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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  • Description
  • Claims
  • Application Information

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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

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

Example Embodiment

[0024]Example 1

[0025]A method for improving the enzymatic hydrolysis efficiency of corn stover by using Rhodococcus biphenylophilus, including the steps:

[0026](1) Preparation of corn stover powder

[0027]Sun-dried and crushed corn stalks to obtain corn stalk powder;

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

[0029]Configure the LB medium supplemented with inorganic salts, inoculate the activated Rhodococcus biphenylophilus colonies, culture for 12 hours in an incubator set at 28°C and 200 rpm, OD600When the value reaches 2.5, prepare an ice box, put the bacteria liquid in a sterile and dry 50mL centrifuge tube in a sterile operating table, and use a low-temperature high-speed centrifuge to separate the bacteria and the bacteria liquid at 4°C and 8000rpm to obtain the supernatant As a crude enzyme solution of Rhodococcus biphenylophilus;

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

[0031]Wherein LB m...

Example Embodiment

[0042]Example 2

[0043]Calculation of Enzymatic Hydrolysis Efficiency of Corn Stalk

[0044]The corn stover powder pretreated with Rhodococcus biphenylophilus in Example 1 and the corn stover powder pretreated with sterile water in the comparative example were respectively subjected to enzymatic hydrolysis with cellulase, and a control group without added cellulase was set. The reducing sugar concentration was measured every 3h, and the reducing sugar content change curve was drawn. The result was that the reducing sugar concentration of cellulase enzymatic hydrolysis reached the highest after the enzyme treatment for 3h, and the control group had almost no reducing sugar production, 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 stover pretreated with sterile water. In order to compare the effects of different t...

Example Embodiment

[0048]Example 3

[0049]Scanning electron microscope observation of pretreated straw

[0050]The corn stalk powder after the pretreatment in step (3) of Example 1 and the corn stalk powder after the pretreatment in step (3) of the comparative example were used to observe the surface structure of the corn stalk material with a scanning electron microscope to compare the microstructure of the corn stalk after different pretreatments. Structural changes.

[0051]The results are shown in Figure 2. The corn stalk pretreated by Rhodococcus biphenylophilus has a significant structural change. The corn stalk material of the comparative example (Figure 2A withFigure 2C ) The surface structure is more complete, with an obvious lamellar structure attached to the surface of the corn stalks. Observing the section, it is found that the broken section is neat and basically without delamination. The holes are evenly distributed and the edges are smooth. On the surface of the corn stover material pretreated ...

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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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