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Pichia pastoris and application thereof

A technology of Pichia pastoris and hyphae, applied in the field of microbial engineering, can solve the problems of complex strain requirements and no related research on fermentation and aroma production of finished tea.

Active Publication Date: 2019-05-14
冯磊
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no relevant research on the use of microorganisms to ferment tea extracts to produce aroma and finished tea
CN108432906A discloses a method for making wine-flavored tea leaves by fermentation of microbial strains, but it needs to be combined with 10-20 parts of lactic acid bacteria, 10-30 parts of cerevisiae, 50-70 parts of yeast, and 5-10 parts of digestive enzymes. The requirements for strains are complex, and the main purpose is to give tea products a bouquet

Method used

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  • Pichia pastoris and application thereof
  • Pichia pastoris and application thereof
  • Pichia pastoris and application thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0029]Wherein, the enzyme treatment liquid of tea refers to the clear liquid obtained by discarding tea residue after tea is treated with glycosidase. For example, the preparation method of the tea enzyme treatment solution may include: mixing tea leaves with water, and then introducing an enzyme auxiliary agent for enzyme treatment. The enzyme auxiliary agent can be various common glycosidases, preferably cellulase and / or pectinase, as described above, and will not be repeated here. With respect to per gram of tea leaves on a dry basis, the amount of the enzyme auxiliary agent can be 80-180U (such as 80U, 100U, 120U, 140U, 160U, 170U, 172U, 175U, 178U, 180U or between the above-mentioned values. Arbitrary value), the consumption of water can be 15-20g. Mixing can be performed under conventional conditions, but preferably, the mixing is performed under ultrasonic conditions. The frequency of ultrasound may be 25-40 kHz. The temperature of mixing may be 40-60°C. The mixing ...

Embodiment 1

[0034] This example is used to illustrate the taxonomic status of the strain FLei-01 of the present invention.

[0035] (1) Strain culture medium: commercial Martin's medium;

[0036] (2) Genome extraction reagents: SDS, Tris base, EDTA;

[0037] (3) Reagents and primers required for PCR amplification: TaqDNA polymerase, dNTP, GoldView nucleic acid dye;

[0038] (4)DNA Marker;

[0039] (5) ITS amplification primers:

[0040] ITS1: 5'-tccgtaggtgaacctgcgg-3'

[0041] ITS4: 5'-tcctccgcttattgatatgc-3'

[0042] (6) Gene extraction and amplification experimental methods and steps:

[0043] Genome extraction; collect the cells; rinse with water for 3 times; fully absorb the residual moisture of the cells with filter paper; put the dried cells into a mortar, add an appropriate amount of 6% SET solution and quartz sand to grind thoroughly, grind as much as possible Sufficiently; absorb the ground liquid, add it to a 2ml centrifuge tube, bathe in 75°C water for 30 minutes, freeze ...

Embodiment 2

[0048] This example is used to illustrate that the strain FLei-01 of the present invention can ferment tea to produce aroma.

[0049] (1) Carry out boiling water extraction to tealeaves according to the mode of Table 1, the tea water extract after boiling water extraction is packed in the triangular bottle of 150ml on average, and packs 40ml in each bottle of tea water. Sterilize. 5% (V / V) inoculation of FLei-01 strain seed liquid (seed liquid adopts Cha's culture medium, and the concentration of FLei-01 in terms of dry cell weight is 2g / L); each kind of tea is used as a blank control (no inoculation strains) and inoculate a sample.

[0050] (2) Cultivate at 30°C and 180r / min for 8 days after inoculation, observe and smell its smell every day, find that the color of the tea becomes darker, and emit a strong aroma, and score it for sensory quality (the scoring criteria are shown in Table 2 below ), were evaluated by 30 adult consumers, and the results were averaged and shown ...

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Abstract

The invention relates to the technical field of microorganisms, and discloses pichia pastoris and application thereof. The preservation number of the pichia pastoris is CCTCC NO:M 2018666. The invention further discloses a preparation capable of enhancing aroma of tea leaves. The preparation contains the pichia pastoris and an enzyme assistant. The invention further discloses application of the pichia pastoris or the preparation in tea product aroma enhancement. In addition, the invention discloses a method for preparing a tea product. The method comprises the steps that the pichia pastoris isinoculated into a tea leaf extracting solution for culturing. By utilizing the pichia pastoris, the aroma of the tea product can be effectively improved on the basis that the original tea fragrance of the tea product is kept.

Description

technical field [0001] The invention relates to the field of microbial engineering, in particular to a strain of Pichia sp. and its application. Background technique [0002] Tea is one of the most consumed beverages in the world. China is the hometown of tea, the first country to discover and utilize tea, and has always been a major tea-producing country. The annual production of tea accounts for about a quarter of the world's total. one. Tea can eliminate food and greasy, reduce fire and improve eyesight, calm the mind and relieve troubles, clear heat and detoxify, promote body fluid and quench thirst. The tea polyphenols contained in tea have strong antioxidant and physiological activities. They are free radical scavengers in the human body and can block the synthesis of carcinogens such as nitrosamines in the body. It can also absorb radioactive substances to achieve the effect of radiation protection (Chen Rui. (2004). Chemical composition and application of tea funct...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/16A23F3/10A23F3/40C12R1/645
Inventor 冯磊赵江源李铭刚文孟良梁宇鹏
Owner 冯磊
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