A method for preparing heat-resistant ferulic acid esterase by submerged liquid fermentation of bacillus licheniformis

A Bacillus licheniformis, submerged liquid fermentation technology, applied in the field of microbial fermentation, can solve the problems of limiting the effective degradation of cellulose and hemicellulose, and achieve the effects of low production cost, simple follow-up treatment, and wide application range

Inactive Publication Date: 2018-02-13
XUZHOU UNIV OF TECH
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Problems solved by technology

On the other hand, in the plant cell wall, ferulic acid or dimeric ferulic acid is mainly connected to the arabinose residue of arabinoxylan in the form of ester bond, which enhances the strength of the arabinoxylan chain, but from the space Limits the effective degradation of cellulose and hemicellulose in plant cell walls by animals and microorganisms

Method used

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  • A method for preparing heat-resistant ferulic acid esterase by submerged liquid fermentation of bacillus licheniformis
  • A method for preparing heat-resistant ferulic acid esterase by submerged liquid fermentation of bacillus licheniformis
  • A method for preparing heat-resistant ferulic acid esterase by submerged liquid fermentation of bacillus licheniformis

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Embodiment 1

[0024] The screening method of bacillus licheniformis DBM12 comprises the following steps:

[0025] Step 1, soil sample collection and processing

[0026] The soil samples were collected from livestock manure compost in the countryside of Yunlong District, Xuzhou City, Jiangsu Province, which was undergoing high-temperature fermentation. The collected soil samples were stored in sealed sterile plastic bags at room temperature, and the target strains were isolated within 1 week.

[0027] Take 5-10g of soil samples and put them into a conical flask filled with 20-40mL sterile water and small glass beads, put them on a shaker, rotate and shake at 120-160r / min for 20-30min, then place the conical flask at 70 Heat in a water bath at -80°C for 15-30 minutes, and shake once every 5 minutes during this period.

[0028] Step 2, enrichment culture

[0029] Take 1-2mL heat-treated soil samples in a Erlenmeyer flask filled with 30-50mL enrichment medium 1, and shake at 160-180r / min at ...

Embodiment 2

[0049] This embodiment illustrates the preparation of heat-resistant ferulic acid esterase preparation by deep liquid ventilation fermentation, including the following steps:

[0050] Step 1, preparation of primary seeds: fill 30mL-50mL of liquid seed medium in a 250mL Erlenmeyer flask, adjust the pH to 7.0, sterilize at 121°C for 20min, and after cooling, the volume ratio is 4-6v / v % of the inoculum amount to inoculate, cultivate on a shaker at 40-45° C. for 18-24 hours, and obtain a first-grade seed liquid.

[0051] Primary seed medium: beef extract 0.4g, peptone 0.8g, yeast powder 1g, NaCl 0.5g, KH 2 PO 4 0.1g, MgSO 4 ·7H 2 O 0.04, distilled water 100mL, pH 7.0, sterilize at 121°C for 20min.

[0052] Step 2, preparation of secondary seeds: Inoculate the cultivated primary seed liquid into the secondary seed medium according to the inoculation amount of 6-10v / v%, and fill 100-120mL secondary seed medium in a 1000mL triangular flask. Cultivate at 32-37°C for 18-24h.

...

Embodiment 3

[0067] This example illustrates the properties of the ferulic acid esterase preparations of the present invention.

[0068] (1) Effect of temperature on storage stability of ferulic acid esterase solid enzyme preparation

[0069] The prepared solid ferulic acid esterase enzyme preparations were stored at five temperature environments of -20°C, 4°C, 28°C, 37°C and 50°C for 100 days, and the enzyme activity was measured every 20 days. The effect of temperature on the stability of ferulic acid esterase was investigated. Figure 4 It is the enzyme activity change curve of ferulic acid esterase solid enzyme preparation under different temperature storage conditions. The enzyme activity is the ratio of the activity of the enzyme preparation after storage at different temperatures to the corresponding enzyme activity of ferulic acid esterase before storage.

[0070] Depend on figure 1 It can be seen that temperature has a certain influence on the stability of ferulic acid esterase...

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Abstract

The invention discloses a method for preparing heat-resistant feruloyl esterase by deep liquid fermentation of Bacillus licheniformis, which comprises the following steps: by using Bacillus licheniformis as a production strain, carrying out strain activation, seed solution culture, fermentation tank deep liquid ventilation fermentation and enzyme preparation production to obtain the heat-resistant feruloyl esterase preparation. The obtained solid feruloyl esterase preparation has the advantages of favorable stability, wide application range, simple preparation technique and high enzyme activity. The enzyme activity of the enzyme preparation can reach 378 U / g, and the overall yield reaches 81.5%. The heat-resistant feruloyl esterase has excellent industrialized 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 ferulic acid esterase by submerged liquid fermentation of Bacillus licheniformis. Background technique [0002] Ferulic acid (ferulic acid, FA) is a natural antioxidant commonly found in plants. It is a phenolic acid in a variety of plants. Aggregation, lowering blood fat and other unique functions. However, ferulic acid exists mostly in the form of cross-linking in plants, and humans and animals cannot directly absorb this ferulic acid, and must rely on microorganisms to produce esterase to release ferulic acid. On the other hand, in the plant cell wall, ferulic acid or dimeric ferulic acid is mainly connected to the arabinose residue of arabinoxylan in the form of ester bonds, which enhances the strength of the arabinoxylan chain, but from the space This limits the effective degradation of cellulose and hemicell...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/18C12N1/20C12R1/10
CPCC12N9/18C12Y301/01073
Inventor 高兆建刘恩岐孙会刚侯进慧曹泽虹徐大伟杨宪瑶
Owner XUZHOU UNIV OF TECH
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