Stenotrophomonas FQ1 strain and its screening method and use

A technology of Stenotrophomonas and screening method, which is applied in the field of Stenotrophomonas FQ1 strain and its screening, can solve the problems of high cost, unsuitability for large-scale industrial production, environmental pollution, etc., and achieve low production cost and easy operation. The effect of simplicity and simple equipment

Active Publication Date: 2014-04-02
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cellulose in Enteromorpha is the main component used to produce bioethanol. Commercial cellulase is generally used to degrade the cellulose in Enteromorpha. However, the cost of commercial cellulase is relatively high, and the enzymatic hydrolysis process of cellulose Pollution to the environment, not suitable for large-scale industrial production

Method used

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  • Stenotrophomonas FQ1 strain and its screening method and use
  • Stenotrophomonas FQ1 strain and its screening method and use
  • Stenotrophomonas FQ1 strain and its screening method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Screening of Stenotrophomonas FQ1 Strain

[0032](1) Collected from rotting Enteromorpha algae growing under natural conditions and densely floating in the green tide sea area. The collection sites are the No. 1 and No. 6 bathing beaches in Qingdao, Shandong. After collection, they were transported in an incubator at low temperature and brought back to the laboratory for bacterial screening.

[0033] Put 1mL of the collected rotting Enteromorpha algae suspension into the Erlenmeyer flask containing 30mL of selective medium, fix the Erlenmeyer flask on a shaker, and culture at 30°C and 140r / min for 2-3 days until the culture The liquid becomes cloudy.

[0034] Wherein the selection medium is: cellulose powder 5g,

[0035] Sodium nitrate 1g,

[0036] Potassium chloride 0.5g,

[0037] Disodium hydrogen phosphate 1.2g,

[0038] Potassium dihydrogen phosphate 0.9g,

[0039] Magnes...

Embodiment 2

[0052] Preparation of crude enzyme solution.

[0053] (1) Culture Stenotrophomonas FQ1 obtained in Example 1 in LB liquid medium at 28°C and 140r / min for 24h, and then add freezing buffer to the bacterial liquid. The volume concentration of the buffer solution is 50% glycerol, the adding volume ratio of the buffer solution and the bacterial solution is 1:1, adding the freezing buffer, in order to better protect the Stenotrophomonas FQ1 bacterial strain, so that the bacterial strain will not be affected by Dehydration and death.

[0054] (2) Activate the above-mentioned Stenotrophomonas FQ1 strain in LB culture medium on a slant surface. After 2 generations of activation, pick a full ring and place it in LB liquid culture medium. After culturing at 28°C and 140r / min for 24 hours, take 1mL In 50mL sterilized and cooled cellulose liquid fermentation medium, shake culture at 35°C and 140r / min for 6 days.

[0055] Wherein the liquid fermentation medium is:

[0056] ...

Embodiment 3

[0065] Preparation of crude enzyme solution.

[0066] Step (1) and step (2) are the same as in Example 2.

[0067] (3) Filtrating the liquid fermentation culture liquid obtained in step (2), and taking the filtered solution as the crude enzyme liquid.

[0068] Activity detection of crude enzyme solution:

[0069] Determination of filter paper enzyme activity (FPA): The determination method is the same as in Example 2, and the measured enzyme activity is 24.90 U / mL.

[0070] Determination of CMC enzyme activity: the assay method is the same as in Example 2, and the enzyme activity measured is 49.28 U / mL.

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Abstract

The invention belongs to the technical field of microorganism and especially relates to a stenotrophomonas FQ1 strain and its screening method and use. The stenotrophomonas FQ1 strain has the accession number of CGMCC No.8272. A use method of the stenotrophomonas FQ1 strain for enteromorpha prolifera enzymolysis comprises the following steps of carrying out activation culture on the frozen stenotrophomonas FQ1 strain by a LB slant culture medium, picking a full ring of the culture, putting it into a LB liquid culture medium, carrying out shaking culture for 24-28h, taking 1-2mL of the bacterial solution, putting the bacterial solution into a sterilized cellulose liquid fermentation medium, carrying out shaking culture for 5-7 days, and carrying out low-temperature centrifugation on the fermentation broth and collecting a supernatant as a crude enzyme liquid, or filtering the fermentation broth and collecting the filtrate as a crude enzyme liquid; and carrying out a process of reaction of the crude enzyme liquid and clean enteromorpha prolifera powder at a temperature of 45-55 DEG C. The stenotrophomonas FQ1 strain has a cellulase production capability and can effectively degrade cellulose materials in enteromorpha prolifera into reducing sugar. A production cost is low, production equipment is simple, reaction conditions are mild, and the stenotrophomonas FQ1 strain is suitable for large-scale industrial production.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a Stenotrophomonas FQ1 strain and a screening method and application thereof. Background technique [0002] At present, the supply of oil resources in the world is becoming increasingly tense, environmental pollution is becoming more and more serious, and energy issues are attracting more and more people's attention. In the past two years, major energy consuming countries have been seeking new energy alternatives to petroleum. Bioethanol fuel has been valued and actively supported by many countries because of its outstanding environmental protection and renewability. As a renewable resource, bioethanol can reduce the consumption of petroleum, and can be directly used as liquid fuel or mixed with gasoline. [0003] Beginning in 2007, large-scale Enteromorpha broke out in the Yellow Sea of ​​my country for 7 consecutive years. The analysis results of the sugar compone...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/04C12P19/14C12P19/02C12R1/01
Inventor 费岚邵飞胡乐琴何培民贾睿
Owner SHANGHAI OCEAN UNIV
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