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Method for producing cellulase by using induction and regulation of substrate

A cellulase and substrate-induced technology is applied in the field of microbial engineering to achieve the effects of improving cellulase activity, improving fermentation level and reducing application costs

Inactive Publication Date: 2013-04-17
河南天冠纤维乙醇有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It adopts the preparation of liquid strains, solid-state thick-layer ventilated fermentation culture to produce high-activity cellulase, but the fermentation products it uses are rice bran and bran, and straw microcrystalline cellulose cannot be used, so it needs a soluble carbon Source A and insoluble straw microcrystalline cellulose are used together as inducers and mass-produced as a substrate-induced production of high-efficiency cellulase

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] ⑴ culture medium

[0034] Fermentation medium: straw microcrystalline cellulose 4.0%, glucose 1.0%, bran 1.0%, corn steep liquor 1.5%, (NH 4 ) 2 SO 4 0.6%, KH 2 PO 4 0.2%, CaCl 2 0.08%, MgSO4 ·7H 2 O 0.1%, MnSO 4 0.02%, FeSO 4 0.02%, ZnSO 4 ·7H 2 O 0.06%, Cocl 2 6H 2 O 0.04%, the balance is water.

[0035] Feed medium: 16% soluble carbon source A, 5% straw microcrystalline cellulose, 2.0% corn steep liquor, (NH 4 ) 2 SO 4 0.4%, KH 2 PO 4 0.4%, CaCl 2 0.1%, MgSO 4 ·7H 2 O0.2%, the balance is water.

[0036] ⑵Inoculate the liquid bacteria in a 50L fermenter at 8% (V / V), the initial pH is 5.0, the stirring speed is 300rpm, the fermentation temperature is 29°C, and the aerated culture is carried out. 3 The feed rate is continuously fed until the end of the fermentation; the pH of the fermentation process is adjusted to 4.2-4.8 by feeding ammonia water; the fermentation lasts for 90-168 hours, the fermentation is stopped, and the cellulase enzyme ...

Embodiment 2

[0038] ⑴ culture medium

[0039] Fermentation medium: straw microcrystalline cellulose 5.0%, glucose 0.8%, bran 1.0%, corn steep liquor 1.2%, (NH 4 ) 2 SO 4 0.4%, KH 2 PO 4 0.2%, CaCl 2 0.08%, MgSO 4 ·7H 2 O 0.1%, MnSO 4 0.02%, FeSO 4 0.02%, ZnSO 4 ·7H 2 O 0.06%, Cocl 2 6H 2 O 0.08%, the balance is water.

[0040] Feed medium: soluble carbon source A 18%, straw microcrystalline cellulose 6%, corn steep liquor 1.6%, (NH 4 ) 2 SO 4 0.4%, KH 2 PO 4 0.4%, CaCl 2 0.1%, MgSO 4 ·7H 2 O0.2%, the balance is water.

[0041] ⑵ Inoculate the liquid bacteria at 10% (V / V) in 6m 3 Fermentation tank, initial pH 4.8, agitation speed 150rpm, fermentation temperature 30°C for aerated culture, after 50 hours of fermentation, 6.4L / h*m 3 The feed rate is continuously fed until the end of fermentation; the pH is adjusted to 4.4-4.8 by feeding ammonia water during the fermentation process; the fermentation lasts for 124 hours, the fermentation is stopped, and the cellula...

Embodiment 3

[0043] ⑴ culture medium

[0044] Fermentation medium: straw microcrystalline cellulose 5.5%, glucose 0.5%, bran 0.5%, corn steep liquor 1.5%, (NH 4 ) 2 SO 4 0.4%, KH 2 PO 4 0.2%, CaCl 2 0.05%, MgSO 4 ·7H 2 O 0.06%, MnSO 4 0.04%, FeSO 4 0.04%, ZnSO 4 ·7H 2 O 0.06%, Cocl 2 6H 2 O 0.08%.

[0045] Feed medium: soluble carbon source A 14%, straw microcrystalline cellulose 8%, corn steep liquor 2.0%, (NH 4 ) 2 SO 4 0.6%, KH 2 PO 4 0.5%, CaCl 2 0.1%, MgSO 4 ·7H 2 O0.1%, the balance is water.

[0046] ⑵ Inoculate the liquid bacteria at 10% (V / V) in 60m 3 Fermentation tank, initial pH 4.8, agitation speed 100rpm, fermentation temperature 30°C for aerated culture, after 40 hours of fermentation, 7.8L / h*m 3 The feed rate is continuously fed until the end of the fermentation; the pH of the fermentation process is adjusted to 4.4-4.8 by feeding ammonia water; the fermentation lasts for 86-144 hours, stop the fermentation, and the cellulase enzyme activity (FPA)...

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Abstract

The present invention provides a method for producing cellulase by using induction and regulation of substrate. Trichoderma viride is used as cellulase-producing bacteria. A soluble carbon source A and insoluble straw microcrystalline cellulose are used as an inducer for regulating metabolic processes. High active cellulase is produced by controlling medium composition, pH regulation, ventilation ratio, inducer adding manner and the like. The method provided by the present invention plays a significant role on improving cellulase activity, and reducing application cost of cellulose; is strong in operability; has no pollution as developing and utilizing straw and other natural cellulosic resources; is an effective means to promote the industrialization of straw ethanol; and has broad application prospects.

Description

technical field [0001] The invention belongs to the technical field of microbial engineering, and in particular relates to a method for inducing and regulating cellulase production by using a substrate. Background technique [0002] Cellulose is a polymer substance with a complex structure, so the enzymes that can degrade cellulose are not single enzymes, but a complex multi-enzyme system. Cellulase refers to the general term for a group of enzymes that can hydrolyze cellulose β-1,4 glucosidic bonds and turn cellulose into cellobiose and glucose. It is a multi-component enzyme system that acts synergistically. Wide range of applications. For example, cellulase can bioconvert industrial and agricultural waste such as crop stalks and bagasse into glucose, which can be further converted into biochemical products such as alcohol and single-cell protein by other microorganisms. In addition, cellulase can also be used in medicine, printing and dyeing, tea processing and other as...

Claims

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Application Information

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IPC IPC(8): C12N9/42C12R1/885
Inventor 赵子高王林风阎振丽
Owner 河南天冠纤维乙醇有限公司
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