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Preparation method of natural plant foliage fertilizer containing chitosan oligosaccharide

A technology of natural plants and chitosan oligosaccharides, which is applied in animal carcass fertilizers, organic fertilizers, fertilization devices, etc., can solve the problems affecting the popularization and application of chitosan oligosaccharides and chitosan oligosaccharides, environmental pollution by strong acid, and high cost, and achieves easy industrialization. The effect of production and promotion and application, making up for deficiencies, and simple equipment

Pending Publication Date: 2018-12-21
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The chemical method has high requirements on equipment, and the use of a large amount of strong acid is likely to cause environmental pollution, and because the degradation process is a random process, the degree of polymerization of the product is difficult to control
The enzymatic hydrolysis method uses chitin enzyme to degrade chitin or chitosan enzyme to degrade chitosan, so as to obtain chitooligosaccharide or chitooligosaccharide with a certain degree of polymerization, but the enzymatic hydrolysis method needs to use chitin or chitosan with high purity At the same time, the price of commercialized chitinase and chitosanase is relatively high, which leads to the high cost of enzymatic hydrolysis preparation process, which makes the price of chitooligosaccharide or chitooligosaccharide products remain high, which affects the quality of chitosan and chitosan. Promotion and Application of Oligosaccharides

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Cloning of chitinase gene saChiA4 and construction of plasmid pP43NMK::saChiA4

[0023] Primers 5'-CCGGAATTCATGGAACGCGTACTACCC-3' and 5'-CCCAAGCTTGCAGGCGCCGTTGTCCGC-3' were designed, and the genome of Streptomyces albolongus was extracted as a template for PCR amplification. After the target fragment was amplified, the target fragment was ligated with the linearized pP43NMK plasmid using a seamless splicing kit, and the ligated product was transformed into E. / ml) on an LB plate, cultivate overnight at 37°C, pick a single colony, and perform colony PCR verification. The successful verification of the single colony contains the plasmid pP43NMK::saChiA4.

Embodiment 2

[0024] Example 2 Preparation of recombinant Bacillus subtilis containing chitinase gene saChiA4

[0025] Take Bacillus subtilis to coat LB plates, pick a single colony with good growth, incubate in LB medium at 37°C for 10 h, take 100 μL of bacteria, and use the Spizizen method to prepare Bacillus subtilis competent cells. The plasmid pP43NMK::saChiA4 was transformed into Bacillus subtilis competent cells by electric shock, and evenly spread on LB plates with kana resistance (50ug / ml), and cultured at 37°C for 10h. Pick a single colony for colony PCR verification, and verify that the correct Bacillus subtilis is the recombinant Bacillus subtilis B.S.saChiA4 containing chitinase saChiA4.

Embodiment 3

[0026] Embodiment 3 uses recombinant Bacillus subtilis to ferment shrimp head

[0027] Take 1kg of shrimp head homogenate, add 3kg of water to sterilize, and put it in a 5L fermenter to sterilize at 121°C for 20min. After cooling to room temperature, the recombinant Bacillus subtilis B.S.saChiA4 seed liquid cultured in LB medium was inoculated with 5% inoculum amount. The temperature of the fermentation tank was controlled at 37° C., the stirring speed was 300 rpm, and the ventilation rate was 0.5 vvm for 5 days of fermentation. After the fermentation is finished, the fermentation broth is centrifuged at 8000rpm for 10min, and the centrifugation supernatant of the fermentation broth is taken. The chitooligosaccharide concentration detected in the supernatant was 15.9g / L.

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Abstract

The invention provides a preparation method of natural plant foliage fertilizer containing chitosan oligosaccharide. The preparation method comprises the steps of fermenting crustacean processing by-products by using recombinant bacillus subtilis, and compounding fermentation liquid as well as traditional Chinese herbal medicine extracts such as astragalus membranaceus and dandelion to obtain thefoliage fertilizer which can be applied to growth of crops. Compared with the chemical method and the enzymolysis method, the preparation method of the natural plant foliage fertilizer containing thechitosan oligosaccharide is low in cost, simple in equipment, environmentally friendly, convenient in industrialized production and popularization application and capable of covering the shortage in the prior art.

Description

technical field [0001] The invention belongs to the technical field of foliar fertilizers, and in particular relates to a preparation method of natural plant foliar fertilizers containing chitooligosaccharides. Background technique [0002] Chitooligosaccharides and chitosan oligosaccharides are oligosaccharides produced by the degradation of chitin or chitosan, which have the characteristics of good water solubility, high biological activity, and strong functions. Oligochitosan and oligochitosan have a similar structure, and the amino group of oligochitosan monomer is replaced with an acetyl amino group to form oligochitosan. Therefore, the two have similar physical and chemical properties and physiological activities in many aspects. Chitooligosaccharides and chitosan oligosaccharides are widely used in agriculture. They can significantly enhance plant disease resistance, promote plant growth, improve the quality of agricultural products, and prolong the freshness of fruit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C05F15/00
CPCC05F1/005C05G5/20C05F11/00C05F11/08
Inventor 毛相朝孙建安周燕
Owner OCEAN UNIV OF CHINA
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