A method for producing cellulase by fermenting corn stalk furfural residue with mixed bacteria

A corn stalk and cellulase technology, which is applied in the field of cellulase production from corn stalk furfural slag mixed with bacteria, can solve the problems of unfavorable production of cellulase by Trichoderma reesei, decreased enzyme activity of filter paper, and low enzyme activity of filter paper. Achieve good economic and social benefits, high substrate concentration, and simple process

Active Publication Date: 2021-11-09
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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Problems solved by technology

CN 101100660 A proposes a method for producing cellulase by mixed fermentation of Trichoderma koningenii and Rhizopus oryzae, but the growth conditions of the two strains are not the same, and Rhizopus oryzae grows faster than Trichoderma koningingii, which easily leads to mixed During the cultivation process, Rhizopus oryzae grows predominantly, which actually reduces the enzyme activity of the filter paper. Therefore, it is necessary to strictly control the cultivation method and conditions. The two strains need to be inoculated in stages, and the operation is more complicated. Problems such as nutrient competition cannot be avoided; although CN 102154243 A uses Trichoderma viride and Aspergillus niger for mixed fermentation to improve the activity of β-glucosidase, it also has the same problem; Gao Xingxing et al. use Trichoderma reesei and Aspergillus niger for mixed fermentation , in addition to the antagonism due to the different growth conditions, the addition of Aspergillus niger changed the pH value of the fermentation system, which was not conducive to the production of cellulase by Trichoderma reesei, resulting in a higher filter paper enzyme activity than single Richter's in mixed fermentation. Trichoderma is still low (Food Science, 2012,33(19):193-198)

Method used

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  • A method for producing cellulase by fermenting corn stalk furfural residue with mixed bacteria
  • A method for producing cellulase by fermenting corn stalk furfural residue with mixed bacteria
  • A method for producing cellulase by fermenting corn stalk furfural residue with mixed bacteria

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Embodiment 1

[0043] Cellulase produced by mixed bacteria fermentation

[0044] The Trichoderma reesei CICC13052 and CICC40360 preserved on the slant were respectively activated, transferred from the spore suspension to the seed culture solution with a 5% inoculation amount, and cultured on a shaker at 28° C. and 180 rpm for 24-48 hours. Then transfer it to the enzyme-producing medium containing 50g / L furfural slag at an inoculation ratio of 1:1 and a total inoculation amount of 5%, and take samples at regular intervals to determine the enzyme activity. The result is as figure 1 It was shown that the cellulase activity of the mixed bacteria could reach 0.82 FPU / mL when 50 g / L corn stalk furfural residue was used as the induced carbon source.

Embodiment 2

[0052] Effects of Different Substrate Concentrations on Enzyme Production by Mixed Bacteria Fermentation

[0053] According to the method described in Example 1, the mixed bacteria were transferred to the enzyme-producing medium with different concentrations of furfural slag at an inoculation ratio of 1:1 and a total inoculation amount of 5%, and samples were taken at regular intervals to determine the enzyme activity to explore its optimal carbon source. content and enzyme production. The result is as Figure 4 As shown, it can be seen from the figure that the mixed bacteria have better substrate tolerance, even at the concentration of 200g / L furfural residue, they can also ferment and produce enzymes. When 100g / L furfural residue is used as the induced carbon source, the cellulose The maximum enzyme activity can reach 0.87FPU / mL, so the substrate concentration in subsequent tests is preferably 100g / L.

Embodiment 3

[0055] Effects of Different Initial pH on Enzyme Production by Mixed Bacteria Fermentation

[0056] The pH of the medium is closely related to the growth and reproduction of cells and the production of enzymes by fermentation. During the fermentation process, with the growth and reproduction of cells and the accumulation of metabolic products, the pH of the medium often changes, so this experiment only controls the initial pH .

[0057] According to the method described in Example 2, the mixed bacteria are transferred to the product containing 100g / L furfural slag with an initial pH of 2.80, 4.32, 4.80, 6.00, 7.00, 8.00 with an inoculation ratio of 1:1 and a total inoculation amount of 5%. Fermentation was carried out in the enzyme medium, samples were taken regularly to measure the enzyme activity, the effect of the initial pH on the enzyme production of the mixed bacteria fermentation was observed, and the optimal initial pH and enzyme production were explored. The result i...

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Abstract

The invention provides a method for producing cellulase by fermenting corn stalk furfural slag with mixed bacteria. In the method, Trichoderma reesei CICC13052 and CICC40360 are used to carry out mixed fermentation in an enzyme production medium using corn stalk furfural slag as an induced carbon source to produce cellulase. Cellulase. The method of the invention has cheap and easy-to-obtain raw materials, overcomes the disadvantage of low enzyme production by a single strain, and improves production efficiency, and both of the mixed bacteria belong to Trichoderma reesei, have the same growth conditions, have no antagonistic effect, and are beneficial to mixed culture. The method of the invention is easy to operate and low in cost, and is expected to provide a simple and economical fermentation process for the industrialization of cellulase production by microbial fermentation.

Description

technical field [0001] The invention belongs to the field of bioenergy utilization, in particular to a method for producing cellulase by fermenting corn stalk furfural residue with mixed bacteria. Background technique [0002] Cellulase has good application prospects in textile industry, feed industry, brewing, food processing and development of new energy. In recent years, with the increasingly severe energy problems, the proposition of using cellulase to degrade straw raw materials to produce fiber fuel ethanol and other new energy sources has been put on the agenda, but the high price has hindered its wide application. Therefore, the production of high-efficiency and low-cost cellulase has epoch-making significance for finally solving the energy shortage problem faced by human beings. [0003] Cellulase comes from a wide range of sources, and bacteria, actinomycetes, fungi, and animals can all produce cellulase. The cellulase synthesized by different microorganisms has ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/42C12R1/885
CPCC12N9/2437C12N9/2445C12Y302/01004C12Y302/01021C12Y302/01091
Inventor 张宗超刘会芳刘秀梅
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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