Study on Enzymatic Properties of Feeding Xylanase

Xylanase refers to a group of enzymes that can degrade xylan into oligosaccharides and xylose, including exo-β-1,4-xylanase and endo-β-1. 4-xylanase and β-xylanase [1]. Xylanase has a wide range of applications in the fields of feed, food, paper, textile, medicine and energy. As a feed enzyme preparation, xylanase can effectively eliminate the anti-nutritional effect of xylan and promote the digestion and absorption of roughage by livestock and poultry; as a new pulp bleaching aid, xylanase can reduce chlorine for bleaching and solve pulp. Environmental pollution problems in industry; xylanase hydrolyzes hemicellulose in plants, and the formed pentose sugar is further used in the production of xylitol, alcohol, organic acids and other products, and plays an important role in the biotransformation of renewable resources [ 2]. Gv8 China Feed Industry Information Network - based on feed, serving animal husbandry

In the early stage of the laboratory, a strain of xylan degrading efficiently was obtained [3]. In this experiment, the enzymatic properties of the xylanase produced by solid fermentation of the strain were studied, including optimum pH and pH stability. Sex, optimal reaction temperature, thermal stability and ability to tolerate pepsin trypsin, etc., in order to have a deeper understanding of the xylanase, and have some guidance on the application of the xylanase effect. Gv8 China Feed Industry Information Network - based on feed, serving animal husbandry

1 Materials and methods Gv8 China feed industry information network - based on feed, service animal husbandry

1.1 Xylanase is obtained by solid fermentation in our laboratory. Gv8 China Feed Industry Information Network - based on feed, serving animal husbandry

1.2 Reagents Gv8 China Feed Industry Information Network - based on feed, service animal husbandry

Oat xylan (X0627), pepsin (P7000), and trypsin (T1005) were purchased from sigma. For DNS reagents, see the DNS configuration method in GB/T 23874-2009. The remaining reagents were of analytical grade. Gv8 China Feed Industry Information Network - based on feed, serving animal husbandry

1.3 enzyme activity determination method Gv8 China feed industry information network - based on feed, service animal husbandry

It is determined by DNS spectrophotometry. For details, see GB/T 23874-2009. Enzyme activity definition: The amount of enzyme required to degrade and release 1 μmol of reducing sugar per minute from a xylan solution at a concentration of 5 mg/mL at 37 ° C and a pH of 5.50 is an enzyme activity unit U. Gv8 China Feed Industry Information Network - based on feed, serving animal husbandry

2 experimental design Gv8 China feed industry information network - based on feed, service animal husbandry

2.1 Optimum reaction of xylanase pH Gv8 China Feed Industry Information Network - based on feed, service animal husbandry

Use 0.1 mol/L buffers of different pH (2.0, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 8.0) (sodium dihydrogen phosphate-citrate buffer with pH ≤ 8.0) The xylan substrate was prepared by glycine-sodium hydroxide buffer (pH>8.0), and the enzyme activity was determined according to the national standard. The highest enzyme activity was 100%, and the enzyme activity under other conditions accounted for the highest enzyme activity. The percentage is the relative enzyme activity at this pH. Gv8 China Feed Industry Information Network - based on feed, serving animal husbandry

2.2 pH stability of xylanase Gv8 China Feed Industry Information Network - based on feed, serving animal husbandry

Add equal amounts of the above buffers with different pH (2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 9.0) and mix them. The mixture was incubated at 37 ° C for 4 h in water bath, and the enzyme activity was determined according to the national standard after cooling in an ice water bath. The highest enzyme activity is 100%. Gv8 China Feed Industry Information Network - based on feed, serving animal husbandry

2.3 Optimum reaction temperature of xylanase Gv8 China Feed Industry Information Network - based on feed, service animal husbandry

The solution was prepared with a buffer of the optimum pH, and reacted at different temperatures of 30, 40, 45, 50, 55, 60, 65 and 70 ° C to measure the enzyme activity according to the national standard method. The temperature at which the enzyme activity is highest is the optimum reaction temperature. Gv8 China Feed Industry Information Network - based on feed, serving animal husbandry

2.4 Thermal stability of xylanase Gv8 China Feed Industry Information Network - based on feed, serving animal husbandry

The solution was prepared with the optimal pH buffer, and treated at 50, 55, 60 ° C for 10, 20, 40, 60, 80, 100, 120 min, and the enzyme activity was determined according to the national standard method. The enzyme activity of the untreated enzyme was 100%. Gv8 China Feed Industry Information Network - based on feed, serving animal husbandry

2.5 The effect of pepsin and trypsin on xylanase activity Gv8 China Feed Industry Information Network - based on feed, serving animal husbandry

In a 0.5 mL xylanase solution, add 0.5 mL of a 0.01 mg/mL pepsin solution (configured with a pH of 2.0, a concentration of 0.01 mol/L in HCl) and a 0.5 mL concentration of 0.2 mg/mL. Trypsin (disposed with potassium phosphate monobasic buffer solution at a pH of 7.0 and a concentration of 0.1 mol/L) was treated in a 37 ° C water bath for 30, 60, and 90 min, respectively, and the enzyme activity was determined according to the national standard method. The enzyme activity of the untreated enzyme was 100%. Gv8 China Feed Industry Information Network - based on feed, serving animal husbandry

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