Preparation method of liquid cellulase

A cellulase and liquid technology, applied in the field of liquid cellulase production, can solve the problems of large power consumption, large consumption of raw material microcrystalline cellulose and lactose, low equipment utilization rate, etc., so as to reduce costs and improve equipment utilization rate. , the effect of high-yield preparation

Active Publication Date: 2010-03-03
ANHUI BBCA FERMENTATION TECH ENG RES
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Problems solved by technology

At present, the fermentation of liquid cellulase mainly adopts batch fermentation, the fermentation period is longer, up to more than 7 days, the utilization rate of equipment is low, the power consumption is large, the preparation cost of seeds is high, and the cost of raw materials microcrystalline cellulose and lactose is high. Consumption is large, so the cost remains high

Method used

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  • Preparation method of liquid cellulase

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Effect test

Embodiment 1

[0029] Trichoderma viride (China Industrial Microorganism Culture Management and Preservation Center, CICC 40635) was used to produce cellulase by semi-continuous fermentation of microcrystalline cellulose in a 250mL Erlenmeyer flask with a liquid volume of 50mL, cyclic fermentation 10 times, and a fermentation temperature of 30 ℃, and the rotation speed is 250 rpm.

[0030] Wherein the fermentation medium contains 3% microcrystalline cellulose, 3% corn steep liquor, 3% urea, 4% ammonium sulfate and 4% dipotassium hydrogen phosphate.

[0031] Release 50% of the culture medium after the first fermentation for 96 hours, and then add an equal amount of fresh culture medium. Release 50% of the culture medium every 24 hours thereafter, and then add an equal amount of fresh culture medium, and stop the fermentation after a total of 10 cycles of fermentation. The ingredients in the supplemented medium are 3% microcrystalline cellulose, 2% corn steep liquor, 1% urea, 2% ammonium sulf...

Embodiment 2

[0034] Use Trichoderma viride (China Industrial Microorganism Culture Collection Center, CICC 41495) in a 50L stainless steel standard fermenter with a liquid volume of 30L. Microcrystalline cellulose is used for semi-continuous fermentation to produce cellulase, and the fermentation cycle is 10 times, the fermentation temperature is 30°C, the stirring speed is 300 rpm, the ventilation is 0.8vvm, and the tank pressure is 0.05Mpa.

[0035] Wherein the fermentation medium contains 2% microcrystalline cellulose, 2% corn steep liquor, 3% urea, 5% ammonium sulfate, 2% dipotassium hydrogen phosphate and 0.001% foam enemy in the fermentation medium.

[0036] After 85 hours of first fermentation, release 30% of the culture medium, and then add an equal amount of fresh medium. Release 20%, 30%, ... 80% of the culture solution every 24 hours, add an equal amount of fresh medium, cycle fermentation for 7 times, then release 60% of the culture solution every 24 hours, and at the same time...

Embodiment 3

[0039] Use Trichoderma viride (China Industrial Microorganism Culture Collection Center, CICC 40492) in a 1T stainless steel standard fermenter with a liquid volume of 750L. The cellulase is produced by semi-continuous fermentation of microcrystalline cellulose, the fermentation cycle is 10 times, the fermentation temperature is 30°C, the stirring speed is 200 rpm, the ventilation is 0.7vvm, and the tank pressure is 0.05Mpa.

[0040] The fermentation medium contains 2% microcrystalline cellulose, 2% corn steep liquor, 12% urea, 2.5% ammonium sulfate, 2.5% dipotassium hydrogen phosphate, and 0.001% foam enemy.

[0041] After 75 hours of first fermentation, release 40% of the culture medium, and then add an equal amount of fresh culture medium. After that, release 40% of the culture medium every 24 hours, and then add an equal amount of fresh culture medium, and finish the fermentation after a total of 12 cycles of fermentation. The ingredients in the supplemented medium are 2%...

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Abstract

The invention relates to a preparation method of liquid cellulase. The invention adopts a semicontinuous fermentation mode to utilize microorganism fermentation so as to produce cellulase in culture medium. The preparation method comprises the following steps of: (I) inoculating the microorganism into a fermentation medium, insulating and culturing; and (II) discharging 10 percent to 80 percent ofculture liquid obtained by the step (I), and simultaneously supplying new fermentation medium with same volume, and culturing; and circling the step (II) for more than 10 to 15 times, and dischargingthe tank. The method is applicable to industrial production, can improve utilization ratio of equipment, and simultaneously reduce the cost of preparing seeds; in addition, the cellulase and microcrystalline cellulose degradation products in the fermentation liquid also provide good external environment for the induction of cellulase of the following semicontinuous fermentation, therefore, the cellulase can be prepared with high yield.

Description

technical field [0001] The invention relates to the field of biological fermentation, in particular to a method for producing liquid cellulase by semi-continuous fermentation. Background technique [0002] As the main polysaccharide product of plant photosynthesis, cellulose is the most abundant renewable natural resource on earth. It is estimated that the energy contained in cellulose on the earth is approximately equivalent to the energy contained in 640 billion tons of petroleum, and the renewability of cellulose is incomparable to mineral energy such as petroleum. Making full use of cellulase to treat cellulose may fundamentally solve the energy and food problems faced by human beings, and its far-reaching impact cannot be underestimated. [0003] However, about 80% of cellulose has not been developed and utilized at present, which has an extremely attractive prospect. In recent years, the application research of cellulase at home and abroad is very active, and a numbe...

Claims

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Application Information

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IPC IPC(8): C12N9/42C12R1/885
Inventor 李荣杰薛培俭穆晓玲李维理
Owner ANHUI BBCA FERMENTATION TECH ENG RES
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