CN102286403A - Bacillus pumilus.Van35 preparation for treating cut tobacco leaves - Google Patents

Abstract

The invention discloses a tobacco microbial preparation and a preparation method and use thereof and belongs to the technical field of tobacco additives. In the invention, the corresponding microbial preparation is obtained by liquid fermentation culture of a productive strain. The productive strain is bacillus pumilus.Van35 preparation. The strain was collected in China General Microbiological Culture Collection Center on November 5, 2009, and the collection number is CGMCC No.3411. The microbial preparation can obviously increase smoke, pleasure and other qualities, reduce mixed smells, irritation, smoke concentration and hardness and the like and make aftertaste comfortable, thereby improving the smoking quality of the cut tobacco leaves.

Description

A kind of bacillus pumilus preparation that is used for the pipe tobacco processing

Technical field

The invention belongs to the technical field of cigarette additive, more particularly, the present invention relates to a kind of microbial preparation.Also relate to the preparation method of described microbial preparation and the application in tobacco industry thereof simultaneously.

Background technology

Tobacco is one of mainstay of China's finance.State Tobacco Monopoly Bureau spells out the 7th of Eleventh Five-Year Plan Chinese cigarette development in science and technology outline, " capture the gordian technique that influences tobacco incense makings, perfume quantity ", dwindles and external quality of tobacco gap, to improve the domestic and international market competitive power.

Main component in the tobacco is carbohydrate and nitrogenous compound.The former mainly comprises cell wall constituents such as carbohydrate, starch and Mierocrystalline cellulose, xylogen, hemicellulose, pectin, and the latter mainly comprises nicotine, protein etc., and this class macromolecular complex confrontation tobacco interior quality and local flavor all have disadvantageous effect.Microorganism passes through growth and breeding, can produce the enzyme of many biologically actives, quicken the degraded of macromolecule substrate and the decomposition of some objectionable impurities, and form a series of useful micromolecular compounds by absorption, utilization, the conversion of specified microorganisms, tobacco component is changed, finally cause the change of cigarette quality.

Existing more at home and abroad at present with the research report of biotechnology applications in cigarette industry.RayF.Dawson studies show that, the thermophilic actinomyces can make the ageing tobacco leaf produce a kind of interesting fragrance, especially fruit fragrance and wine perfume (or spice) etc.Reid etc. studies show that the genus bacillus in the tobacco leaf, thermophilic bacterium and micrococcus are useful and necessary to the fragrance and the quality of tobacco leaf.Koller has carried out the microbial inoculant tobacco leaf the earliest to increase tabacco fragrance, to improve the trial of tobacco leaf smoking quality, and researcher has begun the research of microbial inoculant tobacco leaf and obtained a large amount of achievements afterwards.People such as Izquierdo Tamayoa discover that with the tobacco leaf of microbial inoculant, fragrance and proterties all improve, and especially when adopting micrococcus sp or Bacillaceae or both mixtures inoculation tobacco leaf, effect is more remarkable.China just carries out the research of tabacco fragrance material chemical composition analysis up to the nineties in 20th century, lures fragrant microbe research also to be in the exploratory stage about tobacco leaf.Xie He etc. studies show that, the genus bacillus of tobacco leaf surface is inoculated on the cured tobacco leaf, can obviously improve tabacco fragrance, improve quality of tobacco.The dominant bacteria on flue-cured tobacco blade face has been identified in the research of Chen Fu magnitude, and shows that handling tobacco leaf with dominant bacteria can shorten fermentation time and improve quality of tobacco.

Up to now, the research of using in cigarette industry about microorganism focuses mostly on and reduces tobacco leaf in the specific function that utilizes tobacco leaf self or some environmental microorganism and stimulate, reduce nicotine content or improve physical and chemical quality, attempt to reach the purpose of quickening tobacco ageing, and the using microbe preparation is directly handled pipe tobacco with the report that improves the cigarette smoking quality seldom.By literature search, do not find the report identical as yet with content of the present invention.

Summary of the invention

The objective of the invention is to deficiency, a kind of new microbial preparation is provided at existing pipe tobacco treatment technology.

Another object of the present invention provides the preparation method of described microbial preparation.

Further aim of the present invention provides the application of described microbial preparation in pipe tobacco is handled.

Purpose of the present invention is achieved by following technical proposals.

* except as otherwise noted, percentage ratio of the present invention is mass percent.

A. the invention provides a kind of new microbial preparation, this microbial preparation is to adopt following method to prepare: get the liquid spawn that obtains bacterium Van35 from the separation of flue-cured tobacco blade face, inoculum size according to 0.1% is inoculated in the LB liquid nutrient medium of 200ml, at 37 ℃ of shaking tables, rotating speed 180r/min shaking culture is to OD value=2.0; The centrifugal 10min of 3500rpm/min uses the sterilized water washing precipitation, and is centrifugal, and at last with the vibration of 20mL sterilized water evenly, dilution is required microbial preparation for 10 times during use; Wherein, the liquid spawn of described bacterium Van35 belongs to bacillus pumilus through evaluation, this bacterial strain called after Bacillus pumilus.

B. the invention provides a kind of preparation method of described microbial preparation, this method adopts following step:

(1) separates the liquid spawn that obtains bacterium Van35 on the flue-cured tobacco blade face;

(2) get liquid spawn, the inoculum size according to 0.1% is inoculated in the LB liquid nutrient medium of 200ml;

(3) at 37 ℃ of shaking tables, rotating speed 180r/min shaking culture is to OD value=2.0; The centrifugal 10min of 3500rpm/min;

(4) use the sterilized water washing precipitation, centrifugal, at last with the vibration of 20mL sterilized water evenly, dilution is required microbial inoculum for 10 times during use.

C. the invention provides the application of described microbial preparation in pipe tobacco is handled.

Compared with prior art, the present invention has following outstanding advantage:

(1) microbial preparation provided by the invention can obviously improve the qualities such as perfume quantity, fragrance of pipe tobacco, declines such as assorted gas, pungency, and flue gas concentration and strength reduce, and it is comfortable that pleasant impression becomes.

(2) cost of the present invention is low, controllability is strong, simple to operate, be convenient to realize large-scale industrial production, have the excellent development application prospect.

The explanation of preservation biomaterial

The bacterial strain of the bacillus pumilus Van35 that the present invention relates to has been kept at China Committee for Culture Collection of Microorganisms common micro-organisms center; Address: China. No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City; Preservation date: on November 5th, 2009; Preserving number is CGMCC No.3411.

Embodiment

In order to make purpose of the present invention, technical scheme and advantage clearer, reach testing data by the following examples, the present invention is described in more detail.Should be appreciated that specific embodiment described herein only in order to explanation the present invention, and be not intended to limit the scope of the invention.

Embodiment 1

---the test tube slant is cultivated

Adopt Luria-Bertani (LB) substratum, filling a prescription is: peptone 10g, and sodium-chlor 10g, yeast extract 5g, agar 20g, the distilled water constant volume is to 1000ml.Sterilized 25 minutes, and put into the inclined-plane for 121 ℃.Inoculation Van35 cultivates 24h for 28 ℃, obtains the test tube kind.

Embodiment 2

---seed culture

Adopt Luria-Bertani (LB) substratum, filling a prescription is: peptone 10g, and sodium-chlor 10g, yeast extract 5g, the distilled water constant volume is to 1000ml.Sterilized 25 minutes for 121 ℃.Inoculation Van35 test tube kind is cultivated 12h for 28 ℃, obtains liquid spawn.

Embodiment 3

---the preparation microbial preparation

Get liquid spawn, the inoculum size according to 0.1% is inoculated in the LB liquid nutrient medium of 200ml, and at 37 ℃ of shaking tables, rotating speed 180r/min shaking culture is to OD value=2.0.The centrifugal 10min of 3500rpm/min uses the sterilized water washing precipitation, and is centrifugal, and then with the vibration of 20mL sterilized water evenly, dilution is microbial inoculum for 10 times during use.

Application Example

---microbial preparation is to the effect of pipe tobacco

With the microbial preparation for preparing pipe tobacco is handled, concrete experimental procedure is as follows:

1) takes by weighing the pipe tobacco 100g of water ratio at 11%-13%.

2) get microbial inoculum 20ml, with miniaturised nebuliser it is sprayed onto on the 100g pipe tobacco equably, sealing divides evaporation to prevent water.

3) sterilized water that directly sprays equivalent in contrast.

4) sample is enclosed sealing bag, divide evaporation, 30 ℃ of constant temperature 6h to prevent water.

5) pipe tobacco is placed 120 ℃ of baking ovens dry, make water ratio reach 11%-13%.

6) with pipe tobacco be 60% climatic chamber balance 48h in 22 ℃ of relative humidity.

7) take out the part pipe tobacco, detect main chemical compositions content and flavor matter.

8) remaining pipe tobacco is rolled into cigarette by hand and prop up, sample and contrast random number, the group of smokeing panel test that is made up of the expert that smokes panel test carries out the sensory evaluating smoking.

The detected result of routine chemical components shows: after described microbial preparation is handled, bigger change has taken place in the content of the biomacromolecule compound of pipe tobacco, wherein tangible raising has taken place in reducing sugar and sugar contents, decline has taken place in the content of total nitrogen, Mierocrystalline cellulose and nicotine, volatilization alkali and ligroin extraction total amount descend to some extent, and the variation of the content of these compounds helps the sucking quality of tobacco leaf.

The aroma component detected result shows: after microbial preparation is handled, the aroma component of pipe tobacco comprises that the content of furfuryl alcohol, vanillin food grade,1000.000000ine mesh, dihydroactinidiolide, phenylcarbinol, solanone, Megastigmatrienone, dorinone, western cypress three enediols and neophytadiene etc. significantly improves than contrast.

Sensory evaluating smoking result shows: pipe tobacco is after microbial preparation is handled, and perfume quantity, fragrance contrast significantly promote, declines such as assorted gas, pungency, and flue gas concentration and strength reduce, and it is comfortable that pleasant impression becomes.

Test-results shows that the microbial preparation of this invention has obvious promotion pipe tobacco chemical composition change, thereby reduces pipe tobacco pungency, fine and smooth pure and mild flue gas, can reach the purpose that improves the pipe tobacco suction quality.

Claims (3)

1. microbial preparation, it is characterized in that, this microbial preparation is to adopt following method to prepare: get that to separate the liquid spawn that obtains bacterium Van35 from the flue-cured tobacco blade face be raw material, inoculum size according to 0.1% is inoculated in the LB liquid nutrient medium of 200ml, at 37 ℃ of shaking tables, rotating speed 180r/min shaking culture is to OD value=2.0; The centrifugal 10min of 3500rpm/min uses the sterilized water washing precipitation, and is centrifugal, and then with the vibration of 20mL sterilized water evenly, dilution is required microbial preparation for 10 times during use; Wherein, the liquid spawn of described bacterium Van35 belongs to bacillus pumilus through evaluation, this bacterial strain called after Bacillus pumilus.

2. the preparation method of the described microbial preparation of claim 1, this method adopts following step:

(1) separates the liquid spawn that obtains bacterium Van35 on the flue-cured tobacco blade face;

(2) get liquid spawn, the inoculum size according to 0.1% is inoculated in the LB liquid nutrient medium of 200ml;

(3) at 37 ℃ of shaking tables, rotating speed 180r/min shaking culture is to OD value=2.0; The centrifugal 10min of 3500rpm/min;

(4) use the sterilized water washing precipitation, centrifugal, at last with the vibration of 20mL sterilized water evenly, dilution is required microbial inoculum for 10 times during use.

3. the application of the described microbial preparation of claim 1 in pipe tobacco is handled.