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EP0024152B1 - Procédé en continu pour la dénitrification d'extraits de tabac - Google Patents

Procédé en continu pour la dénitrification d'extraits de tabac Download PDF

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Publication number
EP0024152B1
EP0024152B1 EP19800302646 EP80302646A EP0024152B1 EP 0024152 B1 EP0024152 B1 EP 0024152B1 EP 19800302646 EP19800302646 EP 19800302646 EP 80302646 A EP80302646 A EP 80302646A EP 0024152 B1 EP0024152 B1 EP 0024152B1
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EP
European Patent Office
Prior art keywords
added
mixture
extract
tobacco
microorganisms
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired
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EP19800302646
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German (de)
English (en)
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EP0024152A2 (fr
EP0024152A3 (en
Inventor
Helmut Gaisch
Beth Krasna
Dieter Schulthess
Claude Ruf
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Philip Morris Products SA
Original Assignee
Fabriques de Tabac Reunies SA
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Publication date
Priority claimed from LU81611A external-priority patent/LU81611A1/de
Priority claimed from LU82199A external-priority patent/LU82199A1/de
Priority claimed from US06/156,910 external-priority patent/US4622982A/en
Application filed by Fabriques de Tabac Reunies SA filed Critical Fabriques de Tabac Reunies SA
Publication of EP0024152A2 publication Critical patent/EP0024152A2/fr
Publication of EP0024152A3 publication Critical patent/EP0024152A3/en
Application granted granted Critical
Publication of EP0024152B1 publication Critical patent/EP0024152B1/fr
Expired legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
    • A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
    • A24B15/00—Chemical features or treatment of tobacco; Tobacco substitutes, e.g. in liquid form
    • A24B15/18—Treatment of tobacco products or tobacco substitutes
    • A24B15/20—Biochemical treatment

Definitions

  • This invention relates to a continuous process for reducing the levels of certain nitrogen-containing compounds, notably nitrates, nitrites and ammonium compounds present in tobacco materials using microorganisms.
  • the DE-A-2816427 describes a process for microbial degradation of nitrate, nitrite and other nitrogen containing compounds under nitrogen deficiency or oxygen deficiency conditions.
  • the microorganisms may be selected from the genus Aerobacter, Pseudomonas, Micrococcus or Escherichia, with Enterobacter aerogenes being specifically employed in the examples, and obtain their nitrogen or oxygen requirements respectively from nitrate or nitrite degradation. More specifically, in this latter process, an extract of the tobacco is mixed with a culture of the microorganisms and necessary nutrient additives to form a work mixture, and the mixture is maintained under conditions in which the organisms are in their exponential growth phase.
  • EP-A-0 005 082 (published October 31, 1979) describes a process for microbial reduction of nitrates in tobacco by dissimilatory denitrification whereby nitrogen gas is the end product.
  • the microorganism specifically suggested for use in the process is Paracoccus denitrificans (or Micrococcus denitrificans). Species of the genera Pseudomonas, Alcaligenes, Bacillus and Propionibacterium can also be employed.
  • US-A-3,845,774 to Tso et al. describes a tobacco curing treatment method wherein the tobacco is homogenised and incubated during curing in order to regulate the composition of the final product. Nitrate-nitrogen and total nitrogen are reduced somewhat, but the amount of reduction is not as significant as that of the present process. No specific methods or means for reducing nitrate-nitrogen are suggested.
  • Gravely et al., US-A-3,747,608 relates to a method for aerobic microbial digestion of pectin- bound plant material, specifically tobacco materials, using pectolytic enzyme-producing microorganisms.
  • Examples 11 and 13 disclose data related to concomitant denitration of tobacco using the microorganism Erwinia carotovora, ATCC 495.
  • This microorganism is unsuitable for use in the present invention since pectolytic enzyme-producing microorganisms, such as Erwinia carotovora, destroy the structural integrity of the tobacco.
  • Denitration by means of microorganisms is also known outside the tobacco arts. Representative examples are US-A-3,709,364 to Savage, US-A-3,829,377 to Hashimoto, US-A-4,039,438 to Anderson, and US-A-4,043,936 to Francis et al. which describe denitrification of waste water using anaerobic bacteria to reduce the nitrate to nitrogen gas. Members of the Thiobacillus, Pseudomonas, Chromobacter, Bacillus and Chlostridium genera are among the microorganisms which may be employed.
  • Microorganisms have also been used to modify other tobacco components.
  • US-A-4,037,609 and US­A­4,O38,993 to Geiss et al. disclose methods for reducing the nicotine content of tobacco by microbial treatment using microorganisms obtained from tobacco, including Pseudomonas putida and Cellulomonas sp. Aerobic fermentation techniques are employed wherein nicotine is degraded via microbial action to 3-succinoylpyridine.
  • the latter microorganism is capable of reducing nitrate to nitrite and actively produces nitrogen gas.
  • US-A-2,149,179 relates to an accelerated aging method for tobacco wherein the aging is effected by means of fermentation with exclusion of oxygen employing microorganisms capable of growing in the absence of oxygen.
  • the microorganisms may be those which are bred on noble tobaccos or anaerobic yeasts.
  • fermentation times of only days, rather than months are required.
  • the purpose of the claimed fermentation process is to improve the bouquet of the tobacco. Nicotine content in the tobacco is also reduced.
  • a prior process of Suchsland which used microorganisms to decompose complex organic substances in tobacco into simpler compounds, did not prove practical since the oxidation effected by oxygen during the fermentation was ignored.
  • the present process provides a method whereby nitrates, nitrites and ammonium compounds can be efficiently eliminated from tobacco via an assimilatory metabolic process on a large, technical scale under economical conditions, with a minimal requirement of manpower or energy and minimal addition to or transformations of the tobacco extract components, other than such denitration.
  • the invention is characterised in that sterile tobacco extract and additives are continuously added to the work mixture at a dilution rate which does not exceed the growth rate of the microorganisms, while the pH, temperature and aeration are maintained at levels which promote aerobic assimilation, the extract, phosphate and a carbon source being added in amounts such that the overall addition thereof is 0.1-7.5 g nitrate ions per liter added, 1.0 to 10 g P0 4 ions per liter added and sufficient carbon source to provide at least 16.5 assimilative carbon atoms/N0 3 ion added, and that work mixture is continuously withdrawn at such a rate as to keep the total volume of work mixture constant.
  • microrganisms may subsequently be removed from the withdrawn mixture.
  • Preferred microorganisms for use in the present process are Candida yeasts. Denitration with such yeasts may be effected continuously as above described employing a dilution and withdrawal rate of 0.1 to 0.35 liter of additives and extract per liter of work mixture per hour while maintaining a pH of 3.5 to 7.2, a temperature of 25° to 37°C and an aeration rate of 0.8 to 2.5 liters per liter work mixture per minute. Enterobacter aerogenes may also be used in the present process.
  • Denitration with such microorganisms may be effected on a continuous basis employing a dilution and withdrawal rate of 0.1 to 0.25 liter of additives and extract per liter of work mixture per hour while maintaining a pH of 5.5-8, a temperature of 30°-40°C and an aeration rate of 1.0 to 3 liters air per liter work mixture per minute.
  • Figure 1 is a flow diagram of a tobacco denitration system for the extraction of tobacco through the denitration steps of the present invention up to the reapplication of the denitrated extract to the extracted tobacco. It is possible to run a system employing the denitration method of the invention for extended periods with little or no supervision.
  • Continuous assimilatory denitration is effected according to the present invention by introducing material to be treated into a fermentation vessel while withdrawing treated extract from the fermentor at the same rate, such that the overall volume of material in the fementor, that is, the work mixture, remains constant. Moreover, the rate of extract introduction and withdrawal and the process conditions are such that there is no need to repeatedly inoculate the work mixture; rather after a single inoculation a system employing the present denitration process can be run for extended periods without reinoculation.
  • the metabolic pathway employed in assimilatory denitration can be represented as follows: Such assimilatory denitration thus involves the use of nitrate as a nitrogen source to build up cell material.
  • microorganisms capable of assimilatory denitration must be employed.
  • Various yeasts are capable of nitrate assimilation.
  • the Candida yeasts the Candida utilis NCYC 707, 321 and 359 strains, the Candida utilis DSM 70167 strain, which is the same as the NCYC 359 strain, and the Candida berthetii CBS 5452 strain have been found particularly effective in the practice of the present invention.
  • Candida yeasts and Enterobacter aerogenes ATCC 13048 are characterized in Tables I ⁇ III.
  • the work mixture comprises suitable microorganisms in tobacco extract under conditions which promote aerobic assimilation of nitrogen-containing compounds.
  • suitable microorganisms in tobacco extract under conditions which promote aerobic assimilation of nitrogen-containing compounds.
  • 30-100 g of starter culture mass of microorganisms are inoculated per liter of tobacco extract under conditions favorable to aerobic assimilation.
  • a starter culture which is in exponential growth phase, and preferably late exponential growth phase, and which has been pregrown on tobacco extract is employed.
  • Such a starter culture of Candida yeast can be prepared for example by inoculating tobacco extract with 2 loops of yeast, incubating the inoculated yeast for 9 hours, employing 10 ml of the resultant solution as an inoculum for 200 ml of fresh extract substrate and thereafter incubating for 15 hours.
  • the amount of such a starter culture used to inoculate the work mixture is sufficient to produce a final concentration of culture in mixture of at least O.5­-1 %.
  • Candida yeasts are an aeration rate of 0.8 to 2.5 liters air/liters work mixture/minute, a pH value within the range of 3.5 to 7.2 and a temperature at a point between 25° and 37°C which is favorable for a large proportion of nitrate elimination relative to carbon added plus adequate agitation.
  • a sterile additive mixture made up of tobacco extract and additives is added to the work mixture at a dilution rate which does not exceed the growth rate of the microorganisms employed.
  • This dilution rate is measured as liters of additive solution per liters work mixture per hour. Generally, dilution rates of 0.1 to 0.35 I/l/hr are acceptable. Sterilization of the additive mixture can be accomplished by heating.
  • the additive mixture may be added as a single solution containing the tobacco extract and additives or the individual materials may be separately introduced into the work mixture.
  • Overall the total additions to the work mixture comprise 0.1 to 7.5 grams nitrate/liter of total additive mixture depending on the microorganism employed and 1.0 to 10 grams phosphate/liter of total additive mixture, as well as a carbon source at a concentration sufficient to provide at least 16.5 assimilative carbon atoms per molecule of nitrate added.
  • a portion of the work mixture is continuously removed at a rate such as to keep the volume of the work mixture constant.
  • the withdrawn work mixture may be further treated to remove the biomass therefrom, that is, the microorganisms are removed, whereby denitrated extract is obtained which is of substantially the same composition as the original extract except for the removal of the nitrate.
  • the starter culture it is preferable to allow the starter culture to reach the exponential growth phase in the work mixture prior to commencing continuous operation of the process.
  • the process can be operated on a continuous basis with maintainance and regulation of conditions for the effect desired with little or no supervision.
  • various treatment phases are required in a noncontinuing, charged, so called batch process and if mistakes are made with a batch process, new conditions for assimilation have to be achieved, which could take hours. Production by batch techniques is, thus, costlier and requires more personnel than with the process described in the invention.
  • the aqueous tobacco extract employed in the process of the invention may be obtained in a conventional manner.
  • One method comprises contacting tobacco with water in a 1:10 ratio, commonly at elevated temperature.
  • the insoluble tobacco residue is thereupon separated from the aqueous extract by suitable solid/liquid separation techniques, such as centrifugation, pressing or the like.
  • the insoluble residue may then be dried or subjected to reconstitution. If necessary, the concentration of nitrate is adjusted for addition to the work mixture by evaporation or dilution of the tobacco extract.
  • the additive mixture must contain phosphates, as well as a carbon source, in amounts sufficient for cell growth and total nitrate absorption.
  • a carbon source typically 1.0 to 10 grams of phosphate/liter of additive mixture and enough carbon source to provide at least 16.5 assimilative carbon atoms/molecule of nitrate added are adequate for additive mixtures containing 3-7.5 grams nitrate.
  • concentrations of carbon source and phosphate are such that they are consumed during assimilation of the nitrate and thus do not reach the final denitrated extract. However, higher concentrations of these materials can be tolerated in the practice of the present invention.
  • the carbon source may be any material that will provide the necessary carbons in an organic form usable by the microorganism to assimilate nitrates, nitrites and the like.
  • Various carbon sources have been found suitable in the practice of the invention.
  • glucose, dextrose monohydrate and beet molasses have proven satisfactory.
  • the carbon may also be derived from the acid employed to adjust the pH of the work mixture, for example, from lactic acid.
  • Candida yeasts glucose, sucrose, maltose, cellobiose, ethanol, glycerin or citrate are all suitable carbon sources.
  • lactose may additionally be employed.
  • the amount of carbon source is preferably kept as close as possible to the threshold, since any excess will remain in the final denitrated extract.
  • the threshold is generally about 20 ⁇ 5 carbons/nitrate molecule. With good aeration, a maximum of about 6.2 g/I N0 3 can be assimilated with a 4% glucose solution. In general, with nitrate levels of 3-7.5 g/I added to the work mixture, a concentration of 2.4-6% glucose is required when employing Candida yeasts.
  • temperature affects the amount of carbon required. At temperatures of about 28-30°C, minimal amounts, i.e., about 16.5 carbons, may be used. Outside this temperature range, the amount of carbon must be increased. Further temperature and growth rate of the microorganisms are directly related. Thus higher temperatures favor increased growth of microorganisms. However, increases in temperature also increase the fermentation rate, with resultant alcohol formation rather than growth. The rate of fermentation may be checked by measuring ethanol formation during the process. Temperatures which minimize fermentation while maximizing growth are thus preferred. In the case of Candida utilis 30°C is the preferred temperature.
  • acids and/or bases are employed in the work mixture.
  • Orthophosphoric acid and/or potassium hydroxide are preferred for this purpose.
  • the agent employed to adjust pH may also be the phosphate or carbon source, for example phosphoric, lactic or citric acid or mixtures thereof. Thus, where phosphoric acid is employed to regulate pH, no other addition of phosphate in the additive mixture is required.
  • Aeration of the work mixture is generally at a rate which is sufficient to avoid fermentation while favoring assimilation. Generally, a rate of 0.8 liters air per liter work mixture per minute is the threshold aeration rate required to avoid fermentation where maximum carbon levels are employed. Overall, aeration rates of between 0.5 and 2.5 are suitable in the practice of the process, with rates of 1.0 to 2.0 being particularly effective.
  • a denitration system employing the process of the invention is depicted in the flow diagram of Figure 1.
  • the reference numbers refer to the denitration stages as follows:
  • the material to be treated for example, tobacco stems, is added from tobacco supply 1 and mixed with water from water tank 2 in washer 4.
  • the soluble components are separated from the insoluble tobacco residue.
  • the insoluble residue is passed to drier 23 or reconstitution stage 22.
  • the extracted soluble components are conveyed to 6 where sterilization by heating and thereafter cooling takes place.
  • the treatment in sterilization sections 6 and 7 may consist of preheating to 100°C, sterilization of 110°C for over 40 minutes and cooling to 30°C.
  • fermentor 10 may contain a work mixture, comprising the product solution with the necessary additives and an inoculum of the desired microorganism, from which all the nitrates, nitrites and ammonium compounds will generally be eliminated after about 8-20 hours, whereupon the continuing process can be started by the dosage pump 9 at the rate desired for dilution and regulated in such a way as to keep the volume of the work mixture in fermentor 10 constant.
  • the working conditions are regulated in such a way as to totally eliminate nitrates, nitrites and ammonium compounds contained in the product solution and to completely use all additives from mixer 5 during this assimilation.
  • the treated work mixture in fermentor 10 is removed.
  • the biomass A is removed from the treated work mixture in centrifuge 13 and may be saved for further usage. If necessary, the treated work mixture may be pasteurized in section 12 as shown in Figure 1 or pasteurization of the liquid portion resulting from biomass removal in section 13 may be effected.
  • the remaining treated liquid is conveyed to treatment section 14 as final solution for concentration, as by evaporation.
  • This final solution containing most of the components of the product solution, except the nitrates, nitrites and ammonium compounds may now be used in any way.
  • the solution may for example, be sprayed onto the dried or reconstituted tobacco residue with device 15 for readding materials.
  • the reconstituted product B resembles tobacco sheets.
  • the present invention comprises extracting tobacco with water employing a 10:1 water to tobacco ratio at 90°C for 60 minutes.
  • the extract thus formed is separated from the insoluble tobacco residue.
  • the nitrate concentration in the extract is adjusted to the desired level by conventional means such as dilution or evaporation.
  • the extract at a dilution of 3 to 7.5 g N0 3/ liters, preferably 4.5-5.5 g/I and most preferably 5 g/I, is thereupon combined with sufficient K 2 HP0 4 to give a phosphate concentration of 1.1-1.5, preferably 1.25, and glucose is added to a concentration of 4%, along with 250 ppm antifoam, such as Paracum 24/sw.
  • the pH is adjusted to 5.5 employing KOH.
  • the mixture may then be sterilized at 110°C for forty minutes. Alternatively, the extract and additives may be separately sterilized prior to mixing.
  • the sterilized extract solution containing the additives is thereupon introduced into a fermentation vessel containing a work mixture at a rate of 0.18-0.22 I/I/hr, preferably 0.2 I/I/hr.
  • the work mixture contains a suitable microorganism and is preferably a starter culture of Candida utilis NCYC 707 yeast in exponential, most preferably late exponential, growth phase which has been built up as above described.
  • the pH of the work mixture is maintained at about 5.5 ⁇ 0.3 preferably by addition of a mixture of 9 parts lactic acid to 1 part o-phosphoric acid and/or KOH.
  • the temperature of the mixture is maintained at 30 ⁇ 3°C.
  • the vessel containing the work mixture is aerated at a rate of 1.4-1.6 and preferably 1.5 I/I/min.
  • a portion of the work mixture i.e., treated extract is withdrawn from the fermentation vessel so that the volume of work mixture remains constant.
  • the treated extract is thereupon pasteurized, separated from the biomass and concentrated.
  • the thus denitrated, concentrated extract may then be applied to the dried and/or reconstituted insoluble tobacco residue.
  • the process of the invention has been practiced continuously for five weeks with production of 2400 liters denitrated extract per day which is equal to one-fifth of the volume of the fermentor employed per hour, i.e., 100 liters fermented denitrated extract per hour.
  • Enterobacter aerogenes ATCC 13048 or other bacterium
  • the conditions of the work mixture are adjusted to a pH of 5.5-8.0, preferably 7.0, and a temperature of 30°-40°C, preferably 37°C, and the process is operated at an aeration rate of 1.0-3, preferbaly 2 I/I/min., a dilution rate of 0.1-0.25, preferably 0.2 I/I/hr., with the addition mixture containing 0.1-7.5 g nitrate/I, preferably 5 g/I.
  • the invention is preferably used in treatment of tobacco extracts, but is not limited to that usage. Elimination of nitrates, nitrates and ammonium compounds from foods and other consumer items may also be desirable. Where these materials are in liquid form, they may be used as the nitrate solution for treatment in the practice of the invention. Otherwise an aqueous solution can be obtained by washing, which solution, following denitration, may be recombined with the insoluble fraction of the material to form the final denitrated product.
  • the work conditions can be gauged by the nitrate concentration of the product solution.
  • concentration of the sum of all compounds to be eliminated i.e., nitrates and nitrites and ammonium compounds should be considered.
  • This total concentration of these materials in the overall additive mixture should be between 3 and 7.5 g/liter.
  • the remaining parameters may be the same as in treatment of tobacco.
  • Tobacco stems were extracted with water and the resultant extracts were treated with Candida utilis NCYC 707 according to the process of the invention using the conditions specified in Table IV.
  • the results are set forth in Table IV. "0" indicates an amount, which is not detectable using normal analysis conditions; it is smaller than 10 ppm in the case of carbon and phosphate and is less than 1 ppm for nitrates, nitrites and ammonium compounds.
  • Seven tobacco extracts were prepared by separately washing tobacco stems and by-products with water at a 1:10 tobacco to water ratio and combining a stem extract with a by-product extract. With appropriate installations, it would be possible to effect the extraction, as well as the denitration, on a continuous basis.
  • the nitrate levels of each tobacco extract are set forth in Table V. To each tobacco extract were added glucose, KH 2 p o 4 and Paracum 24/sw antifoam as indicated in Table V. the pH was adjusted to 5.5 with KOH. The extract and additives were sterilized at 110°C for 40 min.
  • the work mixture comprised Candida Utilis 707, lactic acid, KOH and Paracum-24/sw and had a pH of 5.5.
  • the 14 I fermentor which was employed, was equipped with an electromagnetic valve on the incoming air coupled with a timer. The incoming air was shut off for 1 min. every two hours to thus purge the condenser on the outgoing air.
  • the biomass was removed from the denitrated extract by centrifugation.
  • the resulting 1478 g of extract containing 3% tobacco solubles was thereupon concentrated to give an average concentration of 39.06 tobacco solubles and reapplied to dried tobacco stems.
  • the stems were continuously washed in a screw extractor at a stem to water ratio of 1:10.
  • the extraction was carried out at 90°C and the tobacco extract (out of the extractor) to water (into the extractor) ratio was 0.74.
  • the tobacco extract was then sterilized by pumping it through 3 heat exchangers: the first to pre-heat it to 110°C, the second to hold it at that temperature for 40 minutes and the third to cool it down to room temperature. Analytical values are given in Table VIII.
  • a dextrose solution was prepared batchwise, but then continuously pumped through a second line of 3 heat exchangers for sterilization using the above conditions.
  • the two flows i.e., sugar solution and tobacco extract, were then regulated to the desired sugar concentration in the tobacco extract and then pumped into the fermentor.
  • Nitrate and sugar values are given in Table VIII.
  • the fermentor Before the start of continuous operation, the fermentor was filled with 480 kg of tobacco extract, 20.2 kg of dextrose, 2.4 kg of KH 2 p o 4 and 120 ml of an antifoaming agent, and then sterilized at 120°C for 40 minutes. After the fermentor had been cooled down, it was inoculated with 13 I of a starter culture of Candida Utilis 707 grown in tobacco extract. After 12 hours there was no more sugar or nitrate in the batch and the yeasts were in the exponential phase. At this point continuous operation was started. The operating conditions are given in Table VII. The pH regulation was done with phosphoric acid at 25%. The fermentor was equipped with a mechanical foam separator, a turbine aeration/agitation system, and a weight control system.
  • the continuous stream of fermented extract leaving the fermentor was centrifuged to remove the biomass and then pasteurized before being concentrated.

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  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
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Claims (18)

1. Procédé de dénitrification d'un extrait de tabax aqueux au moyen de microorganismes qui sont capables d'assimilation en métabolisme aéobie, de liaisons contentant de l'azote, selon lequel l'extrait est mélangé à une culture des microorganismes et à des additifs nutritifs nécessaires de façon à former un mélange de travail et ce mélange est maintenu dans des conditions dans lesquelles les dits organismes se trouvent dans leur phase de croissance exponentielle, caractérisé en ce que l'extrait de tabac stérile et les additifs sont ajoutés en continu au mélange de travail sous un taux de dilution qui n'excède pas le taux de croissance des microorganismes, tandis que le pH, la température et l'aération sont maintenus à des niveaux tels que l'assimilation aérobie est promue, l'extrait, un phosphate et une source de carbone étant ajoutés en quantités telles que leur somme totale atteint 0,1-7,5 g d'ions nitrate par litre ajouté, 1,0-10 g d'ions P04 par litre ajouté et une quantité suffisante de source de carbone, pour produire au moins 16,5 atomes assimilateurs de carbone par ion N03 ajouté, et en ce que le mélange de travail est retiré en continu à un taux tel que son volume total est maintenu constant.
2. Procédé selon la revendication 1, caractérisé en ce que les microorganismes sont enlevés du mélange retiré.
3. Procédé selon la revendication 1, caractérisé en ce que le microorganisme est une levure de Candida utilis NCYC 707, 321 ou 359, ou Candida Berthetii CBS 5452.
4. Procédé selon la revendication 3, caractérisé en ce que le pH est maintenu entre 3,5 et 7,2.
5. Procédé selon les revendications 3 ou 4, caractérisé en ce que la température est maintenue entre 25° et 37°C.
6. Procédé selon les revendications 3, 4 ou 5, caractérisé en ce que le taux de dilution est compris entre 0,1 et 0,25 I/I/h.
7. Procédé selon l'une quelconque des revendications 3 à 6, caractérisé en ce que le taux d'aération est compris entre 0,5 et 2,5 I/I/min, de préférence entre 1,0 et 2,0 I/I/min.
8. Procédé selon l'une quelconque des revendications 3 à 7, caractérisé en ce que l'ajonction totale de nitrate est de 3 à 7,5 g nitrate par litre ajouté, de préférence 4,5 à 5,5 g par litre ajouté.
9. Procédé selon la revendication 1 ou la revendication 2, caractérisé en ce que le microorganisme est Enterobacter aerogenes et de préférence Enterobacter aerogenes ATCC 13048.
10. Procédé selon la revendication 9, caractérisé en ce que le pH est maintenu entre 5,5 et 8,0, de préférence à 7,0.
11. Procédé selon la revendication 9 ou la revendication 10, caractérisé en ce que la température est maintenue entre 30°C et 40°C, de préférence à 37°C.
12. Procédé selon les revendications 9, 10 ou 11, caractérisé en ce qu le taux de dilution est compris entre 0,1 et 0,25 I/I/h et a de préférence la valeur 0,2 I/I/h.
13. Procédé selon l'une quelconque des revendications 9 à 12, caractérisé en ce que l'adjonction totale de nitrate est de 5,0 g par litre ajouté.
14. Procédé selon l'une quelconque des revendications 9 à 13, caractérisé en ce que le taux d'aération est compris entre 1,0 et 3,0 I/I/min, et a de préférence la valeur 2,0 I/I/min.
15. Procédé selon l'une quelconque des revendications 1 à 14, caractérisé en ce que, comme source de carbone, on utilise une ou plusieurs des substances glucose, dextrose, sucrose, maltose, cellobiose, lactose, ethanol, glycérine et citrate.
16. Procédé selon la revendication 15, caractérisé en ce que la source de carbone est le glucose à une concentration de 2,4 à 6%, de préférence 4%.
17. Procédé selon l'une quelconque des revendications 1 à 16, caractérisé en ce que l'adjonction totale de phosphate est de 1,1 à 1,5 g par litre ajouté, de préférence 1,25 g par litre ajouté.
18. Procédé selon l'une quelconque des revendications 1 à 17, caractérisé en ce qu'un produit de prévention de la mousse est ajouté au mélange de travail, de préférence à une concentration d'au moins 250 ppm.
EP19800302646 1979-08-20 1980-08-04 Procédé en continu pour la dénitrification d'extraits de tabac Expired EP0024152B1 (fr)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
LU81611 1979-08-20
LU81611A LU81611A1 (de) 1979-08-20 1979-08-20 Verfahren zur gewinnung einer von nitraten freien loesung aus einer nitrate enthaltenden produktloesung
LU82199 1980-02-25
LU82199A LU82199A1 (de) 1980-02-25 1980-02-25 Verfahren zur gewinnung einer von nitraten freien loesung aus einer nitrate enthaltenden produktloesung unter verwendung von prokarotischen mikroorganismen
US156910 1980-06-06
US06/156,910 US4622982A (en) 1979-08-20 1980-06-06 Continuous method of denitrating tobacco extracts

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EP0024152A2 EP0024152A2 (fr) 1981-02-25
EP0024152A3 EP0024152A3 (en) 1981-04-08
EP0024152B1 true EP0024152B1 (fr) 1983-11-09

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AU (1) AU534357B2 (fr)
BR (1) BR8005251A (fr)
CA (1) CA1129357A (fr)
DE (1) DE3065528D1 (fr)

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DE3100715A1 (de) * 1981-01-13 1982-07-22 Fabriques de Tabac Réunies S.A., 2003 Neuchâtel Verfahren zur aufbereitung von tabak und tabak, aufbereitet nach diesem verfahren
CA1191673A (fr) * 1981-06-25 1985-08-13 Hernan G. Bravo Methode de denitrification du tabac
DE3136299A1 (de) * 1981-09-12 1983-04-14 Fabriques de Tabac Réunies S.A., 2003 Neuchâtel Durchlaufverfahren zum mikrobiellen abbau von nitrate enthaltenden tabakinhaltsstoffen
DE3370651D1 (en) * 1983-12-09 1987-05-07 Tabac Fab Reunies Sa Continuous fermentation process
FR2596621B1 (fr) * 1986-04-07 1991-02-15 Ltr Ind Procede de preparation de tabac reconstitue aromatise et tabac reconstitue aromatise obtenu par ce procede
WO2012054869A1 (fr) 2010-10-22 2012-04-26 Bepex International, Llc Système et procédé pour le traitement continu de matières solides à une pression non atmosphérique
CN113475744B (zh) * 2021-06-21 2022-05-24 河南中烟工业有限责任公司 一种利用微球菌制备烟草提取物的方法

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DE252121C (fr) *
US3747608A (en) * 1971-06-18 1973-07-24 Brown & Williamson Tobacco Corp Microbial digestion of tobacco materials
ZA763603B (en) * 1976-06-17 1978-03-29 Tobacco Res & Dev Tobacco products and methods for their preparation
DE2816427C2 (de) * 1977-05-06 1982-09-16 Fabriques de Tabac Réunies S.A., 2003 Neuchâtel Verfahren zum Veredeln von Tabak
US4556073A (en) * 1978-06-15 1985-12-03 Brown & Williamson Tobacco Corporation Process for reduction of nitrate content of tobacco by microbial treatment

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CA1129357A (fr) 1982-08-10
EP0024152A2 (fr) 1981-02-25
AU6064380A (en) 1981-03-19
BR8005251A (pt) 1981-03-04
DE3065528D1 (en) 1983-12-15
AU534357B2 (en) 1984-01-26
EP0024152A3 (en) 1981-04-08

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