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Preparation method of bibacterial preparation for producing multiple enzymes and application of bibacterial preparation

An application method and technology of bacterial preparations, which are applied in the field of microorganisms, can solve the problems such as the lack of Bacillus amyloliquefaciens, and achieve the effects of improving food value, strengthening the fermentation process, and improving the quality of tobacco leaves

Inactive Publication Date: 2019-05-24
GUIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no patent application for using Bacillus amyloliquefaciens to improve the quality and flavor of tobacco

Method used

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  • Preparation method of bibacterial preparation for producing multiple enzymes and application of bibacterial preparation
  • Preparation method of bibacterial preparation for producing multiple enzymes and application of bibacterial preparation
  • Preparation method of bibacterial preparation for producing multiple enzymes and application of bibacterial preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] This embodiment is a method for preparing a multi-enzyme-producing double-bacteria preparation of Bacillus amyloliquefaciens GUHP-86 and GZU03, which specifically includes the following steps:

[0023] (1) Prepare and sterilize the seed liquid medium, mix the medium tryptone 1.25g / L, yeast extract 0.625g / L, NaCl 6.25g / L, and glucose 0.625g / L according to the following formula, press triangle 20% of the bottle capacity was divided into conical flasks, sterilized by high-pressure steam at 121°C for 20 minutes, and after cooling, they were respectively inserted into the Bacillus amyloliquefaciens GUHP-86 and GZU03 and cultured at 37°C and 180r / min for 18h±2h. get the seed solution;

[0024] (2) Centrifuge the certain amount of seed liquid at 4°C and 8000r / min for 15min, collect the precipitate and wash the precipitate with sterile physiological saline, then add an equal amount of sterile physiological saline to the precipitate, shake well, Obtain the single preparation of...

Embodiment 2

[0027] In this example, 20 g of tobacco leaves pasteurized at 121°C for 5 min were weighed in a sealed bag of 14 cm × 20 cm, and 1 mL of GZU03 and 3 ml of GUHP-86 were absorbed into the tobacco leaves, and 4 mL of sugar-salt solution In the tobacco leaves, mix thoroughly to obtain sample 1.

[0028] The preparation steps of the sugar-salt solution are as follows: Weigh the MgCl 2 5g, KCl 0.5g, CaCl 2 0.5g, 0.5g NaCl, 2g glucose, add water to 1000mL, dissolve to obtain a sugar-salt solution.

[0029] Weigh 20g of tobacco leaves pasteurized at 121°C for 5min into a sealed bag of 14cm×20cm, draw 4mL of sterile saline and 4mL of sugar-salt solution into the tobacco leaves, and mix well to obtain Comparative Sample 1.

[0030] The sample 1 and the control sample 1 were cultivated in the incubator at 37°C at the same time, and the ventilation was carried out regularly every day, and when the humidity decreased, a sugar-salt solution must be added.

[0031] More than 10 sensory ...

Embodiment 3

[0033] In this example, 20 g of tobacco leaves pasteurized at 121 °C for 5 minutes were weighed in a sealed bag of 14 cm × 20 cm, and 2 mL of GZU03 and 2 mL of GUHP-86 were absorbed into the tobacco leaves, and 4 mL of sugar-salt solution In the tobacco leaves, mix thoroughly to obtain sample 2.

[0034] The preparation steps of the sugar-salt solution are as follows: Weigh the MgCl 2 5g, KCl 0.5g, CaCl 2 0.5g, 0.5g NaCl, 2g glucose, add water to 1000mL, dissolve to obtain a sugar-salt solution.

[0035] Weigh 20g of tobacco leaves pasteurized at 121°C for 5min into a sealed bag of size 14cm×20cm, draw 4mL of sterile normal saline and 4mL of sugar-salt solution into the tobacco leaves, and mix well to obtain Comparative Sample 2.

[0036] The sample 2 and the comparison sample 2 were cultivated in the incubator at 37°C at the same time, and the ventilation was carried out regularly every day, and when the humidity decreased, a sugar-salt solution must be added.

[0037] T...

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Abstract

disclosed is a bibacterial preparation for producing multiple enzymes. The bibacterial preparation is prepared from bacillus amyloliquefaciens GUHP-86 and bacillus amyloliquefaciens GZU03. The preparation method includes 1), preparation and sterilization of a seed solution; 2), preparation of single preparations; 3), preparation of the bibacterial preparation for producing the multiple enzymes. In the method, tobacco leaves are put in a sealed bag, two single preparations are sucked up respectively on the tobacco leaves, the total volume of the bibacterial preparation for producing the multiple enzymes is controlled, a sugar and salt solution is applied to the tobacco leaves with even mixing, the tobacco leaves are put in an incubator for cultivation with regular ventilation every day, and the sugar and salt solution is refilled when the humidity drops. The bibacterial preparation has the advantages that nattokinase, protease and amylase activities can be improved during tobacco fermentation, quality of the tobacco leaves is improved, the edible value of snus is greatly improved, and the fermentation process can be strengthened; the method is suitable for improving the quality ofthe snus.

Description

technical field [0001] The present invention relates to microorganisms, and also relates to tobacco, and furthermore, relates to the application method of the preparation made of two kinds of bacilli in tobacco processing. Background technique [0002] Snus is a nicotine-containing smokeless tobacco product made by mixing powdered tobacco with salt, water, and sometimes flavorings such as bergamot, rose oil, or licorice. Snus is an important supplement form of tobacco, usually placed between the lips and gums for consumption, or chewed in the mouth for use. Bagged snus is generally a moist tobacco powder or shredded tobacco product made from air-dried or open-flame flue-cured tobacco through crushing, heat treatment and packaging, and is contained between the lips and gums when used. [0003] The raw material of snus is the main factor that responds to the quality of snus. The raw material of snus has the disadvantages of high irritation, heavy green miscellaneous gas and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A24B15/20C12R1/07
Inventor 何腊平帅瑶陶菡
Owner GUIZHOU UNIV
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