Liquid-solid two phase fermentation method for producing high activity cellulase for fodder

A cellulase and endocellulase technology, applied in the field of bioengineering, can solve the problems of long fermentation period, easy environmental pollution of fermentation liquid and high volume production efficiency

Inactive Publication Date: 2005-08-03
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Liquid submerged fermentation, high energy consumption, long fermentation cycle, low volumetric production efficiency, complex post-treatment process, high production cost, and the fermentation liquid is easy to pollute the environment
In contrast, solid fermentation technology has high volume production efficiency, low energy consumption, simple post-treatment process, and does not pollute the environment. The difficulty is that the fermentation conditions are difficult to control and easy to pollute
More than a dozen domestic fermentation manufacturers that produce cellulase by solid fermentation are all in a state of unstable enzyme production, serious pollution, and abnormal production due to backward technology, and cannot form industrialization

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: the preparation of the secondary liquid bacterial classification of Trichoderma korning, concrete operating steps are as follows:

[0031] A. Slope bacteria (solid)

[0032] Peel and wash the potatoes, cut them into small pieces, put them in water, potatoes: water = 1:4 weight ratio, soak at 80°C for 1 hour, boil and filter with a filter cloth, dilute the filtrate with water to the original volume. Add 2 parts of sugar, 1.5 parts of agar, natural pH value, and 8 pounds of sterilization. Trichoderma koningii V6 strain was streaked and cultured at 28-30°C.

[0033] B. Secondary strains (liquid)

[0034] The formula of the culture medium is the same as that of the slant, except that agar is not added, the natural pH value is bottled, sterilized at 8 lbs, and the bacterium suspension of the above-mentioned slant strain is 4% by weight, cultured on a shaker at 28-30°C.

Embodiment 2

[0035] Embodiment 2: the preparation of cellulase

[0036] (1) Preparation of liquid strains for production

[0037] Culture medium: 10 parts of bran, 5 parts of potato, 100 parts of water, all are parts by weight, the same below. Boil, filter, add 0.5 parts of peptone, Na 2 HPO 4 0.45 parts and KH 2 PO 40.035 parts, sterilized, cooled, inoculated with 6 parts of the secondary liquid strains of Trichoderma korningen, ventilated and stirred, cultivated for 30 hours, and used as liquid strains for production.

[0038] The preparation of the secondary liquid strain of Trichoderma koningeni is as described in Example 1.

[0039] (2) Preparation of cellulase preparation culture by thick-layer ventilated fermentation

[0040] Raw materials: 25 parts of corn stalk powder and 40 parts of wheat straw powder, 40 parts of bran, 1 part of ammonium sulfate, all are parts by weight, moistened, raw materials: water=1: 2.8 (weight ratio), sterilized, cooled, inoculated Step (1) 6 part...

Embodiment 3

[0046] Embodiment 3: the preparation of cellulase

[0047] As described in Example 2, the difference is:

[0048] (1) Preparation of liquid strains for production

[0049] Culture medium: 11 parts of bran, 6 parts of potato, and 100 parts of water, all in parts by weight, the same below. Boil, filter, add 0.6 parts of peptone, Na 2 HPO 4 0.5 parts and KH 2 PO 4 0.04 parts, sterilized, cooled, inoculated with 10 parts of the secondary liquid strains of Trichoderma koningenii, ventilated and stirred, cultivated for 34 hours, and used as liquid strains for production.

[0050] (2) Preparation of cellulase preparation culture by thick-layer ventilated fermentation

[0051] Raw material: 60 parts of wheat straw powder, 35 parts of bran, 2 parts of ammonium sulfate, all are parts by weight, moisten water, raw material: water=1: 3.0 (weight ratio), sterilization, cooling, the production of inoculation step (1) Use 7 parts (weight) of liquid strains, transfer to aseptic solid...

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PUM

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Abstract

The present invention relates to two-phase liquid-solid fermentation process of producing high activity cellulase for fodder, and belongs to the field of bioengineering technology. Koningic engineering strain with high antimetabolite repression, high cellulase yield and complete endocellulose, exocellulose and glucosidase is selected and used in two-phase liquid-solid fermentation process of stereo fermentation engineering technology to produce cellulase. The fermented product has activity as high as 500-700 IU / g dry leaven, and may be further prepared through serial biochemical engineering process to obtain high purity cellulase powder with activity up to 8000-10000 IU / g. The present invention is used in fodder industry to develop stalk resource while avoiding environmental pollution.

Description

(1) Technical field [0001] The invention relates to a preparation method of high-activity cellulase with complete enzyme spectrum of three types of endo-cellulase, exo-cellulase and glucoglucosidase, which can be used as a special enzyme preparation for feed and belongs to the technical field of bioengineering. (2) Background technology [0002] The amount of crop straw is huge, with 1 billion tons of various types of straw each year. So far, it has not been well utilized. A large amount of straw has been burned, which not only causes environmental pollution, but also greatly wastes resources. Therefore, to seek high-value ecological utilization of straw, It is an urgent problem to be solved at present. [0003] As the main enzyme species for the degradation and transformation of renewable biomass resources, cellulase has long been highly valued by the scientific and technological community, and extensive basic and applied research has been carried out. However, most of the...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N9/42
Inventor 张玉忠孙彩云陈秀兰刘玉庆陈蕾蕾宋桂经
Owner SHANDONG UNIV
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