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Fermentation inoculum for producing cellulase and hemicellulase

A technology of hemicellulase and fermentation agent, 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: 2013-06-19
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] Embodiment 1, the screening of active ingredient in fermented inoculum

[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, agar 15g, switchgrass (the only carbon source) 10g.

[0018] For the liquid medium, crush the switchgrass through a 2mm sieve and add it to the medium, and the rest of the ingredients are the same.

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

[0020] The obtained cultures were studied by establishing 16S rDNA and 26S rDNA D1 / D2 region clone librarie...

Embodiment 2

[0021] Embodiment 2, produce the fermenting bacterial agent of cellulase and hemicellulase.

[0022] 1. Preparation of bacteria agent

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

[0024] Strain activation: Achromobacter xylosoxidans ATCC15173 and Alcaligenes faecalisATCC15554 were respectively inoculated on nutrient agar (composition of medium per liter: 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 inoculated on PDA (per liter medium composition: potato 200g, glucose 20g, agar 15g, distilled water 1000mL, natural pH), and cultured at 27°C for 4 days.

[0025] The activated bacterial strains were inoculated in corresponding ...

Embodiment 3

[0035] Embodiment 3, the fermentation bacterial agent that produces cellulase and hemicellulase.

[0036] 1. Preparation of bacteria agent

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

[0038] Strain activation: see Example 2

[0039] Inoculate the activated strains in the corresponding liquid medium: Achromobacter xylosoxidans ATCC15173 and Alcaligenes faecalis ATCC15554 were inoculated in 50ml of nutrient broth, shaken at 150rpm for 2 days; Fusarium sporotrichioides ATCC26533 and Fusariumpoae ATCC15654 were inoculated in 50ml of PDA liquid medium Medium, 150rpm shaking culture 4d. Take Achromobacter xylosoxidans ATCC15173 6ml, Alcaligenes faecalis ATCC15554 3ml, Fusarium sporotrichioides ATCC26533 10ml, Fusariumpoae ATCC15654 2ml and mix, so that the fi...

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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 lignocellulosic materials to produce cellulase and hemicellulase. Background technique [0002] With the development of the economy, human beings have an increasing demand for non-renewable fossil energy such as oil, natural gas, and coal, which has caused many problems such as energy security, oil consumption, and global warming. These problems have prompted countries all over the world to accelerate research on safe, clean and renewable alternative energy sources. Fuel ethanol is a clean alternative energy source that has made great progress in current research and application. However, its production currently mainly relies on starch and sugar raw materials. The use of these two raw materials involves competition with humans and animals for food and land. The resulting rise in food prices has emerged. The development of cheap raw materials other than grain is of great significan...

Claims

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Application Information

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Patent Type & Authority Patents(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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