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A method for fermenting tobacco with mixed bacteria

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

Active Publication Date: 2022-02-15
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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  • A method for fermenting tobacco with mixed bacteria
  • A method for fermenting tobacco with mixed bacteria

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Experimental program
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Effect test

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 38h;

[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 liquid, ...

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 method for fermenting tobacco with mixed bacteria, which comprises the following steps: inoculating Pichia pastoris FBKL2.0008 and Issa orientalis FBKL2.0002 strains on a slant medium, culturing at 28°C for 24h-48h , until the bacteria are full of the test tube; the strains cultivated on the slant are respectively activated by streaking on the malt juice agar medium, and cultivated at 28°C for 24h-48h; seed liquid preparation: the yeast seed liquid of Pichia pastoris obtained from the cultivation Mix it with the Issaccharomyces orientalis seed liquid to obtain a mixed bacterial liquid, and the Pichia seed liquid accounts for 60%-80% of the mixed bacterial liquid; add the mixed bacterial liquid to tobacco at 0.4ml / g, and add aseptic Water 0.2ml / g tobacco, cultured at 28-30°C for 15-20d. Dry at a low temperature of 40-60°C for 15-30 minutes. The invention has rose fragrance, powder fragrance, honey-like fragrance and wine fragrance, has simple process, is not easy to pollute, and has wide 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 redu...

Claims

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Application Information

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Patent Type & Authority Patents(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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