Fermentation inoculum for producing cellulase and hemicellulase

A technology of hemicellulase and cellulase, applied in the direction of microorganism-based methods, biochemical equipment and methods, enzymes, etc., can solve problems such as technical and cost constraints, low conversion efficiency, and restrictions on large-scale industrial applications

Active Publication Date: 2011-08-24
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there are numerous technical and cost constraints at each step of these transformations
Hydrolysis is one of the key steps. Cellulase and hemicellulase play an important role in the lignocellulosic ethanol conversion process due to their mild action conditions and less pollution. However, the current enzyme production costs are high and the conversion efficiency is low. Low level seriously restricts the large-scale industrial application of cellulosic ethanol conversion and becomes the bottleneck of industrial technology

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1. Screening of active ingredients in fermentation bacteria

[0017] Composition of MCD medium (g / L): NaNO 3 2g, KH 2 PO 4 1g, MgSO 4 ·7H 2 O 1g, KCl 0.5g, FeSO 4 0.01g, 15g agar, 10g switchgrass (the only carbon source).

[0018] For the liquid medium, the switchgrass is crushed through a 2mm sieve and then added to the medium, and the rest of the ingredients are the same.

[0019] 0.5g of the samples initially used for screening (sample sources: rotten dead branches, dead leaves, rotten switchgrass) were respectively inserted into 100ml of Modified Czapek Dox (MCD) agar. After 14 days of cultivation at 30°C, 0.05 g of switchgrass overgrown with microorganisms was transferred to a new medium for subculture. Select the cultures that quickly soften switchgrass and rapidly degrade cellulose and hemicellulose to screen the active ingredients.

[0020] The 16S rDNA and 26S rDNA D1 / D2 region clone libraries were established by molecular ecology methods to study the cultu...

Embodiment 2

[0021] Example 2. Fermentation agent for producing cellulase and hemicellulase.

[0022] 1. Preparation of bacterial agents

[0023] Achromobacter xylosoxidans ATCC15173; Alcaligenes faecalis ATCC15554; Fusarium sporotrichioides ATCC26533; Fusarium poae ATCC15654 were all purchased from the American Type Culture Collection. The sources of the strains were soil, manure, bean hulls and barley seeds.

[0024] Strain activation: Achromobacter xylosoxidans ATCC15173 and Alcaligenes faecalisATCC15554 were respectively inoculated on nutrient agar (per liter of medium composition: peptone 10g, beef extract 3g, NaCl 5g, agar 15g, distilled water 1000ml, pH 7.4), cultured at 27°C for 2 days; Fusarium Sporotrichioides ATCC26533 and Fusariumpoae ATCC15654 were respectively inoculated on PDA (composition per liter of medium: 200g potato, 20g glucose, 15g agar, 1000mL distilled water, natural pH), and cultivated at 27°C for 4 days.

[0025] The activated strains were respectively inoculated into t...

Embodiment 3

[0035] Example 3. Fermentation agent for producing cellulase and hemicellulase.

[0036] 1. Preparation of bacterial agents

[0037] Achromobacter xylosoxidans ATCC15173; Alcaligenes faecalis ATCC15554; Fusarium sporotrichioides ATCC26533; Fusarium poae ATCC15654 were all purchased from the American Type Culture Collection. The sources of the strains were soil, manure, bean hulls and barley seeds.

[0038] Strain activation: see Example 2

[0039] The activated strains were respectively inoculated into the corresponding liquid medium: Achromobacter xylosoxidans ATCC15173 and Alcaligenes faecalis ATCC15554 were respectively inoculated in 50ml nutrient broth, shaking at 150rpm for 2 days; Fusarium sporotrichioides ATCC26533 and Fusariumpoae ATCC15654 were respectively inoculated into 50ml PDA liquid medium Culturing with shaking at 150 rpm for 4 days. Take Achromobacter xylosoxidans ATCC15173 6ml, Alcaligenes faecalis ATCC15554 3ml, Fusarium sporotrichioides ATCC26533 10ml, Fusariumpo...

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PUM

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Abstract

The invention discloses a microbial inoculum for producing cellulase and hemicellulase by a natural lignocellulose material. The microbial inoculum for producing the cellulase and the hemicellulase comprises the following active ingredients: Achromobacter xylosoxidans, Alcaligenes faecalis, Fusarium sporotrichioides and Fusarium poae. In the invention, microbes in the microbial inoculum for producing the cellulase and the hemicellulase have a strong synergistic effect so that a stable ecological system can be formed in an autologous mode. The microbial inoculum is especially developed for an energy crop-Panicum virgatum but is not limited to production of the cellulase and the hemicellulase by use of the Panicum virgatum, and meanwhile the inoculum can be also used for producing the cellulase and the hemicellulase by waste crop straw.

Description

Technical field [0001] The invention relates to a bacterial agent for fermenting switchgrass lignocellulose materials to produce cellulase and hemicellulase. Background technique [0002] With the development of economy, mankind's demand for non-renewable fossil energy such as oil, natural gas and coal is increasing, which has caused many problems such as energy security, oil consumption, and global warming. These problems have prompted countries around the world to accelerate research on safe, clean, and renewable alternative energy sources. Fuel ethanol is a clean alternative energy that has made great progress in current research and application, but currently its production mainly relies on starch and sugar raw materials. The use of these two raw materials involves the problem of competing with humans and animals for food. The resulting increase in food prices has already emerged. The development of cheap raw materials other than grain is of great significance to the produc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/00C12N1/14C12N1/20C12N9/42C12R1/025C12R1/05C12R1/77
Inventor 杨洪岩李玉花吴昊
Owner NORTHEAST FORESTRY UNIVERSITY
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