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Culture medium and method for preparing polyphenol oxidase through fungus fermentation process

A polyphenol oxidase and culture medium technology, applied in the field of microbial fermentation, can solve the problems of difficult large-scale production, difficult to expand application, low expression activity, etc., and achieve the effect of increasing production, abundant sources, and low price

Inactive Publication Date: 2015-11-25
FUJIAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, most of the polyphenol oxidases used in tea processing research and application are derived from plants. Due to the limitation of season and extraction cost, it is difficult to achieve large-scale production
At the same time, polyphenol oxidases from different plants are difficult to fully exert their catalytic performance in tea processing because of the differences in their substrates.
Many units are trying to express polyphenol oxidase from different plant sources through genetic engineering technology, but it is also difficult to expand the application due to the low expression activity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 1. Culture medium formula

[0037] Fermentation in a final volume of 1 L Medium Hydration Mixture Calculate, the formula of its each component is:

[0038] Bran 30g

[0039] Tea 6g

[0040] Peptone 2.5g

[0041] K H 2 PO 4 2.0g

[0042] MgSO 4 .7H 2 O0.2g

[0043] MnSO 4 .5H 2 O0.5g

[0044] CuSO 4 .5H 2 O0.02g.

[0045] 2. The strain and culture of S. coronoidis

[0046] The use of Cystis anterior coronarius ( Eurotium cristatum ) were stored on potato dextrose agar slant medium at 4°C. Under aseptic conditions, transfer the strain to fresh potato dextrose agar medium, culture in the dark at 28°C for 5 days, wash the mature conidia with sterile water, prepare spore suspension, spore suspension The concentration is 1~2×10 8 spores / mL.

[0047] The formula of described potato dextrose agar slant medium, with final volume being the medium aqueous solution of 1L, the formula of its each component is:

[0048] Potatoes 200g

[0049] Glucose 18~20g ...

Embodiment 2

[0073] 1. Based on the aqueous medium solution with a final volume of 1L, the formula of each component is:

[0074] Bran 48.9g

[0075] Tea 3.5g

[0076] Ammonium nitrate 10.5g

[0077] K H 2 PO 4 2.0g

[0078] MgSO 4 .7H 2 O0.5g

[0079] Anhydrous CaCl 2 0.075g

[0080] CuSO 4 .5H 2 O0.01g

[0081] 2. The strain and culture of S. coronoidis

[0082] The use of Cystis anterior coronarius ( Eurotium cristatum ) were stored on potato dextrose agar slant medium at 4°C. Under aseptic conditions, transfer the strain to fresh potato dextrose agar medium, culture in the dark at 26°C for 7 days, wash the mature conidia with sterile water, prepare spore suspension, spore suspension The concentration is 1~2×10 8 spores / mL.

[0083] The formula of described potato dextrose agar slant medium and its preparation method are the same as in Example 1.

[0084] 3. Preparation of Polyphenol Oxidase

[0085] Prepare the fermentation medium according to the above formula, and ...

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Abstract

The invention relates to a method for producing polyphenol oxidase through fungus fermentation process. The method comprises the following steps: mixing wheat bran, tea leaves, a nitrogen source additive, KH2PO4, MgSO4.7H2O, MnSO4.5H2O and CuSO4.5H2O, and diluting to 1L; under the aseptic operating condition, adding fungi spore suspending liquid, performing shaking-flask culture under the temperature of 26-28 DEG C and at the rotating speed of 120 r / min for 3-5 days, and performing centrifugation, so as to obtain an aqueous solution of the polyphenol oxidase. According to the invention, the tea leaves are used as main components of a culture medium, and polyphenol compounds contained in the tea leaves are used as induction ingredients to facilitate the increase of yield of the polyphenol oxidase; besides, the wheat bran and tea leaves as the raw materials are rich in resources and low in price; through the adoption of the culture medium in the method, the enzyme activity can reach 241-256.6 U / mL.min.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a medium for preparing polyphenol oxidase by fungal fermentation and a preparation method thereof. Background technique [0002] Polyphenol oxidase (Polyphenoloxidase, PPO) is a class of copper proteins widely present in bacteria, fungi, plants and mammals, including laccase (Laccase), tyrosinase (Tyrosinase) and catechol oxidase ( Catecholase). Polyphenol oxidase can effectively catalyze the oxidation of polyphenolic compounds to form corresponding quinones, and is a key enzyme in melanin metabolism and catecholamine synthesis. Polyphenol oxidase is the main enzyme that causes browning of plant leaves, fruits and vegetables; at the same time, because it participates in the degradation process of polyphenolic compounds, it has been well received in the fields of phenolic wastewater treatment, lignin degradation and dye decolorization. research and app...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12R1/645
CPCC12N9/0059C12Y110/03001
Inventor 杨民和张婉蓉巫婷玉黄鹭强
Owner FUJIAN NORMAL UNIV
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