Method for high-yield production of cellulase by trichoderma reesei fermentation
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A technology of Trichoderma reesei and cellulase, which is applied in the field of fermentation engineering and can solve problems that do not involve nitrogen sources
Active Publication Date: 2021-01-05
上海汉禾生物新材料科技有限公司 +1
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Although the method discloses the source of carbon source in the production process of cellulase, it does not involve nitrogen source and the applicable ratio of carbon source and nitrogen source in the production process
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preparation example 1
[0054] Preparation Example 1 Preparation of Induced Carbon Source
[0055] S1: Take 17% (w / v) pretreated biomass material, add 14FPU / g cellulase solution according to the mass of the material, react at 50°C, pH 4.8, and 180rpm for 24 hours, and obtain the enzymatic hydrolysis product of biomass material;
[0056] S2: Take 50% (w / v) glucose solution, add: 25CBU / g β-glucosidase, react under the conditions of 60°C, pH 4.8, and 180rpm for 24h to obtain a sophorose inducer.
[0057] S3: The enzymatic hydrolyzate of the biomass material and the sophorose inducer are mixed according to the mass ratio of 3:7 to obtain the induced carbon source.
preparation example 2
[0058] Preparation Example 2: Preparation of Induced Carbon Source
[0059] S1: Take 17% (w / v) pretreated biomass material, add betaine with a material quality of 1‰ and stir, then add 14FPU / g cellulase solution according to the material quality, and react under the conditions of 50°C, pH 4.8, and 180rpm After 24 hours, the enzymatic hydrolysis product of biomass material was obtained; the preparation method of S2-S3 was the same as that of Preparation Example 1.
preparation example 3
[0060] Preparation Example 3: Preparation of Induced Carbon Source
[0061] S1: Take 17% (w / v) pretreated biomass material, add betaine with a material quality of 3‰ and stir, then add 14FPU / g cellulase solution according to the material quality, and react under the conditions of 50°C, pH 4.8, and 180rpm After 24 hours, the enzymatic hydrolysis product of biomass material was obtained; the preparation method of S2-S3 was the same as that of Preparation Example 1.
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Abstract
The invention discloses a method for producing cellulase by trichoderma reesei fermentation. The method is characterized in that in a trichoderma reesei fermentation culture stage, a mixture of an induction carbon source and a nitrogen source is fed in a fed-batch manner, and preferably, the fed-batch feeding time is 24-26h after fermentation. The induction carbon source is obtained by mixing a biomass material enzymolysis product and a sophorose inducer according to a ratio of (3-5): (7-9), and the nitrogen source is a mixture of corn steep liquor and ammonium sulfate. The method has the advantages that the adding amount of phosphate in a trichoderma reesei fermentation culture medium is optimized, and production of the cellulase by the trichoderma reesei through permentation is performedwith low culture medium cost; the optimal nitrogen source feeding ratio is selected, so that the nitrogen source cost is saved; and the carbon-nitrogen ratio in the feeding process is strictly controlled, so that the trichoderma reesei can synthesize the cellulase at high yield.
Description
technical field [0001] The invention belongs to the technical field of fermentation engineering, and in particular relates to a method for fermenting Trichoderma reesei to high-yield cellulase by dynamically adjusting the carbon-to-nitrogen ratio and the carbon-to-phosphorus ratio in the fermentation process. Background technique [0002] The development of the energy industry affects the growth of the national economy and the progress of society. Today, commercial energy is mainly composed of fossil fuels. However, with the growth of the world population and the rapid increase of human needs, the depletion of fossil fuels has become the focus of attention. Developing the new energy industry to meet the ever-increasing energy demand has become an important development direction for all countries. [0003] As a large agricultural country with rich agricultural resources, China is rich in lignocellulosic biomass resources. Lignocellulosic biomass is a kind of clean energy, wh...
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