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

Streptomyces lutetsis lt-2 producing keratinase and its application method

A technology of keratinase and application method, which is applied in the field of microorganisms and achieves the effects of mild degradation conditions, broad application prospects and simple operation process

Active Publication Date: 2018-04-03
HUNAN INST OF MICROBIOLOGY
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the disulfide bond in the structure of keratin makes it resistant to degradation and is not degraded by most proteases such as papain, pepsin and trypsin.

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
  • Streptomyces lutetsis lt-2 producing keratinase and its application method
  • Streptomyces lutetsis lt-2 producing keratinase and its application method
  • Streptomyces lutetsis lt-2 producing keratinase and its application method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Screening and identification of keratinase-producing strains

[0022] 1. Culture medium formula

[0023] (1) Preliminary screening of solid medium: every 1000 ml contains 10-15 grams of pig hair powder, K 2 HPO 4 1.0~1.4g, KH 2 PO 4 0.5~1.0g, NaCl 0.1~0.5g, MgSO 4 0.05 to 0.1 grams, 15 to 20 grams of agar, and water as the balance. pH7.5~8.0.

[0024] The production of pig hair powder on the screening medium: remove the pig skin, wash the pig hair, dry to constant weight, then cut the pig hair to about 1cm with scissors, then crush it with a pulverizer, and pass through a 100-mesh sieve.

[0025] (2) Preservation medium on slant

[0026] Gao's medium: every 1000 ml contains 15-20 grams of soluble starch, KNO 3 0.8~1.2g, NaCl0.5~1.0g, K 2 HPO 4 0.5~1.0g, MgSO 4 0.5~1.0g, FeSO 4 ·7H 2 O 0.005-0.01 grams. pH7.2-7.5, 15-20 grams of agar, and water as the balance.

[0027] LB solid medium: 8-12 grams of protein per 1000 ml, 3-7 grams of yeast po...

Embodiment 2

[0047] Degradation effect determination of embodiment 2 bacterial strain

[0048] 1. Determination of keratinase activity

[0049] Add 5 mg of pig hair powder to 1 ml of 50 mmol / L Tris buffer (pH8.0) to suspend; add 1 ml of crude enzyme solution diluted 10 times with the same Tris buffer; add 2 ml of TCA ( 0.4mol / L) stop solution; react at 50°C, 150rpm warm bath for 45min; add 2ml of TCA (0.4mol / L) to stop the reaction; centrifuge at 6000rpm for 10min; OD value.

[0050] Definition of enzyme activity unit: Under specific reaction conditions, the amount of enzyme produced by decomposing keratin per milliliter increases the OD value by 0.01, which is regarded as 1 U of keratinase activity unit.

[0051] Enzyme reaction substrate pig hair powder production: the method is the same as that of the screening medium pig hair powder, except that the pig hair powder that has been crushed and passed through a 100-mesh sieve is then ground with liquid nitrogen to make it finer and finer...

Embodiment 3

[0056] The expanded culture method of embodiment 3 bacterial strain LT-2

[0057] 3. The expansion culture method of bacterial strain LT-2 is

[0058] Strain LT-2 was inoculated on the slant of solid seed medium, activated and cultured at 30-35°C for 2-3 days, and a large number of spores were formed on the slant.

[0059] Scrape the spores from the activated slope, use sterile water to make a uniform spore suspension, then inoculate the spore suspension into the spore-forming medium, and culture it on a shaking table at 30-35 ° C for 3-5 days at 200 rpm. The bottle culture solution is the seed solution.

[0060] Fermentation tank culture: The amount of medium in the fermenter is 50-70% of the total volume, the inoculum volume accounts for 1-3% of the medium volume, the fermentation temperature is 30-35°C, sterile air is introduced, and the culture is stirred at 180-200rpm , the ratio of the ventilation volume to the volume of the fermentation broth is 0.9:1-1:1, the tank pr...

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

PropertyMeasurementUnit
particle diameteraaaaaaaaaa
Login to View More

Abstract

The invention relates to a keratinase-producing Streptomyces lutetes LT-2 and an application method thereof, belonging to the technical field of microorganisms. The bacterial strain LT‑2 is classified as: Streptomyces rutgersensis, and the preservation number is: CCTCC NO: M 2015440. The invention discloses that the morphological characteristics of the fungus on Gao's No. 1 medium are: yellow and white air filaments, yellow when the spores are piled up, light yellow-brown base filaments, and brown pigment is produced over time; the spore filaments are straight or wavy, and divided The spores are spherical to ellipsoid, and the surface of the spores is smooth. Applied to the degradation of α-keratin such as human and animal hair, hoof, claw, horn, nail, etc.; applied to the degradation of β-keratin such as feathers, beaks, toenails, scales and claws of reptiles, etc. Compared with other bacterial strains degrading keratin, the bacterial strain of the present invention has mild fermentation conditions, convenient application, high enzymatic activity of the keratinase produced and relatively stable.

Description

technical field [0001] The invention belongs to the technical field of microbes, and relates to a novel bacterial strain Streptomyces lutetsis LT-2 producing keratinase and an application method thereof. Background technique [0002] Keratin can be divided into α-keratin and β-keratin. α-keratin is mainly found in mammalian hair, feathers, animal horns, fingernails, claws and hoofs. The harder beta keratin is found in the toenails, shells, scales, beaks, claws and setae of reptiles. Its crude protein content is more than 80%, and the total amount of various amino acids is more than 70%. It also contains more macroelements, trace elements and unknown growth factors. It is a good source of feed protein and fertilizer, and has great significance for its development and utilization. important application prospects. [0003] In recent years, with the rapid development of large-scale breeding industry, a large amount of livestock and poultry waste has been produced, mainly in th...

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): C12N1/20C12N9/52C12R1/465
Inventor 杜东霞许隽贺月林尹红梅刘标汪彬陈薇吴迎奔王震许丽娟
Owner HUNAN INST OF MICROBIOLOGY
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