Penicillium oxalicum FH6 strain, screening method and application thereof

A Penicillium oxalicum, screening method technology, applied in the field of microorganisms, can solve the problems of high cost, unsuitable for large-scale industrial production, environmental pollution, etc., and achieve the effects of low production cost, easy operation and simple equipment

Active Publication Date: 2014-04-23
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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  • Penicillium oxalicum FH6 strain, screening method and application thereof
  • Penicillium oxalicum FH6 strain, screening method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Screening of Penicillium oxalicum FH6 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 to 3 days until cultured 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] Magnesium sulfate 0.5g,

[0040] Yeast paste 0.5g,

[0041] Hydrolyzed casein 0.5g,

[0...

Embodiment 2

[0052] Preparation of crude enzyme solution.

[0053] (1) Cultivate the Penicillium oxalicum FH6 obtained in Example 1 in Martin 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, and the volume ratio of the buffer solution and the bacterial solution is 1:1. The freezing buffer is added to better protect the Penicillium oxalicum FH6 strain, so that the bacterial strain will not be dehydrated. die.

[0054] (2) Activate the above-mentioned Penicillium oxalicum FH6 strain in Martin medium for slant activation. After activation for 2 generations, pick a full circle and put it in Martin liquid medium. After culturing for 24 hours at 28°C and 140r / min, take 1mL into 50mL In the 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] Yeast extract 1g,

[0057...

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 13.42 U / mL.

[0070] Determination of CMC enzyme activity: the assay method is the same as in Example 2, and the enzyme activity measured is 28.01 U / mL.

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Abstract

Belonging to the technical field of microorganisms, the invention in particular relates to a Penicillium oxalicum FH6 strain, a screening method and application thereof. The preservation number of the Penicillium oxalicum FH6 strain is CGMCC No.8270. The method for applying the strain to enzymolysis of enteromorpha includes: subjecting a cryopreserved FH6 strain to activation culture in a Martin slant culture medium, then picking a full ring and placing it in a Martin liquid culture medium to perform shaking culture for 24-28h, then taking 1-2mL of a bacterial solution to conduct shaking culture for 5-7d in a sterilized cellulose liquid fermentation medium; then carrying out low temperature centrifugation on the fermented liquid, taking the supernatant as a crude enzyme solution or filtering the fermented liquid, and taking the filtrate as a crude enzyme solution; and reacting the crude enzyme solution with clean enteromorpha powder at 45-55DEG C. The Penicillium oxalicum FH6 strain has a cellulose production ability, can effectively degrade cellulose materials in enteromorpha into reducing sugar. Also, the production cost is low, the equipment is simple, and the reaction is mild, thus being 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 Penicillium oxalicum FH6 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 com...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12Q1/04C12P19/14C12P19/02C12R1/80
Inventor 费岚邵飞胡乐琴何培民贾睿
Owner SHANGHAI OCEAN UNIV
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