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Compound enzyme for degrading tobacco stems and application of compound enzyme in degrading tobacco stems

A compound enzyme and compound enzyme preparation technology, applied in the biological field, can solve the problems of difficult wide application, low enzymatic hydrolysis efficiency, large enzyme dosage, etc.

Active Publication Date: 2021-05-28
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First of all, it is difficult to maximize the utility of each enzyme, resulting in a large amount of enzymes and waste of resources; on the other hand, the enzymatic hydrolysis efficiency is low, the cycle is long, and the requirements for environmental conditions are relatively high, which makes this method difficult to obtain. wide application

Method used

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  • Compound enzyme for degrading tobacco stems and application of compound enzyme in degrading tobacco stems
  • Compound enzyme for degrading tobacco stems and application of compound enzyme in degrading tobacco stems
  • Compound enzyme for degrading tobacco stems and application of compound enzyme in degrading tobacco stems

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1: Isolation and identification of Pycnoporus sanguineus (Pycnoporus sanguineus) WYS377 high-yielding laccase strain

[0059] 1. Collection and isolation of strains: In October 2006, the inventor Li Wei isolated and purified a strain of bacteria from decayed wood samples collected in the Wuyishan National Nature Reserve in Fujian Province through PDA medium, and recorded the strain as strain WYS377.

[0060] 2. Morphological identification of the strain: Inoculate the above-mentioned obtained strain WYS377 on PDA medium, culture at 30°C for about 4-6 days to cover a 90mm petri dish, and red pigments begin to accumulate in about 8 days. The hyphae are white fluffy at first, the edges are radial, and the center of the colony has an obvious concentric structure. As the number of days of cultivation increases, these hyphae will be covered by a layer of white powdery substance. The white powdery substance is analyzed by microscopic observation. The powdery substance ...

Embodiment 2

[0064] Embodiment 2: the preparation of laccase

[0065] Microporosa haemophilus WYS377 is an aerobic bacterium with a growth temperature of 28-30°C. During fermentation, it can secrete various substances including laccase into the medium. In this example, the lacquer enzyme. Proceed as follows:

[0066] 1. Enzyme-producing fermentation: Inoculate the Pyroporositum WYS377 of Example 1 on the PDA medium, cultivate it at 30°C for 4 days, and use a sterile punch to make the growth medium into a bacterium with a length and width of 1 cm. Take 8 pieces of fungus cake and place them in 100mL enzyme-producing fermentation medium, then culture them in the dark at 28°C and 180r / min for 10 days, add the inducer 2,5-dimethylaniline on the 4th day of culture, 2, The concentration of 5-dimethylaniline in the culture system was 10 μM. After the culture was completed, it was centrifuged at 5000 r / min for 20 min, and the supernatant was collected. The obtained supernatant was WYS377 laccase...

Embodiment 3

[0070] Embodiment 3: Utilize compound enzyme preparation to degrade tobacco stem

[0071] 1. Preparation of compound enzyme preparation

[0072] According to the ratio of the enzymatic activity (U) of laccase, cellulase, hemicellulase and pectinase according to the dry powder, cellulase, hemicellulase and pectinase of D. Mixed at a ratio of 0.146:0.01:0.01:0.0753 to obtain WYS377 compound enzyme preparation;

[0073] The commercially available laccase, cellulase, hemicellulase, and pectinase of Example 2 are 0.146:0.01 according to the enzyme activity (U) ratio of laccase, cellulase, hemicellulase and pectinase : 0.01: 0.0753 ratio mixed to obtain commercially available laccase compound enzyme preparation.

[0074] 2. Using compound enzyme preparation to degrade tobacco stems

[0075] (1) Prepare WYS377 compound enzyme preparation into compound enzyme liquid by using distilled water (pH value is neutral). Dry the tobacco stems at 60°C to constant weight, cut them into piec...

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Abstract

The invention discloses a compound enzyme for degrading tobacco stems and an application of the compound enzyme in degrading the tobacco stems. The invention discloses a compound enzyme for degrading tobacco stems, which is prepared from laccase, cellulase, hemicellulase and pectinase, the laccase is prepared from Pycnoporus sanguineus WYS377, The preservation number of the Pycnoporus sanguineus WYS377 in the General Microbiological Culture Collection Center of the China Committee for Culture Collection of Microorganisms is CGMCC No. 18573. The compound enzyme provided by the invention can be used for realizing effective degradation of tobacco stem biomass. The compound enzyme disclosed by the invention not only can be used for degrading tobacco stem biomass, but also can be used for degrading other waste biomass, such as corn straw, wheat straw, rice straw and other biomass materials containing lignocellulose structures, and has a wide application prospect.

Description

technical field [0001] The invention relates to a complex enzyme for degrading tobacco stems and its application in the field of biotechnology. Background technique [0002] China is a large agricultural country, and about 4 billion tons of biomass materials are discarded every year, such as forestry waste, poultry manure, domestic garbage and straw. These waste biomass materials are generally utilized through direct incineration, biotransformation, thermochemical conversion, liquefaction, etc., but for straw, due to its rich lignocellulose structure, its utilization efficiency is low and it is difficult to are degraded, so pretreatment of straw to improve its utilization efficiency is an urgent problem to be solved at present. [0003] Tobacco stem is an important part of tobacco raw materials, accounting for about 30% of the total weight of tobacco leaves, but it usually exists as a waste biomass in the cigarette industry, which wastes resources and causes serious environ...

Claims

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Application Information

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IPC IPC(8): C12N9/88C12N9/42C12N9/18C12N9/02A24B15/20
CPCC12N9/2402C12N9/0061C12Y110/03002C12Y302/01004C12N9/18C12N9/88C12Y301/01011C12Y302/01015C12Y402/02002A24B15/20C12P2203/00
Inventor 尹文兵李伟宋自力
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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