Method for increasing yield of cellulase produced from fungus
A cellulase and fungus technology is applied in the field of increasing the yield of cellulase produced by fungi, which can solve the problems of high cost, complicated operation and low efficiency, and achieve the effects of low cost, convenient operation and high efficiency.
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Embodiment 1
[0026] A method for improving fungal cellulase activity according to the present invention comprises the following steps:
[0027] S1, providing spore liquid of Trichoderma reesei Rut-C30 strain. Specifically, this step is: inoculate Trichoderma reesei strain Rut-C30 on a potato medium (PDA) plate, grow at 28°C for 4 days, use methods known in the art to collect spores, dilute in sterile water, Make the concentration of spores in the spore liquid be 10 7 -10 8 pieces / ml.
[0028] S2, inoculating the spores into the seed medium to obtain seeds. Specifically, in this embodiment, this step includes: inoculating the spores in step S1 into a seed medium, and vibrating at 28° C. and 220 rpm for 30 hours to obtain seeds. Among them, the seed medium includes: glucose 15g / L, corn steep liquor 20g / L, KH 2 PO 4 15g / L, (NH4) 2 SO 4 5g / L, CaCl 2 0.5g / L, MgSO 4 ·7H 2 O 1g / L, FeSO 4 ·7H 2 O5mg / L, MnSO 4 ·H 2 O 1.6mg / L, ZnSO 4 ·7H 2 O 1.4mg / L, CoCl 2 ·6H 2 O 2mg / L.
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Embodiment 2
[0033] Example 2 is similar to Example 1, the difference is that step S3 is to inoculate the seeds in the fermentation medium containing dimethylformamide, so that the volume ratio of the seeds to the fermentation medium containing dimethylformamide is 1:10, shake culture at 28°C and 220rpm for 4 days, centrifuge to take the supernatant, and obtain the crude enzyme solution. Specifically, in this embodiment, the fermentation medium containing dimethylformamide is added by adding dimethylformamide to the fermentation medium, and the volume ratio of dimethylformamide and fermentation medium is 0.5: 100. Among them, the fermentation medium includes: glucose 5g / L, lactose 37g / L, corn steep liquor 27g / L, KH 2 PO 4 6g / L, (NH 4 ) 2 SO 4 5g / L, CaCl 2 0.5g / L, MgSO 4 ·7H 2 O 1g / L, FeSO 4 ·7H 2 O 5mg / L, MnSO 4 ·H 2O 1.6mg / L, ZnSO 4 ·7H 2 O 1.4mg / L, CoCl 2 ·6H 2 O 2mg / L.
[0034] The obtained crude enzyme solution was subjected to SDS-PAGE protein electrophoresis. Tak...
Embodiment 3
[0037] Example 3 is similar to Example 2, the difference is that in the fermentation medium containing dimethylformamide in step S3, dimethylformamide is added to the fermentation medium, and dimethylformamide and fermentation The volume ratio of the medium is 1:100.
[0038] The obtained crude enzyme solution was subjected to SDS-PAGE protein electrophoresis, taking the experimental results on the third day as an example, such as figure 1 As shown in column 4, the protein expression of Trichoderma reesei Rut-C30 was increased by using the fermentation medium containing dimethylformamide provided in this example.
[0039] The obtained crude enzyme solution was tested for CMCNa endonuclease activity, pNPC exonuclease activity, filter paper enzyme activity and hemicellulase activity respectively. Take the experimental results on the third day as an example, such as figure 2 As shown, the CMCNa endonuclease activity has increased by about 60% compared with the first control gr...
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