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A bacterial-enzyme combination preparation containing Bacillus subtilis strain xp and its application in accelerating starch degradation in tobacco flakes

A combination technology of Bacillus subtilis and bacterial enzymes, applied in the field of microorganisms, can solve the problems of reagent consumption error, unsatisfactory starch effect, large error, etc., and achieve the effect of improving the grade of flakes

Active Publication Date: 2016-04-27
CHINA TOBACCO HENAN IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The artificial aging speed is fast, but the aging effect is poor; the natural aging is to stack the tobacco leaves in the storage according to certain requirements, and carry out the aging under a certain humidity (sometimes need to control the temperature), and the aging time is 1-3 years
Although the effect of natural aging is better, there are still some disadvantages, such as: 1) Because tobacco leaves are rich in nutrients and have long-term close contact with the external environment, tobacco leaves are prone to insects
2) During the changing seasons of spring, summer, autumn and winter, tobacco leaves are prone to mildew due to large changes in temperature and humidity
Among them, the iodine chromogenic method uses directly boiling the tobacco powder and then measuring its filtrate. The method is relatively simple, but the effect of simple heating will destroy the cell wall of the tobacco leaf and expose the starch, so the measured value is relatively small and the error is large.
Fehling's reagent method and phenol method are used to indirectly measure the content of reducing sugars in starch production products. They are not used due to time-consuming, reagent-consuming and large errors.

Method used

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  • A bacterial-enzyme combination preparation containing Bacillus subtilis strain xp and its application in accelerating starch degradation in tobacco flakes
  • A bacterial-enzyme combination preparation containing Bacillus subtilis strain xp and its application in accelerating starch degradation in tobacco flakes
  • A bacterial-enzyme combination preparation containing Bacillus subtilis strain xp and its application in accelerating starch degradation in tobacco flakes

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Experimental program
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Effect test

Embodiment 1

[0038] Preparation of microorganism-enzyme preparation of the present invention and method for measuring enzyme activity

[0039] 1. Preparation of microbial preparations

[0040] 1) Slant culture: suspend the strain xp in 0.5ml LB liquid medium, then pick up the bacterial liquid with an inoculation loop and culture it on a slant at 37°C for 14 hours;

[0041] 2) Seed culture: Take the bacteria obtained in step 1) and inoculate a single colony with an inoculation loop in a 300ml Erlenmeyer flask containing 100ml LB liquid medium under aseptic conditions, and culture it on a shaker at 37°C until OD 600 The value is 5.0 (about 10h, the rotation speed is 200rpm), and the seed solution is obtained. Collect the seed solution, put it into a 250ml bacterial solution collection bottle, and centrifuge at 4°C, the centrifugation condition used is 8000rpm, 20min. The supernatant was taken for enzyme activity assay. The enzyme activity was determined by the starch-iodine chromogenic me...

Embodiment 2

[0060] A bacterium-enzyme joint preparation containing bacillus subtilis strain xp, which is prepared by the following steps:

[0061] 1) Slant culture: suspend the strain xp in LB liquid medium, then pick up the bacterial liquid with an inoculation loop and culture it on a slant at 37°C for 14 hours;

[0062] 2) Seed culture: Take the bacteria obtained in step 1) and pick a single colony with an inoculation loop under sterile conditions to inoculate in LB liquid culture based on a shaker at 37°C to OD 600 The value is 5.0 (about 10h), and the seed solution is obtained;

[0063] 3) Mix the seed liquid with amylase, cellulase and glucoamylase in a volume ratio of 1:1:1:1. The enzyme activity concentrations of the amylase, cellulase and glucoamylase are 8U / ml, 800U / ml and 100U / ml respectively.

[0064] At a temperature of 30°C and a humidity of 60%, 100ml of the above bacteria-enzyme joint preparation was evenly sprayed on the surface of 500g of tobacco flakes, and samples wer...

Embodiment 3

[0069] A bacterium-enzyme joint preparation containing bacillus subtilis strain xp, which is prepared by the following steps:

[0070] 1) Slant culture: suspend the strain xp in LB liquid medium, then pick up the bacterial liquid with an inoculation loop and culture it on a slant at 37°C for 14 hours;

[0071] 2) Seed culture: Take the bacteria obtained in step 1) and pick a single colony with an inoculation loop under sterile conditions to inoculate in LB liquid culture based on a shaker at 37°C to OD 600 The value is 5.0 (about 10h), and the seed solution is obtained;

[0072] 3) Mix the seed liquid with amylase, cellulase and glucoamylase in a volume ratio of 1:1:1:1. The enzyme activity concentrations of the amylase, cellulase and glucoamylase are 8U / ml, 800U / ml and 100U / ml respectively.

[0073] At a temperature of 30°C, 100ml of the above bacteria-enzyme joint preparation was applied to 10L of the water-soluble extract solution (pH value of natural, about 6.0) of the p...

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Abstract

The invention belongs to the technical field of microorganisms, and in particular relates to an amylase-producing Bacillus subtilis strain xp, which is classified as Bacillus? subtilis , the strain's preservation number in the China Center for Type Culture Collection is: CCTCC? No: M2011452. The invention also discloses a bacteria-enzyme joint preparation prepared by using the strain xp and its application in accelerating starch degradation in tobacco flakes.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a bacteria-enzyme joint preparation containing bacillus subtilis strain xp and its application in accelerating starch degradation in tobacco flakes. Background technique [0002] Tobacco flakes, also known as reconstituted tobacco leaves or homogeneous tobacco leaves, are comprehensively utilized and finely processed products in the tobacco industry, mainly composed of tobacco powder, chips, tobacco stems or low-grade tobacco leaves with adhesives and other additives. In fact, all tobacco substances can be used in the production of tobacco sheets. The discarded tobacco stems, stem sticks, fragments, and some low-grade tobacco leaves and short stems in the entire cigarette production process are processed by different methods to make them with properties close to or even The tobacco sheet is superior to natural tobacco leaves, and the manufactured tobacco sheet ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/26C12N9/42C12N9/34A24B15/20C12R1/125
Inventor 马宇平周浩王墨染宋金勇彭玉富唐鸿志
Owner CHINA TOBACCO HENAN IND
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