD). Pigs were housed individually in metabolism cages (0.7 x 1.5m) with woven wire flooring in an environmentally controlled room (22–24ºC) and were given ad libitum access to water through a water nipple. Pigs were hand-fed four times per d (08.00, 12.00, 16.00 and 20.00 hours) in bowl feeders to make sure fresh feed was available.

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.

It is concluded that the results obtained from the present study indicated that the recombinant Xyn2 is more suitable for use in the feed industry. A diet supplemented with the unprocessed enzyme at a lower level improves both the performance and nutrient digestibilities in weaned pigs. However, a proper process is needed to improve its potential benefits as a feed additive for piglet nutrition.

J He, J Yin, L Wang, B Yu and D Chen, 2010. British Journal of Nutrition. 103: 1507–1513

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