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A strain of Aspergillus clavinus producing ferulic acid esterase and its application

A technology of ferulic acid esterase and Aspergillus clavatus, applied in the biological field, can solve problems such as unfavorable sustainable development, and achieve the effects of easy culture and preservation, high enzyme activity and strong growth ability

Active Publication Date: 2016-09-21
华工利亚(佛山)生物科技有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although the chemical method is simple to operate and has high efficiency, the subsequent treatment of the extraction waste liquid is not conducive to sustainable development

Method used

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  • A strain of Aspergillus clavinus producing ferulic acid esterase and its application
  • A strain of Aspergillus clavinus producing ferulic acid esterase and its application
  • A strain of Aspergillus clavinus producing ferulic acid esterase and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Primary screening of Aspergillus clavus, high-yielding strains were screened by Oxford cup transparent circle screening experiment.

[0041] Collect soil samples from the South China Botanical Garden, weigh 10g of the sample, put it into a 300mL Erlenmeyer flask filled with 90mL of sterile saline containing glass beads, shake for 20 minutes, and make a 10-fold incremental serial dilution of the sample homogeneous solution 1:10, 1:100 , 1:1000, and 1:10000, take 1.0mL each of the diluted sample homogeneous solution 1:100, 1:1000, and 1:10000 and add it to a sterile plate; add 15mL of PDA medium and mix evenly, let it stand for 1min, and put it at 28 Cultivate upside down at ℃ for 72 hours, transfer the single colony on the culture medium to the slant PDA medium and culture for 72 hours at 28℃, then transfer to the primary screening medium containing ethyl ferulic acid and culture at 28℃ for 60 hours, and screen for the production of ferulic acid ester strains, spare;

...

Embodiment 2

[0044] Microbiological characteristics and identification of strains

[0045] The colony of Aspergillus clavus grows slowly on Cha's culture medium, and the diameter is 30mm after 7 days of culture; the texture is velvety to powdery, the mycelium is white, the conidia are dark blue-green in structure, and there is no exudate. The growth rate of colony on malt juice agar is relatively fast, and the diameter is 45mm after 7 days of cultivation. The colony was cultured on Saccharomyces chagheri paste agar for 7 days, with a diameter of 40 mm, velvety texture, and a slightly thick bottle; conidia had many structures, grayish blue, thinner, white edges, and no exudate.

[0046] Under the microscope, it can be observed that the conidia heads are rod-shaped when they are young, and the conidiophores occur in the matrix, and the lengths of the spores vary; the apical capsule gradually expands from the top of the spore to a club shape, with a diameter of 50-60 mm, and a sporulation str...

Embodiment 3

[0049] Determination of the Release Amount of Bound Ferulic Acid by Solid State Fermentation

[0050] According to the formula of the seed medium, add 10g of bran and 10g of water and mix them into a 500mL Erlenmeyer flask. After the seed medium is sterilized, inoculate the Aspergillus clavatus (Aspergillus clavatus) slant strain with an inoculation amount of 0.5%. After culturing at 28°C for 3 days, the sticks Aspergillus seeds;

[0051] According to the formula of the fermentation medium, add 200g of bran and 200g of water, mix and sterilize, put it on a porcelain plate to cool, control the thickness of wheat bran within 2.5cm, and the inoculation amount of Aspergillus clavus seeds is 0.5% (the bacterial content is 1~2×10 9 Individual / g), culture temperature is 28 ℃, carry out ventilated culture, fermentation time is 48h. Solid-state fermented bran was leached in a water bath at 40°C for 60 min at the ratio of 15 g to 90 mL of water, centrifuged at 12 000 r / min for 20 min a...

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Abstract

The present invention provides a ferulic esterase-producing Aspergillus clavus and application thereof, and the classification of the Aspergillus clavus is named as Aspergillus clavus ( Aspergillus clavatus ) Tobacco green, preserved by the China Center for Type Culture Collection, referred to as CCTCC, the preservation number is CCTCC NO: M2013641, and the preservation date is December 8, 2013. Aspergillus clavus is used to produce ferulic acid esterase. The strain of the present invention uses wheat bran as the fermentation substrate to ferment and produce ferulic acid esterase, and uses ferulic acid esterase to break the ester bond between plant cell wall polysaccharides and lignin and ferulic acid to obtain the bound phenolic acid ferulic acid method, and the use of microbial growth cycle is short, not subject to climate restrictions, easy to achieve large-scale production, providing a new biological method for the extraction of bound phenolic acids such as ferulic acid.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a ferulic esterase-producing Aspergillus clavus and application thereof. Background technique [0002] Wheat is one of the crops with the most extensive planting area in the world, accounting for about 26% of the planting area of ​​grain crops. The annual output of wheat in my country is high, and the annual output of wheat bran can reach more than 20 million tons. However, the development of bran has always been Utilization is seldom, and it is basically only used as feed raw materials. Wheat bran mainly contains non-starch polysaccharides, starch, fat, lignin, protein, vitamins and phenolic substances, among which arabinoxylan-based non-starch polysaccharides contain 46%, which is the main component of wheat cell walls. Element. Bran also contains a certain amount of phenolic acids, such as ferulic acid, p-coumaric acid, caffeic acid, sinapic acid and syringic acid, et...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N9/18C12R1/66
Inventor 吴晖胡博涵赖富饶刘素纯曾岚
Owner 华工利亚(佛山)生物科技有限责任公司
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