Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Bacteriostatic protease CmYC1 prepared by eukaryotic fermentation and application of bacteriostatic protease CmYC1

A technology of protease and sclerotinia, applied in the biological field, can solve the problems of chemical fungicide restrictions and other problems

Active Publication Date: 2020-03-17
HUAZHONG AGRI UNIV
View PDF10 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, with people's concern about food safety and environmental pollution, some chemical fungicides have been severely restricted in many developed countries and regions

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Bacteriostatic protease CmYC1 prepared by eukaryotic fermentation and application of bacteriostatic protease CmYC1
  • Bacteriostatic protease CmYC1 prepared by eukaryotic fermentation and application of bacteriostatic protease CmYC1
  • Bacteriostatic protease CmYC1 prepared by eukaryotic fermentation and application of bacteriostatic protease CmYC1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] A method utilizing eukaryotic fermentation to prepare antibacterial protease CmYC1:

[0017] The nucleotide sequence shown in SEQ ID NO.1 was artificially synthesized, and the sequence shown in SEQ ID NO.1 was used as a template (primer F: atcgaattcATGGCTCCCGTTGTCACTCCT, R: aaatatgcggccgcTTAGTGGTGGTGGTGGTGGTGTGCGCCGTTGCCGTTGTAGA), and the amplified sequence was constructed on the eukaryotic expression vector pPIC9. The plasmid with correct sequencing was transferred to yeast expression strain GS115, and positive transformants were screened with MD medium (1% yeast extract, 2% peptone, 1×YNB, 2% glucose, 1% agar) to obtain expression inhibitory The engineering bacteria of bacterial protease CmYC1.

[0018] Using BMGY medium (1% yeast extract, 2% peptone, 1×YNB, 1% glycerol, 0.0002% biotin, pH=7) the engineering bacteria of aprotinase CmYC1 were expanded and shaken, centrifuged to collect the thalline, and The cells were resuspended with BMMY (1% yeast extract, 2% pepton...

Embodiment 2

[0026] Analysis experiment of protease CmYC1 enzymatic characteristics.

[0027] 1) Definition of protease activity: 1 enzyme activity unit (1U) is the amount of enzyme required to produce 1 microgram of tyrosine in 1mL of enzyme solution at 40°C and pH 7.2 for 1min.

[0028] 2) Drawing of standard curve. Use distilled water to prepare solutions containing 10, 20, 30, 40, 50, 60, 70, 80, 90, and 100 μg / mL tyrosine respectively, absorb 1 mL of the above-mentioned tyrosine solutions with different concentrations, and add 0.55 mol / L sodium carbonate solution 5mL, and then add 1mL of diluted Folin's reagent. Shake well and place in a water bath, keep warm at 40°C for 15 minutes to develop color, after cooling, use No. 1 tube as a blank, measure the absorbance at a wavelength of 660nm with a spectrophotometer, do 3 parallel runs, and take the average value. Draw the standard curve with the concentration of tyrosine as the abscissa and the absorbance as the ordinate.

[0029] 3) ...

Embodiment 3

[0044] Effect of protease CmYC1 on the mycelial growth of Sclerotinia botrytis, Botrytis cinerea, Sclerotinia sclerotiorum, Mucor, Rhizoctonia, Fusarium graminearum, Fusarium oxysporum, Pythium melon and fruit:

[0045] Induce 48h CmYC1 yeast fermented liquid (8.26U / ml) to add in PDA culture medium (every 100mlPDA adds 10ml fermented liquid), boil and inactivate fermented liquid as control; In the culture medium, the bacteriostatic rate was calculated after 24 hours (bacterial inhibition rate=(control colony radius-treated colony radius)*100%). Protease Cm YC1 has a certain inhibitory effect on the mycelial growth of Sclerotinia, Botrytis cinerea, Sclerotinia micromyces, Mucor, Rhizoctonia, Gibberella, Fusarium oxysporum, and Pythium melon.

[0046] strain Antibacterial rate (%) Sclerotinia sclerotiorum 22.21±2.35 Sclerotinia minor 22.54±2.38 Botrytis cinerea 29.88±3.67 Mucor hiemails 10.62±1.49 Rhizoctonia solani 13.98±0.81 Fusar...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the technical field of biology and relates to bacteriostatic protease CmYC1 prepared through eukaryotic fermentation and application of the bacteriostatic protease CmYC1. Thenovel bacteriostatic protease CmYC1 can be used for inhibiting the growth of sclerotinia sclerotiorum, botrytis cinerea, sclerotinia minor, mucor hiemails, rhizoctonia solani, fusarium graminearum, fusarium oxysporum and pythium aphanidermatum, inhibit the infection of the ascospores of the sclerotinia sclerotiorum on leaves of oilseed rapes and has a great potential of being developed into a bacteriostatic agent.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to an antibacterial protease CmYC1 prepared by eukaryotic fermentation and an application thereof. Background technique [0002] Sclerotinia sclerotiorum (Lib.) de Bary] is an important plant pathogenic fungus, which can cause sclerotinia in many crops. In actual production, the prevention and control of sclerotinia mainly adopts the comprehensive control strategy of agricultural measures, supplemented by chemical control (Hou Mingsheng and Huang Junbin, 2006). Sclerotinia minor is an important plant pathogenic fungus worldwide, which can cause basal rot of plants, resulting in serious economic losses (Subbarao, 1998). Botrytis cinerea can damage more than 1400 species of plants, including various economic crops such as fruits, vegetables and horticultural crops (Elad et al., 2016). The harmful parts include almost all parts of the plant, including seeds, seedlings...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N9/50A01N63/50A01P3/00
CPCA01N63/00C12N9/50
Inventor 杨龙王永春余涵李国庆吴明德张静
Owner HUAZHONG AGRI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com