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Method for preparing heat-resistant acid-resistant feruloyl esterase by using trichoderma atroviride

A technology of ferulic acid esterase and Trichoderma dark viridans, which is applied in the field of microbial fermentation, can solve the problems of chemical synthesis with many reaction steps, low oryzanol content, and limited output, and achieve low water consumption and short fermentation cycle , the effect of low environmental pollution

Inactive Publication Date: 2015-09-09
XUZHOU UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The chemical synthesis method has many reaction steps and a long cycle; the raw material oryzanol content of the alkaline hydrolysis method is low in rice bran, and the yield is limited; in addition, these two methods are polluting, so the enzymatic method to produce ferulic acid has become a research hotspot in recent years

Method used

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  • Method for preparing heat-resistant acid-resistant feruloyl esterase by using trichoderma atroviride
  • Method for preparing heat-resistant acid-resistant feruloyl esterase by using trichoderma atroviride
  • Method for preparing heat-resistant acid-resistant feruloyl esterase by using trichoderma atroviride

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Embodiment 1

[0023] The present embodiment illustrates the screening method of Trichoderma dark green AWS26, and concrete steps are as follows:

[0024] Step 1, sampling and sample processing:

[0025] 10 soil samples were taken from the high-temperature fermented compost in Dahan Village, Yunlong District, Xuzhou City, Jiangsu Province. Weigh 5g of solid soil sample, put it into a triangular flask filled with glass beads and 50mL of sterilized saline, vibrate at 160r / min for 30min to make a suspension, and then put the triangular flask into a constant temperature shaking water bath at 60-70°C In the pot, shake at 100r / min for 20-30min to obtain a soil-like turbid solution.

[0026] Step 2, enrichment culture:

[0027] Take 2-4mL of the soil sample turbid solution obtained in step 1, add it to a 250mL Erlenmeyer flask containing 30-60mL enrichment medium 1, and place it in a constant temperature shaker at 45-50°C and a speed of 160-200r / min After culturing for 5-7 days, an enriched cult...

Embodiment 2

[0057] This embodiment illustrates the method for preparing heat-resistant and acid-resistant ferulic acid esterase by using Trichoderma dark green as bacterial strain solid-state fermentation method, comprising the following steps:

[0058] Step 1, strain preparation: first, under aseptic conditions, pick the spores of the test tube strains stored in two rings and inoculate them on the activated slant medium for activation, and cultivate them in an incubator at 30-35°C for 3-5 days; Inject 1mL of sterile saline into the slant of Trichoderma viridis, and repeatedly blow the spores on the slant with a pipette gun to further dilute the spore concentration to 10 7 pc / mL, then inoculate 2mL of spore suspension with the above concentration into a 250mL Erlenmeyer flask containing 50g of solid spore culture medium, shake the Erlenmeyer flask fully to mix the spores with the culture medium, and place in a 30-40°C incubator Cultivate for 3-4 days, turn over the solid medium once a day...

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Abstract

The invention discloses a method for preparing heat-resistant acid-resistant feruloyl esterase by using trichoderma atroviride. According to the method, trichoderma atroviride serves as a production strain, heat-resistant feruloyl esterase is prepared through the processing steps of preparing fermenting strains, carrying out triangular flask solid-state fermentation, carrying out koji tray solid-state fermentation, preparing a crude enzyme preparation and purifying the crude enzyme preparation, and the obtained enzyme preparation is high in enzyme activity, good in acid resistance and high in heat resistance; the preparation method is short in fermentation cycle, low in energy consumption, low in water consumption, low in environmental pollution degree and high in specific yield and has excellent industrialization application prospects.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a method for preparing heat-resistant and acid-resistant ferulic acid esterase from Trichoderma dark viride. Background technique [0002] Ferulic acid esterase (E.C13.1.1.73, feruloyl esterase, FAE) is a carboxylesterase that has been studied more recently. It is an extracellular enzyme that can hydrolyze methyl ferulic acid, polysaccharide The ester bond in ferulic acid ester, etc., will release free ferulic acid monomer or dimer. In the field of food and medicine, ferulic acid esterase can be used to degrade the ferulic acid ester bond in the plant cell wall to obtain free ferulic acid with medicinal value and health care function. Ferulic acid is a derivative of cinnamic acid, its chemical name is 4-hydroxy-3-methoxycinnamic acid, and it is also a natural antioxidant, which is effective for diseases such as cardiovascular diseases, diabetes, Alzhei...

Claims

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Application Information

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IPC IPC(8): C12N9/18C12N1/14C12R1/885
CPCC12N9/18C12Y301/01073
Inventor 高兆建刘恩岐唐仕荣张建萍孙会刚徐大伟杨宪瑶
Owner XUZHOU UNIV OF TECH
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