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Method for producing cellulose by microorganism mixing fermentation

A technology of cellulase and mixed fermentation, applied in the field of microbial engineering, can solve the problems of incomplete composition of cellulase system, lack of β-glucosidase, and improvement of enzyme production activity, so as to shorten the time of enzyme production and improve The effect of vitality and good application prospects

Inactive Publication Date: 2008-01-09
DONGGUAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
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Problems solved by technology

At present, the source of industrial cellulase is mainly produced by the aerobic filamentous fungus Trichoderma, such as Trichoderma reesei, Trichoderma viride, Trichoderma koningii, etc., and its production application There are two common problems in cellulite: one is that the enzyme activity of the existing production technology needs to be improved; the other is that the cellulase system produced by Trichoderma is not complete, and often lacks β-glucosidase, which will seriously affect the enzymatic efficiency

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1: The preparation of slant strains and secondary liquid strains, the specific operation steps are as follows.

[0037] (1) Production strain

[0038] Trichoderma koningii QF-2 and Rhizopus oryzae NRRL395HS99 (both preserved by the Microbiology Laboratory of our school).

[0039] (2) Preparation of Inclined Strains

[0040] Prepare potato agar medium (PDA) slant first, then streak and inoculate Trichoderma korningii strain and Rhizopus oryzae strain respectively, and form a large number of spores after culturing at 28°C for 5 days and 7 days respectively, which can be used as secondary seed liquid and solid-state fermentation time inoculation.

[0041] (3) Preparation of secondary liquid strains

[0042] The culture medium formula is the same as the slant bacterial classification, but does not add agar, bottling is sterilized, connects the spore liquid of the slant bacterial classification by 3~5% weight percentage (the number of spores per milliliter is 1...

Embodiment 2

[0043] Embodiment 2: preparation of solid-state fermentation medium and liquid fermentation medium

[0044] (1) Solid-state fermentation medium: 3 parts of bagasse powder, 2 parts of bran, 10 parts of Mandels nutrient solution, adjusted to pH 4.5-5.0, sterilized at 121°C for 25 minutes.

[0045] (2) Liquid fermentation medium: 3 parts of rice straw powder, 100 parts of Mandels nutrient solution, adjusted to pH 4.5-5.0, put into an automatic fermenter, and sterilized at 121° C. for 20 minutes.

Embodiment 3

[0046] Embodiment 3: the process steps of producing cellulase by microbial mixed liquid state fermentation

[0047] (1) liquid seed production: according to the method for embodiment 1 seed production;

[0048] (2) earlier with the secondary liquid bacterial classification of Trichoderma korningii, inoculate in the liquid fermentation medium that makes according to the method for embodiment 2 by the 5% inoculum size of liquid fermentation medium volume, after 48 hours aerated culture under 30 ℃, Then inoculate 4% of the volume of Rhizopus oryzae secondary liquid strain, continue aeration and culture at 30°C for 72 hours, filter the culture solution, and the filtrate is the original enzyme solution;

[0049] (3) The above filtrate is concentrated to 3 / 10 of the original volume, and the concentrated solution is spray-dried to make a powdery high-activity cellulase preparation.

[0050] The filter paper enzyme activity in the original enzyme solution is 2.2IU / ml, and the β-gluco...

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Abstract

Production cellulase by microbe mixed fermentation is carried out by taking Trichoderma koningii and Rhizopus oryzae as cellulase production bacterium, inoculating spore fluid or secondary seed fluid into solid or liquefied culture medium and controlling culture medium and amount of Trichoderma koningii and Rhizopus oryzae and other culture conditions during mixed bacterium culture. The filter-paper activity, beta-glucosidese activity are increased by 60-100% and 400-500% and fermentation time is shortened. It's benefit to secrete cellulase and improves enzyme system. It's cheap and simple.

Description

technical field [0001] The invention belongs to the technical field of microbial engineering, and in particular relates to a method for producing cellulase by microbial mixed fermentation. Background technique [0002] Cellulase is a general term for a group of enzymes that act synergistically to catalyze the hydrolysis of cellulose to finally produce glucose. They are widely used in many fields of industry and agriculture. For example, cellulase can biotransform industrial and agricultural waste such as straw and bagasse into glucose, which can be further transformed by other microorganisms to produce biochemical products such as alcohol, lactic acid, and single-cell protein; using cellulase as a feed additive can improve the utilization of feed Rate; cellulase can also be used in waste paper deinking, fabric polishing and many other aspects. In the research work of plant biotechnology, cellulase is often used to hydrolyze the cell wall for genetic engineering and cell eng...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12N1/14
Inventor 邹水洋吴清林邓韵王尚武
Owner DONGGUAN UNIV OF TECH
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