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Preparation method of penicillium group compounded cellulose synergistic degradation compound enzyme

A technology of synergistic degradation and cellulose, applied in the field of microorganisms, to improve the degradation efficiency, reduce the amount of added enzymes, and improve the efficiency of enzymatic hydrolysis

Inactive Publication Date: 2022-03-11
GUANGZHOU INST OF ENERGY CONVERSION - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the enzymatic hydrolysis efficiency of cellulase on the pretreated fiber substrate still needs to be further improved.

Method used

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  • Preparation method of penicillium group compounded cellulose synergistic degradation compound enzyme
  • Preparation method of penicillium group compounded cellulose synergistic degradation compound enzyme
  • Preparation method of penicillium group compounded cellulose synergistic degradation compound enzyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Preparation of compound enzyme combination preparation with synergistic cellulose degradation

[0027] A method for preparing a compound enzyme of penicillium flora to synergistically degrade cellulose, comprising the following steps:

[0028] 1. Medium:

[0029] PDA liquid medium: Weigh 200g of peeled potatoes, add water to boil, filter through gauze to obtain potato juice, then add 20g of glucose, dissolve and add water to 1L, pH value is natural, sterilized at 115°C for 30min, and made into grade one Seed medium.

[0030] Solid-state fermentation medium: composed of 5g rice straw and 10.5mL nutrient salt solution, the composition of nutrient salt solution is: each liter of nutrient salt solution contains (NH 4 ) 2 SO 4 10.0g, KH 2 PO 4 4.0g, MgSO 4 ·7H 2 O 0.5g, CaCl 22H 2 O 0.5g, the balance is 1000mL water. The initial pH was adjusted to 6 (adjusted with 1M NaOH), and sterilized at 115°C for 30 minutes to make a solid-state fermentation medi...

Embodiment 2

[0045] Example 2: Compound enzyme combination preparation applied to enzymatic hydrolysis of poplar wood cellulose

[0046] 1. Reaction substrate: crush poplar to 60 mesh, dry at 105°C to constant weight, accurately weigh (10g) poplar into a pressure bottle, first add 0.5% v / v sulfuric acid solution to make the solid-liquid ratio 1 :20(g:L), use a sterilizer for 1 hour at 121°C. After the pretreatment, use a G3 sand core funnel to separate the mixture from solid to liquid, then wash the residue with deionized water to a neutral pH value, and dry it in an oven at 105°C to constant weight; then weigh the acid pretreated bottom Add 2% w / v sodium hydroxide solution to make the solid-to-liquid ratio 1:20 (g:L), and use a sterilizing pot to treat at 121°C for 1h. After pretreatment, use G3 sand core funnel to separate the mixture from solid to liquid, then wash the residue with deionized water to a neutral pH value, place it in an oven at 105°C and dry it to constant weight, and fi...

Embodiment 3

[0049] Embodiment 3: Comparison of different enzymolysis formulations

[0050] The poplar wood with acid-base composite pretreatment was prepared according to the method of Example 2.

[0051] Take 0.1g poplar pretreated by acid-base compound, and carry out enzymatic hydrolysis in 5mL different solution formulation systems. Among them, formula 1: do not add CTec2 cellulase, M1 compound enzyme combination preparation, protein amount 40 mg, make up 5 mL with acetic acid-sodium acetate buffer solution with a concentration of 0.2M and pH=4.8; formula 2: do not add CTec2 cellulase , M18 compound enzyme combination preparation, the amount of protein is 40mg, supplement 5mL with acetic acid-sodium acetate buffer solution with a concentration of 0.2M and pH=4.8; formula 3: CTec2 cellulase 10FPU / g substrate, M1 compound enzyme combination preparation, protein Amount 40mg, make up 5mL with acetic acid-sodium acetate buffer solution with a concentration of 0.2M and pH=4.8; formula 4: CT...

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Abstract

The invention discloses a preparation method of a penicillium group compounded cellulose synergistic degradation compound enzyme. The compound enzyme is prepared by adding a bacterial liquid M18, which is prepared by compounding penicillium oxalicum XZH-2, penicillium XZH-6, penicillium citrinum XZH-16, penicillium XZH-22, neurospora crassa N1 and orange thermoascomycetes T2 according to a volume ratio of 1: 8: 1: 1: 1: 1, into a solid-state fermentation culture medium, and carrying out mixed fermentation. When the compound enzyme combined preparation is applied to wood cellulose enzymolysis, the enzymolysis efficiency is improved by 21.5%, and the highest cellulose conversion rate can reach 88.7%. The compound enzyme combined preparation is an oxidation hydrolase with a synergistic cellulose degradation effect, and can break glucosidic bonds of cellulose through an oxidation effect and promote combination of cellulase and a fiber substrate so as to improve the enzymolysis efficiency, so that the enzyme addition amount in the enzymolysis process is effectively reduced. The production cost of the cellulosic ethanol can be further reduced, and the industrialization process of the cellulosic ethanol is accelerated.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a preparation method of a compound enzyme of Penicillium flora compounding and synergistically degrading cellulose. Background technique [0002] Implement renewable energy substitution actions, vigorously develop wind energy, solar energy, biomass energy, ocean energy, geothermal energy, etc., and continuously increase the proportion of non-fossil energy consumption. Among them, biomass energy is a green renewable resource with great potential for development, and using it as a raw material to prepare alcohol fuels is an important direction for future development. However, how to saccharify lignocellulosic biomass efficiently, environmentally friendly and at low cost is the key to the production of cellulosic alcohol fuels. [0003] The efficient utilization of biomass requires the biotransformation of cellulase. However, cellulase is not a single-component ...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N9/42C12N9/02C12P7/10C12P19/14C12R1/80C12R1/645
CPCC12N1/14C12N9/2437C12N9/0083C12P7/10C12P19/14C12Y302/01004C12Y302/01091C12Y302/01021C12P2201/00C12P2203/00Y02E50/10
Inventor 梁翠谊亓伟胡云子王闻王琼王忠铭
Owner GUANGZHOU INST OF ENERGY CONVERSION - CHINESE ACAD OF SCI
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