Method for preparing complex enzyme preparation of heat-resistant ferulic acid esterase and cellulose by fermenting trichoderma atroviride through airlift fermentation tank

The technology of an air-lift type fermenter and a compound enzyme preparation is applied in the field of microbial fermentation, and can solve the problems of loss of ferulic acid esterase enzyme activity, limited application of ferulic acid esterase, insufficient tolerance, etc. Strong catalytic activity and good thermal stability

Inactive Publication Date: 2015-08-19
XUZHOU UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] At present, there is a great demand for ferulic acid esterase in the feed industry, and the ferulic acid esterase used in industry is mainly derived from Aspergillus niger, because Aspergillus niger ferulic acid esterase is resistant to high temperature, acid or alkali and other conditions Insufficient performance, in the process of industrial production and use, the enzyme activity of ferulic acid esterase is seriously lost, which largely limits the application of ferulic acid esterase in industry

Method used

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  • Method for preparing complex enzyme preparation of heat-resistant ferulic acid esterase and cellulose by fermenting trichoderma atroviride through airlift fermentation tank
  • Method for preparing complex enzyme preparation of heat-resistant ferulic acid esterase and cellulose by fermenting trichoderma atroviride through airlift fermentation tank
  • Method for preparing complex enzyme preparation of heat-resistant ferulic acid esterase and cellulose by fermenting trichoderma atroviride through airlift fermentation tank

Examples

Experimental program
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Effect test

Embodiment 1

[0025] The present embodiment illustrates the screening method of Trichoderma dark green AWS26, and concrete steps are as follows:

[0026] Step 1, sampling and sample processing:

[0027] 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.

[0028] Step 2, enrichment culture:

[0029]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 cultu...

Embodiment 2

[0058] This example illustrates the preparation of ferulic acid esterase and cellulase composite enzyme preparation by Trichoderma dark green AWS26 fermented in an air-lift fermenter. Specific steps are as follows:

[0059] Step 1, activation of fermentation strains: Streak and activate Trichoderma dark green Aws26 with the preservation number CGMCC No.8673 on the PDA medium plate, and then place it upside down in a constant temperature incubator at 40-42°C for 24-48h for activation culture .

[0060] Step 2, prepare Trichoderma dark green spore suspension: Scrape Trichoderma dark green mycelium from the activated strain in step 1 with an inoculation loop, transfer to Trichoderma dark green spore medium, and place it at 37-40 ° C for biochemical Cultivate in the incubator for 72 hours until the mycelium grows spores, recover its vigor after two subcultures, wash and collect the spores with sterile water containing 0.01% Tween 80 several times, and prepare the spore content of...

Embodiment 3

[0076] This example illustrates the properties of the compound enzyme preparation of ferulic acid esterase and cellulase obtained in the present invention.

[0077] 1. Determination of the optimum temperature for enzyme reaction

[0078] Place the reaction system in a water bath at 20°C, 25°C, 30°C, 35°C, 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, 75°C, 80°C, and after appropriate dilution , to measure the activity of ferulic acid esterase and cellulase respectively, and study the optimum temperature of enzymatic reaction. The enzyme activity measured at the optimum temperature is 100%, and the activity measured at other temperatures is its relative activity. The result is as figure 1 As shown, ferulic acid esterase and cellulase have basically the same change trend with the increase of temperature. From 20°C to 65°C, the enzyme activity of ferulic acid esterase increased gradually with the increase of temperature, and the enzyme activity reached the maximum when the tempera...

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Abstract

The invention discloses a method for preparing a complex enzyme preparation of heat-resistant ferulic acid esterase and cellulose by fermenting trichoderma atroviride through an airlift fermentation tank. According to the method, the trichoderma atroviride is taken as a production strain, wherein the method comprises the following steps: preparing a fermentation strain, preparing a spore suspension, preparing a seed solution, carrying out liquid fermentation through a shaking flask, preparing the complex enzyme preparation and preparing the complex enzyme preparation of heat-resistant ferulic acid esterase and cellulose. The prepared complex enzyme preparation is high in activity, soluble in water, free of peculiar smell, high in heat stability and high in catalytic activity, and has a good prospect of industrialization application.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a method for preparing a heat-resistant ferulic acid esterase and cellulase composite enzyme preparation by fermenting an air-lift fermenter of Trichoderma dark greens. Background technique [0002] Ferulic acid esterase (E.C.3.1.1.73, ferulicacid esterase, FAE), also known as cinnamic acid esterase, is a new type of enzyme preparation, which mainly uses methyl ferulic acid, oligosaccharide ferulic acid ester, polysaccharide ferulic acid Ester and lignin ferulic acid ester as substrates, destroy the ester bond between ferulic acid and polysaccharide, release ferulic acid from the plant cell wall, make the remaining polysaccharide main chain easy to be degraded, and effectively improve the content of a large amount of wood fiber Nutritional value of ferulic acid by-products. Ferulic acid has a variety of physiological functions such as scavenging free r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/18C12N9/42C12R1/885
Inventor 刘恩岐高兆建张建萍侯进慧唐仕荣孙会刚徐大伟
Owner XUZHOU UNIV OF TECH
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