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Aspergillus niger and applications thereof in preparation of Pu'er tea theine compounds

A technology of Aspergillus niger and compound, applied in the application field of production and biotransformation, can solve the problems of polluting the environment, high consumption, high difficulty and the like

Active Publication Date: 2017-07-25
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If chemical synthesis is to be carried out, there are disadvantages such as difficulty, low yield, consumption of a large amount of organic solvents, and environmental pollution.

Method used

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  • Aspergillus niger and applications thereof in preparation of Pu'er tea theine compounds
  • Aspergillus niger and applications thereof in preparation of Pu'er tea theine compounds
  • Aspergillus niger and applications thereof in preparation of Pu'er tea theine compounds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Example 1 Isolation and identification of Aspergillus niger M03

[0081] 1 primary screening

[0082] Preparation of bacteria solution: Take a Pu'er cooked tea sample, soak it in sterilized distilled water for 10 minutes, let it stand, and take the supernatant as the bacteria solution. Take 1 mL of the bacterial solution and add it to a test tube filled with 9 mL of sterile water, and so on, to make 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 Use a sterilized pipette to draw one drop of the diluted bacterial solution with different concentrations into the primary screening medium, and spread it evenly with a coating rod. Place the petri dish upside down in a constant temperature incubator at 28°C and cultivate for a certain period of time, so that colonies appear, pick a single colony, inoculate it into the screening medium by streaking on a plate, continue to cultivate until a single colony appears, pick each single colony for preservation On the slant of the primary scr...

Embodiment 2

[0097] Example 2 Fermentation strain preparation

[0098] Take the low-temperature-preserved strain M03, inoculate it on the PDA solid medium, culture it in a 28°C constant temperature incubator for 72 hours until the spores mature, wash the spores on the PDA slant medium with sterile saline, and make 1× 10 6 CFU / mL spore suspension, take the spore suspension and inoculate it in PDB liquid medium at an inoculum size of 2% (volume), place it in a shaker at 28° C. and 200 rpm for 18 hours, and obtain a seed liquid.

[0099] Composition of PDA medium: 5.0 g of potato extract powder, 15.0 g of glucose, 15.0 g of agar, and 1000 mL of distilled water.

[0100] Composition of PDB medium: 5.0 g of potato extract powder, 15.0 g of peptone, 15.0 g of glucose, 5.0 g of sodium chloride, and 1000 mL of distilled water.

Embodiment 3

[0101] Example 3 Biotransformation of Pu-erh tea compounds by solid fermented sun-dried green hair tea

[0102] Take the Aspergillus niger seed solution cultivated at 35°C and 200rpm for 24h, and add the mycelia of Aspergillus niger evenly to the sun-dried green maocha with 35% sterilized water in a proportion of 2% by mass percentage, and at 40°C, Under the condition of 75% humidity, after "stacking" for 28 days, the piles were turned every 7 days.

[0103] Take fermented Pu-erh tea, ultrasonically extract with methanol, concentrate to obtain extract, extract with chloroform to remove caffeine, extract with ethyl acetate, extract 3 times, combine extracts, concentrate under reduced pressure to obtain extract, extract with a small amount of After dissolved in methanol, it was separated by Sephadex LH-20 column chromatography. A total of 40 fractions were collected and analyzed by LC-MS. The same fractions were combined to obtain 12 fractions containing puerhin. Then these f...

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Abstract

The invention relates to aspergillus niger M03 and applications of the aspergillus niger in preparation of Pu'er tea theine through microbial conversion. The aspergillus niger M03 strain capable of realizing bioconversion for preparing Pu'er tea theine is separated from Pu'er ripe tea, is preserved in the China General Microbiological Culture Collection Center (CGMCC), is assigned with the accession number of CGMCC No.10930. According to the aspergillus niger M03 and the applications, the aspergillus niger M03 strain is taken as a fermenting strain, the piling solid fermentation is adopted, and the sun-withering crude tea leaves of Yunnan large-leaf tea are adopted as the raw material; or liquid fermentation is adopted, catechin compounds and theanine are adopted as substrates, piling solid fermentation and liquid fermatneion are carried out, and thus the Pu'er tea theine compounds are prepared. The aspergillus niger M03 can be used for specifically preparing the Pu'er tea theine compounds, has the advantages that the environment is protected, the operation is simple, and the popularization is convenient, and thus the application prospect is very good.

Description

technical field [0001] The invention relates to the field of microbial transformation, relates to the application of Aspergillus niger, in particular to the production and biotransformation of a strain of Aspergillus niger and its puer tea compounds. Background technique [0002] Pu-erh tea is made from the dried young leaves of Camellia sinensis var.assamica (Masters) Kitamura, a unique large-leaf tea in Yunnan (commonly known as sun-dried green hair tea) through a special post-fermentation process. It is a famous post-fermentation tea. Tea. Pu'er tea has a unique flavor and quality, and has many health benefits, and the effect is more significant than other teas. Studies have shown that this is closely related to the subsequent fermentation process. After the post-fermentation process of Pu-erh tea, under the enzymatic or non-enzymatic action of microorganisms, the content of catechins contained in it is greatly reduced, and many new compounds different from other teas ar...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P17/16C12R1/685
CPCC12P17/16C12R2001/685C12N1/145Y02P20/59Y02P20/133
Inventor 屠鹏飞姜勇叶静王伟楠
Owner PEKING UNIV
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