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

Method for producing low-temperature alkaline protease by alternative pseudomonad

A technology of Pseudomonas alternans and protease, which is applied in the field of low-temperature alkaline protease and products produced by Pseudomonas alternans, and can solve the problem of not producing low-temperature alkaline protease

Inactive Publication Date: 2007-11-14
HUAIHAI INST OF TECH
View PDF0 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no literature report on low-temperature alkaline protease produced by Pseudoalteromonas.flavipulchra at home and abroad

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
  • Method for producing low-temperature alkaline protease by alternative pseudomonad
  • Method for producing low-temperature alkaline protease by alternative pseudomonad
  • Method for producing low-temperature alkaline protease by alternative pseudomonad

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1. A Pseudomonas alternata HH407 (Pseudoalteromonas flavipulchra HH407) CCTCC NO: M 207009. The strain has the following characteristics: the bacterium can grow at 0°C, the growth temperature range of the bacterium is 0-35°C, and the optimum growth temperature is 20°C; the growth pH range is 5.5-11, and the optimum growth pH is 8.5; The NaCl concentration for growth is 0.5%-13%, and the optimum NaCl concentration for growth is 4%. Without NaCl, it does not grow. It is a typical marine psychrophilic moderate halophilic microorganism; the bacteria can use tryptone, yeast extract, peptone, Cellobiose, starch, gelatin, acetic acid, citric acid, methanol and ethanol.

Embodiment 2

[0073] Example 2. A kind of method that utilizes a kind of Pseudomonas alternata HH407 CCTCCNO described in embodiment 1: M 207009 to produce low temperature alkaline protease, its steps are as follows,

[0074] (1) Inoculate the slant 1 ring of the strain into a 250ml Erlenmeyer flask with 50ml of fermentation medium, 180r / min, 20°C, and cultivate for 16h; insert 2% inoculum into the fermentation medium, 180r / min, 20°C, and cultivate 36h;

[0075] (2) Centrifuge the cultured fermented liquid at 10000r / min for 10min, add 60% ammonium sulfate to the supernatant, let it stand for 3h, centrifuge at 10000r / min for 30min, dissolve it with an appropriate amount of Tris buffer with a pH of 8.0, and then dissolve it at 4°C Dialyze for 24 hours; then obtain low-temperature alkaline protease through DEAE-sepharose Flat Flow and SephacrylS-200 methods.

Embodiment 3

[0076] Example 3. The low-temperature alkaline protease produced by the method described in a kind of embodiment 2, this low-temperature alkaline protease has the following characteristics:

[0077] (1) The optimum action temperature of the enzyme is 35°C, and the catalytic efficiency is still 22% at 0°C; the enzyme is stable at 25°C, 35°C, and 45°C, and the enzyme activity is above 80% at 55°C for 30 minutes after incubation for 60 minutes Loss of 55% enzyme activity; it is a low temperature protease;

[0078] (2) The optimal pH of the enzyme is 10.0, and the relative enzyme activity is more than 80% at pH 9.0-11, and there is enzyme activity at pH 5-12; the most stable pH of the enzyme is 9.0; it is an alkaline protease;

[0079] (3)Sr 2+ 、Ba 2+ 、Cu 2+ , Mn 2+ 、K +Activation of enzymes, Hg 2+ 、Al 3+ , Zn 2+ 、Ni 2+ 、Co 2+ , Fe 3+ , Li 2+ Has an inhibitory effect;

[0080] (4) PMSF inhibits enzyme activity, and EDTA still has 106.6% enzyme activity, showing that ...

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

A alternately false single afterbirth bacterium HH407( Pseudoalteromonas flavipulchra HH407) CCTCC N0: M207009. This bacterium can grow up under 0degree C, the growth temperature of this bacterium scope is 0-35 degree C, the most suitable growth temperature is 20degree C; Growth pH scope is 5.5-11, most suitable growth pH is 8.5; The NaCl density of growth is 0.5%-13%, the optimal growth of NaCl concentration of 4%, no growth without NaCl, it is a typically moderate marine Psychrophilic Halophilic microorganisms; bacteria can use pancreatic peptone, yeast extract, peptone, cellobiose, starch, gelatin, acetic acid, citric acid, methanol and ethanol. The present invention also opens an alternate use of low-temperature alkaline-producing Pseudomonas methods and by the method of low-temperature alkaline protease products. The low-temperature alkaline protease can be used as the main enzyme detergent additives can be directly room temperature washing, saves energy; and in food manufacturing, leather, silk, environmental protection, medicine and other fields also has broad application prospects.

Description

technical field [0001] The present invention relates to a Pseudomonas alternata HH407 strain CCTCC NO: M 2070009 isolated from Lianyungang sea area (preserved in China Center for Type Culture Collection CCTCC on January 29, 2007, and the preservation number is CCTCC NO: M 207009); The invention also relates to a method and a product for producing low-temperature alkaline protease by the bacterial strain. Background technique [0002] Low-temperature enzymes have higher catalytic efficiency than medium-temperature enzymes at low temperatures, and can play a role in reducing energy consumption in industrial processing. Biological governance and other aspects have broad application prospects. Screening low-temperature enzymes with application value from marine microorganisms has become an important way to develop marine biological resources. [0003] Alkaline protease is the main additive of enzyme-added detergents. At present, medium-temperature alkaline protease is mainly a...

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): C12N1/20C12N9/52C12R1/38
Inventor 王淑军陈丽吕明生李丹李华钟陈建安杨锦宇
Owner HUAIHAI INST OF TECH
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