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Bacterium-enzyme combined preparation containing bacillussubtilis strain xp and application of bacterium-enzyme combined preparation in accelerating starch degradation in tobacco stem

A combination technology of Bacillus subtilis and bacterial enzymes, applied in the field of microorganisms, can solve the problems of poor alcoholization effect, large error, and easy mildew of tobacco stems, etc.

Active Publication Date: 2014-04-30
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 stems 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 alcoholization is better, there are still some disadvantages, such as: 1) Because the tobacco stems are rich in nutrients and have long-term close contact with the external environment, the tobacco stems are prone to insects
2) During the changing seasons of spring, summer, autumn and winter, tobacco stems 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 tobacco cell wall 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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  • Bacterium-enzyme combined preparation containing bacillussubtilis strain xp and application of bacterium-enzyme combined preparation in accelerating starch degradation in tobacco stem
  • Bacterium-enzyme combined preparation containing bacillussubtilis strain xp and application of bacterium-enzyme combined preparation in accelerating starch degradation in tobacco stem
  • Bacterium-enzyme combined preparation containing bacillussubtilis strain xp and application of bacterium-enzyme combined preparation in accelerating starch degradation in tobacco stem

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Preparation of microorganism-enzyme preparation of the present invention and method for measuring enzyme activity

[0042] 1. Preparation of microbial preparations

[0043] 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;

[0044] 2) Seed culture: Take the bacteria obtained in step 1) and inoculate a single colony with an inoculation loop in a 300 ml Erlenmeyer flask containing 100 ml LB liquid medium under aseptic conditions, and culture it on a shaker at 37°C until OD 600 The value is 5.0 (about 10 h, the rotation speed is 200 rpm), and the seed solution is obtained. Collect the seed liquid, put it into a 250 ml bacterial liquid collection bottle, and centrifuge at 4°C, and the centrifugation condition used is 8000 rpm for 20 min. The supernatant was taken for enzyme activity assay. The enzyme activity was determined by the starch-iodine chro...

Embodiment 2

[0064] A bacterium-enzyme joint preparation containing bacillus subtilis strain xp, which is prepared by the following steps:

[0065] 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;

[0066] 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 10 h), and the seed liquid is obtained;

[0067] 3) Mix the seed liquid with amylase, cellulase, laccase and glucoamylase in a volume ratio of 1:1:1:1:1. The enzyme activity concentrations of the amylase, cellulase, laccase and glucoamylase are 8 U / ml, 800 U / ml, 900 U / ml and 100 U / ml respectively.

[0068] 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...

Embodiment 3

[0072] A kind of bacterial enzyme joint preparation containing bacillus subtilis bacterial strain xp, it is made by following steps:

[0073] 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;

[0074] 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 10 h), and the seed liquid is obtained;

[0075] 3) Mix the seed liquid with amylase, cellulase, laccase and glucoamylase in a volume ratio of 1:1:1:1:1. The enzyme activity concentrations of the amylase, cellulase, laccase and glucoamylase are 8 U / ml, 800 U / ml, 900 U / ml and 100 U / ml respectively.

[0076] Add 100ml of the above bacteria-enzyme combination preparation to 1000ml of water soaked with 500g of dried tobacco stems at a temperature...

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Abstract

The invention belongs to the technical field of microorganisms and particularly relates to a bacillussubtilis strain xp producing amylase. A classification designation of the bacillussubtilis strain xp is Bacillussubtilis. The collection number of the strain in China Center for Type Culture Collection (CCTCC) is CCTCC NO: M2011452. The invention further discloses a bacterium-enzyme combined preparation prepared by the strain xp, and an application of the bacterium-enzyme combined preparation in accelerating starch degradation in a tobacco stem.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a bacteria-enzyme joint preparation containing bacillus subtilis strain xp and its application in accelerating starch degradation in tobacco stems. Background technique [0002] Tobacco stems are an important part of tobacco leaves. After being separated by threshing and redrying, tobacco stems accounting for about 25% of the total weight of tobacco leaves can be obtained. The nature of the chemical composition of tobacco stems is basically the same as that of leaves, but there are differences in content, which leads to insufficient sound in the process of burning and smoking the shredded stems made of tobacco stems, flat taste, less irritation and energy, and more miscellaneous gas. Heavy, especially woody miscellaneous gas. At present, the utilization of tobacco stems is mainly to make them into high-filled cut stems, which are used as filling raw materials for ci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/26C12N9/42C12N9/02C12N9/34A24B5/16C12R1/125
Inventor 马宇平周浩张俊岭王墨染张东豫许平
Owner CHINA TOBACCO HENAN IND
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