Liquid-solid two phase fermentation method for producing high activity cellulase for fodder
A cellulase and endocellulase technology, applied in the field of bioengineering, can solve the problems of long fermentation period, easy environmental pollution of fermentation liquid and high volume production efficiency
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Embodiment 1
[0030] Embodiment 1: the preparation of the secondary liquid bacterial classification of Trichoderma korning, concrete operating steps are as follows:
[0031] A. Slope bacteria (solid)
[0032] Peel and wash the potatoes, cut them into small pieces, put them in water, potatoes: water = 1:4 weight ratio, soak at 80°C for 1 hour, boil and filter with a filter cloth, dilute the filtrate with water to the original volume. Add 2 parts of sugar, 1.5 parts of agar, natural pH value, and 8 pounds of sterilization. Trichoderma koningii V6 strain was streaked and cultured at 28-30°C.
[0033] B. Secondary strains (liquid)
[0034] The formula of the culture medium is the same as that of the slant, except that agar is not added, the natural pH value is bottled, sterilized at 8 lbs, and the bacterium suspension of the above-mentioned slant strain is 4% by weight, cultured on a shaker at 28-30°C.
Embodiment 2
[0035] Embodiment 2: the preparation of cellulase
[0036] (1) Preparation of liquid strains for production
[0037] Culture medium: 10 parts of bran, 5 parts of potato, 100 parts of water, all are parts by weight, the same below. Boil, filter, add 0.5 parts of peptone, Na 2 HPO 4 0.45 parts and KH 2 PO 40.035 parts, sterilized, cooled, inoculated with 6 parts of the secondary liquid strains of Trichoderma korningen, ventilated and stirred, cultivated for 30 hours, and used as liquid strains for production.
[0038] The preparation of the secondary liquid strain of Trichoderma koningeni is as described in Example 1.
[0039] (2) Preparation of cellulase preparation culture by thick-layer ventilated fermentation
[0040] Raw materials: 25 parts of corn stalk powder and 40 parts of wheat straw powder, 40 parts of bran, 1 part of ammonium sulfate, all are parts by weight, moistened, raw materials: water=1: 2.8 (weight ratio), sterilized, cooled, inoculated Step (1) 6 part...
Embodiment 3
[0046] Embodiment 3: the preparation of cellulase
[0047] As described in Example 2, the difference is:
[0048] (1) Preparation of liquid strains for production
[0049] Culture medium: 11 parts of bran, 6 parts of potato, and 100 parts of water, all in parts by weight, the same below. Boil, filter, add 0.6 parts of peptone, Na 2 HPO 4 0.5 parts and KH 2 PO 4 0.04 parts, sterilized, cooled, inoculated with 10 parts of the secondary liquid strains of Trichoderma koningenii, ventilated and stirred, cultivated for 34 hours, and used as liquid strains for production.
[0050] (2) Preparation of cellulase preparation culture by thick-layer ventilated fermentation
[0051] Raw material: 60 parts of wheat straw powder, 35 parts of bran, 2 parts of ammonium sulfate, all are parts by weight, moisten water, raw material: water=1: 3.0 (weight ratio), sterilization, cooling, the production of inoculation step (1) Use 7 parts (weight) of liquid strains, transfer to aseptic solid...
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