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Method for producing feruloyl esterase

A ferulic acid esterase and production method technology, applied in the field of biological enzyme preparations, can solve the problems of difficult enzyme extraction and purification, low yield of ferulic esterase, unstable product quality, etc., shorten the production cycle, reduce Production cost and operation cost reduction effect

Inactive Publication Date: 2015-06-03
HENAN UNIVERSITY OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of original strains to produce ferulic acid esterase has the characteristics of low yield, difficult enzyme extraction and purification, and unstable product quality.

Method used

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  • Method for producing feruloyl esterase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Strain preparation:

[0027] (1.1) Inoculate 1% by weight of the Pichia pastoris strain constitutively expressing ferulic acid esterase A into 500mL of sterilized medium, cultivate it at 28°C and 220 rpm for 60 hours, and use it as a seed solution for later use; The medium is composed of the following raw materials in mass percentage: 1% yeast powder, 2% peptone, 2% glucose, and the balance is water;

[0028] (1.2) Prepare 30 L of sterilized medium in a 50 L fermenter: take industrial-grade raw materials according to mass percentage: 2% starch hydrolyzate, 1% corn steep liquor, 2% ammonia water, and the balance is water; adjust the pH with phosphoric acid After reaching 6.8, add 1mL / m 3 Trace element mother liquid, sterilized at 124°C for 30 minutes, and used as seed liquid culture medium for later use;

[0029] (1.3) Transfer the seed solution prepared in step (1.1) into the 50L fermenter prepared in step (1.2) containing 30 L of sterilized medium, and cultivate ...

Embodiment 2

[0039] (1) Strain preparation:

[0040] (1.1) Inoculate 1% by weight of the Pichia pastoris strain constitutively expressing ferulic acid esterase A into 500mL of sterilized medium, cultivate it at 28°C and 200 rpm for 48 hours, and use it as a seed solution for later use; The medium is composed of the following raw materials in mass percentage: 1% yeast powder, 2% peptone, 2% glucose, and the balance is water;

[0041] (1.2) Prepare 30 L of sterilized medium in a 50 L fermenter: take industrial-grade raw materials according to mass percentage: 2% starch hydrolyzate, 1% corn steep liquor, 2% ammonia water, and the balance is water; adjust the pH with phosphoric acid After reaching 6.8, add 1mL / m 3 Trace element mother liquid, sterilized at 120°C for 30 minutes, and used as seed liquid culture medium for later use;

[0042] (1.3) Transfer the seed solution prepared in step (1.1) into the 50L fermenter prepared in step (1.2) containing 30 L of sterilized medium, and cultivate ...

Embodiment 3

[0052] (1) Strain preparation:

[0053] (1.1) Inoculate 1% by weight of Pichia pastoris strains constitutively expressing ferulic acid esterase A into 500mL of sterilized medium, cultivate them at 30°C and 240 rpm for 60 hours, and use them as seed liquid for later use; The medium is composed of the following raw materials in mass percentage: 1% yeast powder, 2% peptone, 2% glucose, and the balance is water;

[0054] (1.2) Prepare 30 L of sterilized medium in a 50 L fermenter: take industrial-grade raw materials according to mass percentage: 2% starch hydrolyzate, 1% corn steep liquor, 2% ammonia water, and the balance is water; adjust the pH with phosphoric acid After reaching 6.8, add 1mL / m 3 Trace element mother liquid, sterilized at 121°C for 30 minutes, and used as seed liquid culture medium for later use;

[0055] (1.3) Transfer the seed solution prepared in step (3.1) into the 50L fermenter prepared in step (3.2) containing 30 L of sterilized medium, and cultivate at ...

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Abstract

The invention discloses a method for producing feruloyl esterase and belongs to the technical field of bio-enzyme preparation production. The method comprises the following steps: fermenting by utilizing a genetic engineering strain of the constitutive expressed feruloyl esterase, and by taking the starch saccharification liquid, the maize slurry and the ammonia water as main raw materials, preparing the feruloyl esterase, continuously adding a fermentation culture medium and discharging part of the fermenting fluid in bathes during the fermentation engineering, continuously extending the fermenting period under a no-sundry bacterium contamination condition, and filtering by a plate frame filter, and performing ultrafiltration concentration and spray drying on the fermenting fluid to obtain the high-activity feruloyl esterase preparation. The specific steps are as follows: (1) strain preparation; (2) culture expansion; (3) submerged fermentation and feruloyl esterase production; and (4) feruloyl esterase purification. By adopting the method under the conditions of no serious sundry bacterium contamination and no obvious reduction of the enzyme activity in unit volume of the fermenting fluid, the feruloyl esterase can be continuously fermented and produced, the production period can be shortened and the production cost can be reduced.

Description

technical field [0001] The invention belongs to the technical field of production of biological enzyme preparations, and in particular relates to a production method of ferulic acid esterase A. Background technique [0002] Ferulic acid esterase belongs to carboxyl ester hydrolase, which can hydrolyze the ester bond between ferulic acid and arabinose or galactose in hemicellulose, making other hemicellulose hydrolysis-related enzymes more accessible to their substrates , to promote the complete hydrolysis of hemicellulose. The enzyme has great potential application value in lignocellulose degradation, paper making, organic food, feed, medicine and other industries. [0003] Ferulic acid esterase is mainly fermented by original strains and genetically engineered strains. At present, since ferulic acid esterase has not been included in the "Food Additives" catalogue, the large-scale production process of ferulic acid esterase has not been reported; people's theoretical and a...

Claims

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Application Information

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IPC IPC(8): C12N9/18C12R1/84
CPCC12N9/18C12Y301/01073
Inventor 张帅兵翟焕趁胡元森蔡静平贾峰王金水屈建航李海峰翟丹丹
Owner HENAN UNIVERSITY OF TECHNOLOGY
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