DE2941680A1 - Continuous fermentative denitrification of tobacco extract - under aerobic conditions, by adding, with phosphate and carbon source, to microorganism culture - Google Patents

Abstract

An aq. tobacco extract is denitrated by adding it to a mixt. of tobacco extract and microorganisms, in the exponential growth phase, which are capable of aerobic assimilation of N-contg. cpds., at a dilution rate not greater than the microorganisms' growth rate. Phosphate and carbon sources are also added to provide in total 0.1-7.5g. nitrate and 1-10g. PO4 per l., and C source to supply at least 16.5 assimilable C atoms per NO3. Appropriate pH, temp. and aeration conditions are maintained. The extract, phosphate and C sources are sterile when added and the reaction mixt. is withdrawn at a rate which maintains a constant vol. Opt. the microorganisms are recovered from the withdrawn mixt.. The nitrite, nitrate and ammonium cpd. contents are reduced in a continuous system which is more productive than batch methods and requires little or no supervision. The N-oxide content of the resulting tobacco smoke is reduced.

Description

Name: Process for obtaining a

nitrate-free solution from a nitrate-containing product solution PROCEDURE FOR THE OBTAINING A NITRATE-FREE SOLUTION FROM A NITRATE CONTAINING PRODUCT SOLUTION The invention relates to a method for obtaining one of nitrates, nitrites and ammonium compounds substantially free target solution from a washout aqueous product solution obtained from tobacco plant parts as tobacco extract Eliminate nitrates, nitrites and ammonium compounds caused by microorganisms evoked, metabolic, aerobic assimilation under largely gentle Transfer of the remaining solution components of the product solution into the target solution.

Nitrates found in tobacco perform in some cases Conversion to substances undesirable for human consumption, for example nitrogen oxides in tobacco smoke. This can be avoided by removing the nitrates before theirs can develop undesirable effects. To this end, the undesirable Compounds washed out and eliminated in the aqueous product solution. The at Wash out with water-soluble substances which pass into the aqueous solution can then, after the nitrates are no longer contained in the aqueous solution are to be added again to the washed-out starting substances. The withdrawal of nitrates should be done as completely as possible.

What applies to nitrates also applies to a large extent to nitrites and ammonium compounds, and therefore it is desirable that in addition to the nitrates also the nitrites and ammonium compounds should be removed. On the other hand, others should be in the aqueous product solution contained substances are not degraded or converted as far as possible.

The object of the invention is to provide a method of the type mentioned at the beginning Art to be designed in such a way that it is economical under these conditions - especially with the lowest possible consumption of additives and energy - to operate on a large scale is.

The invention is characterized in that a predetermined working mix portion one obtained from product solution and additives and mixed with the microorganisms Working mixture in a state of assimilation that degrades nitrate, nitrite and ammonium compounds is kept by this working mix portion with one out of the on a nitrate concentration from 3 to 7.5 g. 1-1 (grams of nitrate per liter of additive solution), a phosphate concentration from 1.0 to 10 g. 1-1 (grams of phosphate per liter of additional solution) and a carbon source in a concentration of 16.5 - 10 assimilable carbon atoms per each a nitrate molecule adjusted additional solution obtained from the product solution and additives continuously while maintaining a dilution rate of 0.1 to 0.35 1 1 1-1. h-1 (liter Additional solution per liter of working mixture portion per hour) and in the process Ventilation range from 0.8 to 2.5 l. l 1. min. 1 (liters of air per liter of working mixture portion per minute), in the pH range from 3.5 to 6 and in the temperature range of 250C (degrees Celsius) up to 370C in each case on a triplet of values, in terms of a high ratio of degraded nitrate to carbon used is, and by continuously treated mixture removed from the Arbeitsn'ischungsportion with such a nbnahlner @ te, that>. to volume of the working mix portion remains constant; and that the target solution by withdrawing the biomass from the removed, treated mixture is obtained.

Experience has shown that if two values are optimized for the value triple very variable with the third size. So you can, for example, with a ventilation with 1.5 1. 1 5. min. 1 and a pH value of 5.5 the temperature between 260C and 37 ° C and you can, for example, with a ventilation of 1> 5 1. 1-1 . min. 1 and a temperature of 300C select the pH value between 3.9 and 5.5 and you can as a further example at a pH of 5.5 and a temperature of 300C the ventilation in the range 0.5 to 1.0 1. l 1. Select min. -1 and find it are still in the range of very favorable operating conditions.

The invention comes to a continuous process and only this makes it possible (apart from the one-off start-up phase, whose products are can discard if necessary) constantly within a certain tolerance limits the for to maintain the desired effect as optimally recognized operating conditions and readjust if necessary. In contrast, with a discontinuous, batch-wise, so-called batch processes, the various stages of treatment run through and you must discard the entire batch if an error is made.

In the batch process, the assimilation conditions must be specified for each batch bring about anew and that takes hours.

This limits the throughput and increases the workload Investment and manpower required than in the inventive process.

The prescribed nitrate concentration is when it is in the product solution is not present from the start, adjusted by narrowing or diluting the same. Carbon and phosphate are added in the required amount. Should be in the Produnktlösung a higher concentration of phosphate and / or carbon from the start be present, this can be harmless 1. I will be accepted. The concentrations are coordinated in such a way that the carbon and the phosphate during assimilation of the nitrate are used up by the microorganisms, ^ (I ' Remnants of these additives get into the target solution.

The pH is preferably adjusted with orthophosphoric acid respectively Potassium hydroxide adjusted.

Since the product solutions to be treated are made from natural products The solutions obtained are the other solution components of which differ from case to case are, even the optimal operating conditions are not always in all circumstances the same, but within the stated ranges.

The invention is preferably applicable to tobacco, but it is not limited to this application. There are also foodstuffs and other luxury items known to contain nitrates, nitrites and / or ammonium compounds by Conversion to substances undesirable for human consumption, e.g. nitrosamines, to lead. As with tobacco, this can be counteracted by using the eliminates unwanted connections. If this is an aqueous solution from the start this aqueous solution is used as the product solution, otherwise it gets as with tobacco, by washing out to a product solution in which you eliminate the unwanted connections and then you then the Can add the starting product again.

For tobacco you can refer to the Orientate nitrate concentration of the product solution because the percentage of the Nitrates accompany nitrites and ammonium compounds in the main tobacco types is about the same within the accuracy required here. That is at food and other luxury foods by no means the case.

This is why operating conditions are used instead of a nitrate concentration on the concentration of the total of the compounds to be eliminated, i.e. the concentration the sum of nitrates, nitrites and ammonium compounds, which are contained in the additive solution 3 to 7.5 g. Should be 1-1. All other ren parallels can be the same be like the tobacco treatment ilin.

The following explanations relate to the application of the Invention to tobacco, but can be applied analogously with the restrictions just made on the treatment of relevant foods and other luxury items.

The assimilation produced during the preparation takes place preferentially using an inoculum of 30 to 100 g. 1-1 (grams of culture volume per liter Working mix portion). The assimilation is preferably carried out below application of a microorganism culture from the standard strains listed below: Candida utilis NCYC 707; Candida Berthetii CBS 5452; Candida utilis NCYC 321; Candida utilis NCYC 359, corresponds to DSM 70167; Enterobacter aerogenes ATCC 13048 DSM 30053.

These strains are at the depository locations indicated by the abbreviations in the form required for patent disclosure under the specified Number available.

The abbreviations mean: NCYC National Collection of Yeast Cultures, Brewing Industry Research Foundation; CBS Centraal Bureau for Mold Cultures; ATCC American Type Culture Collection; DSM German collection of microorganisms.

The description of the trunks results from the following lists I, II and III. In these lists: "+" means good; "~" weak; "-" absent.

TABLE I: Characterization of Candida yeast Plasmodium or Pseudoplasmodium -; mobile cells -; Ballistospores -; monopolar bud -; bipolar sprouting -; Sprouts on stalk -; triangular cells -; moon-shaped Cells -; short-lived cells with slow growth on malt agar and strong acetic acid production -; Formation of a real mycelius -; Formation of pseudomycelia +; Cultures red or orange -.

TABLE II: Characterization of Candida utilis NCYC 707, NCYC 359 and CBS 321 by the characters before the slash and Candida berthetii CBS 5452 with the characters after the slash.

Fermentation: glucose + / +; Galactose - / -; Sucrose +/-; Maltose - / -; Cellobiose - / -; Trehalose - / -; Lactose - / -; Melibiosis - / -; Raffinose +/-; Melezitosis - / -; Inulin - / -.

Assimilation: glucose + / +; Galactose - / -; L-sorbose - / -; Sucrose +/-; Maltose +/-; Cellobiose + / +; Trehalose +, - / -; Lactose - / -; Melibiosis - / -; Raffinose +/-; Melezitosis +/-; Inulin +/-; soluble starch - / -; D-xylose +, - / -; L-arabinose - / -; D-arabinose - / -; D-ribose - / -; L-rhamnose - / -; Ethanol +, - / +; Glycerin + / +; Erythrol - / -; Ribitol - / -; Galactitol - / -; D-mannitol +, -, - / -; D-glucitol - / -; i -methyl-D-glucoside +. ~ / -; Salicin + / +; DL-lactate +/-; Succinate +, - / +, -; Citrate + / + ,; Inositol - / -; Assimilation of potassium nitrate + / +; Growth in vitamin-free medium +, / +; growth-promoting Vitamins thiamine / lacking; NaCl tolerance% (W / V) 6-8 / 6-7; maximum growth temperature in ° C 39-43 / 40-41.

SETTING UP III: Characterization of the ATCC 13048 cell shape short rods; Flagellation peritrich; Agility +; Sporulation -; Pigment -; Grief reaction 02 behavior aerobic +; anaerobic +; Catalase +; Oxidase -; Formation of nitrite from nitrate +; Indole -; Methyl red -; Vosges-Proskauer +; Citrate +; H2S -; Urease -; gelatin Lysine decarboxylase +; Arginine dihydrolase -; Ornithine decarboylase +; Phenylalanine deaminase -; Malonate +; Gas from glucose +; Lactose +; Sucrose +; Mannitol +; Dulcit -; Salicin +; Adonit +; Inositol +; Sorbitol +; Arabinose +; Raffinose +; Rhamnose +.

The invention will now be explained with reference to the accompanying drawings and some Examples explained in more detail.

For example, the drawing shows a generalized flow sheet to the inventive method. The reference numbers used in the index drawing the meaning apparent from the following table IV.

LISTING IV: Product supply 1; Water tank 2; Stock for accessories 3; Washer with separator 4; Mixer 5; Treatment branch 6; Branch of treatment 7; Mixer 8; Metering pump 9; Work tank 10; Centrifuge 13; Branch of treatment 14; Reproducing device 15; Control device 16; pH regulator 17; Temperature controller 18; Aerator 19; Stirrer 20; Antifoam dispenser 21; Reconstitution Facility 22 and dryer 23.

From the product supply 1 comes product, for example tobacco ribs, with water from the water tank 2 in the washer 4. There the soluble components washed out and arrive as a product solution in the treatment branch 6. In the treatment branch is sterilized by heating and then cooled again.

The necessary additives, primarily phosphates and glucose, arrive from the supply 3 with water in the mixer 5 and as a solution in the treatment branch 7, where they can be sterilized by heating and then cooled, In the mixer 8 then the solutions from the two treatment branches 6 and 7 are mixed and now reach the working container 10 via the metering pump 9 through the devices 17 to 20 is the working mixture portion located in the working container 10 with regard to pH and temperature continuously adjusted, ventilated in a metered manner and stirred intensively.

Treated working mixture is continuously dosed from the working container 10 peeled off, possibly pasteurized, which is not shown in the drawing is, and then arrives at the centrifuge 13. The biomass is extracted and further applied, nourishing the remaining liquid reaches the treatment branch 14 as a target solution, where it is concentrated. These Target solution, which contains essentially all components of the product solution, except the nitrates, nitrites and ammonium compounds, which have been completely removed, can now can be reused as required, for example the washed-out, in the dryer 23 on the original moisture content of the product the product supply 1 dried or alternatively in the reconstitution device 22 the product reconstituted to form pieces of tobacco leaf-like film in the reproducing device 15 can be added by spraying or in some other way.

To start up, a working mixture portion is placed in the working container 10 given from the product solution with the necessary additives and an inoculum of the microorganism in question. After about 8 to 20 hours are in this working mix portion all nitrates, nitrites and ammonium compounds are broken down and the continuous Operation can begin by now switching on the metering pump 9, accordingly the desired dilution rate and controlled so that the volume of the filling of the Working container 10, so that the working mixture portion remains constant.

The treatment in treatment branches 6 and 7 is carried out by preheating to 1000C, sterilize at 1100C for 40 minutes and then cool 300C. In the treatment branch 14, vacuum evaporation takes place. The metering pump 9 has a Delivery rate from 50 to 200 1.1 1.h 1. The working container 10 has a capacity of 750 1 and is 2/3 full during operation. l) the volume is accordingly a working mix portion 500 1.

The operating conditions are thereby, depending on the to be processed Products and microorganisms used, adjusted according to the in the product solution contained nitrates, nitrates and ammonium compounds are completely eliminated and the additives added in ischr 5 have just been used up in this assimilation will.

Advantageous operating conditions result from the following List V of the examples listed. These examples are from tobacco stems started out and Candida utilis NCYC 707 was used as a microorganism. With others Tobacco plant parts as a starting product and / or other microorganisms are the Modify operating conditions within the scope of the information given at the beginning.

o in the list as concentration means a chemically under normal analysis conditions no longer noticeable content that is less than 10 ppm each for carbon and phosphate and less than 1 ppm each for Nitrate, nitrite and ammonium compounds.

POSITION V: Example 1; 2; 3; 4; 5; 6; 7; 8th; 9; 10; Concentration of the additional solution in g.1-1 at: Nitrate 5.0; 5.0; 5.0; 5.0; 5.0; 4.1; 4.9; 7.5; 3.0; 1.0 Nitrite 0; 0; 0; 0; 0; 0; 0; 0; 0; 0; Ammonium ver- 0.08; 0.08; 0.08; 0.08; 0.08; 0.1; 0.1; 0.1; 0.1; 0.1 ties Phosphate 1.25; 1.25; 10.0; 1.25; 1.25; 1.25; 1.25; 1.6; 0.75; 0.75 Glucose 40; 40; 40; 40; 40; 40; 40; 60; 2.4; 0.8 Assimilation operating conditions: Temperature (° C) 30; 30; 30; 30; 30; 37; 26; 30; 30; 30; pH 5.5; 3.9; 5.5; 5.5; 5.5; 5.5; 5.5; 5.5; 5.5; 5.5 Ventilation degree 1.5; 1.5; 1.5; 1.5; 1.5; 1.5; 1.5; 2.0; 1.0; 0.5 (1.1-1.min.-1) Dilution rate 0.2; 0.2; 0.2, 0.01, 0.34; 0.2; 0.2; 0.2; 0.2; 0.2 (1.1 .h concentration of the target solution in g.1-1 at: Nitrate o; O; O; O; O; O; O; O; O; O; Nitrite 0; 0; 0; 0; 0; 0; 0; 0; 0; 0; Ammonium 0; 0; 0; 0; 0; 0; 0; 0; 0; 0; ties Phosphate 0.1; 0.1; 8.4; 0.1; 0.1; 0.1; 0.1; O; o; 0; Glucose o; O; O; O; O; O; 0; 0; O; O;

Claims (9)

Claims: 1. Process for the production of one of nitrates, nitrites and ammonium compounds substantially free target solution from a washout aqueous product solution obtained from tobacco plant parts as tobacco extract Eliminate nitrates, nitrites and ammonium compounds caused by microorganisms evoked, metabolic, aerobic assimilation under largely gentle Transfer of the remaining solution components of the product solution to the target soldering, characterized in that a predetermined working mix portion of one Working mixture obtained from the product solution and additives and mixed with the microorganisms into a state of assimilation that degrades nitrate, nitrite and ammonium compounds is kept by this working mix portion with one out of the on a nitrate concentration from 3 to 7.5 g. 1 1, a phosphate concentration of 1.0 to 10 g. 1 1 as well as a A source of carbon in a concentration of 16.5-10 assimilable each Carbon atoms each set from the product solution and a nitrate molecule Additions obtained additional solution continuously while maintaining a dilution rate from 0.1 to 0.35 1. l1 charged and thereby in the ventilation range from 0.8 to 2.5 1 . 1-1. min. 1, in the pH range from 3.5 to 6 and in the temperature range of 25 ° C to 370C is held at a value triple, which with respect to a large Ratio of degraded nitrate to carbon used is favorable, and by continuously treated mixture is removed from the working mixture portion with such a rate of decrease that the volume of the working mix portion becomes constant remain; and that the Ziellosunc elurc) l withdrawal of the biomass from the removed, treated mixture is obtained. 2. The method according to claim 1, characterized in that the assimilation by using a Candida from the group consisting of Candida utilis NCYC 707, Candida Berthetii CBS 5452, Candida utilis NCYC 321, and Candida utilis NCYC 359 takes place and that the working mixture portion with one of the on a nitrate concentration from 4.5 to 5.5 g. 1-1 and a phosphate concentration of 1.1 to 1.5 g. 1-1 set Additional solution continuously while maintaining a dilution rate of 0.18 to 0.22 1 . l 1. h-1 charged, continuously with 1.4 to 1.6 1. 1-1. at least 1 ventilated and open a pH of 5.5 + 0.3 and a temperature of 30 ° C # 30C is maintained. 3. The method according to claim 2, characterized in that the working mixture portion with one of the to a nitrate concentration of 5.0 g. 1-1 and to a phosphate concentration of 1.25 g. 1 1 and a glucose concentration of 40 g. 1-1 set additional solution continuously while maintaining a dilution rate of 0.2 l 1-1. h-1 loaded, running with 1.5 1. 1-1. at least 1 aerated and at a pH value of 5.5 and a temperature is held at 300C. 4. The method according to claim 1, characterized in that the assimilation by using a Candida from the group consisting of Candida utilis NCYC 707, Candida Berthetii CBS 5452, Candida utilis NCYC 321, and Candida utilis NCYC 359 takes place and as a carbon source a carbon is used from cter Group consisting of glucose, sucrose, maltose, cellobiose, ethctnol, and clycerin Citrate. 5. The method according to claim 1, characterized in that the assimilation by using the Enterobacter aerogenes ATCC 13048 and that the working mixture portion with one from the on (a nitrate concentration of 4.5 to 5.5 g. 1-1 and one Phosphate concentration from 1.1 to 1.5 g. 1-1 set additional solution continuously in compliance with a dilution rate of 0.18 bic 0.22 1 * 1 1 h-1, continuously with 1.4 to 1.6 1. 1-1. min.-1 ventilated and at a pH value between 5.5 t, £) and a temperature between 350C and 370C is kept. 6. The method according to claim 5, characterized in that the Arbeitsmischun <isportion with one of the to a nitrate concentration of 5.0 g. 1-1 and to a phosphate concentration of 1.2 g. 1-1 and a glucose concentration of 40 y. 1-1 set additional solution continuously while maintaining a dilution rate of 0.2 1. 1-1. h 1 loaded, continuously with 1.5 l. 1-1. min.-1 aerated and at a pH value of 6.0 and one Temperature of 37 ° C is maintained. 7. The method according to claim 5, characterized in that the assimilation by using the Enterobacter aerogenes ATCC 13048 and that as a carbon source a carbon is used from the group consisting of glucose, sucrose, matlose, Cellobiose, ethanol, glycerine, citrate and lactose. 8. The method according to claim 2 or 5, characterized in that the Additional solution a phosphate concentration of 1.2 g. Has 1-1. 9. Process for obtaining one of nitrates, nitrites and ammonium compounds essentially free target solution from a nitrate, nitrite and / or ammonium compound containing product solution by eliminating the nitrates, nitrites and ammonium compounds with metabolic, aerobic assimilation caused by microorganisms largely more gentle transfer of the remaining components of the product solution in the target solution, characterized in that based on claim 1 a predetermined working mixture portion of one obtained from product solution and additives and working mixture mixed with the microorganisms in a nitrate, nitrite etc. Ammonium compounds-degrading assimilation state is kept by this Working mixture portion with a concentration of the compounds to be eliminated from 3 to 7.5 a 1-1, a phosphate concentration of 1.0 to 10 g. 1 1 like one Carbon source in a concentration of 16.5-10 assimilable carbon atoms each to one molecule each of the compound to be eliminated from the product solution and additives obtained additional solution continuously in compliance one Dilution rate from 0.1 to 0.35 1. 1-1. h-l loaded and thereby in the ventilation area from 0.8 to 2.5 1. l 1. min.-1, in the pH range from 3.5 to 6 and in the temperature range from 25 ° C to 370 is held in each case on a value triplet, which with regard to a high ratio of degraded compound to be eliminated to the compound used Carbon is beneficial, and by continuously treating mixture from the working mixture portion is decreased at such a rate that the volume of the working mix portion remains constant; and that the target solution by withdrawing the biomass from the removed, treated mixture is obtained.