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A kind of pectin esterase and its preparation method and application

A pectin esterase and protease inhibitor technology, applied in the field of pectin esterase and its preparation, can solve the problems of prolonging the production cycle and increasing production costs, and achieve low cost, short fermentation cycle, and great theoretical and practical significance Effect

Active Publication Date: 2020-12-04
INST OF BAST FIBER CROPS CHINESE ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

If the pectin esterase produced by natural microbial fermentation needs to be purified and separated to obtain a single pectin esterase, this process adds new production equipment, prolongs the production cycle, and increases production costs

Method used

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  • A kind of pectin esterase and its preparation method and application
  • A kind of pectin esterase and its preparation method and application

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Embodiment 1

[0030] A kind of preparation method of embodiment 1 pectin esterase

[0031] Such as figure 1 As shown, 1 mL of the genetically engineered bacteria pEASY-E1-243 / BL21 was inoculated into 200 mL of the improved LB culture medium (containing Amp 120 μg / mL), mixed thoroughly, 200 r / min, 37 ° C for 10-12 hours, that is, the seed solution.

[0032] The mass ratio formula of the above-mentioned improved LB culture solution is: glucose 0.5% + tryptone 1% + yeast extract 0.5% + NaCl 1%, and use NH 3 ·H 2 O to adjust the pH value to 7.5.

[0033] 600mL seed solution was directly inoculated into a 50L fermenter (Bio, Germany) filled with 30L modified LB culture solution (containing Amp120μg / mL), and ammonia water (NH 3 ·H 2 O), the pH is maintained to 7.0, the ventilation rate is 0.8m 3 / h, 37℃, 200r / min culture 3-4h, make OD 600 When it reaches about 0.5, add IPTG to a final concentration of 1.2mmol / L, change to 28°C for induction and culture for 12-18h, and then obtain a mature ...

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Abstract

The invention discloses pectin esterase and a preparation method and application thereof. The preparation method comprises the steps of inoculating a genetically engineered strain pEASY-E1-243 / BL21 into an improved LB fermentation culture solution, carrying out expanding culture fermentation, then adding IPTG to induce production of pectinase, adding a protease inhibitor after obtaining a mature fermentation solution, and carrying out ultrafiltration to collect an enzyme solution with the molecular weight of 10 to 50 kD. The method for removing ester from high-ester pectin by utilizing the pectin esterase comprises the steps of adding the pectin esterase into a high-ester pectin solution with the concentration of 10 to 12w / v%, controlling the adding amount to be 100 to 200U / mL, and carrying out enzymolysis at 40 to 45 DEG C for 2 to 3h. The preparation method of the pectin esterase, which is provided by the invention, is simple, short in fermentation period and low in cost, and the product contains few exogenous enzymes. High-ester pectin is subjected to enzymolysis, so that the main chain of pectin in a raw material can be prevented from being damaged, and high-molecular-weight low-ester pectin with uniform quality can be obtained.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation, in particular to a pectin esterase and its preparation method and application. Background technique [0002] Pectin is a high-molecular sugar polymer existing in the plant cell wall, and its relative molecular weight is between 5×10 5 -30×10 5 between. Usually, pectin with a degree of esterification higher than 50% (equivalent to a methoxyl content of 7%-16.3%) is called a high-ester pectin, and a pectin with a degree of esterification lower than 50% (equivalent to a methoxyl content of less than 7%) is called a high-ester pectin. %) pectin is called low ester pectin. [0003] In the process of food processing, high-ester pectin can form gel when heated under the conditions of sugar concentration higher than 55°Brix and pH value lower than 3.5, which is mainly used in the production of high-sugar food; low-ester pectin only needs a lower The concentration of sugar or even under...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/18C12P19/04C12P21/02C12R1/19
CPCC12N9/18C12P19/04C12P21/02C12Y301/01011
Inventor 成莉凤彭源德段盛文冯湘沅杨琦郑科刘志远
Owner INST OF BAST FIBER CROPS CHINESE ACADEMY OF AGRI SCI
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