Identification, Cloning, Purification and Activity Determination of a Recombinant Xylanase Enzyme Identified from Bovine Rumen Metagenome

Poultry, like many animals, uses enzymes for the digestion of feed. To overcome the undesirable effects of deleterious factors and improve digestion of feed, enzymes are used as supplements in feed formulation. Supplementation of feed with definite enzymes enhances its nutritive value, thereby increasing the effectiveness of digestion. Enzymes are the most important and useful additives in the animal feed industry. Xylanases are widely used in the animal feed industry. Pretreatment of animal feed with xylanases increases the nutritional value and digestion in ruminants. In this study, metagenomic data of bovine rumen were explored and then a thermostable xylanase gene was identified using TAXyl software and named PersiXyn9. The laboratory tests were carried out to determine the enzymatic activity. Protein expression was done in E. coli (BL21) and a Nickel column was used for protein purification. The biochemical parameters of the enzyme showed the optimal activity at 50℃, and the optimum pH of 6. Kinetic values, Km and Vmax, for this enzyme were calculated to be 1.18 mg/ml and 206.78 μmol min-1mg-1 respectively. Scanning electron microscopy (SEM) images of the activity of the enzyme on poultry feed at 50℃ represent the high efficiency of the xylanase as an additive in the poultry feed industry.