A kind of Pseudomonas aeruginosa producing keratinase and its application

A technology of Pseudomonas aeruginosa and keratinase, applied in the direction of bacteria, microorganism-based methods, microorganisms, etc., can solve the problems of energy and environmental pollution, difficult direct utilization, difficult biodegradation, etc., and achieve high degradation rate and good stability , The effect of simple degradation conditions

Active Publication Date: 2019-05-07
GUANGDONG WENS DAHUANONG BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because its main component, keratin, is a hard protein with strong resistance and difficult to biodegrade, it is difficult to use directly, and traditional physical and chemical processing methods: high temperature and high pressure, acid-base treatment, etc. not only consume a lot of energy but also cause serious problems. environmental pollution problems

Method used

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  • A kind of Pseudomonas aeruginosa producing keratinase and its application
  • A kind of Pseudomonas aeruginosa producing keratinase and its application
  • A kind of Pseudomonas aeruginosa producing keratinase and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1 is used to illustrate the screening and identification process of bacterial classification:

[0048] 1. Screening method for strains producing keratinase:

[0049] 1) Soil samples were collected from a chicken farm of Guangdong Wens Food Group Co., Ltd.;

[0050] 2) Mix the collected samples in sterile distilled water, take the supernatant after standing, press 10 -6 、10 -7 、10 -8 After performing gradient dilution, spread on the agar plate medium containing milk; then place it in a 37°C incubator and incubate it upside down for 24 hours;

[0051] 3) Select a single colony that can produce a transparent degradation circle on the agar plate medium, inoculate it in a liquid inorganic salt medium containing feathers, and culture it with shaking at a temperature of 37 ° C and 180 rpm for 72 hours to obtain a culture medium for the strain; finally select The culture solution of strains with obvious feather degradation is inoculated into a new feather-contain...

Embodiment 2

[0077] Example 2 is used to illustrate the degradation conditions of Pseudomonas aeruginosa producing keratinase to keratin:

[0078] 1) Prepare 50 mL of liquid inorganic salt medium with a pH of 4 to 11 according to the gradient;

[0079] 2) Add 0.5 mL of Pseudomonas aeruginosa strains and 0.5 g of feathers to each liquid inorganic salt medium prepared in step 1), and ferment for 24 hours under the conditions of 220 rpm and 37° C., and pass The degradation rate of feathers is detected by weight loss method;

[0080] 3) The degradation rate of different pH to feather is as follows: image 3 Shown: the results show that when the pH of the liquid inorganic salt medium=11, the degradation rate of feathers is the highest;

[0081] 4) Take the strain of Pseudomonas aeruginosa, add it to the liquid inorganic salt medium with pH = 11, and ferment under the temperature conditions of 27°C, 32°C, 37°C, 42°C and 47°C respectively at a rotation speed of 220rpm 24h, then detect the degr...

Embodiment 3

[0084] Embodiment 3 is used to illustrate the influence of Pseudomonas aeruginosa bacterial age on keratin degradation rate:

[0085] Inject Pseudomonas aeruginosa strains with ages of 4h, 8h, 12h, 16h, 20h and 24h into the liquid inorganic salt medium respectively, the inoculation amount of the strains is 1%, and the feather concentration is 10g / L, pH =11, temperature is 24h under the condition of 37 ℃ of fermentation; The degradation rate of feather is as follows Figure 5 Shown: when the bacterial age is 12h, the degradation rate of feathers is the highest, followed by 16h; before 12h, with the increase of bacterial age, the activity of Pseudomonas aeruginosa strains gradually increases, and the degradation rate of feathers also increases. After 16 hours, the degradation rate of feather decreased rapidly with the increase of bacterial age.

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Abstract

The invention discloses a keratinase-producing Pseudomonas aeruginosa and its application. The Pseudomonas aeruginosa was preserved in the China General Microorganism Culture Collection and Management Center on March 7, 2016, and the preservation number is CGMCC No. 12185; said keratinase-producing Pseudomonas aeruginosa degrades keratin-containing substances, comprising the following steps: 1) preparing a liquid inorganic salt medium; 2) taking Pseudomonas aeruginosa strains and keratin-containing substances, Added to the liquid inorganic salt medium prepared in step 1), the bacterial age of the Pseudomonas aeruginosa strains is t, 4h≤t≤24h, and the Pseudomonas aeruginosa strains in the liquid inorganic salts The inoculum amount in the culture medium is a, 0.5%≤a≤5%, and the fermentation is carried out under the conditions of 150-220 rpm and a temperature of 27-47° C. to degrade keratin-containing substances; the Pseudomonas aeruginosa is in Keratin can be efficiently degraded in liquid medium with feather as the sole carbon and nitrogen source.

Description

technical field [0001] The invention relates to the field of microbial strains and applications, in particular to a keratinase-producing Pseudomonas aeruginosa and applications thereof. Background technique [0002] Feather is an important protein resource, and its protein content is as high as 85-90%. However, because its main component, keratin, is a hard protein with strong resistance and refractory biodegradation, it is difficult to use directly, and the traditional physical and chemical processing methods: high temperature and high pressure, acid-base treatment, etc. not only consume a lot of energy but also cause severe environmental pollution problems. At present, the annual production of feather waste in my country's aquaculture industry has reached more than one million tons. With the continuous development of my country's aquaculture industry, this amount is still growing. It is an inevitable choice to realize high-value transformation and recycling of waste resou...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/385
CPCC12N1/205C12R2001/385
Inventor 黄妙容钟楚红任广彩刘传高陈瑞爱叶俊贤
Owner GUANGDONG WENS DAHUANONG BIOTECH
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