A kind of Cladosporium producing sterol esterase and method for producing enzyme

A technology for producing sterol esterase and sterol esterase crude enzyme is applied in the field of bioengineering, which can solve the problems of long operation time, high equipment requirements and high construction cost, and achieves good ability to hydrolyze sterol esters, huge application potential and fermentation cost. low effect

Active Publication Date: 2022-06-24
OCEAN UNIV OF CHINA
View PDF10 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can be mainly classified into two categories. The first category is to use large and medium-sized professional equipment to detect blood samples using the principle of optical absorption. This type of detection method has high requirements for equipment, and the measurement takes a long time and costs high; The second category is to use a special medical hand-held detector for detection, mainly by means of immunological methods, electrochemical methods and some optical methods for detection, which also has the disadvantages of long operation time and high cost.

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
  • A kind of Cladosporium producing sterol esterase and method for producing enzyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Sterol esterase Cladosporium Screening and identification methods of sp. strains.

[0025] (1) Preliminary screening: The samples with higher content of sterol esters were selected as the screening raw materials, and some samples were taken and dissolved in sterilized physiological saline to make their tissue distribution uniform. The samples were inoculated and spread on the primary screening plate medium with sterol ester as the main carbon source after aseptic operation. Invert at 28-30°C for 2-3 days, observe the colony morphology and the transparent circle of the medium.

[0026] (2) Re-screening: Sort out the strains with transparent circles in the primary screening medium, carry out liquid shake flask re-screening, the fermentation conditions are 28-30 ° C, 150-200 rpm, culture for 1-2 days, and then collect the fermentation broth, Enzyme activity was determined.

[0027] (3) Identification of strains: identification of strains is carried out throug...

Embodiment 2

[0028] Example 2 Cladosporium sp. DZ16 strain activation and enzyme production culture.

[0029] (1) Activation of strains.

[0030] The first step is to preserve and activate the strains. Cladosporium The sp. DZ16 strain was transferred from the strain storage tube frozen at -80°C to PDA medium in a 100mL conical flask for growth, with a liquid volume of 20mL, and cultured at 28-32°C for 24-48h.

[0031] The second step is to spread the strains on a plate, pick the seed solution with vigorous growth, dilute and separate, and spread it on the PDA solid medium with a coating rod, cultivate at 28-32 ° C for 48 hours, pick out the large colonies that grow well, and save them for future use. .

[0032] (2) Fermentation broth culture.

[0033] In the first step of low-salt liquid activation culture, the large colonies that grow well in the flat coating are inoculated into the low-salt liquid activation medium for cultivation. The medium contains 0.2% yeast extract, 0.3% pepto...

Embodiment 3

[0035] Example 3 Determination of enzyme activity of crude enzyme solution.

[0036]Using 0.25mL 0.48mg / mL stigmaster acetate solution as substrate, add 0.5mL 4-AA phenol working solution, 2U / mL cholesterol oxidase 0.25mL and 12U / mL horseradish peroxidase 0.25mL, at 40 After preheating at ℃ for 5 min, add 25 µL of the enzyme solution to be tested, react for 30 min, terminate the reaction with 0.5 mL of 0.1 mol / L HCl, measure the absorbance at OD500, and use the inactivated enzyme solution as a blank control group to measure the enzyme activity. At 0.5-0.8 U / mL.

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 discloses a sterol esterase-producing Cladosporium obtained by screening ( Cladosporium sp.) DZ16, deposit number CCTCC M 2016685. The bacteria colony of the present invention is in the shape of velvet, and the bacteria are activated through the first step of bacteria preservation and the second step of bacteria plate coating. In the early stage of fermentation, the first step of low-salt liquid activation culture is used to increase the biomass of the strain, and in the later stage, the second step of multi-factor complex action is used to induce the strain to secrete enzyme protein and increase the enzyme activity. The sterol esterase produced by the fermentation of the bacterial strain of the invention has good application potential in the fields of medical detection, food, pulp and paper industry, environmental protection and the like.

Description

technical field [0001] The invention relates to a sterol esterase-producing Cladosporium strain obtained by screening and an enzyme-producing method, belonging to the technical field of bioengineering. Background technique [0002] Sterols are derivatives of alkylene oxide polyhydrophenanthrene, widely distributed in the biological world, play a variety of physiological functions in animals and plants, and play an important role in the body's metabolic process. Cholesterol is the most abundant steroid compound in animals, and it is mostly found in internal organs such as liver, kidney and intestine, skin and adipose tissue. It is the precursor of five types of steroid hormones, vitamin D and bile acid substance. Ensuring the supply of cholesterol and clarifying its specific content is necessary to maintain the normal life activities of the body. Therefore, the determination of sterol content in the body has important guiding significance in medical and clinical aspects. ...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N9/18C12R1/645
CPCC12N9/18C12Y301/01013C12R2001/645C12N1/145
Inventor 王鹏江晓路乔乐克杜春影
Owner OCEAN UNIV OF CHINA
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap