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Method for improving Zea mays L. straw enzymolysis efficiency by Rhodococcus biphenylivorans

A technology for Rhodococcus biphenophila and corn stalks is applied in the field of improving the enzymatic hydrolysis efficiency of corn stalks by using Rhodococcus biphenophila, and can solve the problems of low enzymatic hydrolysis efficiency, hindering the contact between cellulase and cellulose, and difficulty in production, etc. To achieve the effect of improving hydrolysis effect and increasing accessibility

Pending Publication Date: 2021-01-15
HUBEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cellulose can be converted into glucose by enzymatic hydrolysis for the production of ethanol and organic acids, but lignin in lignocellulose hinders the contact between cellulase and cellulose, resulting in low enzymatic hydrolysis efficiency
In order to improve the efficiency of enzymatic hydrolysis, current studies have shown that pretreatment of corn stalks with lignin peroxidase extracted from Trichoderma reesei can specifically degrade lignin components, thereby improving the effect of cellulase. Lignin peroxidase is too expensive and difficult to produce, making it difficult to apply it to large-scale biological pretreatment

Method used

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  • Method for improving Zea mays L. straw enzymolysis efficiency by Rhodococcus biphenylivorans
  • Method for improving Zea mays L. straw enzymolysis efficiency by Rhodococcus biphenylivorans
  • Method for improving Zea mays L. straw enzymolysis efficiency by Rhodococcus biphenylivorans

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A method for improving the enzymatic hydrolysis efficiency of corn stalks by using Rhodococcus biphenylophilum, comprising the steps of:

[0026] (1) Preparation of corn stalk powder

[0027] drying and crushing corn stalks to obtain corn stalk powder;

[0028] (2) Preparation of Rhodococcus biphenylphila crude enzyme solution

[0029] Configure LB medium with added inorganic salts, inoculate activated Rhodococcus biphenylophilus colonies, and culture in an incubator set at 28°C and 200rpm for 12h, OD 600 When the value reaches 2.5, prepare an ice box, put the bacterial liquid into a sterilized and dry 50mL centrifuge tube in a sterile operating table, use a low-temperature high-speed centrifuge to separate the bacterial cells and bacterial liquid at 4°C and 8000rpm, and obtain the supernatant As the crude enzyme solution of Rhodococcus biphenylophilus;

[0030] LB medium with inorganic salts added: 50 mL of LB medium + 50 mL of inorganic salt medium;

[0031] Where...

Embodiment 2

[0043] Calculation of Enzymatic Hydrolysis Efficiency of Corn Stalk

[0044] The corn stalk powder pretreated by Rhodococcus biphenylphila in Example 1 and the corn stalk powder pretreated by sterile water in the comparative example were subjected to enzymatic hydrolysis treatment with cellulase, and a control group without cellulase was set. The concentration of reducing sugar was measured every 3 hours, and the curve of reducing sugar content was drawn. The result was that the concentration of reducing sugar hydrolyzed by cellulase reached the highest after 3 hours of enzyme treatment, and basically no reducing sugar was produced in the control group, indicating that there was basically no dissolution of straw components. , after pretreatment of straw raw materials with Rhodococcus biphenylophilus, the hydrolysis effect of cellulase is significantly higher than that of corn straw pretreated with sterile water. In order to compare the effects of different treatments on cellulo...

Embodiment 3

[0049] Observation of pretreated straw by scanning electron microscope

[0050] The corn stalk powder after the pretreatment of the step (3) of Example 1 and the corn stalk powder after the pretreatment of the step (3) of the comparative example were respectively used scanning electron microscopy to observe the surface structure of the corn stalk material, and the microscopic structure of the corn stalk after different pretreatments was compared. Structural changes.

[0051] The results are shown in Figure 2, the corn stalks pretreated by Rhodococcus biphenylophilus had obvious structural changes, and the comparative corn stalk materials ( Figure 2A and Figure 2C) The surface structure is more complete, with obvious lamellar structure attached to the surface of corn stalks. Observation of the section shows that the broken section is neat and basically without delamination, the holes are evenly distributed and the edges are smooth. However, on the surface of corn stover mat...

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Abstract

The invention relates to the field of biotransformation, in particular to a method for improving Zea mays L. straw enzymolysis efficiency by Rhodococcus biphenylivorans. The method for improving the Zea mays L. straw enzymolysis efficiency by the Rhodococcus biphenylivorans comprises the following steps of: 1) drying Zea mays L. straws in the sun, smashing the dried Zea mays L. straws to obtain Zea mays L. straw powders; 2) inoculating activated Rhodococcus biphenylivorans in culture solution, culturing until OD600 is greater than or equal to 2.0, and collecting the supernate of the culture solution to obtain Rhodococcus biphenylivorans crude enzyme liquor; 3) adding the Rhodococcus biphenylivorans crude enzyme liquor into the Zea mays L. straw powders, and carrying out processing for 3-10days; and 4) adding cellulase into the pre-processed Zea mays L. straw powders, and carrying out enzymolysis reaction, wherein the bacterial strain of the Rhodococcus biphenylivorans is named Rhodococcus biphenylivorans B403, the collection number of the Rhodococcus biphenylivorans B403 is CCTCC NO: M 2019087, and the collection date is January 25th, 2019. The method disclosed by the invention has the characteristics of being simple to operate, low in cost and suitable for centralized processing of the straws on a large scale.

Description

technical field [0001] The invention relates to the field of biotransformation, in particular to a method for improving the enzymatic hydrolysis efficiency of corn stalks by using Rhodococcus biphenylophilum. Background technique [0002] In recent years, energy crisis has appeared worldwide, environmental pollution has become increasingly serious, and people have to look for new green energy to replace fossil fuels. Among many new energy sources, biomass energy has always been favored by people. According to estimates, the world produces about 150 billion tons of biomass annually, including plant-based starch, agricultural waste and lignocellulose. In my country, the annual output of crop corn stalks is about 700 million tons, and the use of cellulose-containing corn stalks to produce fuel has attracted more and more attention from researchers. [0003] The main component of corn stover is lignocellulose, which usually contains three main components: cellulose, hemicellulo...

Claims

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Application Information

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IPC IPC(8): C12P19/14C12P19/02C12R1/01
CPCC12P19/02C12P19/14C12P2201/00C12P2203/00
Inventor 江正兵宋慧婷肖文静
Owner HUBEI UNIV
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