US20080220121A1 - Microbiologically Stabilised Beer - Google Patents
Microbiologically Stabilised Beer Download PDFInfo
- Publication number
- US20080220121A1 US20080220121A1 US12/091,419 US9141906A US2008220121A1 US 20080220121 A1 US20080220121 A1 US 20080220121A1 US 9141906 A US9141906 A US 9141906A US 2008220121 A1 US2008220121 A1 US 2008220121A1
- Authority
- US
- United States
- Prior art keywords
- beer
- isomaltulose
- mixed
- turbidity
- alcohol
- 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.)
- Abandoned
Links
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Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
- C12C11/00—Fermentation processes for beer
- C12C11/003—Fermentation of beerwort
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
- C12C5/00—Other raw materials for the preparation of beer
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/04—Preparation of other alcoholic beverages by mixing, e.g. for preparation of liqueurs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Definitions
- the present invention relates to agents and processes for the production of microbiologically stabilised beer.
- a beer produced to be low in organisms harmful to beer is suitable for storage for longer periods and can even in an environment not completely germ free easily be bottled or racked in kegs without a negative effect having to be expected.
- a risk of contamination with germs, i.e. of microbiological destabilisation, of the beer arises above all in connection with bottling of the beer, racking in kegs or filling into similar vessels.
- “Low germ” should not be understood to mean complete freedom from germs.
- the number of germs harmful to beer, in particular micro-organisms that are present in beer, such as bacteria and fungi, should be so low that the beer does not spoil during a prolonged storage period of weeks and months. If the number of germs harmful to beer is kept low, the probability that cultures harmful to beer develop is reduced.
- bottling/racking takes place in a beverage-sterile or beer-sterile manner.
- HTST high temperature short time processing
- beer is heated by means of a high-temperature short-time pasteuriser, usually a plate heat exchanger by the continuous flow method and subsequently cooled again.
- the HTST-facility is installed directly in front of the filling machine. Nevertheless, there is a risk of secondary contamination, for example during the bottling/racking process or by filling into containers which have already been contaminated with germs.
- HTST processing is a frequently used measure; however, it is not safe enough for bottling/racking of sugar-containing beer beverages.
- a further measure for low germ bottling/racking of beer is membrane filtration or cold disinfection.
- the process corresponds to the HTST process, however, a membrane filtration system is used for germ reduction instead of a high-temperature short-term pasteuriser, which system allows micro-organisms present in the beer to be separated off as a result of the pore size of the filter. In this case, too, there is a risk of secondary contamination.
- the process is again not sufficiently reliable for bottling/racking of sugar-containing beer beverages.
- a further measure is full pasteurising of bottles or cans which have already been filled and closed, preferably in a tunnel pasteuriser or chamber pasteuriser.
- the containers into which the beer has been filled are heated e.g. by means of hot water or steam and subsequently cooled again.
- the process is frequently used in breweries with long distribution channels, e.g. when exporting beer to countries overseas.
- the danger of secondary contamination during bottling/racking is thus controllable.
- sugar-containing beverages such as Malzbier or mixed beer beverages can be stabilised biologically with sufficient reliability.
- This process is cost intensive as a result of the large amount of equipment required. Apart from the high operating costs such as increased water and energy consumption, the high investment costs for plant and the high space requirement are a disadvantage.
- the containers and closures (in particular the temperature and pressure stability) need to satisfy high requirements regularly.
- Plastic bottles made of PET currently commonly used in the beverage sector cannot be pasteurised according to known methods.
- a further disadvantage is the impairment of the taste caused by the past
- a further possibility for low germ bottling/racking of beer or for providing bottled/racked microbiologically stabilised beer are processes known from the juice and lemonade industry. These include chemical sterilisation. In the case of some lemonades, it is possible according to the law on food products to use the preservation dimethyl dicarbonate (DMDC) shortly before bottling. As a rule, this requires prior HTST treatment. When correctly used, the substance added continues to take effect in the filled and closed bottle and degrades after a certain period. A disadvantage is the very difficult technical application of the substance in the brewing plant since it is harmful to health and has a high freezing point.
- DMDC dimethyl dicarbonate
- a further type of bottling/racking known from the field of the juice and lemonade industry is aseptic, i.e. sterile bottling/racking. This is associated with a considerable expenditure on equipment and plant facilities such as clean room and isolation technology. In addition, this process presupposes a changeover of the quality assurance concept including validation and employee qualification.
- the use for industrial scale beer bottling/racking as well as for bottling/racking from pub breweries is unrealistic because of the high technical and organisational effort/expenditure.
- the technical problem on which the present invention is based consists essentially of providing processes and means for the production and/or bottling/racking of beer or mixed beer beverages which can be carried out easily and cost effectively and which more effectively reduce or stabilise the number of micro-organisms harmful to beer during and/or after production. In this way, a beer or mixed beer beverage microbiologically stabilised in the bottled/racked state is to be obtained.
- the processes and means must further be suitable for use in known beer production processes and brewing facilities without essential adaptation of the process steps.
- the technical problem is solved by providing a process for the production of beer or mixed beer beverage from brewing liquor, hops and at least one source of carbohydrate, wherein in a first step (a) brewing liquor, hops and the source of carbohydrate are mixed to form wort. In a subsequent step (b) the wort is boiled. In a subsequent step (c) the wort is microbially fermented.
- the process according to the invention is characterised in that the source of carbohydrate contains isomaltulose or an isomaltulose-containing mixture as germ-stabilising agent.
- the invention thus teaches the production of beer or mixed beer beverages from brewing liquor, hops and a source of carbohydrate, wherein isomaltulose or an isomaltulose-containing mixture is contained in the source of carbohydrate as germ-stabilising agent or the source of carbohydrate preferably consists of it.
- the inventors have surprisingly found that isomaltulose or an isomaltulose-containing mixture in the source of carbohydrate reduces the germ count of micro-organisms recognised as disadvantageous, i.e. harmful to beer, and/or microbially stabilises the beer obtained.
- isomaltulose or an isomaltulose-containing mixture By using isomaltulose or an isomaltulose-containing mixture, the number of these micro-organisms does not increase in the further production process and during bottling/racking such that, subsequently, a beer low in germs harmful to beer or a beer essentially free from germs harmful to beer is obtained which is microbially stabilised. Beer obtained and bottled/racked in this way is then suitable in particular for prolonged storage.
- Micro-organisms which are also referred to as germs consist above all of bacteria and fungi such as moulds or yeasts. These include not only those organisms which detrimentally influence quality and taste of the food as a result or their metabolic activity but also potentially pathogenic germs which may endanger the health of human beings.
- the germ stabilisation according to the invention leads to the number of such micro-organisms not rising above a certain threshold in appropriate storage periods such that the food is not spoilt or does not pose a risk to health.
- Beer should be understood to mean here also a mixed beer beverage which is obtained when at least one further carbohydrate component which is not or only partially fermented, is added to the beer before, during or after its production.
- beer should be understood here to mean a mixed beer beverage in the case of which isomaltulose is added to conventionally produced beer during or after production.
- the germ-stabilising agent is added to a source of carbohydrate containing malted grain (malt), raw grain or a mixture of malted grain and raw grain.
- malt and/or raw grain is thus partially replaced by isomaltulose or by an isomaltulose-containing mixture.
- the ratio of the other components of the source of carbohydrate, i.e. in particular malt and/or the raw grain, to the isomaltulose added is 6:1 to 1:1, preferably 4:1 to 2:1, particularly preferably, depending on the field of application, precisely 6:1, 5:1, 4:1, 3:1, 2:1 or 1:1.
- isomaltulose or the isomaltulose-containing mixture is added to the source of carbohydrate, preferably before mixing with brewing liquor and hops, preferably as a syrup, as a solution and/or as a crystalline solid.
- isomaltulose or the isomaltulose-containing mixture is added to the wort together with the other source of carbohydrate, the brewing liquor and hops.
- isomaltulose is added to the beer in the brewhouse and subsequently passes through the entire main and secondary fermentation process.
- isomaltulose is added exclusively or additionally after main fermentation.
- the addition of isomaltulose after main fermentation ensures that the isomaltulose cannot be metabolised during main fermentation although, as a rule, a residual activity of the yeast is still present during secondary fermentation.
- isomaltulose is added to the beer additionally or exclusively only after filtration. If the isomaltulose is added to the beer after filtration, the mixture is not subjected to any modifications by the desired fermentation process as a result of the yeast being separated off.
- the isomaltulose is added additionally or exclusively directly before bottling/racking or before storage of the beer or mixed beer beverage.
- isomaltulose acts according to the invention as a germ-stabilising, germ-growth inhibiting or antigerminative agent.
- the isomaltulose addition be it as a syrup, as a solution or as a crystalline substance, can be accommodated without difficulties into known beer production processes such that any additional effort in terms of process technology or equipment is avoided.
- the known detrimental effect of taste deterioration which is known from other sugar substitutes or sweetening agents, does not occur in the case of the use, preferred according to the invention, of isomaltulose in mixed beer beverages.
- the addition of isomaltulose does not or only insignificantly change taste aspects such as palatefulness in beer or mixed beer beverages.
- a further subject matter of the present invention is consequently also a beer or mixed beer beverage in which isomaltulose is contained as sweetening agent.
- isomaltulose is contained as the only sweetening agent.
- isomaltulose is contained as the only body-providing sweetening agent.
- a subject matter according to the invention is consequently also the use of isomaltulose as germ-stabilising or germ-inhibiting agent for low germ production, above all bottling/racking of beers or mixed beer beverages, in particular according to the process according to the invention described here and its preferred modifications according to the invention.
- Isomaltulose is a reducing sugar which surprisingly can be assimilated and metabolised by beer yeasts such as Saccharomyces cerevisiae or Saccharomyces carlsbergensis either not at all or only with great difficulty.
- Isomaltulose (6-O- ⁇ -D-glucopyranosyl fructose), known by the name of PalatinoseTM, is a disaccharide ketose which occurs naturally e.g. in honey.
- isomaltulose can be produced from sucrose on an industrial scale by enzymatic rearrangement, using for example immobilised bacteria cells, in particular of the species Protaminobacter rubrum, Erwinia rhapontici and Serratia plymuthica or a sucrose isomerase isolated therefrom.
- An “isomaltulose-containing mixture” is a combination of isomaltulose with at least one further carbohydrate, in particular fructose, glucose, sucrose, trehalulose, leucrose, tagatose, turanose, isomaltose, isomelizitose, oligosaccharides with a degree of polymerisation of 3 or 4 or more or mixtures thereof.
- the mixture contains isomaltulose and fructose, in a further modification, the mixture contains isomaltulose and glucose, in a further modification the mixture contains isomaltulose and sucrose, in a further modification the mixture contains isomaltulose and trehalulose, in a further modification the mixture contains isomaltulose and leucrose, in a further modification the mixture contains isomaltulose and tagatose, in a further modification the mixture contains isomaltulose and turanose, in a further modification the mixture contains isomaltulose and isomaltose, in a further modification the mixture contains isomaltulose and isomelizitose, in a further modification the mixture contains isomaltulose and oligosaccharides with a degree of polymerisation of 3 or 4 or more.
- the isomaltulose-containing mixture is the sucrose-isomerisation product which has been obtained by transglucosidation of sucrose, preferably using dead or living cells of Protaminobacter rubrum or enzyme extracts produced therefrom.
- isomaltulose-containing mixtures contain approximately 79-85% isomaltulose, 8-10% trehalulose, 0.5-2% sucrose, 1-1.5% isomaltose, oligosaccharide, 2.5-3.5% fructose and 2.0-2.5% glucose or they consist thereof, these details relating to the solids content as a percentage.
- “Wort” should be understood to mean the extract liberated from insoluble components which consists of a carbohydrate source, e.g. malt, to which water and preferably hops have been added and which has been boiled. After boiling with hops, the so-called finished wort is obtained. After cooling, the boiled wort is available as pitching wort. Preferably, the wort is produced by mashing, lautering, wort boiling and wort treatment. The production of the wort has the particular aim of converting initially undissolved components of the source of carbohydrate, in particular malt, into soluble fermentable substances, separating off the remaining solid components and finally adding seasoning, i.e. the hop.
- a carbohydrate source e.g. malt
- the so-called finished wort After boiling with hops, the so-called finished wort is obtained. After cooling, the boiled wort is available as pitching wort.
- the wort is produced by mashing, lautering, wort boiling and wort treatment.
- the milled source of carbohydrate in particular malt
- the brewing liquor is preferably first mixed with the brewing liquor.
- a targeted enzymatic conversion of constituents of the source of carbohydrate takes place in a specific temperature-time programme, the most important process being the complete decomposition of starch to fermentable sugars such as glucose, maltose or maltotriose and non-fermentable dextrins.
- the temperature optimum of maltose formation is 60° C.-65° C., that of dextrin formation 70° C.-75° C. The temperature determines the final fermentation of the wort depending on the type of beer.
- the wort is preferably boiled for 60 min to 100 min, preferably with the addition of hops, preferably approximately 150 to 500 g/hl of hops being added, depending on the type of beer to be produced.
- hops preferably approximately 150 to 500 g/hl of hops being added, depending on the type of beer to be produced.
- the content of original wort is adjusted.
- sterilisation additionally occurs, a coagulation of proteins takes place, hop bitter compounds are isomerised and aroma compounds are formed and partially evaporated.
- the boiled and hopped wort is subsequently preferably liberated from sediment particles in the whirlpool and/or by filtration.
- the cold break is preferably partially removed and an intensive aeration takes place to supply the micro-organisms used for fermentation with oxygen.
- at least one suitable, fermentation-intensive micro-organism for example yeast, is added to the wort. Since the wort used for fermentation may contain different sources of carbohydrate, pale or dark microbiologically stabilised beers can be produced by using the process according to the invention.
- part of the extract of the wort is preferably replaced by isomaltulose.
- the proportion of metabolisable carbohydrates in the wort is reduced such that in addition the alcohol content of the beverage produced is preferably reduced compared with that of a normal beer.
- the alcohol content of the beers produced according to the invention can, if necessary, be further reduced by using alcohol removal processes.
- An “alcohol-free beer” should be understood to mean a beer with an alcohol content of less than 0.5% which preferably contains approximately 7 to 8% of original wort (if not indicated otherwise, % values are to be understood as % by vol.).
- a “low-alcohol beer” should be understood to mean, according to the invention, a beer which has an alcohol content of less than 5%, in particular less than 4%.
- a “source of carbohydrate” should be understood to mean materials containing carbohydrates such as grain products in the case of which the carbohydrates can be converted at least partially during the production of the wort into fermentable soluble sugars such as glucose, maltose or maltotriose which are then utilised as source of carbohydrates by micro-organisms, in particular yeasts, during fermentation.
- the source of carbohydrate used is malted grain, raw grain or a mixture thereof.
- Malted grain preferably consists of grains and seeds of barley, wheat, rye, oats, millet, triticale, rice, sorghum and/or sweet corn which have been subjected to a malt production process.
- Raw grain preferably consists of grains and seeds of barley, wheat, rye, oats, millet, sorghum, triticale, rice and/or sweet corn, which have been milled, but not malted.
- the starting materials are saccharified before fermentation.
- the malt-inherent, hydrolytically active enzymes such as amylases, maltases etc., which convert starch into non-fermentable dextrins and fermentable glucose, maltose and maltotriose, are used.
- the steeped cereals are allowed to germinate preferably at 12° C. to 18° C. and the germination process is interrupted as soon as the enzyme formation and dissolution processes have reached the desired degree. This takes place preferably by using elevated temperatures with a high throughput of air.
- the temperature can preferably be raised to approximately 80 to 85° C. in order to adjust the water content of the malt to preferably approximately 4 to 5%. This process is referred to as kilning.
- the fermentation process takes place preferably in two stages.
- the main fermentation is initiated by adding micro-organisms, in particular yeasts, bottom-fermenting yeasts or top-fermenting yeasts.
- the yeasts separates off at the bottom or in the hopper of the fermentation vessel.
- the green beer obtained during main fermentation is preferably cooled down and subjected to secondary fermentation whereby the residual extract is fermented and the beer preferably clarified.
- the taste of wort also disappears as a result of which the pure beer taste is formed in particular during secondary fermentation. This process is also referred to as maturation. Fermentation can be influenced e.g. by different fermentation temperatures, top-fermenting and bottom-fermenting methods of production, open fermentation or closed fermentation etc.
- top-fermented or bottom-fermented beer is produced using the process according to the invention.
- Bottom-fermented beer is obtained in the case of bottom fermentation, wherein the yeast settles on the bottom of the vessel after fermentation and can be separated off from there.
- Top-fermented beer is a beer which is obtained by top fermentation, wherein the yeast rises upwards at the end of fermentation and is separated off at the top as far as possible.
- the fermentation process is provided for the fermentation process to be carried out using at least one yeast and at least one acidogen selected from the group consisting of representatives of Lactobacillus sp., Acetobacter sp. and Gluconobacter sp.
- the fermentation is, for example, provided for the fermentation to be carried out using S. cerevisiae and/or S. diastaticus and/or Schizosaccharomyces pombe and a representative of Lactobacillus .
- Lactobacilli which are also known as lactic acid bacteria are capable of fermenting lactic acid.
- Low-alcohol or alcohol-free Beers or beer-like beverages produced as a result of such fermentation are characterised by a mild acidic taste which corresponds approximately to that of Hopkins Wei ⁇ e beer.
- the fermentation to be carried out using S. cerevisiae and/or S. diastaticus and/or Schizosaccharomyces pombe and a representative of Acetobacter .
- the species of Acetobacter comprises, in the narrower sense of the word, the acetic acid bacteria which are capable of forming acetic acid by the oxidation of ethanol.
- the low-alcohol or alcohol-free beers or beer-like beverages thus produced are given an acidic taste which is markedly different from the taste of the beverages obtained using Lactobacillus.
- the fermentation is carried out using S. cerevisiae and/or S. diastaticus and/or Schizosaccharomyces pombe and representative of Gluconobacter.
- Gluconobacter is capable, on the one hand, of oxidising ethanol to acetic acid and, on the other hand, glucose to gluconic acid.
- the low-alcohol or alcohol-free beers or beer-like beverages produced by this mixed fermentation also have a pleasant acidic taste.
- subject matter of the present invention is also a microbiologically stabilised beer which can be produced according to the process described above and which is preferably produced according to this process.
- a subject matter is also a microbiologically stabilised low-alcohol or alcohol-free beer, dietetic beer, malted drink, “Malzbier” or alcohol-free beer-like soft drink produced according to the invention.
- it is a pale microbiologically stabilised low-alcohol or alcohol-free beer or a dark microbiologically stabilised low-alcohol or alcohol free beer.
- “Malted drink” should be understood to be a slightly hopped, carbon dioxide-containing and dark beverage with a predominantly malt-aromatic taste of malt sweetness which, moreover, is low in alcohol to free from alcohol.
- the malted drink is brewed with approximately 7-8% original wort from the malt content. After filtering, it is adjusted preferably with sweetening sugars (glucose, sucrose) to 12% of original wort (approximately one third of the original wort).
- isomaltulose in comparison with conventional sugar, i.e. sucrose, such as little sweetening strength, higher microbiological stability, suitability for diabetics, anticariogenic properties, an even higher proportion of isomaltulose can be chosen in the original wort.
- a further subject matter is also a biologically stabilised mixed beer beverage which contains the microbiologically stabilised beer according to the invention and at least one further component selected from: extracts of herbs, aroma compounds, caffeine, colorants, amino acids, culinary acids, fruit components such as fruit juice, fruit flesh, fruit pulp or fruit extracts, sugar, sugar substitutes such as sugar alcohols, intensive sweeteners, water, distilled spirits (ethanol).
- the mixed beer beverage consists of the microbiologically stabilised beer according to the invention and the at least one further component.
- Herb components should be understood to mean: extracts, solutions, essences from parts of plants, preferably anise, valerian root, stinging nettle, blackberry leaves, strawberry leaves, fennel, lady's mantle, goose grass, ginseng, rosehip, hibiscus flowers, raspberry leaves, elderberry, hops, ginger, St.-John's wort, camomile, coriander, curled mint, lapacho plant, lavender, lemon grass, marjoram, mallow, balm, mistletoe, peppermint, marigold, rosemary, gentian, milfoil, thyme, hyssop, cinnamon etc.
- “Fruit components” should be understood to mean in particular: fruit extracts, preferably from apples, bananas, pears, pineapple, oranges, grapefruit, cherry, sour cherry, limes, lemons, passion fruit, peaches, sea buckthorn, raspberries, strawberries, blackberries, redcurrants, gooseberries, kiwi fruits etc.
- the mixed beer beverage to contain natural or nature-identical odour-bearing substances and/or flavourings as aroma components, such as essential oils from plants or fruit such as citrus oil, peppermint oil or clove oil, fruit essences, aroma-conferring fruit juices, anise, menthol, eucalyptus etc.
- natural or nature-identical odour-bearing substances and/or flavourings as aroma components such as essential oils from plants or fruit such as citrus oil, peppermint oil or clove oil, fruit essences, aroma-conferring fruit juices, anise, menthol, eucalyptus etc.
- the colorant components are preferably colorants of plant origin such as carotinoids, flavonoids or anthocyans, colorants of animal origin, inorganic pigments such as iron oxide pigments, products from enzymatic and non-enzymatic browning, products formed by heating such as caramel, sugar colour or synthetic colorants such as azo compounds, triphenylmethane compounds, indigoid compounds, xanthene compounds or quinoline compounds.
- Suitable synthetic colorants are for example erythrosine, indigo carmine or tartrazine which are used for colour correction and/or for producing a pleasing appearance of the mixed beer beverage according to the invention.
- amino acid components are preferably mixtures of essential amino acids.
- Preferred amino acids are his, lle, leu, lys, thr, trp, val and taurin.
- the acid components are preferably culinary acids.
- the beverages according to the invention are available as carbonated drinks, in other words, they may contain carbonic acid/carbon dioxide.
- the mixed beer beverages according to the invention also contain caffeine components such as extracts, preparations or essences from coffee beans, tea plant or parts thereof, mate tea plant or parts thereof, cola nut, cocoa bean or guarana.
- caffeine components such as extracts, preparations or essences from coffee beans, tea plant or parts thereof, mate tea plant or parts thereof, cola nut, cocoa bean or guarana.
- FIG. 1 Isomaltulose concentrations before and after incubation with organisms harmful to beer
- FIG. 2 Isomaltulose concentration as a function of different stability factors
- FIG. 3 Isomaltulose concentration in samples incubated with S. cerevisiae MJJ 2;
- FIG. 4 Sugar spectrum analysis after seven day incubation
- FIG. 5 Acid concentration as a function of the micro-organisms selected
- FIG. 7 Proportion of aroma components after fermentation of real worts.
- FIG. 8 Taste assessment of beers made from real worts.
- FIG. 9 Isomaltulose content after fermentation of model media. The range of values of 5% above and below the initial value is highlighted.
- FIG. 10 Isomaltulose content after fermentation of model media with bacteria. The value range of 5% above and below the initial value is highlighted.
- FIG. 11 Turbidity development in contaminated mixed beer beverages with sweetening agents sucrose (Suc), isomaltulose (Pal) or sweetener mixture (Sm) at angles of measurement of 900 and 250.
- sucrose sucrose
- Pal isomaltulose
- Sm sweetener mixture
- FIG. 12 Swelling of PET bottles with contaminated mixed beer beverages with the sweeteners sucrose (Suc), Isomaltulose (Pal) or sweetener mixture (Sm).
- the model medium was prepared as follows: 50 g of isomaltulose were dissolved in 500 ml of doubly distilled water; 6.7 g of yeast nitrogen base (YNB) were dissolved in 500 ml of doubly distilled water; 5 ml of the isomaltulose solution were autoclaved in test tubes with Durham tubes; the YNB solution was autoclaved individually, subsequently 5 ml each were pipetted into the autoclaved test tubes that had already been filled with 5 ml of isomaltulose solution.
- YNB yeast nitrogen base
- the untreated model medium has a pH value of 5.1.
- the quantities of 0.1 normal sulphuric acid were determined as being 0.05 mole/l which were required to adjust the necessary pH values. Sulphuric acid was chosen so as not to supply an additional source of C to the medium.
- a 20% isohumulone solution was diluted in a ratio of 1:20 such that 10 mg of the solution contained approximately 0.1 mg of isohumulones:
- the alcohol content was adjusted to the respective concentrations using 96% non-denatured alcohol.
- test tubes incubated in an aerobic atmosphere were closed with a cotton wool stopper, anaerobic samples were also closed with cotton wool stoppers but incubated in an anaerobic vessel.
- micro-organisms A group of micro-organisms was selected which are either known to be harmful to beer or exhibited the ability in preliminary tests of utilising isomaltulose.
- the micro-organisms are shown in Table 2.
- micro-organisms were cultivated in 100 ml of nutrient broth, subsequently washed with isomaltulose solution and resuspended in 5 ml of isomaltulose solution.
- the test tubes were incubated with 0.5 ml of the resuspended solution. Incubation took place at 26° C.
- the occurrence of turbidity and gas formation was evaluated in the test tubes as an indicator of growth.
- the extract was measured before and after incubation and the content of reducing sugars was determined by extinction measurement during the reaction with dinitrosalicylic acid (DNS) before and after incubation.
- DNS dinitrosalicylic acid
- the extract measurement of the solution was effected by density determination using an U-tube densimeter (Anton Paar).
- the content of isomaltulose was determined by the reduction of 3,5-dinitrosalicylic acid to 3-amino-5-nitrosalicylic acid. This produces a colour change in the solution from yellow to reddish brown.
- the concentration of a reducing sugar can be determined quantitatively by photometry on the basis of this colour change. Since isomaltulose is the only sugar in the test medium (in contrast to real media such as wort), it is possible by means of the DNS method to determine the concentration of isomaltulose before and after incubation.
- Table 3 shows the results of the visual growth assessment on the basis of the parameters of turbidity development and gas formation (n.o.: not observed, +: turbidity and/or gas formation, ⁇ : no turbidity or gas formation).
- This yeast formed a slight turbidity in the course of the first week which increased in intensity in the second week. Obviously, the formation of turbidity was delayed by the alcohol concentrations of 6% and 6.5%. Similarly, at the pH value of 3.6, a developing turbidity was noted only three days after the formation of turbidity was observed at higher pH values. It appeared remarkable that large parts of the forming cell mass settled on the surface of the liquid. This applied both to the aerobically and anaerobically incubated batches. In none of the batches did the yeast form gas within three weeks. In the course of the second and third week, the yeast had sedimented completely except for the cell substance adhering to the glass at the surface of the liquid.
- Saccharomyces diastaticus has a tendency towards stunted growth in the case of isomaltulose present as only available source of carbohydrate.
- FIG. 1 the isomaltulose concentrations without incubation and those after three weeks' anaerobic incubation without preservation factors, determined by DNS assay, are illustrated for all the batches (anaerobic incubation, without preservation factors, three weeks incubation at 26° C.).
- Saccharomyces cerevisiae MJJ 2 and Schizosaccharomyces pombe were capable of degrading isomaltulose in an anaerobic atmosphere without added preservation factors within the period under consideration.
- Saccharomyces cerevisiae MJJ 2 reduced the isomaltulose content in the model solution by 26%, whereas Schizosaccharomyces pombe was able to completely metabolise isomaltulose.
- the bacteria investigated were incubated only anaerobically, a degradation of isomaltulose was not detected in any batch.
- the yeasts were also incubated aerobically. Under aerobic conditions, Schizosaccharomyces pombe metabolises isomaltulose completely under all preservation parameters and in all concentrations.
- FIG. 2 shows isomaltulose concentrations measured by the DNS method in the samples incubated with Saccharomyces diastaticus (mean values of the series after three weeks incubation at 26° C.). The results are illustrated as mean values of the different preservation factors.
- Saccharomyces diastaticus exhibited the ability of metabolising isomaltulose also under aerobic conditions. In this series of measurement, deviations in the measured concentration occurred which do not fit into a range of errors of 5%. In the case of measured concentrations of more than 5% isomaltulose, it is assumed here that lower proportions of water have evaporated from the solution in the course of the incubation time. For this reason, these measured values should be considered only as a trend.
- FIG. 3 shows isomaltulose concentrations measured by the DNS method in the samples incubated with Saccharomyces cerevisiae MJJ 2 (mean values of the selective factor series, aerobically incubated, in comparison with anaerobic incubation at 26° C.).
- Saccharomyces cerevisiae MJJ 2 exhibited a better isomaltulose utilisation in an aerobic atmosphere than in the absence of oxygen. It can be clearly recognised in the figure that the variation of the pH value and the addition of alcohol within the scale examined here are not capable of inhibiting this yeast during the metabolisation of isomaltulose. However, the isomaltulose utilisation is obviously rendered more difficult for Saccharomyces cerevisiae MJJ 2 in the presence of hop bitter compounds and in the absence of oxygen.
- Isomaltulose was added to commercial dietetic beer (Henninger Di decisiv-Pils) in a quantity such that the content of the real residual extract was 4% wt./wt. In this beer, all beer-inherent selective factors are present in combination.
- the isomaltulose-containing dietetic beer was then incubated with the test micro-organisms (Example 1, Table 2) for seven days at 26° C.
- the test series was accompanied by analysis, by analyzing the worts and the beers according to the rules for methods of analysis for brewing technology of the Mittelbin Analysenkommission e.V. (Central European analytical committee) (MEBAK) and the saccharide spectra were measured additionally.
- the isomaltulose utilisation was determined by analysing the saccharide spectrum and acid spectrum since further reducing sugars, apart from isomaltulose, occur in the actual solution of beer. Before the analysis, the test micro-organisms were separated off by membrane filtration (0.45 ⁇ m) in order to produce a stable state. Measuring the acid spectrum and saccharide spectrum was effected by the HPLC/GC method in a manner known per se.
- FIG. 4 shows the results of the sugar spectrum analysis of the dietetic beers after seven days' incubation at 26° C.
- Measurable concentrations of glucose and fructose were formed again only in the beer incubated with Schizosaccharomyces pombe .
- Table 5 the isomaltulose concentrations of the individual beers and the corresponding ratio to the initial concentration are illustrated.
- lactate was metabolised as a result of the lack of utilisable substrate, a process described in the literature, in the course of which lactate is utilised as substrate for gluconeogenesis.
- lactobacillus brevis the same content was measured as in the non-incubated sample. Consequently, neither lactate was metabolised nor was it newly formed by the metabolism of the bacterium. This is taken as a further indication of the lack of metabolic activity of this bacterium.
- Dietetic beer should be understood to mean beer in the case of which, ideally, only isomaltulose is present as carbohydrate in the finished product.
- the malt grist consisted, in a proportion of 100%, of Pilsen malt, hops were added in a dose of 90 mg ⁇ -acid/l finished wort, consisting of CO 2 extract.
- the main fermentation took place without pressure using the top-fermenting yeast Saccharomyces carlsbergensis MJJ 11, at 15 to 19° C.
- the temperature was raised in order to degrade the extract content of the wort as far as possible.
- the hose transfer took place after reaching the predetermined attenuation limit.
- the test series was accompanied by analyses by which the worts and beers were analysed according to the methods of analysis for brewing technology of the Mittelbridge Analysenkommission e.V. (MEBAK) and the saccharide spectra were additionally measured.
- the contents of selected aroma-active esters and higher alcohols as well as acetaldehyde were measured.
- the acid contents of the finished beers were determined analytically and the finished beers subjected to a taste assessment both in comparison with each other and as an evaluation.
- the main fermentation process took place without pressure using the bottom-fermenting yeast Saccharomyces carlsbergensis MJJ 11, at 12° C.
- the hose transfer took place with an extract content which was approximately 1.5% above the expected extract on final fermentation.
- the alcohol-free beer produced according to the invention exhibits no impairment of the sensory characteristics. In many cases, the alcohol-free beer according to the invention was preferred to known alcohol-free beer.
- the original wort was adjusted in such a way that half of the extract was obtained from the malt whereas the other half was added in the form of isomaltulose (crystal sugar) to the boil. In total, a 12% original wort was adjusted. The addition of hops took place to 15 mg ⁇ -acid/l finished wort.
- the malted drink was filled into a keg and, after cooling, a little yeast ( Saccharomyces carlsbergensis MJJ 11) was added. After a contact time of half a day at 0° C., the yeast was largely separated off by transferring it under pressure to another storage vessel. This type of fermentation is modelled on the so-called cold contact process. After two weeks storage, filtration and bottling/racking took place.
- the malted drinks thus obtained were assessed both from the sensory point of view and analytically (according to MEBAK and aroma-specific) as in Example 3 and 4 and compared with commercial maltbeer.
- the malted drink produced according to the invention also showed no impairment of the sensory characteristics.
- the malted drink according to the invention was preferred to known, commercial Malzbier.
- the lemonade part was made from a flavour system with lemon-lime aroma from Wild, item no. 3-110050331 and citric acid. The following were used as sweeteners:
- the sweetener mixture was chosen since this product is used on an industrial scale for the production of mixed beer beverages and is consequently a practical product. Moreover, as a result of the mixture of the sweeteners, taste disadvantages of the individual components are compensated for. Citric acid is used for acidification.
- the weighed ingredients were brought to solution in brewing liquor.
- Sucrose and sweetener mixture were metered in a manner known per se, and corresponded to the recommendations of Wild Flavours.
- the isomaltulose was metered by adjusting, after tasting, the same sweetening strength as in the other lemonades. Beer and lemonade together were filled into a 30 l keg, mixed and subsequently carbonated. Carbonation took place by introducing CO 2 through the liquid channel of the dispensing head. To bond CO 2 , the kegs subjected to a gas pressure of 1.8 bar were stored at 0° C. for 24 hours. The keg pressure after cold storage was 0.8-0.9 bar.
- Triangular taste assessments with preferences were carried out with 10 tasters. Using the same basic beer, the variants with different sweeteners were tasted in comparison with each other. This had the purpose of finding out which sweetener was preferred from the sensory point of view. The deviating sample was to be determined and the respective preference recorded. The preference was taken into account only in the case of the right allocation of the deviating sample.
- the water After treatment, the water is characterised by a very low hardness which is ideally required for producing acidified mixed beer beverages.
- dietetic beer contains considerably less apparent residual extract (carbohydrates), the alcohol-free beer less alcohol with a similar residual extract content and the Doppelbock both clearly more alcohol as well as residual extract.
- the dietetic beer contains fewer bitterness units which could have a negative effect in the case of contamination with micro-organisms harmful to beer.
- the bitterness units are approximately half as high as those of the basic beers.
- the bitterness values of the mixed beer beverages are all of a similar order of magnitude.
- the alcohol contents of the mixed beer beverages were measured in line with the basic beers; the beverages produced with the alcohol-free beer contain less than 0.1 vol. % of alcohol and should therefore be regarded as alcohol-free, like the basic beer.
- the beverages with the Pilsen or dietetic beer used have an alcohol content of approximately 2.5 and slightly less than 2.7 vol. % respectively, and the value of the mixed beverages produced with Doppelbock is slightly less than 4 vol. %.
- the extract contents measured also correspond to the basic beers, the beverages sweetened with isomaltulose having the highest extract content due to the increased addition in comparison with sucrose.
- the sucrose-containing beverages have the second highest extracts, the mixtures sweetened with the sweeteners the lowest.
- the beverages based on dietetic beer have the lowest extract content, followed by the mixed beverages with the Pilsen beer.
- the basic beers of Doppelbock and the alcohol-free beer caused approximately the same residual extract content, since the alcohol-free beer obviously still contained unfermented residual extract.
- the suitability for diabetics of the dietetic beer also leads to the mixed beverages with the sweetener mixture (almost no calorific value) and the isomaltulose (low glycemic index) being suitable for diabetics.
- the taste assessments were carried out by ten people in each case; since the three sweeteners were the subject matter of the test, a total of thirty taste assessments to be evaluated per basic beer resulted.
- the sweetener-containing beverage was preferred 8 times, that containing isomaltulose 5 times. 2 tasters preferred the mixed beer beverage sweetened with sucrose with Pilsen as basic beer.
- the isomaltulose-containing beverage was preferred most frequently, the sweetener-containing mixed beer beverage was least frequently found to be the best.
- sugars isomaltulose and sucrose were found to be the most agreeable method of sweetening about equally frequently in the case of the four different basic beers (isomaltulose was preferred slightly more frequently), the use of the sweetener mixture was clearly preferred most rarely.
- FIG. 6 the results of the evaluating taste assessment are illustrated.
- the aroma profiles are similar also for mixed beer beverages with dietetic Pilsen as basic beer ( FIG. 6 b ). Again, the taste impressions of bitterness and acidity are most prevalent in the case of sweetening with sweeteners, these parameters being perceived as least intensive in the case of beverages sweetened with isomaltulose.
- the beer sweetened with sucrose received just about the poorest assessment regarding the parameter of palatefulness. Overall, the palatefulness was assessed substantially more poorly, as was to be expected, than in the case of the mixed beer beverages with Pilsen basic beer.
- the mixed beer beverages with Pilsen as basic beer were rated with the highest value in the case of this assessment parameter.
- the beverages based on Doppelbock on the other hand, were assessed as being least refreshing. No clear verdict was given regarding the different sweeteners, isomaltulose having been assessed as least refreshing, for example, in the Pilsen mixed beverage but best in the case of dietetic mixed beer beverage.
- the assessments of the other sweeteners are similar.
- Sweetening of the mixed beer beverages with sucrose and isomaltulose should be regarded as being approximately equivalent on the basis of the taste assessments carried out, the use of the sweetener mixture losing out clearly in comparison.
- the finished mixed beverages were filled by means of a manual filling facility into 0.5 l PET bottles. Before closing them with a screw cap, 50 ⁇ l of the washed pure culture suspension were pipetted into the bottles in the stream of CO 2 .
- Incubation was effected at 20° C. with exclusion of light in closed bottles.
- the turbidity in the bottles was measured with a process photometer (Sigrist KTL 30-21) at angles of measurement of 90° and 25°. The measurement at two different angles of measurement was carried out in order to take into account the different cell sizes of the yeasts and bacteria.
- the detectable turbidity maximum was at 20 EBC.
- Swelling of the bottle was determined on the basis of the deformation at selected points of the PET bottles used:
- a digital slide gauge was used for the measurement.
- FIG. 11 a shows the turbidity developments of mixed beer beverage with the basic Pilsen beer, contaminated with Saccharomyces diastaticus .
- the beverage with the sucrose sweetening reaches the maximum turbidity value as early as on the second day of incubation.
- the turbidity increases slowly in order to reach the maximum value also within the period under consideration.
- Saccharomyces diastaticus is capable of growing on the basis of the residual extract of the basic beer and of rendering the beverage turbid.
- the turbidity in the isomaltulose-containing beverage increases most slowly; it can be assumed that the growth takes place essentially on the basis of the beer extract and isomaltulose does not accelerate the beverage becoming spoiled.
- FIG. 11 b shows the turbidity developments of the mixed beer beverage with the dietetic Pilsen basic beer, contaminated with Saccharomyces diastaticus .
- a clear turbidity arises only in the case of conventional sucrose sweetening.
- the dietetic beer does not contribute any extract of its own such that the conclusion can be drawn that Saccharomyces diastaticus is incapable of utilising either the sweetener mixture or the isomaltulose as substrate.
- FIG. 11 c shows the turbidity developments of the mixed beer beverage with the alcohol-free Pilsen basic beer, contaminated with Saccharomyces diastaticus .
- the batches with the alcohol-free beer all batches reach the turbidity maximum in the period considered.
- Sucrose produces the most rapid increase in turbidity, in the two other batches the yeast is capable of growing, unhampered by alcohol, as a result of the residual extract from the beer content. The slowest spoilage occurs in the case of the isomaltulose-containing batches.
- FIG. 11 d shows the turbidity developments of the mixed beer beverage with the Doppelbock basic beer, contaminated with Saccharomyces diastaticus .
- the basic beer used moreover, contained large quantities of fermentable residual extract on the basis of which cell growth occurred.
- FIG. 11 e shows the turbidity developments of the mixed beer beverage with the Pilsen basic beer, contaminated by Saccharomyces cerevisiae MJJ2.
- the increase in turbidity occurs in the case of the isomaltulose batch.
- the turbidity which does not increase in the case of sweetener sweetening shows that neither the sweeteners nor the residual extract of the Pilsen beer can be utilised as a basis for growth.
- FIG. 11 f shows the turbidity developments of the mixed beer beverage with the dietetic Pilsen basic beer, contaminated by Saccharomyces cerevisiae .
- the sucrose-containing beverage becomes turbid very rapidly. Isomaltulose is metabolised more slowly than sucrose but more rapidly than when using Pilsen beer as basic beer, this being essentially attributable to the fact that dietetic beer contains less hops and slightly less alcohol. These two substances inhibit the growth of yeast less strongly in this trial batch. In the batch with the sweeteners, no increase in turbidity occurs.
- FIG. 11 g shows the turbidity developments of the mixed beer beverage with the alcohol-free Pilsen basic beer, contaminated with Saccharomyces cerevisiae .
- this yeast is capable of utilising at least part of the residual extract present in the beer, shown by the increase in turbidity in the case of sweetener sweetening.
- sucrose sweetening the turbidity maximum is reached as early as on the third day.
- the increase in turbidity takes place more slowly. In all batches, parts of the residual extract present in the beer are utilised.
- FIG. 11 h shows the turbidity developments of the mixed beer beverage with the Doppelbock basic beer, contaminated with Saccharomyces cerevisiae .
- the yeast contemplated here, too, is capable of growing very rapidly in the case of all the batches with Doppelbock as basic beer. This is attributed to the large quantity of fermentable residual extract and the relatively low influences of the selective factors of pH value and content of bitter compounds.
- FIG. 11 i shows the turbidity developments of the mixed beer beverage with the Pilsen basic beer, contaminated with Schizosaccharomyces pombe .
- the batches with the Pilsen basic beer and the yeast Schizosaccharomyces pombe exhibit a highly non-typical development. Since the batches with the sweeteners on the one hand and sucrose on the other exhibit a considerable turbidity right from the beginning, but which remains almost constant over the entire period, the conclusion is drawn that a colloidal turbidity of the beer portion is present, possibly caused by an excessive absorption of oxygen during bottling/racking. This is supported by the fact that the values are high mainly in the case of the turbidity measurement of 25°. Measurements at 25° show a trend towards smaller turbidity particles, whereas yeast cells should be recognisable even at 90°.
- FIG. 11 j shows the turbidity developments of the mixed beer beverage with the dietetic Pilsen basic beer, contaminated with Schizosaccharomyces pombe .
- the mixed beverage with the dietetic beer as beer portion using Schizosaccharomyces pombe no marked increase in turbidity can be seen in any of the batches. This is assessed as being due to the yeast being prevented from growing by the combined effect of the exclusion of oxygen, presence of alcohol and hops.
- these yeasts had been capable of utilising isomaltulose in a similar series of experiments in pure dietetic beer, the pH value reduced as a result of the lemonade probably plays a decisive part regarding the non-occurrence of cell growth in these batches.
- Table 10 shows that the pH values of the mixed beer beverages amount to 3.5 or even slightly less, whereas the pH value of a beer is slightly above 4.
- FIG. 11 k shows the turbidity developments of the mixed beer beverage with the alcohol-free Pilsen basic beer, contaminated with Schizosaccharomyces pombe .
- Schizosaccharomyces pombe is obviously capable of developing a slight activity in spite of the low pH value. This applies at least to the conversion with sucrose.
- the batches were sweetened with isomaltulose and sweeteners which are more difficult to utilise, only very slight and ambiguous increases in turbidity can be noted in the period under consideration.
- FIG. 11 l shows the turbidity developments of the mixed beer beverage with the Doppelbock basic beer, contaminated with Schizosaccharomyces pombe .
- the beverages which have been produced with Doppelbock exhibit a fairly rapid increase in turbidity in the case of all batches. This is attributed to the fact that not only the above-mentioned high proportions of fermentable sugars are introduced from the beer, but also that these mixed beer beverages additionally have the highest pH values of all the batches (compare Table 12). These are just above 3.8.
- FIG. 11 m shows the turbidity developments of the mixed beer beverage with the Pilsen basic beer, contaminated with Pediococcus damnosus .
- Pilsen beer When using Pilsen beer, all the batches remained stable for a long period after contamination with Pediococcus damnosus .
- the batches sweetened with sucrose exhibited a marked turbidity and consequently spoilage by the end of the period under consideration.
- FIG. 11 n shows the turbidity developments of the mixed beer beverage with the dietetic Pilsen basic beer, contaminated with Pediococcus damnosus .
- the turbidity developments detected when using the dietetic basic beer correlate well with those of the Pilsen basic beer.
- the more rapid increase in turbidity in the case of sucrose-containing batches is due to the slightly lower contents of alcohol and bitter compounds.
- FIG. 11 o shows the turbidity development of the mixed beer beverage with the alcohol-free Pilsen basic beer, contaminated with Pediococcus damnosus.
- FIG. 11 p shows the turbidity developments of the mixed beer beverage with the Doppelbock basic beer, contaminated with Pediococcus damnosus .
- the formation of turbidity observed in the other batches is delayed here by the higher proportion of alcohol.
- a slight increase in turbidity arises in the case of all the batches. From this, the conclusion can be drawn that a slight growth takes place on the basis of the residual sugar of the beer portion.
- FIG. 11 q shows the turbidity developments of the mixed beer beverage with Pilsen basic beer, contaminated with Lactobacillus brevis .
- the incubation of the batches with the Pilsen basic beer with Lactobacillus brevis no increase in turbidity is observed.
- Constant, slightly elevated values are regarded as basic turbidity which may be the result, among other things, also of the introduction of the cell suspension during inoculation.
- FIG. 11 r shows the turbidity developments of the mixed beer beverage with the dietetic Pilsen basic beer, contaminated with Lactobacillus brevis . All the batches considered remained constant for a prolonged period. A slight increase in turbidity can be seen in the batches with sucrose. This effect may be due to the slightly lower concentrations of alcohol and bitter compounds of the dietetic beer.
- FIG. 11 s shows the turbidity developments of the mixed beer beverage with the alcohol-free Pilsen basic beer, contaminated with Lactobacillus brevis .
- the increase in turbidity observable in the batch containing sucrose proves that L. brevis is capable, in the absence of alcohol, of spoiling sucrose-containing beverages.
- the two other batches remain constant; no utilisation of the isomaltulose or the sweetener takes place.
- FIG. 11 t shows the turbidity developments of the mixed beer beverage with the Doppelbock basic beer, contaminated with Lactobacillus brevis .
- the batches of the mixed beer beverages produced with Doppelbock as basic beer no increase in turbidity can be seen.
- the pH value which is lower than is typical for beer no growth activity can be detected.
- FIG. 11 u shows the turbidity developments of the mixed beer beverage with the Pilsen basic beer, contaminated with Megasphera cerevisiae .
- the test series on the mixtures with Pilsen basic beer will be illustrated here exemplarily for all the batches with Megasphera cerevisiae as a whole. In none of the batches has an increase in turbidity been observed. In all probability, this is due to the pH values which are markedly below 4 even in the case of the Doppelbock mixed beverage.
- FIG. 12 a shows the changes in height of the contaminated bottles following incubation with Saccharomyces diastaticus in comparison with the non-deformed bottle (zero bottle). It can be seen that, following incubation of the filled bottles with Megasphera cerevisiae , slight deviations in the filled bottles in comparison with the zero bottle arise. On the basis of the turbidity measured, however, no growth was determined. Moreover, it should be noted that Megasphera forms only minimum quantities of CO 2 , if any at all, when growth occurs and thus is hardly capable of contributing to an increase in the internal pressure of the bottle.
- FIG. 12 b shows the changes in height of the contaminated bottles after incubation with Saccharomyces diastaticus in comparison with the non-deformed bottle (zero bottle).
- the deformations of the bottle that have developed correlate with the developments determined already for the formation of turbidity. Slight variations in the dimensions may have been caused by an increase in the internal pressure which may arise as a result of storage of a carbonated beverage at 20° C. Substantial changes in the absolute height of the bottles inoculated with Saccharomyces diastaticus can be observed only in the case of sucrose-containing batches.
- FIG. 12 c shows the changes in the diameter of the contaminated bottles following incubation with Saccharomyces diastaticus in comparison with the non-deformed bottle (zero bottle). As in the case of the height measurements, marked changes in the bottle dimensions are recognisable only when Saccharomyces diastaticus was able to utilise sucrose.
- FIG. 12 d shows the changes in the shoulder bases of the contaminated bottles following incubation with Saccharomyces diastaticus in comparison with the non-deformed bottle (zero bottle). Again, marked changes can be noted after incubation with Saccharomyces diastaticus only in the beverages sweetened with sucrose. Interestingly, the strongest deviations occur in the batch with the dietetic beer in which the carbohydrate portion originates solely from the sweetening of the lemonade portion. In all the batches inoculated with Saccharomyces diastaticus , only sucrose was utilised as a basis for metabolism.
- FIG. 12 e shows the changes in height of the contaminated bottles after incubation with Saccharomyces cerevisiae MJJ2 in comparison with the non-deformed bottle (zero bottle).
- Saccharomyces cerevisiae MJJ2 the strongest deviations occurred in the beverages sweetened with sucrose.
- FIG. 12 f shows the changes in diameter of the contaminated bottles following incubation with Saccharomyces cerevisiae MJJ2 in comparison with the non deformed bottle (zero bottle). The same applies to the changes in diameter of the bottles following incubation with Saccharomyces cerevisiae MJJ2 as for the height expansion in that the most marked deformations occurred in the case of beverages sweetened with sucrose, while the expansions in the case of the isomaltulose-containing and sweetener-containing batches are in the region of normal expansion.
- FIG. 12 g shows the changes in the shoulder base heights of the contaminated bottles following incubation with Saccharomyces cerevisiae MJJ2 in comparison with the non-deformed bottle (zero bottle). It can be clearly seen that the relevant deformation of the bottle regarding the parameters of height of the shoulder base of the bottles incubated with Saccharomyces cerevisiae MJJ2 occurred only in the trial batches sweetened with sucrose.
- FIG. 12 h shows the changes in height of the contaminated bottles following incubation with Schizosaccharomyces pombe in comparison with the non-deformed bottle (zero bottle). In none of the batches incubated with Schizosaccharomyces does any notable growth occur, correspondingly, no expansions of the bottles exceeding normal expansion were detected. The same applies to FIGS. 12 i and j.
- FIG. 12 i shows the changes in diameter of the contaminated bottles following incubation with Schizosaccharomyces pombe in comparison with the non-deformed bottle (zero bottle).
- FIG. 12 j shows the changes in shoulder base heights of the contaminated bottles following incubation with Schizosaccharomyces pombe in comparison with the non-deformed bottle (zero bottle).
- Pediococcus damnosus was capable (exception: Doppelbock as basic beer) of forming turbidity in the mixed beverages sweetened with sucrose. In the mixtures with dietetic beer and the alcohol-free beer, this occurred even substantially before the end of the period considered. Slight deviations in the height expansion can be detected.
- FIG. 12 k shows the changes in height of the contaminated bottles following incubation with Pediococcus damnosus in comparison with the non-deformed bottle (zero bottle). This expansion is only slight and was not verified at the other measuring points. The reason for this may be that Pediococcus belongs to the homofermentative lactic acid bacteria which are capable of forming only small quantities of CO 2 even in the case of strong metabolic activity.
- FIG. 12 l shows the changes in height of the contaminated bottles following incubation with Lactobacillus brevis in comparison with the non-deformed bottle (zero bottle).
- the turbidity the growth of L. brevis was detected only in the mixed beer beverage produced with alcohol-free beer and the lemonade portion sweetened with sucrose.
- Lactobacillus brevis is heterofermentative, the growth occurred only very slowly such that a deviation was detected only in the parameter of height expansion which was only slightly above the normal expansion.
- the yeast Saccharomyces diastaticus was selected because it is highly significant as a substance harmful to beer, since it is, apart from the normal ability of attenuating low-molecular carbohydrates, also capable of attenuating longer carbohydrates chains, the so-called dextrins.
- the yeasts Schizosaccharomyces pombe and Saccharomyces cerevisiae MJJ2 were used. They were capable of utilising isomaltulose in preliminary tests; Schizosaccharomyces pombe was capable of doing this also in bottled beer.
- bacteria were also used which have great significance as spoilage germs for bottled beer.
- Pediococcus damnosus Lactobacillus brevis and Megasphera cerevisiae were chosen.
- Saccharomyces diastaticus and Saccharomyces cerevisiae MJJ2 were rapidly capable of spoiling most of the contaminated beverages. This occurred both by forming a strong turbidity and by a strong deformation of the bottles (swelling). But whereas Saccharomyces diastaticus was capable of growing in the trial batch with dietetic beer (introduction of carbohydrates only by the lemonade portion) only in the presence of sucrose, Saccharomyces cerevisiae MJJ2 also grew in the batch sweetened with isomaltulose, though surprisingly more slowly than in the case of sweetening with sucrose.
- Saccharomyces cerevisiae MJJ2 was capable of utilising also isomaltulose under the circumstances concerned, though surprisingly more slowly than sucrose.
- Saccharomyces diastaticus was not able to utilise isomaltulose in any of the beverages produced.
- this yeast is not capable of utilising isomaltulose as a substrate.
- the proven growth of Saccharomyces yeasts in the batches which had been produced with basic beers other than dietetic beer occurred in the case of all sweetening variants in an almost identical manner and is attributed to the utilisation of large quantities of residual extract which had been co-introduced by the beer portion into the finished mixed beverage.
- Schizosaccharomyces pombe did not exhibit any noticeable activity in any of the batches investigated. It is assumed that this is due above all to the pH values which, as a result of the lemonade admixture, are considerably below that of normal beers.
- Saccharomyces cerevisiae MJJ2 is capable of utilising the isomaltulose used as sweetener, though surprisingly more slowly than the sucrose present alternatively. Surprisingly, all other organisms are incapable of utilising isomaltulose.
- isomaltulose In contrast to the use of sucrose for sweetening the lemonade portion of mixed beer beverages, isomaltulose is also capable of substantially improving the biological stability.
- the batches with sweetener mixture have proved just as stable as the beverages sweetened according to the invention with isomaltulose. However, these show poorer tasting results which, on the whole, are unacceptable.
- a Pilsen wort was produced. A part of this wort was treated in such a way that approximately one quarter of the extract consisted of isomaltulose.
- the untreated original wort and the isomaltulose-containing wort were fermented under the same conditions (without pressure, 12° C.) by the same yeasts as those used in the model worts.
- a beer wort (Pilsen type) was produced which was adjusted, by redilution with water and addition of isomaltulose, in such a way that approximately 25% of the extract content of the wort consisted of isomaltulose.
- the unchanged wort and the isomaltulose-containing wort were fermented under the same conditions in parallel by different yeasts.
- the worts were adjusted to a very similar extract content.
- the addition of isomaltulose caused the degree of fermentation to fall since the proportion of non-fermentable carbohydrates (in the analysis of final fermentation) increases as a result of the substitution of wort-inherent extract by isomaltulose.
- the other analytical values change in line with the redilution, i.e. the content such as bitter compounds or protein fractions, falls.
- Table 17 shows the analytical values of worts fermented by Saccharomyces carlsbergensis MJJ 11 and Saccharomyces cerevisiae MJJ 25 (with and without isomaltulose)
- Table 18 shows the analytical values of the worts fermented with Schizosaccharomyces pombe and Saccharomyces cerevisiae MJJ 2 (with and without isomaltulose)
- FIG. 7 a shows the content of aroma components following fermentation of the real worts by Saccharomyces carlsbergensis MJJ 11 and Saccharomyces cerevisiae MJJ 25.
- FIG. 7 b shows the content of aroma components after fermentation of real worts by Schizosaccharomyces pombe and Saccharomyces cerevisiae MJJ 2.
- isomaltulose has no influence worth mentioning on the development of substances from the group of the esters and higher aliphatic alcohols (and acetaldehyde).
- An exception in this respect is the yeast Schizosaccharomyces pombe which obviously forms more acetaldehyde if isomaltulose is also present as substrate, instead of maltose.
- This yeast is also the exception for the formation of vicinal diketones. In the case of the yeasts typical for breweries, the formation of these substances is obviously slowed by the presence of isomaltulose.
- beers were subjected to a taste assessment on completion of the main and secondary fermentation. On a scale of 1 to 5, the following parameters were assessed: impression of sweetness, impression of bitterness, hop aroma, maltiness, fruitiness, liveliness, palatefulness and overall impression.
- FIG. 8 a shows the results of the taste assessment of the beers made from the real worts fermented with Saccharomyces carlsbergensis MJJ 11 (10 tasters): the aroma profiles resulting from the tasting scheme matched in an almost identical manner after fermentation by Saccharomyces carlsbergensis MJJ 11, irrespective of whether isomaltulose was contained in the wort or not. Only with respect to the overall impression was the isomaltulose beer assessed as being better. A frequently mentioned reason was a “more rounded” taste impression although the individual parameters were assessed identically.
- FIG. 8 b shows the results of the taste assessment of the beers made from the real worts; fermented with Saccharomyces cerevisiae MJJ 25 (10 tasters): After fermentation with Saccharomyces cerevisiae MJJ 25, too, the aroma profiles matched in an almost identical manner. Again, the isomaltulose-containing beer was assessed as being slightly better in the overall assessment, although the individual parameters were assessed identically. After fermentation with Saccharomyces cerevisiae MJJ 25, however, both the bitterness impression and the fruitiness of the beers was perceived more strongly.
- FIG. 8 c shows the results of the taste assessment of the beers made from the real worts; fermented with Saccharomyces cerevisiae MJJ 2 (10 tasters): Following fermentation with Saccharomyces cerevisiae MJJ 2, the beers were again assessed fairly similarly. The beer without isomaltulose was perceived as less sweet, instead the impression of bitterness became more dominant though, on the other hand, it was obviously somewhat compensated by the presence of isomaltulose. Both beers were assessed identically for overall quality.
- FIG. 8 d shows the results of the taste assessment of the beers made from the real worts; fermented with Schizosaccharomyces pombe (10 tasters): Following fermentation by Schizosaccharomyces pombe , the beers clearly differed substantially.
- the isomaltulose-containing beer was perceived as sweeter, though obviously this sweetness was perceived as being malty. Although the intensity of bitterness was perceived equally strongly, this bitterness was perceived as slightly more hop-aromatic in the case of beers without isomaltulose, an impression which was obviously compensated by the isomaltulose contained therein. However, when assessing the overall quality, the isomaltulose-containing beer was again assessed as being slightly better.
- the isomaltulose-containing beers were perceived in the majority of cases not as substantially different from the reference beers. Regarding the overall quality, the addition may cause the impression of the beer to appear somewhat “rounder” by compensating for negative influences on the taste such as e.g. strong bitterness.
- a model solution containing 5% isomaltulose and 6.7 g/l YNB (yeast nitrogen base) was produced in a sterile manner. Incubation took place at 26° C. on a 10 ml scale in test tubes with Durham tubes.
- the isomaltulose utilisation was measured by checking the gas development (visually), measurement by means of the DSN method (photometrically) and analysis of the sugar spectrum (HPLC).
- wild strain means that it is a strain isolated from contaminated beer which does not have a DSM number (DSM: Deutsche Sammlung von Mikroorganismen—German collection of micro-organisms).
- Lactobacilli were examined because of the importance of Lactobacilli as substances harmful to beer and as probiotic cultures in the food industry:
- the concentration of isomaltulose was determined by HPLC as being 42.3 g/l. This value was compared with the residual contents after incubation given below, as initial value. The model medium was incubated unchanged and with an addition of beer-typical selective factors.
- FIG. 9 a shows isomaltulose contents of the non-incubated model solution and after an incubation period of 14 days (anaerobic, 26° C.) with Saccharomyces carlsbergensis MJJ 11 (measured by means of HPLC).
- FIG. 9 b shows isomaltulose contents of the non-incubated model solution and after an incubation period of 14 days (anaerobic, 26° C.) with Saccharomyces cerevisiae MJJ 2 (measured by means of HPLC).
- FIG. 9 c shows isomaltulose contents of the non-incubated model solution and after an incubation period of 14 days (aerobic 26° C.) with Saccharomyces cerevisiae MJJ 2 (measured by means of HPLC).
- Saccharomyces cerevisiae MJJ 2 exhibited clearly a better ability of utilising isomaltulose under aerobic conditions.
- approximately 75% of isomaltulose had been metabolised after 14 days.
- a slight degradation of isomaltulose was determined even at a pH value of 4; however, no isomaltulose utilisation took place with a further decrease in the pH value.
- FIG. 9 d shows the sugar contents of the non-incubated model solution and after an incubation period of 14 days (anaerobic, 26° C.) with Saccharomyces pombe (measured by means of HPLC).
- this yeast is under all experimental conditions capable of utilising the isomaltulose offered as substrate. Only in the batch with the lowest adjusted pH value is a measurable residual concentration of isomaltulose still present such that, on further reducing the pH value, the isomaltulose utilisation can possibly be prevented; however, pH values below 3 can be found only in very few beverages and even there they are not far below this value. Selective factors typical for beer, even in combination, such as the batch with 5% of alcohol and existing hop bitter compounds, are not capable of preventing isomaltulose utilisation.
- Schizosaccharomyces pombe is capable of utilising isomaltulose effectively.
- the values measured for the individual sugars of glucose and fructose suggest that this yeast splits isomaltulose in an extracellular manner before the simple sugars are then assimilated.
- FIG. 9 e shows the isomaltulose contents of the non-incubated model solution and after an incubation period of 14 days (anaerobic, 26° C.) with Saccharomyces diastaticus (measured by means of HPLC).
- FIG. 10 the results of the incubation tests with four known bacteria harmful to beer are illustrated.
- FIG. 10 a shows the isomaltulose contents of the non-incubated model solution and after an incubation period of 21 days (anaerobic, 28° C.) with selected bacteria harmful to beer (measured by means of HPLC).
- Lactobacilli In view of the significance of the group of Lactobacilli not only as a substance harmful to beer but also as an organism in the intestinal and dental flora of mammals, and in view of their suitability for use as probiotic cultures in the food industry, a group of various Lactobacilli was examined additionally for its ability to utilise isomaltulose.
- FIG. 10 b shows isomaltulose contents after an incubation period of 21 days (anaerobic, 28° C.) with different Lactobacilli (measured by means of DNS assay).
- FIG. 10 c shows isomaltulose contents after an incubation period of 21 days (anaerobic, 28° C.) with various Lactobacilli, with an additional addition of peptone (measured by means of DNS assay).
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US12/091,419 Abandoned US20080220121A1 (en) | 2005-10-26 | 2006-05-17 | Microbiologically Stabilised Beer |
US12/091,431 Active 2026-11-05 US9359586B2 (en) | 2005-10-26 | 2006-05-17 | Manufacture of beer |
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US12/091,431 Active 2026-11-05 US9359586B2 (en) | 2005-10-26 | 2006-05-17 | Manufacture of beer |
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US (2) | US20080220121A1 (ru) |
EP (2) | EP1943327B1 (ru) |
JP (2) | JP4791549B2 (ru) |
KR (2) | KR101189644B1 (ru) |
CN (2) | CN101321855B (ru) |
AU (2) | AU2006308258B2 (ru) |
BR (2) | BRPI0617877B1 (ru) |
CA (2) | CA2627062C (ru) |
DE (1) | DE102005052210A1 (ru) |
ES (2) | ES2392348T3 (ru) |
MY (2) | MY147478A (ru) |
NO (2) | NO336322B1 (ru) |
RU (2) | RU2380400C1 (ru) |
WO (2) | WO2007048450A1 (ru) |
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US20080248158A1 (en) * | 2005-10-26 | 2008-10-09 | Tillmann Dorr | Manufacture of Beer |
US20100174001A1 (en) * | 2007-06-01 | 2010-07-08 | Suedzucker Aktiengesellschaft Mannheim/Ochsenfurt | Antioxidant for food |
US20130273201A1 (en) * | 2010-12-20 | 2013-10-17 | Nestec S.A. | Flavour modulation by bio-processing using flavour forming bacteria strains |
US20150250223A1 (en) * | 2012-06-20 | 2015-09-10 | Kirin Kabushiki Kaisha | Effervescent beverage comprising an extract of hop oxidation-reaction products |
US20150335696A1 (en) * | 2014-05-26 | 2015-11-26 | Arkray, Inc. | METHODS OF DEGRADING AGEs AND USES THEREOF |
EP2990473A4 (en) * | 2013-04-25 | 2016-11-30 | Suntory Holdings Ltd | FERMENTED MALT DRINK |
CN108753616A (zh) * | 2018-05-30 | 2018-11-06 | 贵州茅台酒股份有限公司 | 一种基于全基因组信息学分析定向筛选角鲨烯菌株的办法 |
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JP5542745B2 (ja) * | 2010-06-07 | 2014-07-09 | キリンビバレッジ株式会社 | 高甘味度甘味料含有飲料およびその製造方法 |
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JP6571913B2 (ja) * | 2014-01-17 | 2019-09-04 | サッポロビール株式会社 | 低アルコールビールテイスト飲料の製造方法 |
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US20190281874A1 (en) * | 2016-12-16 | 2019-09-19 | Nestec S.A. | Oligosaccharides for flavour generation |
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RU2648159C1 (ru) * | 2018-01-19 | 2018-03-22 | Писарницкий Александр Фомич | Композиция ингредиентов для мятного коктейля |
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RU2647766C1 (ru) * | 2018-01-19 | 2018-03-19 | Писарницкий Александр Фомич | Композиция ингредиентов для мятного коктейля |
BE1028037B1 (nl) | 2019-12-23 | 2021-09-06 | Brouwerij De Brabandere Nv | Werkwijze voor het produceren van een microbiologisch stabiel bier zonder pasteurisatie, en een microbiologisch stabiel bier |
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KR102367878B1 (ko) * | 2021-10-29 | 2022-02-25 | 김영도 | 히비스커스 건조 꽃잎, 장미 꽃잎 원액을 이용한 맥주의 제조방법 |
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2006
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- 2006-05-17 BR BRPI0617877-4A patent/BRPI0617877B1/pt active IP Right Grant
- 2006-05-17 KR KR1020087011724A patent/KR101189644B1/ko active IP Right Grant
- 2006-05-17 ES ES06753686T patent/ES2392348T3/es active Active
- 2006-05-17 RU RU2008120678/13A patent/RU2380400C1/ru active
- 2006-05-17 AU AU2006308259A patent/AU2006308259B2/en active Active
- 2006-05-17 CN CN2006800452464A patent/CN101321855B/zh active Active
- 2006-05-17 EP EP06753686A patent/EP1943327B1/de active Active
- 2006-05-17 JP JP2008536946A patent/JP4791549B2/ja not_active Expired - Fee Related
- 2006-05-17 ES ES06753687.0T patent/ES2537306T3/es active Active
- 2006-05-17 EP EP06753687.0A patent/EP1943328B1/de active Active
- 2006-05-17 CN CN2006800490381A patent/CN101346458B/zh active Active
- 2006-05-17 US US12/091,419 patent/US20080220121A1/en not_active Abandoned
- 2006-05-17 WO PCT/EP2006/004683 patent/WO2007048450A1/de active Application Filing
- 2006-05-17 CA CA2627062A patent/CA2627062C/en active Active
- 2006-05-17 RU RU2008120638/10A patent/RU2418848C2/ru active
- 2006-05-17 KR KR1020087011732A patent/KR101189651B1/ko active IP Right Grant
- 2006-05-17 BR BRPI0617878A patent/BRPI0617878B1/pt active IP Right Grant
- 2006-05-17 WO PCT/EP2006/004682 patent/WO2007048449A1/de active Application Filing
- 2006-05-17 US US12/091,431 patent/US9359586B2/en active Active
- 2006-05-17 JP JP2008536945A patent/JP4864977B2/ja active Active
- 2006-05-17 CA CA2627159A patent/CA2627159C/en active Active
- 2006-10-19 MY MYPI20064359A patent/MY147478A/en unknown
- 2006-10-19 MY MYPI20064358A patent/MY154932A/en unknown
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2008
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- 2008-05-20 NO NO20082306A patent/NO336583B1/no unknown
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Cited By (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20080248158A1 (en) * | 2005-10-26 | 2008-10-09 | Tillmann Dorr | Manufacture of Beer |
US9359586B2 (en) | 2005-10-26 | 2016-06-07 | Südzucker Aktiengesellschaft | Manufacture of beer |
US20100174001A1 (en) * | 2007-06-01 | 2010-07-08 | Suedzucker Aktiengesellschaft Mannheim/Ochsenfurt | Antioxidant for food |
US20130273201A1 (en) * | 2010-12-20 | 2013-10-17 | Nestec S.A. | Flavour modulation by bio-processing using flavour forming bacteria strains |
US20150250223A1 (en) * | 2012-06-20 | 2015-09-10 | Kirin Kabushiki Kaisha | Effervescent beverage comprising an extract of hop oxidation-reaction products |
EP2990473A4 (en) * | 2013-04-25 | 2016-11-30 | Suntory Holdings Ltd | FERMENTED MALT DRINK |
US10597619B2 (en) | 2013-04-25 | 2020-03-24 | Suntory Holdings Limited | Fermented malt beverage |
US20150335696A1 (en) * | 2014-05-26 | 2015-11-26 | Arkray, Inc. | METHODS OF DEGRADING AGEs AND USES THEREOF |
CN108753616A (zh) * | 2018-05-30 | 2018-11-06 | 贵州茅台酒股份有限公司 | 一种基于全基因组信息学分析定向筛选角鲨烯菌株的办法 |
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