A kind of method and application of Chaetomium globosa fermentative production of chitinase
A technology of chitinase and Chaetomium globosa, applied in the biological field, can solve the problems of low enzyme activity, lack of efficient degradation, poor stability, etc., and achieve the effects of simple culture method, low nutritional requirements and easy operation.
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[0022] Preparation of the colloidal chitin used: Weigh 10 g of chitin, add 40 mL of acetone to fully infiltrate, add 300 mL of concentrated HCl while grinding at 4 °C, and slowly add to 1.5 L of 50% ethanol after standing for 5 h Stir vigorously while adding, and let stand for 2 h to precipitate colloidal chitin. Pour off the supernatant, collect the colloidal chitin precipitate, and wash it repeatedly with distilled water until neutral.
[0023] The configuration of the DNS reagent used: add 91g potassium sodium tartrate into 500 mL hot water, stir and dissolve in a water bath at 45°C, then add 20g NaOH and 3.15g 3,5-dinitrosalicylic acid (DNS) in sequence, and then add 2.5g Crystalline phenol and 2.5 g of anhydrous sodium sulfite, stirred to dissolve, cooled, and distilled water to 1000mL, stored in a brown bottle in a dark place for one week before use.
Embodiment 1
[0024] Example 1: Fermentation of Chaetomium globosa HY1 to produce chitinase on a shaking table (1)
[0025] Pick a ring from the strains stored on the slant of the test tube and transfer to the activation medium, and activate the turbidity of the bacteria at 28°C and 200 rpm for 25 hours to obtain the activated seed solution. Activation medium: 8 g / L glucose, 3 g / L peptone, 1 g / L potassium dihydrogen phosphate, 1 g / L disodium hydrogen phosphate, pH 6.5.
[0026] Using the preferred fermentation medium: 15 g / L soluble starch, 10 g / L glycerol, 5 g / L tryptone, 2 g / L yeast powder, 4 g / L corn steep liquor, 2 g / L NH 4 NO 3 , 0.5 g / L MnSO 4 , 2 g / L MgSO 4 , 1 g / L Na 2 HPO 4 , 1 g / LK 2 HPO 4 , 4 g / L colloidal chitin, initial pH 6.5. The optimal fermentation medium after sterilization was inserted into the activated seed culture solution according to the inoculum amount of 3% (v / v), cultivated in a rotary shaker at 200 rpm, the culture temperature was 28°C, and the culture wa...
Embodiment 2
[0027] Example 2: Fermentation of Chaetomium globosa HY1 to produce chitinase on a shaking table (2)
[0028] The strains stored on the slant of the test tube were transferred to the activation medium, and activated at 28°C and 200rpm for 30h until the bacteria became turbid to obtain the activated seed solution. Activation medium: 15 g / L glucose, 6 g / L peptone, 3 g / L potassium dihydrogen phosphate, 2.5 g / L disodium hydrogen phosphate, pH 7.0.
[0029] Optimal fermentation medium: 20 g / L soluble starch, 15 g / L glycerol, 10 g / L tryptone, 4 g / L yeast powder, 8 g / L corn steep liquor, 5 g / L NH 4 NO 3 , 1.5 g / L MnSO 4 , 4 g / L MgSO 4 , 2.5 g / L Na 2 HPO 4 , 3 g / LK 2 HPO 4 , 8 g / L colloidal chitin, initial pH 7.5. The optimal fermentation medium after sterilization was inserted into the activated seed culture solution according to the inoculation amount of 6% (v / v), and cultivated in a rotary shaker at 200 rpm at a temperature of 30°C. After 5 days of fermentation, the cultiva...
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