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Method for producing pectinase preparation through solid-state fermentation of penicillium

A technology of pectinase preparation and solid-state fermentation, applied in the direction of microorganism-based methods, biochemical equipment and methods, enzymes, etc., can solve the problem of limiting the application range and effect of pectinase, the inability to achieve large-scale production, and the ability of strains to accumulate products Low-level problems, to achieve the effect of not being easy to pollute a large area, easy to control the fermentation process, and easy to operate

Pending Publication Date: 2021-10-15
江苏悠恒生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

my country has a great demand for pectinase, but the development of pectinase in China is relatively slow, and pectinase is usually used alone, which limits the application range and effect of pectinase
[0004] In the microbial fermentation industry, the strain is the key factor that directly determines whether the product quality and yield are up to standard. Generally, the strains isolated from nature have a very low ability to accumulate products and cannot meet the requirements of large-scale production.

Method used

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  • Method for producing pectinase preparation through solid-state fermentation of penicillium
  • Method for producing pectinase preparation through solid-state fermentation of penicillium
  • Method for producing pectinase preparation through solid-state fermentation of penicillium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Isolation, Screening and Identification of Penicillium sp. Y

[0034] (1) Isolation of strains

[0035] 5-10 g of soil samples collected from different farmland sampling points in the southern suburbs of Changzhou City, Jiangsu Province, 5-10 cm below the soil surface were stored in sterile 20 mL sealed bags, and stored in a refrigerator at 4 °C for subsequent screening of strains. Weigh 3-5g of soil samples from different sampling points and add them into 250ml Erlenmeyer flasks filled with 50ml sterile normal saline, mix well, shake fully and let stand for 30-60min, take 2-5ml of supernatant and add them into 50ml conical flasks In the 250ml Erlenmeyer flask of the initial enrichment medium, culture it on a constant temperature shaker at 120rpm min-1 at 38-40°C for 3-5 days, then transfer 5ml of the liquid medium to 45ml of fresh inorganic salt screening flask. A 250ml Erlenmeyer flask of culture medium was used for subsequent subcultures. Subculture for several tim...

Embodiment 2

[0048] By selecting the optimal solid substrate for the growth of Penicillium sp.Y, different solid-state fermentation culture substrates were used: rice bran, soybean cake powder, bran, soybean meal, cottonseed cake powder, rapeseed cake powder, and each group of experiments was compared three times. The operation is as follows:

[0049] In the ultra-clean workbench, use an inoculation loop to pick a few bacteria from the plate and insert them into the sterilized seed medium, and insert them into the above-mentioned rice bran at an inoculation amount of 2% after culturing to the logarithmic growth phase. , bean cake powder, bran, soybean meal, cottonseed cake powder, rapeseed cake powder as solid matrix fermentation medium, cultivated in a constant temperature incubator at 38°C for 3 days, and took samples to determine the enzyme activity produced by the strain.

[0050] This example shows that when soybean meal is used as the fermentation substrate, the pectinase activity pr...

Embodiment 3

[0052] By determining the optimum carbon source and its concentration required by the Penicillium sp.Y strain in the fermentation process, corn flour, starch, sucrose, glucose and fructose were selected as the fermentation carbon source, and 1%, 2%, 4%, 6% and 8% carbon source concentrations were added to the fermentation medium, and each group of experiments was compared for 3 times. The specific experimental operations were as follows:

[0053] During the preparation of the fermentation medium, corn flour, starch, sucrose, glucose and fructose were selected as the single carbon source and other variables were kept unchanged. After the medium was prepared, it was sterilized at 115°C for 30 minutes. In the ultra-clean workbench, use a sterilized gun tip to draw 2mL of seed liquid into the fermentation medium, then use a sterile glass rod to stir the medium evenly and place it in a 38°C constant temperature incubator for 3 days, then take a sample for measurement. enzyme activi...

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Abstract

The invention discloses a method for producing a pectinase preparation through solid-state fermentation of penicillium, which comprises the following steps: screening a pectinase high-yield strain Penicilliumsp.Y through high-throughput screening, and optimizing solid-state fermentation conditions of the pectinase high-yield strain Penicilliumsp.Y, so that the activity of the finally produced pectinase preparation is as high as 13800U / g; according to the method for producing the pectinase preparation through solid-state fermentation of the penicillium, an enzyme-producing strain with the enzyme activity as high as 19000U / g is successfully screened out under the condition that the fatality rate is 85.8% by adopting an ethyl methanesulfonate (EMS) mutant strain, and the optimal culture medium components and growth conditions of the strain are determined through a subsequent optimization test: 20g of soybean meal, 68% of initial water content, 3% of glucose content, the content of peptone as 1%, the content of CaCl2 as 0.04%, the content of FeCl3 as 0.04%, the inoculum size as 4%, and the culture temperature as 38 DEG C; and the enzyme activity is up to 2200U / g after the strain is cultured for 5 days, so that the enzyme production capacity of the strain is greatly improved, and an ideal target is achieved.

Description

technical field [0001] The invention relates to the field of biological fermentation engineering, in particular to a method for producing a pectinase preparation by solid-state fermentation of a penicillium strain. Background technique [0002] Enzymes play a vital role in the digestion and absorption of animals and the elimination of anti-nutritional factors in the feed industry. Among them, the compound enzyme preparation is a product that can realize multi-enzyme catalytic effects, and has a wider application field (chemicals, aquaculture, food and environmental protection, etc.), while reducing the production cost of the feed industry, it also plays an important role in environmental protection. play a certain role. And with the rapid development of biotechnology in the domestic and foreign enzyme preparation markets in the 21st century, people will pay more and more attention to the performance of enzyme preparations and their application value in the market. [0003]...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/88C12N1/14C12R1/80
CPCC12N9/88C12Y301/01011C12Y302/01015C12Y402/02002C12N1/14
Inventor 蔡志强邢新荣胡卫华
Owner 江苏悠恒生物技术有限公司
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