SE448932B - PROCEDURE FOR THE MANUFACTURE OF HISTAMINE-CONTAINED FOODS, DRINKS AND FEEDS - Google Patents

Description

448,932 Biogenic amines enter most of the food during the processing of agricultural products. One such biogenic amine is histamine.

It has now been found that numerous microbes are capable of decarboxylating the histine diet present in agricultural products to the biogenic amine histamine.

Histamine is a substance which, even in very small amounts, triggers strong physiological reactions in humans. such as headaches (migraines), nausea and diarrhea. In addition, histamine is a significant factor in allergies and anaphylaxis.

Histamine is a strong base, which forms salts with acids.

Histamine is found in plants, for example in spinach and stinging nettles, and is found in various tissues in animals and humans. Basophilic leukocytes collect histamine in inactive form, but it can be released from them by allergic reactions. ionizing radiation. in case of tissue disorders or through liberators.

It can also be assumed that there is a close connection between the histamine content in the gastric mucosa. ulcer recurrence and stomach acid. By administering histamine receptor antagonists, chronic stomach problems can be significantly improved. The same goes for migraines. For migraine prophylaxis, a healthy histramine-free diet is of great importance.

People with chronic duodenal ulcers store significantly less histamine in the mucous membranes of the body than a healthy person.

While healthy people break down the nutrient-supplied histamine through a histaminase, lack of this enzyme leads to histamine headaches. (Horton's syndrome), also known as histamine phalgia. Also Bing's headache syndrome and Harris' neuralgia. one mostly at certain times of the day. preferably at night. often occurring disorder through half-sided pain attacks in the eye, forehead. the temple area with tear flow, redness of eyes and face. Swelling of the nose ü 'n 448 932 3 mucous membranes etc. is triggered by histamine and can be successfully treated with antihistamines.

Desensitization and induction of an increased resistance to histamine is achieved by the addition of histamine isoprotein. Normally, an antihistamine factor is present in serum. whose absence can be perceived as a sign of a latent allergy.

Histamine is excreted in the faeces and urine.

Histamine increases the contraction of the smooth muscles and relaxes the vascular muscles. In this way, blood pressure is lowered while capillary permeability is increased, leading to the formation of rashes and edema in the skin. The human serum can bind free histamine. Only in allergy sufferers is the ability to bind strongly submerged, it is inhibited by a so-called antipexin.

Marquardt (see Marquardt, "Die Weinwirtschaft", p. 127. 1978) 2 mg histamine per kg of food or stimulant, such as wine, is considered as a permissible tolerance limit. be.

Various nutrients and stimulants have been controlled for their content of histamine. For dairy products, the results compiled in Table 1 below were obtained: lalaellll Histamine content in various dairy products 448 952 4 Product Histamine mg / kg Pasteurized skimmed milk 0.3 n 0.5 n 0.7 extra durable skimmed milk (sperised) 0.8 IN! 0.8 film milk 1.2 Yoghurt 2.1 n 1.7 Tilsitost half 50.0 "60.2 Gouda cheese half 54.0 Gouda cheese half Austrian Nl .0 Gouda cheese half Dutch 180.0 Camembert half 35.0 Camembert half 55, 0 Stilton helfet English 158.0 Harzost 390.0 Harzer Bauernost 383.0 As can be seen from this table, the histamine content of various skimmed milk varieties is well below the tolerance limit set by Marquardt, and it is in itself surprising that fresh milk has a certain degree of It can be assumed that at least a part of this histamine content of the fresh milk can be explained by the cows taking up histamine with the feed, in particular with silage, and that this is transferred to the milk.

However, as further shown in Table 1, the histamine content of sour milk products such as yoghurt and cheese is significantly higher than that of fresh milk. Since, as previously mentioned, microorganisms can convert the histidine present in the milk protein into histamine by decarboxylation, it occurs during the acidification of the milk and during the cheese production this _ _ .. ...__-........ , ._.-____._......_...-. 448,932 drastic increase in histamine content. When consuming only 50 g of Harzer-Bauernost, almost 20 mg of histamine is taken up, ie 10 times the amount that Marquardt considers to be the tolerance limit for one kg of drink, eg wine. Wine also has a fairly high histamine content, especially red wine and champagne. Also in this case, the histamine is formed by converting histidine from protein substances present in fruit juice or in the mash to histamine by microorganisms. The results obtained from wine surveys are summarized in the following Table II.

Histamine content of different wines Product Histamine mg / kg Table wine (Moselle) Ö u \ J1 Table wine (France) u Quality wine (Moselle) u Cabinet wine (Rheinhessen) Cabinet wine (") Auslese (Rheingau) Red wine (Ahr) Tokajer (Hungary) uuuu Tokajer, dry (Hungary) u Red wine (Austria) Sect, dry (Germany) Champagne (France) u '\ 1 \ J' | '\ 1 \) J l-'UT F * J = “\) Å F' F 'U II \ I æi-J JIIU F * O \% IU'1 CDI- 'C' \ In beer studies, it has been established, as previously clarified, that protein-rich beers are more harmful than those with low protein or amino acid content.

The follow-up control of the histamine content in these beers resulted in the fact that the protein-rich beers are also particularly rich in histamine, and that even non-alcoholic beers can also have a relatively high histamine content. 448 952 In the following Table III, the histamine levels of different beers have been compiled, and the trial numbers are also given in this table.

Histamine content of different beers Nr. beer type Histamine mg / l 1 Alt U, 5 2 Alt 9.8 Ä Alt 8.0 Ä Alt 6.8 5 Alt 5.0 6 Weiss 5.3 T Weiss H, 9 3 Weiss Ä, O 9 Weíss H, 7 10 malt 6.8 l Alcohol-free 7.5 12 Alcohol-free Ä .6 15 Export 3.2 lü Export 2.9 lJ PJ 15 Export, 16 Export 7.0 17 Export 6.6 18 Export 7.2 19 Arrow: 3, 5 20 Pils 0.2 21 Pílß 7, H 22 Pils 5.8 23 Píls 5.2 QH Pils 5.9 25 Piia 8.7 448 932 It has now surprisingly been shown that the content of harmful biogenic amines can be significantly reduced. in particular histamine. in food. stimulants and feed. by adding a microorganism or enzyme during any step of the manufacturing process. by means of which the histidine present in the starting material is converted into a harmless product and cannot pass into the harmful histamine. The object of the present invention is to provide a process for the preparation of food, beverages and feed. what years on histamine. The process according to the invention is characterized by the poor either a) carrying out the conversion using such a microorganism species which, in addition to the desired conversion, breaks with the histidine present in the starting material without forming histamine, or that b) before the conversion carried out by the microorganisms is eliminated the histidine from the starting material using a microorganism species. which breaks down histidine without forming histamine, or adding an enzyme isolated from such a microorganism, which breaks down the histidine present in the starting material without forming histamine. and then performs the conversion.

The microorganism added for carrying out the process according to the invention may be a bacterium, preferably from the classes Lactobacilli. Pseudomonas, and micrococci. a fungus or a yeast fungus, this which causes transamination or desamination of histidine. preferably a species of Saccaromyces, or an enzyme derived from microorganisms.

According to a preferred embodiment of the process according to the invention, a foodstuff is prepared. beverages or feed, a microorganism being added during manufacture. or an enzyme. which causes a degradation of the histidine present in the nutrient to form glutamic acid resp. ketoglutaric acid or equivalent salts. In this way, nutrients can be prepared. stimulants or feedstuffs, which show a greatly reduced content of biogenic amines. 448,932 in particular histamine, or which are practically completely free of such biogenic amines.

The process according to the invention is of particular importance where a significant step in the production consists of a conversion effected by microorganisms.

Examples of such transformations effected by microorganisms are alcohol fermentation, for example in wine production or beer production, as well as further lactic acid fermentation, and also such processes that take place in cheese production.

In this context, it can be pointed out in particular that the lactic acid fermentation takes place not only in connection with the production of sour milk products, such as film milk, yoghurt or quark, but also in the production of sauerkraut and in the production of silage.

When using the process according to the invention in the production of a food, stimulant or feed, in which an essential step of the production process consists of a conversion induced by microorganisms, two embodiments of the process according to the invention are particularly advantageous.

According to one preferred embodiment, the flora of microorganisms present in the starting material used is killed and then the conversion effected by microorganisms is carried out by inoculating in the starting material a corresponding pure culture of microorganisms, which produces a transamination or desamination of the starting material. .

When, for example, a lactic acid fermentation is to be carried out, one can also isolate single-cell cultures from the technical cultures of Lactobacillus helveticus or Lactobacillus bulgaricus or Lactobaciller acidophilus and propagate them in a suitable nutrient solution with the addition of pantothenic acid, niacin, riboflavin and calciumcoflavin. The actual pure cultures of the microorganisms thus obtained have an optimum of 0. at fermentation temperatures between # 0- # 4 C. wherein the maximum temperatures at which fermentation is possible with these micro-organisms are in the range between 50-52 C. The above-mentioned reindeer cultures of Lactobacillus helveticus. Lactobacillus bulgaricus and Lactobacillus acidophilus, respectively, ferment glucose and maltose homofermentatively to form d (-) lactic acid. however, without the formation of carbon dioxide. The above microorganisms further desaminate some amino acids, more specifically histidine, arginine. citrulline. ornithine. proline and hydroxyproline to form ammonia.

In this context, it can be explicitly pointed out that a number of large microorganism strains. for example, Lactobacillus strains which are deposited at ordinary deposition sites. has the ability to perform the desired desamination of histidine to form ammonia. Insofar as pure cultures of these deposited strains exhibit the properties mentioned, they are suitable for carrying out the process according to the invention. An example of one of these strains of microorganisms is M. Rogosa, which in the German Samlung von Mikroorganismen (DSM). Griesebachstrasse 8. 34 Göttingen-DT. has the deposit number DSM 20074. This microorganism strain is also deposited at The American Typeculture Collection (ATCC). 12301 Parklawn Drive, Rockville. Maryland 20852-US. and has there the deposit number ATCC 9649.

In the same way as described above for Lactobacillus helveticus and the other mentioned Lactobacillus strains, a single cell culture of Lactobacillus delbrueckii can also be prepared. 448 932 11 In order to carry out a lactic acid fermentation with the single-cell culture of Lactobacillus thus obtained, in addition to the formation of lactic acid, a conversion of the histidine present in the starting material to glutamic acid will also take place according to a primary ammonia cleavage, and no undesired conversion takes place. the histidine to the unwanted histamine during carbon dioxide cleavage.

If the sterilized whole milk is inoculated with the previously described single cell cultures of Lactobacillus helveticus, Lactobacillus bulgaricus, or Lactobacillus acidophilus and the lactic acid fermentation is carried out at a temperature between 40- # 500, a sour milk product is obtained, which is practically free of histamine. than that in the fresh milk used for the production of sour milk. In general, during lactic acid fermentation, it is advantageous to add additional pantothenic acid, niacin and riboflavin to the milk used as starting material to ensure an optimization of the growth conditions. A significant mineral component required for the growth of the pure cultures of Lactobacillus helveticus, Lactobacillus bularicus or Lactobacillus acidophilus is calcium.

However, fresh milk already contains a sufficient amount of calcium, so that an addition of calcium is not necessary in the sour milk production described here.

From the sour milk thus obtained, if desired, a histamine-free quark can also be produced and this can either be sold as such, or it can be used for the production of certain cheeses, for example for the production of Mainzer-Handost, or Olmützer Quargeln or also for Harzost. . The single cell cultures of Lactobacillus delbrueckii obtained according to the procedure described above were used to produce sauerkraut, sour cucumbers and silage with reduced histamine content or even completely histamine-free.

In the production of sauerkraut, the chopped cabbage is seasoned in the usual way, but is not then fermented by the effect of those on the existing lactic acid bacteria. Thermal treatment instead destroys the existing microorganism flora and in connection with this the artificial grafting of a Lactobacillus delbrueckiistam takes place, which converts histidine to glutamic acid, after which the grafted material is then further processed into sauerkraut.

In the same way, even in the production of pickles, a sterilization of the starting material first takes place and then a grafting.

As previously mentioned, the process according to the invention is of great importance in the production of fermented feed, ie so-called silage feed. In this case, the green fodder used as starting material, for example grass, alfalfa, green maize and the like, is first made free of microorganisms by heat treatment, and then artificially inoculated with the mentioned single-cell reindeer culture of Lactobacillus delbrueokii and fermented in the usual manner for this purpose. arranged waterproof containers, ie ordinary feed silos. In a similar way, silage is made from insured silage, whereby after the addition of sugary substances, for example molasses or beet pulp, to the silage this mixture is first made free of microorganisms and then artificially inoculated with the said single cell pure culture of Lactobacillus delbrueckii.

The histamine-free or histamine-poor silage produced in this way is highly desirable, while ...__ ~., -.-...._._, _._.......____. 7 ._.._...._. _, ..,. f. V .f ......._.... H »H 448 932 13 thus prevents the histamine ingested with the feed from passing into the milk of cows. By using such a silage it is possible to produce a practically histamine-free fresh milk.

If it is converted by microorganisms into an alcoholic fermentation, the formation of a histamine-free alcoholic fermentation product can be achieved by first sterilizing the starting material and then carrying out a corresponding fermentation with such a Saccaromyces strain which produces a transamination or desamination of the histide present in the starting material.

It is possible to produce a wine in this way, by first sterilizing the fruit juice and then inoculating a single-cell pure culture of Saccaromyces. As a result, during the alcohol fermentation, the histidine present in the starting material is transaminated or desaminated and is not converted into harmful histamine. However, as the said single-cell pure culture of Saccaromyces is often not optimally suited for carrying out the alcohol fermentation, the previously described methods of winemaking, according to which first the histidine present in the starting material is removed by other microorganisms and only then the alcohol fermentation takes place, are more advantageous. The secondary fermentation in champagne production is also generally not carried out with a specially selected yeast strain but by the addition of pure cultures of Lactobacilli, which will also be described in more detail below.

As shown in Table I, many cheeses are particularly rich in histamine. The difficulty in cheese-making according to the process of the invention consists in the fact that in many cheeses not a single microorganism participates in the formation of the special hard or soft cheese, but that several different micro-organisms can participate in the production of a particular type of cheese, 'ß 448 952 14 If one now wants to produce a histamine-poor cheese according to the process of the invention, one must produce from all of the microorganisms responsible for the production of the particular type of cheese such single-cell cultures which effect a transamination or desamination of histidine.

Blue cheese is produced by the action of a certain blue mold on a mass, which is produced from sheep's milk by precipitation with rennet. While in the traditional Roquefort production an infection of the curd with the blue mold occurs by the addition of moldy bread, according to the invention a single cell culture of the Roquefort cheese blue mold was refined, which caused a desamination of the histidine. The curd was then inoculated with this pure culture of blue mold and thereby succeeded in producing a Roquefort cheese, which contained only very small amounts of histamine and which in taste terms after corresponding maturation was very similar to the Roquefort cheese produced in the traditional way.

When it is desired to produce a nutrient, stimulant or feed for which a manufacturing process consists of a step of induced conversion of microorganisms which is essential, it is not absolutely necessary to use for this conversion such an organism which does not produce decarboxylation of histidine underestimation of the harmful histamine. . It is also possible to convert the histidine present in the starting material into harmless products, so that no histidine is present at all in the subsequent conversion of microorganisms, whereby no histamine can be formed by a decarboxylation process. According to this second preferred embodiment of the process according to the invention, a food, stimulant or feed is prepared according to a process, in which an essential step of the production process consists in a conversion induced by microorganisms, for example an alcoholic fermentation or a single milk. acid fermentation, removing the histidine present in the starting material by the action of the microorganisms by the action of a microorganism or an enzyme which produces a desamination or transamination of histamine, so that in the subsequent microorganism induced the main conversion of the starting material does not contain any starting material. more in this, which could be converted by this second microorganism to histamine by a histidine decarboxylation. If, for example, the product produced is an alcoholic beverage, a practically histamine-free end product can also be obtained, by killing the microorganism flora in the starting material before the alcoholic fermentation, and subsequently adding a microorganism or enzyme, which causes a transamination or desamination of the starting material. the material containing the histidine and then as soon as the existing histidine has been degraded to harmless conversion products, the alcohol fermentation is carried out by inoculating a yeast culture. The yeast strain used can in this case also be one which in itself causes a decarboxylation of histidine during the formation of histamine, since at this time no histidine is present in the fermentation material and therefore no undesired conversion of histidine to histamine can take place. This latter embodiment of the process according to the invention is of particular importance in beer production, since in the working steps which take place before the alcohol fermentation, a conversion of histidine to histamine induced by microorganisms can take place. Histidine is present in considerable amounts in grain protein, for example in barley. According to Souci, Fachmann and Kraut, Nahrwerttabellen (Wissenschaftliche Verlagsgesellschaft, Stuttgart, 1977), the histidine content of barley flour is between 1.42.5 g / kg barley flour and the histidine content of threshed barley with a protein content between 97-113 g / kg was about 1.8 g / kg. During malt preparation, a proteolysis of the grain protein takes place in the mash vessel and in the brewing pan. Furthermore, at this time, microorganisms already exist, such as, for example, saccaromyces diastaticus, lactobacillus fermentum, Lactobacillus pastorianus and Pediococcus cerevisiae, which can cause a decarboxylation of the histidine to form histamine. The microorganisms in question in this case are, of course, not particularly pure single-cell cultures, but the strains present in the natural environment, which, as already mentioned, generally induce this undesired decarboxylation of the histidine.

This microbially decarboxylation of the histidine during the formation of harmful histamine can be stopped in the beer preparation only at the end of the saccharification process by thermal inactivation, but not during the primary fermentation or during ripening. The microbes present in the fermenter, which produce a histidine decarboxylation, to which numerous breeds of Saccaromyces cerevisiae are to be included, thus form considerable histamine amounts. 448 932 It has now surprisingly turned out. that a histamine-free beer can be produced by performing a thermal inactivation of the microorganism flora in the malt and in the mash even before the alcohol fermentation. and then degrades the histidine present in this starting material by adding to the worm a single-cell pure culture of Lactobacillus delbrueckii, as described above in connection with lactic acid fermentation.

The sterilization of the mash Before the addition of the reindeer culture of Lactobacillus delbrueckii is conveniently accomplished by heating the mask for 30-40 minutes to a temperature of between 82 and BSOC. In connection with this the mash is then cooled to 4506 and at this temperature the reindeer culture of Lactobacillus delbrueckii is inoculated. In the mash thus treated, under the action of 'Lactobacillus delbrueckii', a strong homofermentative conversion of glucose and maltose takes place with the formation of lactic acid and at the same time the histidine present is converted to glutamic acid during primary ammonia cleavage.

During this step, the progression of lactic acid formation and the parallel conversion of histidine to glutamic acid is constantly monitored.

As soon as the large-scale histidine 1 is converted into a glutamic acid and the mash is acidified, the main fermentation is carried out. ie the alcoholic fermentation of the oil by the addition of yeast, ie by the addition of Saccaromyces cerviciae.

This alcoholic fermentation is conveniently carried out by inoculating a pure culture of yeast, which partially metabolizes both the ammonia formed during the histidine degradation and the glutamic acid formed from the histidine and also the lactic acid formed by Lactobacillus delbureckii. The alcohol fermentation can thus be carried out with such a culture of saccaromyces oerevisiae. which optimally degrades the by-products previously formed by Lactobacillus 448 932 18 delbrueckii. In the manner described, it has been possible to produce a histamine-free beer. If this seduction is used for the production of an alcohol-free beer, this can also be made histamine-free.

The mentioned conversion of histidine to ammonia and glutamic acid can also be used to great advantage in the production of other alcoholic products, for example wine and spirits, and histamine-free alcoholic products or from these secondary products, for example histamine-free vinegar, are also obtained.

In the production of histamine-free wine according to this last-mentioned embodiment of the process according to the invention, analogous to the beer production, a thermal sterilization of the fruit juice, resp. also the mash in red wine production, to kill the unwanted primary flora of microorganisms. In connection with this, a small amount of a single-cell pure culture of Lactobacillus delbrueckií or other suitable Lactobacilli is then inoculated into the fruit juice or mash, and the material is then kept at the optimum fermentation temperature for these microorganisms in the range between HO-H5 ° C. the fruit juice or histidine present in the mash is converted to ammonia and glutamic acid. Also in this case, lactobacillus delbrueckii or other added Lactobacillus species simultaneously form lactic acid through the fermentation of carbohydrates.

As soon as the desired histidine degradation has taken place, the main fermentation takes place, ie a pure culture of yeast is inoculated, in which case also a culture of Saccaromyces cerevisiae is used, which optimally metabolises the formed ammonia as well as glutamic acid and lactic acid. wo 448 952 19 The above-mentioned procedure can also be used to advantage in the preparation of spirits, ie roasted alcoholic beverages. Although in a properly performed distillation generally no significant amounts of histamine are included in the distillate, it has been found that the commercially available spirits, such as Cognac and the like, often contain large amounts of histamine. Also in this case, the histidine present can be converted into harmless by-products by the addition of a special yeast culture, or generally even better by treating the starting material before alcohol fermentation with a pure culture of Lactobacillus delbrueckii or other pure cultures of Lactobacilli.

According to a further embodiment of the process according to the invention, the formation of histamine in microorganism-induced conversions can also be prevented in cases where normally no significant amounts of histidine are present in the starting material, but where histidine can be released from dead cells of microorganisms. , the feed or nutrient is carried out.

A typical example where histidine is first formed during the preparation process is sect preparation. In the production of sect, as is well known, a primary fermentation is carried out and a mixture of sucrose and yeast is added to the wine thus obtained, after which the secondary fermentation takes place in a closed vessel, for example a pressure tank or a closed bottle. The sucrose is then fermented with the aid of yeast into alcohol and the carbon dioxide thus formed remains loose in the raw sect. An undesirable process which takes place during sect production is that the egg white derived from the yeast fungus or the histidine derived from the egg white is decarboxylated by the living yeast cells to histamine. As can be seen from Table II previously shown, due to this egg white degradation during the formation of histamine, champagne has a significantly higher histamine content than the average values for other wines.

When using the process according to the invention for producing a histidine-free sect, a mixture of sucrose and a yeast fungus is added to the wine obtained from the primary fermentation, as well as a pure culture of the Lactobacilli described above, which produces a desamination of histidine. During the secondary fermentation carried out in the pressure vessel, the histidine derived from the yeast is immediately desaminated by the Laetobacilli present. As a result, there is no histidine left, which can be converted by the living yeast cells present into harmful histamine. According to this procedure, it has been possible to produce a practically histamine-free sect.

From the pure cultures of microorganisms which effect a transamination or desamination of histidine, the enzyme complex responsible for this conversion was isolated.

The cells were mechanically destroyed, the enzyme complex containing the liquid was isolated and lyophilized.

It was found here that 10 ~ 3 g of the enzyme material thus obtained is capable of breaking down 1 gram of histidine. The enzyme complex thus obtained was used in the same manner as the previously described pure cultures of Lactobacillus delbrueckii to degrade the histidine present in the starting material into harmless products. Analogous results were obtained with those using pure cultures of Lactobacillus delbrueckii. 448 932 21 ïlintixlirn is in itself an ozzsenticll zlminoo acid, i.e. an amino acid which must be absorbed by man with the nutrient. Unless the foods prepared according to the process of the invention are rich in protein, such as, for example, sour milk products or cheese, a completely histidine-free food is not obtained by the use of a special microorganism culture, but only the microorganism culture is prevented from performing undesired decarboxylation of histidine to form histamine. . When histidine is degraded by the microorganism culture, it is converted, as has already been explained in detail, for example to harmless glutamic acid.

However, if the food prepared according to the invention is not a significant supplier of protein, as is the case, for example, with alcoholic beverages, there is, of course, no concern at all about completely removing the small amounts of histidine in these products, as they are necessary for nutrition. The egg whites must be supplied to the human organism through other sources such as meat, eggs and dairy products.

Description of an exemplary embodiment The method according to the invention will now be described in more detail with an example.

Example: Production of a histamine-free beer In the production of a beer according to the procedure used hitherto, three significant work steps are performed (see, for example, the section on beer production in "Lehrbuch der chemischen Technologie" by Ost-Rassow, Ausgabe 1965, pp. 155H-lÄ05, Johann Ambrosius Barth, Verlag Leipzig).

Wiiñíñw uuAuTv 448 932 22 l. Malt preparation The main raw material for beer, namely barley, first contains no fermentable extract. The barley must undergo a germination process in the malting plant, whereby enzymes are formed, which transfer the water-insoluble starch in the barley to soluble sugar. The enzymes formed in the germination process also convert other high-molecular substances into low-molecular ones.

The germinated grain is then dried, forming dyes and flavors. 2- âllxesaiae The malt is decomposed and treated with warm water (mash).

Existing soluble compounds are thereby dissolved.

Additional starch is enzymatically converted to soluble maltose. After filtering the wort, it is boiled with the addition of hops, with more or less water evaporating. The warm wort is separated from the hops. 3. Fermentation: After the hop separation, the hot wort must be cooled to a fermentation temperature of between 5-7 ° C in the production of underfermented beers or to a temperature of between lü-16 ° C in the production of superfermented beers.

The brewer's yeast is added and fermented at the indicated temperatures, generally for 8-10 days. The beer thus obtained is subjected to post-fermentation in a storage cellar and is finally freed from its yeast by filtration and bottled.

According to the invention, the hitherto known method for producing beer has been modified in the following way in order to obtain a histamine-free beer: It has surprisingly been found that there are already large amounts of microbes on the surface of the grain converted to malt. of the water, not only the desired transfer of the insoluble starch to soluble sugar takes place, but also a decarboxylation of the histidine abundantly present in the cereal protein vitan to histamine through these microbes. It was further found that these undesirable microbes are heat sensitive.

In the process according to the invention, therefore, the surface of the germinated grain is freed from the interfering microbes by blanching it or immersing it in water at a temperature of 75-1000 for 10 minutes.

The water is removed and rinsing is done several times with 500-g water.

In connection with this, the malt is crushed in a known manner and the mashing is carried out in the usual manner, and the hop addition and the boiling of the wort also take place according to the traditional procedure.

A further essential step of the process according to the invention is that the cooling of the wort separated by hops by filtration is carried out in an absolutely clean, disinfected open or closed cooler, whereby during the cooling a new infection of microbes is prevented, which could bring about a decarboxylation of still present histidine to form histamine. Absolutely disinfected water must also be used when adjusting the wort to the desired extract content.

Vaccination is performed using an approved single-cell culture of Saccaromyces cereviciae, which does not produce histidine decarboxylation. 448 952 24 Supplementation of the description text Example gel 2 Preparation of histamine-free pickled cucumbers 50 kg of freshly harvested cucumbers of the usual smaller size used to make pickled cucumbers are placed in a closable container. In said containers, the cucumbers were subjected to a treatment with water vapor for half to one minute because the product water was boiled with salt and the usual spices, such as chopped onion, dill, etc. remove any microorganisms that were present surface. was added and boiling was continued for a further 2 minutes to ensure that no naturally occurring bacteria were present in said aqueous medium. Then the aqueous solution was cooled to approx. 45oC and the container was filled with the cucumbers while maintaining sterile conditions. Subsequently, a single pure cell culture of Lactobacillus delbrücki was inoculated into the material. which deaminates histidine to release ammonia. At a temperature in the range 35-45UC the fermentation was carried out for 2 weeks.

The pickled cucumbers were then filled into containers, which were sealed while maintaining sterile conditions.

Said product was tested for the presence of histamine en. The histamine content was so detectable by the procedure described above. low that one could not know any histamine at all, i.e. the histamine content of the product was clearly less than 0.1 mg per kg. An additional batch of 50 kg of freshly harvested cucumbers was treated. was excluded and after the addition of said product with water vapor. the same treatment batch of the aqueous solution was not inoculated but the lactic acid fermentation was carried out with Lactobacillus species which had been present on the cucumbers.

The lactic acid fermentation was carried out in the same manner as described above, and the product was filled into containers. This batch had a histamine content of 9.8 mg / kg.

Preparation of n1stamin ¥ in sauerkraut Freshly sliced cabbage ah¿x 1 pieces and salt and common spices ti] l ~ bdttu, now UaruFte1 it was treated for 1 minute with water vapor ¥ nr to kill all naturally Forekommandæ microorganisms. The material was allowed to rest for 2 hours to ensure that its temperature was below ASOC and then a single pure cell culture cage was inoculated with Lactobacillus dnlhrüukl, which deaminates the half-life during the release of ammon1aL. The lactic acid fermentation was carried out for 2 weeks at a temperature in the range 35 ~ 45UC. In the resulting product, no histamine could be detected according to the mutine described above.

An additional batch of sliced cabbage was treated in essentially the same manner, however, the product was not treated with hot water vapor nor was any labeling carried out with Laktobacillu fi dælbrücki. but the fermentation took place with ne lantm- bacillus species which were present in the starting material. The so r ult sultærandn nrødukten showed a histamine content of | U, 2 mg / kg.

ExempQl_¿ Preparation of a histaminized wine The grapes were pressed in the usual way and the sludge was removed from the must. The must is then immediately pasteurized through a heat treatment at a temperature of b? ~ GBÛC under ca. 30 minutes. or by exposing a thin film of the must to a heat treatment at a temperature of approx. 71 ~ ïåu unnar endaat 1 minuï.

Thereafter, the pasteurized must was immediately cooled to a temperature of between 15 DEG-20 DEG C. and inoculated with a single pure cell culture of Lactobacillus delbrücki. which deaminates histidine while releasing ammonia.

The milking of the lymph node was carried out for 24 to 8 hours at a temperature of between 15 DEG-20 DEGC. During said lactic acid fermentation, all of the must present in the must was removed from Lactobacillus delbrücki, and even a small quantity of the core constituents of the must was methanolized by said Lactobacillus uelbrücki to form lactic acid.

The must was then inoculated with yeast and the alcono fermentation was carried out in the usual manner. During the alcoholic fermentation, some of the lactic acid formed by Lactobacillus delbrücki was metabolized by the yeast, but some of the lactic acid remained in the final wine and enhanced its taste.

The yeast used to carry out the alcoholic fermentation was not a single cell culture, but naturally occurring yeast, which would decarboxylate histidine to form histamine if any hlstldin were still present in the mule.

When the alcoholic fermentation was completed, the wine was subjected to ordinary treatment, ie. cooled, filtered »and bottled. The wine was examined for histamine and no histamine could be detected at all. Taste. smell but color of the wine was excellent.

In a second experiment, fresh must was nascentized in the same manner as described above and cooled, but thereafter no inoculation was performed with a single cell culture of Lactobacillus delbrücki. but it was immediately inoculated with the same "layer of yeast" used in the first experiment. The alcohol fermentation was carried out in the same manner as described above and after the alcohol fermentation the wine was separated. filtered and bottled.

This wine had a histamine content of 2.5 mg / kg, ie. a histamine content that is an average value for a white wine. The color orh smell of the wine was excellent. however, the taste was slightly less satisfactory than the taste of the wine which had been produced according to the process of the invention, i.e. in which the histidine was first removed by the lactic acid fermentation. Examination of a histamine-free vohurt Fresh milk was pasteurized and cooled to a temperature between 35-4506. It was then inoculated with a single cell culture of a species of Lactobacillus yogurt, which deaminates histidine to release ammonia. In addition, it was inoculated with such a single cell culture of Streptococcus thermophilus. which also deaminates histidine to release ammonia.

The milk inoculated with both cell cultures was kept at a temperature of 35-45 ° C for 12 hours.

Then the yoghurt was filled into containers and cooled.

No histamine could be detected in the yoghurt at all and it had an excellent taste.

Claims (6)

. 442 952 i za P A T'E N T K R A_! 1. Procedure For the production of food. beverages and feedingstuffs with a reduced histamine content or free from hnnfamxn. in which process a transformation is carried out with the aid of microorganisms and in which the naturally occurring microorganisms in the starting material are killed. characterized in that either a) the conversion is carried out using such a microorganism which precedes the desired conversion with the histidine present in the starting material without forming histamine, or that I b) before the conversion carried out by the microorganisms is eliminated the histidine from the starting material using a microorganism species, which degrades histidine without forming histamine, or adding an enzyme isolated from such a microorganism species. which breaks down the histidine present in the starting material without forming histamine. and then performs the conversion. 2. A method according to claim 1, characterized in that said conversion in which a microorganism is used which is fed to the starting material is an alcoholic fermentation or a lactic acid fermentation. 3. A method according to claim 2, characterized in that after the killing of the naturally occurring microorganisms in the starting material, an alcoholic fermentation is initiated by inoculation with a single pure cell culture of a species of Saccaromyces, which carries out said alcoholic fermentation and which also degrades histidine without forming histamine. 4. A method according to claim 2, characterized in that after killing the naturally occurring microorganisms in the starting material, a single cell is added to a culture of a microorganism which degrades the histidine present in the starting material without forming histamine. . and that after the removal of the histidine from the starting material, an alcoholic fermentation is carried out by inoculation with a yeast culture which would degrade any existing histidine to unwanted histamine. G. 5. A method according to claim 1, characterized in that the only cell culture of the microorganism which removes histidine from the starting material is a single cell culture of a Lactobacillus. which also performs a lactic acid fermentation. and that after the removal of the histidine, the alcoholic fermentation is carried out by inoculation with a yeast culture. which at least partially metabolizes the lactic acid produced by Lactobacillus. 6. A method according to claim 1 or 2, characterized in that after the naturally occurring microorganisms in the starting material have been killed, the lactic acid fermentation is carried out by inoculation with a single cell culture of a Lactobacillus species. which in addition to the lactic acid fermentation causes a breakdown of the histidine present in the starting material without forming histamine. T. Process according to claim 6, characterized in that milk is exposed to a lactic acid fermentation to form a sour milk product. a yogurt or a quark or that a vegetable product as feed. cabbage or small cucumbers are exposed To a lactic acid fermentation to form silage, sauerkraut or pickled cucumbers. B. A method according to claim 1, characterized in that a cheese is prepared and that in a quark or a milk coagulate prepared by the addition of rennet all naturally occurring microorganisms are killed and that at least one microorgan smart is then inoculated in the starting material. . which carries out the conversion of the quark or milk coagulate to form the cheese and which in addition breaks down the histidine present in the quark or milk coagulate without forming histamine.