A kind of Penicillium citrinum strain and application thereof

A technology of Penicillium citrinum and microbial strains, which is applied in the direction of fungi, enzymes, biochemical equipment and methods, etc., can solve the problems such as the lack of reports of Penicillium citrinum cellulase, and achieve broad industrial and agricultural applications and market prospects, The effect of rapid growth and reproduction and simple fermentation process

Inactive Publication Date: 2011-12-14
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There are few reports about cellulase from Penicillium citrinum

Method used

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  • A kind of Penicillium citrinum strain and application thereof
  • A kind of Penicillium citrinum strain and application thereof
  • A kind of Penicillium citrinum strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046]Example 1 Isolation and screening of Penicillium citrinum CR-2

[0047] culture medium

[0048] PDA comprehensive medium: potato 200g, glucose 20g, peptone 10g, KH 2 PO 4 3g, MgSO 4 1.5g, VB1 10mg, agar 20g, water 1000ml, pH natural (do not control pH value). Peel fresh potatoes and cut into small cubes with a side length of 0.5cm, add 1000ml of water, boil and then heat for 10 minutes, then filter through four layers of gauze to obtain potato nutrient solution, add other ingredients except VB1, make up to 1000ml of water , sterilized at 121°C for 20 minutes, VB1 was made into a separate solution, sterilized by filtration with a 0.22μm microporous membrane, and then added to make the final concentration 10mg / L. After mixing, plate or make a slant medium.

[0049] Enrichment medium: CMCNa 1%; K 2 HPO 4 0.1%; Na2CO 3 0.5%; MgSO 4 ·7H 2 O 0.01%; FeSO 4 ·7H 2 O 0.015%; MnSO 4 5×10-5%; peptone 1.0%; yeast extract 1.0%, sterilized at 121°C for 20min. Plate af...

Embodiment 2

[0063] Example 2 Fermentation of Penicillium citrinum CR-2

[0064] Utilize Penicillium citrinum (Penicillium citrinum) CR-2 to produce cellulase, comprising the following steps:

[0065] 1. Spore culture of Penicillium citrinum CR-2

[0066] Sporulation medium: yeast extract powder 10g, glucose 20g, water 1000ml, pH natural, sterilized at 121°C for 20min.

[0067] Use an inoculation shovel to dig out a piece of mycelium with a side length of about 0.5 cm from the slope and inoculate it on a sporulation medium plate, and culture it at 30°C for 3 to 4 days to produce a large number of green spores.

[0068] 2. Production of cellulose degrading enzymes by fermentation

[0069] Enzyme production medium ( / L): 2.3% corn stalk powder, (NH 4 ) 2 SO 4 0.25%, KH 2 PO 4 0.3%, MgSO 4 ·7H 2 O 0.05%, CaCO 3 0.06%, sterilized at 121°C for 20min, natural pH.

[0070] Determination method of endoglucanase activity (carboxymethyl cellulose enzyme activity (CMCase)): DNS method - ...

Embodiment 3

[0077] Example 3 Fermentative production of Penicillium citrinum CR-2 (1)

[0078] Utilize Penicillium citrinum (Penicillium citrinum) CR-2 to produce cellulosic endoglucanase, comprise the following steps:

[0079]1. Spore culture of Penicillium citrinum CR-2

[0080] With embodiment 2.

[0081] 2. Production of cellulose degrading enzymes by fermentation

[0082] The formulation of the enzyme-producing medium and the determination methods of endoglucanase activity and exoglucanase activity are the same as in Example 2. Inoculate the spore suspension into 200mL enzyme-producing medium according to the method and concentration of Example 2, cultivate for 5 days at 30°C at 200rpm, measure the highest activity of CMCase in the fermentation broth to be 49.91IU / ml, and the highest activity of FPA to be 26.9IU / ml (as shown in the figure below).

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Abstract

The present invention relates to Penicillium citrinum CR-2 CGMCC No.3024 and application thereof. The bacterium belongs to the genus Penicillium, has a short culture period and a fast growth and reproduction speed, and can cultivate a large amount of uniform cellulose degrading enzyme-producing bacteria culture solution in a short period of time. The present invention also provides a method for producing endoglucanase and exoglucanase by fermenting and culturing the bacterial strain in a liquid enzyme-producing medium, which can produce endoglucanase and exoglucanase with higher activity in a short period of time. Exoglucanase, and its fermentation process is simple, stable, low in cost and high in yield.

Description

technical field [0001] The invention relates to the field of microorganisms and applications thereof, in particular to a new strain of Penicillium citrinum and its application in the production of cellulose degrading enzymes. Background technique [0002] Cellulosic fuel ethanol is the second generation of fuel ethanol, which is a renewable clean energy. The cellulolytic bacteria capable of producing natural cellulase with high activity and good stability is one of the key factors for obtaining cellulosic fuel ethanol. [0003] Recent studies have shown that some species of Penicillium fungi can not only secrete lignocellulose-degrading enzymes with complete composition and high enzyme activity, but also have the advantages of easy cultivation and fast growth (Henning etc. 2006. Production of cells by Penicillium brasilianum IBT 20888-Effect of substrate onhydrolytic performance. Enzyme and Microbial Technology, 38: 381-390). [0004] Penicillium citrinum is an airborne f...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N9/46C12R1/80
Inventor 顿宝庆曲小爽李桂英张保明路明
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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