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Mixed-strain tobacco fermentation method

A technology of mixed bacteria fermentation and tobacco, applied in the direction of tobacco, tobacco treatment, application, etc., to achieve the effect of outstanding aroma, low production environment requirements, and improved taste

Active Publication Date: 2018-12-28
GUIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report on the method of directly using liquor-producing aroma-producing yeast to ferment tobacco to prepare tobacco with liquor-style characteristics

Method used

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Embodiment 1

[0017] A method for fermenting tobacco with mixed bacteria, comprising the following steps:

[0018] (1) Slant culture: inoculate Pichia kudriavzevii FBKL2.0008 and Issatchenkia orientalis FBKL2.0002 strains on the slant medium and culture at 28°C 48h, until the bacteria overgrow the test tube;

[0019] (2) Activation of strains: the strains cultured on the slant were streaked and activated on the wort agar medium respectively, and cultured at 28°C for 24 hours;

[0020] (3) Preparation of seed liquid: prepare liquid medium (malt extract powder 130g / L, chloramphenicol 0.1g / L, pH5.8-6.2), fill it in a triangular flask, sterilize at 121°C for 20min, and then put it on a clean bench Insert the activated strains into the liquid medium respectively, and cultivate them on a shaker at 28°C for 48 hours;

[0021] (4) Preparation of mixed bacterial liquid: mixing the cultured yeast seed liquid of Pichia pastoris and I. orientalis seed liquid to obtain a mixed bacterial liquid, and th...

Embodiment 2

[0024] A method for fermenting tobacco with mixed bacteria, comprising the following steps:

[0025] (1) Slant culture: inoculate the Pichia pastoris FBKL2.0008 and Issahia orientalis FBKL2.0002 strains on the slant medium, culture at 28°C for 35 hours, and wait until the bacteria are fully grown in the test tube;

[0026] (2) Activation of strains: the strains cultured on the slant were streaked and activated on the wort agar medium respectively, and cultured at 28°C for 30h;

[0027] (3) Preparation of seed liquid: prepare liquid medium (malt extract powder 130g / L, chloramphenicol 0.1g / L, pH5.8-6.2), fill it in a triangular flask, sterilize at 121°C for 20min, and then put it on a clean bench Insert the activated strains into the liquid medium respectively, and cultivate them on a shaker at 28°C for 38 hours;

[0028] (4) Preparation of mixed bacterial liquid: mixing the yeast seed liquid of Pichia orientalis seed liquid obtained by culturing to obtain a mixed bacterial liq...

Embodiment 3

[0031] A method for fermenting tobacco with mixed bacteria, comprising the following steps:

[0032] (1) Incline culture: Inoculate Pichia kudriavzevii (Pichia kudriavzevii) and Issatchenkia orientalis FBKL2.0002 (Issatchenkia orientalis) strains on the slant medium, and cultivate them at 28°C for 24 hours until the bacteria are fully grown. test tube;

[0033] (2) Activation of strains: the strains cultured on the slant were streaked and activated on the wort agar medium respectively, and cultured at 28°C for 24 hours;

[0034] (3) Preparation of seed liquid: prepare liquid medium (malt extract powder 130g / L, chloramphenicol 0.1g / L, pH5.8-6.2), fill it in a triangular flask, sterilize at 121°C for 20min, and then put it on a clean bench Insert the activated strains into the liquid medium respectively, and cultivate them on a shaker at 28°C for 24 hours;

[0035] (4) Preparation of mixed bacterial liquid: mixing the cultured yeast seed liquid of Pichia pastoris and I. orient...

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Abstract

The invention discloses a mixed-strain tobacco fermentation method. The method comprises the steps that strains that Pichia pastoris FBKL2.0008 and Issatchenkia orientalis FBKL2.0002 are inoculated toa slope culture medium to be cultured for 24 h-48 h at the temperature of 28 DEG C until a test tube is full of strains; slope cultured strains are activated by drawing lines on a beer wort agar culture medium and cultured for 24 h-48 h at the temperature of 28 DEG C; seed liquid is prepared, mixed-strain liquid is obtained by mixing the yeast seed liquid of the Pichia pastoris and the yeast seedliquid of Issatchenkia orientalis which are obtained by culturing, and the yeast seed liquid of the Pichia pastoris takes up 60%-80% of the mixed strain liquid; the mixed-strain liquid is added to tobaccos according to the concentration of 0.4 ml / g, sterilized water with the concentration of 0.2 ml / g is added every 3 days, the tobaccos are cultured for 15-20 days at the temperature of 28 DEG C-30DEG C, drying is conducted for 15-30 minutes at the low temperature of 40-60 DEG C, and the tobaccos are obtained. The tobaccos have rose fragrance, powder fragrance, sweet fragrance and bouquet, andthe method has the advantages of simple process, difficult contamination and wide product application.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for fermenting tobacco with Issaccharomyces orientalis and Pichia pastoris. Background technique [0002] After the tobacco leaves are harvested, they need to be processed in different ways according to their characteristics and processing traditions, including preliminary saccharification, aging, and further processing. Fermentation of the tobacco leaves changes the chemical composition of the tobacco, softening its flavor and reducing its nicotine content. The new processing technology of tobacco has shifted from the improvement of planting technology to the research of tobacco flavor. At present, it is reported that the culture solution prepared by actinomycetes isolated from tobacco leaves is used for root irrigation and leaf spraying treatment. The results prove that in the later stage of air curing The content of NNN was significantly reduced, thereby re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A24B15/20
CPCA24B15/20
Inventor 王晓丹邱树毅李朝云
Owner GUIZHOU UNIV
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