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Composition for disaggregating lignocelluloses and method for disaggregating lignocelluloses by virtue of composition

A technology of lignocellulose and depolymerization, applied in the fields of biochemical equipment and methods, enzymes, oxidoreductases, etc., can solve the problems of little guiding significance, unclear mechanism of action, and less obvious synergy.

Active Publication Date: 2016-05-18
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Swollenin protein only exists in Trichoderma reesei, and has little guiding significance for the transformation of other high-producing cellulase fungal enzyme systems
And it has been found that the synergy between cellulase degradation factors and commercial cellulase is not obvious, and the mechanism of action is not yet clear.

Method used

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  • Composition for disaggregating lignocelluloses and method for disaggregating lignocelluloses by virtue of composition
  • Composition for disaggregating lignocelluloses and method for disaggregating lignocelluloses by virtue of composition
  • Composition for disaggregating lignocelluloses and method for disaggregating lignocelluloses by virtue of composition

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Isolate and purify the dehydrogenase in the composition of the present invention from the extracellular enzyme liquid of Penicillium juniper:

[0038] Penicillium piceum (Penicillium piceum) H16, preservation number: CGMCC No. 8339, preservation unit China General Microbiological Culture Collection Center (china general microbiological culture collection center).

[0039] For Penicillium juniper, the most suitable medium conditions are as follows: 3.3% microcrystallinecellulose, 1.7% corncobsteepliquor, 0.5% (NH4) 2 SO 4 , 0.6% KH 2 PO 4 , 0.1% MgSO 4 , 0.25% CaCO 3 , and 0.2% Tween-80. The glass container used for fermentation generally chooses a 300mL Erlenmeyer flask, cultivates 50mL of the culture solution, and shakes it horizontally at 180rpm in a 28-degree incubator for 5 days.

[0040] The instrument used for dehydrogenase separation and purification is AKTApurifier (GE, Sweden). Using acetic acid-sodium acetate solution with a pH value of 4.8-5.0 and a sub...

Embodiment 2

[0043] Enzyme activity assay of dehydrogenase:

[0044] Application of the conventional methods in the art cellulase and hemicellulase national standard assay method composition-dehydrogenase to carboxymethylcellulose (CMC-Na), salicin (Salicin), PNPC, microcrystalline cellulose ( Avicel), xylan (Xylan), PNPG, and cellobiose were tested for enzyme activity, and the test results are shown in Table 1 below: the composition was found to have no glycoside hydrolase activity.

[0045] Table 1. Preliminary analysis of enzymatic properties of dehydrogenases

[0046]

Embodiment 3

[0048] Research on the mechanism of action of the dehydrogenase of the present invention

[0049] When dehydrogenase was added to the crude enzyme solutions of Trichoderma reesei and Penicillium juniperus at a protein concentration of 40 μg / mL, the hydrolysis efficiencies of the crude enzyme solutions of Trichoderma reesei and Penicillium juniperi increased by 27% and 35%, respectively. %. This is the first time that a dehydrogenase has been found in a cellulase system. The dehydrogenase itself has no enzymatic activity, and there is a strong synergistic effect with the cellulase, so the dehydrogenase belongs to the cellulase synergistic composition, such as figure 2 shown.

[0050] The dehydrogenase protein did not detect hydrolase activity, but when it treated four kinds of lignocellulosic materials with different components, when the hydrolyzate was analyzed by HPLC-42A, it was found that there was a small amount of D-glucose and D-glucose in the hydrolyzate - Sorbitol, ...

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Abstract

The invention relates to the technical field of lignocellulose degradation, in particular to a composition for disaggregating lignocelluloses and a method for disaggregating lignocelluloses by virtue of the composition. The composition for disaggregating lignocelluloses disclosed by the invention consists of dehydrogenase and cellulase. Under the circumstance of the existence of the same cellulase content, the degradation efficiency of the lignocellulose is increased to a great extent. The invention also discloses the method for disaggregating the lignocellulose by virtue of the composition, wherein the method is achieved by adding the cellulase and the dehydrogenase in the composition to the lignocellulose for degradation, wherein 1g of the substrate is added with 10FPU of the cellulase and 1g of the substrate is added with 80-120[mu]g of the dehydrogenase. The inventor, on the basis of a great amount of tests, aims at determining relatively appropriate dehydrogenase and cellulase dosages in the composition and relatively appropriate reaction conditions for degrading the lignocellulose in practical production, so that the full use of the composition is achieved to the greatest extent, the yield of saccharides from lignocellulose degradation is improved and production cost is reduced.

Description

technical field [0001] The invention relates to the technical field of lignocellulose degradation, in particular to a composition for lignocellulose depolymerization and a depolymerization method thereof. Background technique [0002] In recent years, based on the in-depth study of the main components of cellulase, the discovery of some non-hydrolytic enzyme-active cellulose-degrading factors has become a current research hotspot. In the biodegradation process of cellulose in nature, besides the main enzymatic mechanism, are there any other novel synergistic degradation factors of cellulase? As early as half a century ago, Reese et al. (Reese et al., 1950) proposed that there should be a hydrogen bond enzyme that destroys crystalline cellulose, but it has not been well confirmed in microorganisms so far (Gao Peiji, 2003). When Yan Boxu and others studied the CBD region of CBHI of Trichoderma pseudoconii and Penicillium violaceum, they found that the adsorption of CBD on cel...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N9/42C12P19/14
Inventor 高乐李晨张东远陈树林
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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