Novel thermal stability enhanced xylanase improves the performance and digestibility parameters in broilers

Xylanases require thermal stability to withstand the pelleting process, pH stability to function in the gastrointestinal tract, and resistance to xylanase inhibitors in raw materials to be effective in animal feed. A GH11 family xylanase originating from an anaerobic fungus, Orpinomyces sp. strain PC-2, has high specific activity and resistance to xylanase inhibitors intrinsically. It was engineered using rational protein design methods to obtain a thermal and pH stable enzyme, OXynA-M. OXynA-M showed resistance to three types of xylanase inhibitors, Triticum aestivum xylanase inhibitors TAXI-IB and TAXI-IIA and xylanase inhibitor protein XIP and showed melting temperature of 87.2°C when measured using differential scanning calorimetry. It was stable at all pH between 2.0-10.0 incubated up to 4 h. Xylo-oligosaccharides (XOS) production profile using a wheat arabinoxylan substrate revealed the production of xylobioses up to xylohexaoses, which are known to have prebiotic functionalities. An animal trial was conducted in broiler chickens to evaluate the in vivo efficacy of the xylanase. In total, 600 1-day-old chickens were divided into six dietary treatments, including a positive control (PC) (T1) without the addition of exogenous enzyme and the rest where exogenous xylanase was added at the rates of 1200, 2400, 4800, 9600, and 240000 U/kg of feed from T2-T6. An increase in OXynA-M xylanase improved the performance parameters in the enzyme-treated groups compared with the control. The viscosity of ileal digesta decreased with increasing enzyme dosage. A significantly lower viscosity of 6.54 cP was determined for the minimum dose in T2 (1200 U/kg), and the viscosity was further reduced in T6 (240000 U/kg) (P<0.05) compared to the control treatment. The apparent ileal digestibility of crude protein, fat, and starch improved as the xylanase dosage increased. The application of OXynA-M xylanase improved the apparent ileal digestibility of crude protein when the dose was higher than that of T2 (1200 U/kg). Furthermore, the AME_n of the diets improved when xylanase was supplemented at a rate of 9600 U/kg (T5) compared with the control treatment (P<0.05).