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Environment-friendly enzyme as well as rapid preparation method and application thereof

An environmentally friendly enzyme and fast technology, applied in the biological field, can solve the problems of high fermentation cost, high price and long time, ranging from 1 month to half a year.

Active Publication Date: 2022-05-17
ZHEJIANG FORESTRY ACAD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditionally, the production of enzymes needs to provide fruits, vegetables and various sugars as fermentation substrates. The price of these raw materials is high, so the fermentation cost is high, and the time is long, ranging from one month to half a year.

Method used

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  • Environment-friendly enzyme as well as rapid preparation method and application thereof
  • Environment-friendly enzyme as well as rapid preparation method and application thereof
  • Environment-friendly enzyme as well as rapid preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Embodiment 1: Carbon source test of Sj18 bacterial strain

[0013] The Sj18 strain has the ability to utilize various sugars. Carbon source test: First, the sj18 strain was obtained by punching 0.5 cm holes on the PDA plate with sj18 colonies, and inoculated the 0.5 cm bacterial block on a blank medium plate containing only agar for 3 days at 32 ° C, and then from the long-term Punch holes at the edge of the grown mycelium, take out the bacterial block and inoculate it on plates of various carbon source media, culture at 32°C for 2 days, and compare the differences in their mycelial growth;

[0014] The carbon sources are respectively: lactose, soluble starch, sucrose, glucose, maltose, fructose, xylose, blank uninoculated, and blank inoculated. The Sj18 strain grows well in different carbon sources, and can make full use of all kinds of carbon sources. For specific growth characteristics, see figure 1 ;

[0015] All the reagents mentioned above are of domestic anal...

Embodiment 2

[0017] Embodiment 2: Utilize rice bran and sawdust to carry out the sequential fermentation of sj18 and 15B1 bacteria

[0018] Prepare 500 mL of culture solution containing 5% sawdust and 5% rice bran, put it into a 1L Erlenmeyer flask, and sterilize at 121°C for 20 minutes. Use a 1cm puncher to punch 2-3 holes on the culture plate of sj18 bacteria to obtain strains, and inoculate the strains into the above-mentioned 1L Erlenmeyer flask culture solution, shake at 200rpm at 32°C for 5 days, and ferment the broth through 200 mesh vacuum filtration to obtain the filtrate;

[0019] Prepare the PDA plate of 15B1 bacteria, pick the clones and inoculate them in 5mL PDB culture medium for overnight culture, then inoculate the 5mL culture medium into the above-mentioned filtrate after sj18 fermentation, culture with shaking at 200rpm at 32°C for 2 days, filter, and the filtrate 4-15 ℃ for 1 day, centrifuged to take the supernatant, and treated at 70 ℃ for 1 hour to obtain inactivated ...

Embodiment 3

[0020] Embodiment 3: Analysis of volatile components detected by GC-MS of fungal enzymes

[0021] This analysis revealed some key components in the enzyme that are effective in repelling mosquitoes. specific method:

[0022] Use a 0.5cm hole punch to take a piece of bacteria from the edge of the colony above 5cm from the culture plate of sj18 bacteria, inoculate it into a headspace glass bottle (with a sealing plug) equipped with a slant of the medium, and cultivate it at 32ºC for three days. The desorption-pretreated SPME tube penetrates the septum of the sample bottle, inserts it into the bottle, and pushes the handle so that the fiber head extends out of the needle tube and is placed in the upper space of the sample (headspace method) for extraction. The extraction time is about 30 minutes. Next, insert the SPME needle tube into the injection port of the GC-MS instrument (Agilent Technologies 7890AGC System, Agilent Technologies 5975C MS), push the handle rod, extend the f...

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PUM

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Abstract

The Xylaria radius sj18 strain and the Pichia fermentans 15B1 strain are sequentially fermented by taking residual wastes of agriculture and animal husbandry production and processing as fermentation substrates, the obtained fermentation liquor is inactivated, and the enzyme prepared by processing and production has the effect of repelling mosquitoes. The method is low in production cost, free of pollution and residues, environmentally friendly and free of toxic and side effects on animals and plants, and has important guiding significance in processing and production of biological insect repellents.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to fungi of the family Xyloceraaceae ( Xylaria radix ) sj18 strain and Pichia fermentum ( Pichia fermentans ) The application of the sequential fermentation product of 15B1 strain in repelling mosquitoes. Background technique [0002] Enzymes are made from animals, plants, fungi, etc., with or without supplementary materials, and contain specific biologically active ingredients (including polysaccharides, oligosaccharides, proteins and peptides, amino acids, vitamins) through microbial fermentation. The product. Enzyme has the effect of repelling mosquitoes, and because it is mostly produced by fermenting sugars with various probiotics, it has better biological safety. Traditionally, the production of enzymes needs to provide fruits, vegetables and various sugars as fermentation substrates. The price of these raw materials is high, so the fermentation cost is high, and t...

Claims

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Application Information

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IPC IPC(8): A01N63/30A01N63/32A01P17/00
CPCA01N63/30A01N63/32Y02W30/40
Inventor 彭华正朱汤军金群英
Owner ZHEJIANG FORESTRY ACAD
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