FI94875C - Process for processing industrial germinated seed material for food use - Google Patents

Description

94875

METHOD FOR THE TREATMENT OF INDUSTRIALLY SEEDABLE SEED MATERIAL FOR FOOD USE

The invention relates to a method for treating industrially germinable seed material for food use.

In this context, the germination process generally refers to the process steps required to produce a germinated product from stock-dry 10 seeds. For example, in the brewing and distillery industry, the germination process, i.e. malting process, used to produce an important raw material for beer or other alcoholic beverages, cereal soils such as barley milk, rye or any malt. The germination process 15 is further used to produce various commercial germ products, e.g. bean sprouts or sprouts for any human consumption.

Germination processes are generally performed under non-aseptic conditions. The seeds to be treated contain microbes from the growing medium or storage. The conditions of the germination process are generally favorable for microbes present in the seed, which usually multiply during the process. Microbes can adversely affect the germinated product or the end product that may be made from the product, but they can also have a beneficial effect on the germinated product.

The main stages of beer production are malting, wort production, main and storage fermentation and post-processing. The purpose of malting is to produce enzymes in the grain which, during the mash, decompose the endosperm substances of the crop into a wort-soluble form. For example, the malting process of barley grains is known to involve three stages, soaking, germination and drying. The cleaned and screened barley grains are soaked in water until the desired moisture is reached, e.g. of the order of n.

43 - 44%. Part of the soaking can be performed so-called. as- 2 94875 as a champion. The barley is allowed to germinate under controlled conditions and the germinated barley is dried in a stream of hot air until germination is complete. At the end of the drying, the root germs are removed from the malt. The control of the malting stages 5 is based on the control of temperature, air flow and humidity.

In addition to malting technology and conditions, the quality of malt is affected by the microbial flora of malt grain, which varies significantly e.g. according to cereal variety, weather-10 conditions, place of production, length of growing season and storage conditions.

The most commonly used grain in malting is barley. The natural microbial flora of barley can be grouped into field and storage fungi, bacteria and hii-15 butters. The most common genera of barley fungi are Fusarium. Alternaria. Cladosporium. Cephalosporium. Epicoccum and Helminthosporium. The occurrence of molds is different in different countries and in different years. Humid weather conditions during grain growth and especially at harvest time favor 20 growths of Fusarium mold. During rainy growing seasons, Fusarium contamination can be very intense. One of the most common bacterial species in cereals is Enterobacter aaolomerans. Other bacteria include Escherichia coli and Pseudomonas. Bacteria of the genus Micrococcus and Bacillus as well as 25 lactic acid bacteria. The number of bacteria in barley is about 10®! - 1 O8 cfu / g (colony forming units / g).

The molds and bacteria contained in barley increase during malting and peak concentrations are usually reached during the germination phase. In this case, Fusarium molds and lactic acid bacteria in particular increase the most. Yeasts also increase during malting. In the drying phase, on the other hand, the mold, yeast and bacterial contents usually decrease. Some of the microbes present in barley have a beneficial effect on malting 35 and the malt product, e.g. beer. It has been estimated that up to 40-50% of some malt enzymes are of microbial origin. Some of the microbes 3 94875 have a corresponding adverse effect on barley and / or malt, respectively.

Among the unfavorable microbes are Fusarium molds, which have been found to cause particularly harmful beer overheating (gushing phenomenon) more often than other molds, whereby the peptides produced by the molds are embryos for gas bubbles that escape from the beer bottle into a strong foam. When more than 50% of malted grains are contaminated with Fusarium molds, the risk of more than 10 effervescence is clearly increased.

Furthermore, species of gram-negative bacteria contained in barley, such as those of the genera Leuconostoc, Pseudomonas and Flavo bacterium, have been found to slow the filterability of mash during wort filtration. The various microbes present in barley can also cause other adverse effects, e.g. reduced germination, taste defects or unfavorable changes in the analytical values of wort and beer.

The quality requirements for malting barley may be specified in the cultivation contract and procurement terms laid down each year. Molds are a microbial group often mentioned in quality standards. In addition, several malting plants have set an upper limit for certain molds. If the proportion of grains contaminated with Fusarium molds is more than 65% or the corresponding proportion of 25 Asoeroillus and Penicillium molds is ♦ more than 50%, the barley can be classified as of poor quality or even unfit for malting.

Efforts have been made to prevent mold overgrowth by using good quality barley or by mixing barley, malt or beer blades. In rainy years, almost the entire barley harvest may be of poor quality, which may make it impossible to obtain good quality barley.

Microbicidal chemicals have also been tried to reduce mold levels, but no safe, generally accepted chemical has been found.

The germination process for producing sprouts for human consumption also provides favorable conditions, e.g.

4 94875 for the growth of molds and bacteria. Such sprout products spoil quickly. Furthermore, in connection with seed germination, e.g. food pathogens causing food poisoning, such as Salmonella, Yersinia and / or Listeria, can increase.

The object of the present invention is to eliminate the above-mentioned drawbacks.

In particular, it is an object of the invention to provide a method by which the quantity and quality of microbial flora 10 can be controlled in a controlled manner during the germination process without adversely affecting the quality of the germinated product or any end product made from the product.

With respect to the features characteristic of the invention, reference is made to the claims.

The invention is based on studies in which it has surprisingly been found that lactic acid bacteria can be used to improve the quality of a germinating product. The substances contained in and / or produced by the preparation according to the invention, i.e. microbicides, inhibit the growth of harmful microbes during the germination process.

The use of lactic acid bacteria in the food and feed industry is well known. Under fermentation conditions, they produce compounds which affect the composition and taste of the products but also inhibit the growth of pathogenic microbes which spoil the products. Lactic acid bacteria have been commonly used in dairy products, meat products, vegetable fermentation and bakery products, as well as in feed preservation.

The addition of lactic acid bacteria or lactic acid has .. been used for a type of malt, the so-called Sauermalz, in the manufacture. The addition to the malt is carried out in the drying step before or during the mash. The purpose of the addition 35 is only to bring about a lowering of the pH of the wort and thus to affect the course of the brewing process and the quality of the finished beer. However, 5,94875 lactic acid bacteria have not previously been used in accordance with the invention to inhibit unwanted microbial growth during the germination process.

The preparation according to the invention can be added to the germinating product at any stage of the germination process.

In a particularly preferred embodiment, a lactic acid bacterial preparation or a preparation produced by a lactic acid bacterium having an antimicrobial growth effect is added to a cereal material such as barley grains during the malting process. The added preparation limits the growth of molds, especially harmful Fusarium molds, and bacteria, as a result of which, for example, the risk of beer overheating due to Fusarium molds is reduced. However, the preparation does not significantly affect the activity of the beneficial microbiota in terms of the quality of the malt obtained or the beer made from it. The added preparation has also not been found to have adverse effects on malt quality and has not been found to contain or produce compounds harmful to malt or beer.

In the malting process, the lactic acid bacteria preparation or the lactic acid bacteria preparation can be added to the barley grains before soaking, in the soaking stage or in the germination stage. The addition is preferably carried out in the soaking or germination step. The malting process can be carried out in other respects in a manner known per se. If desired, e.g. nutrients can be added to the malted barley or conditions can be adjusted, e.g. lactic acid-20 poa, to optimize the growth conditions of lactic acid bacteria.

Any commonly available lactic acid bacterium having an antimicrobial growth effect can be used in the invention. Of the 35 lactic acid bacterial families that can be used are Lactococcus, Leuconostoc, Pediococcus and Lactobacillus. Preferred species include Lactococcus lactis.

6,94875

Leuconostoc mesenteroides. Pediococcus damnosus, Pe-diococcus parvulus. Pediococcus pentosaceus, Lactobacillus curvatus and Lactobacillus plantarum or mixtures thereof, with Lactobacillus plantarum and Pediococcus pentosaceus or mixtures thereof being particularly preferred. The use of genetically modified lactic acid bacteria is also possible.

The lactic acid bacterial preparation may consist of a growth solution with cells, cells or a concentrated tumor solution with cells. The preparation produced by lactic acid bacteria may consist of a cell-free medium, a concentrated medium, a fractionated medium or a fully or partially purified microbicidal product.

According to a particularly preferred embodiment, the treatment is performed with a concentrated or fractionated growth solution, which may be cell-free or contain cells. The concentration can be carried out, for example, by lyophilization or evaporation. The growth solution is concentrated 20, e.g. 2-20 to 40-fold.

Fractionation, i.e. purification of the microbicidal products, can be carried out in a manner known per se, e.g. by chromatographic methods or by ultrafiltration.

In the method according to the invention, the antimicrobial activity of a preparation containing lactic acid bacteria or a preparation produced by lactic acid bacteria added to the seed material corresponds, for example, to a growth solution of about 10 to 10,000 ml / kg of seed vision material, suitably 30 to 7,000 ml / kg of seed to be treated. material, e.g. 40 to 5000 ml / kg of seed material to be treated. The preparation of growth solutions is described in the Examples section. It should be noted that the activity of the preparation has been determined using the growth solutions used in this application. It will be apparent to those skilled in the art that preparations of the invention having similar antimicrobial activity may also be produced using other growth solutions and / or methods.

According to the invention, the preparation contains micro-biocidal compounds or the preparation produces microbicidal compounds during the germination process. When using a cell preparation, cell growth can be promoted, if necessary, e.g. by adjusting the conditions during the germination process or by adding nutrients. The preparation may also accelerate the growth of other lactic acid bacteria present in the germinating material.

As the use of lactic acid bacteria in food is permitted and generally accepted, it is also safe to use a product derived from the cultivation of lactic acid bacteria. Lactic acid bacteria belong to the natural microbial flora of 15 commonly germinated seeds, such as barley grains. Thus, the method according to the invention is as natural as possible. The strain naturally present in the seeds can also be used as the lactic acid bacterial strain.

Thanks to the invention, it is possible to reduce the disadvantages caused by Fusarium contaminants, for example in malting, such as the risk of beer overheating.

Furthermore, the process according to the invention has surprisingly been found to improve the filterability in the brewing process. This has been found to be due to the fact that the preparation according to the invention also limits the harmful mash infiltration retardants, e.g. Leuconostoe, present in malting. The number of species belonging to the genera Pseudomonas and Flavobacterium.

MO According to another preferred embodiment, a lactic acid bacterial preparation or a preparation produced by lactic acid bacteria is added to the seed material in the production of edible sprouts.

Thanks to the present invention, the growth of harmful microbes 35 can be limited during the germination process. The invention makes it possible, for the first time, to use biological means to prevent the growth of harmful microbes present in germinating seed 8 94875 during the industrial germination process.

The method according to the invention generally improves the general level of hygiene of the germination process.

In the following, the invention is illustrated by application examples, which are intended only to illustrate the invention without limiting it.

The treatment according to the invention can also be applied to other types of germination processes.

Figure 1 shows a graph of the microbicidal activity in a lactic acid bacterial growth solution as determined by the turbidometric method. Normal growth curve of the test organism E.aaalomerans E-396 and inhibitory effect of growth solutions of L. plantarum E-76 and P. pentosaceus 15 E-390 on the growth of the test organism.

Figure 2 shows the effect of P. pentosaceus E-390 growth solutions added to the different stages of malting on the total bacterial concentrations in malting.

Figure 3 shows the effect of P. pentosaceus E-390 cells added to different stages of malting on total bacterial concentrations in malting.

Figure 4 shows the total bacterial concentrations at different stages of the experimental maltings when P 2 was added.

25 pentosaceus E-390 and L. plantarum E-76 growth solutions i or concentrated growth solutions in barley leachate.

Figure 5 shows the total bacterial concentrations at different stages of the experimental maltings when P. pentosaceus E-390 and L. plantarum E-76 growth solutions 30 or concentrated and fractionated growth solutions are added to barley leachate.

Figure 6 shows the effect of lactic acid bacterial cultures added to malting on mash filterability (Tepral filtration).

Example 1: MICROBISIC EFFECT OF DIFFERENT LACTIC ACID BACTERIAL STRAINS ON MICROBES IN MALTA

35 9 94875

The experiment investigated the microbicidal effect of preparations produced by different strains of lactic acid bacteria on the microbes present in malting. Sterile filtered medium was used as preparations.

5 1. Yield stocks:

Lactic acid bacterial strains were used as yield strains:

Lactobacillus lactis ssp. lactis VTT-E-90414 (E-414) 10 ssp. diacytilactis VTT-E-90423 (E-423)

Leuconostoc mesenteroides ssp. mesenteroides VTT-E-90389 (E-389) ssp. mesenteroides VTT-E-90415 (E-415) ssp. mesenteroides VTT-E-90466 (E-466) 15 Pediococcus damnosus VTT-E-76065 (E-65)

Pediococcus parvulus VTT-E-88315 (E-315)

Pediococcus pentosaceus VTT-E-76067 (E-67)

Pediococcus pentosaceus VTT-E-76068 (E-68)

Pediococcus pentosaceus VTT-E-88317 (E-317) 20 Pediococcus pentosaceus VTT-E-90390 (E-390) (DSM deposit)

Lactobacillus curvatus VTT-E-90391 (E-391)

Lactobacillus plantarum VTT-E-78076 (E-76) (DSM deposit) 25 Lactobacillus plantarum VTT-E-79098 (E-98) i Strains were obtained from the strain collection of VTT's biotechnology laboratory.

2. Test strains:

As test strains, 30 different harmful microbial species present in malting were used, as well as e.g. lactic acid bacterial strains used as yield strains.

Harmful molds in the experiment were represented by Fusarium molds causing beer overgrowth Γ Gibberella avenacea (formerly Fusarium avenaceum) VTT-D-80141 (D-141) 35 and VTT-D-80147 (D-147) and Fusarium culmorum VTT-D-80148 ( D-148) and VTT-D-80149 (D-149), VTT Biotechnology Laboratory Industrial Microbial Collection] and one 10 94875

Aspergillus laii.

Harmful gram-negative bacteria were represented by two strains of the genus Enterobacter and one species from both the genera Flavobacterium and Pseudomonas.

5 In addition to the strains used as yield strains, the lactic acid bacteria consisted of the strain Lactococcus sp.

E-416.

3. Growth of credit stocks and preparation of sterile filtered growth solution: 10 Lactic acid bacteria were grown in MRS broth (MRS BROTH, Oxoid). Strains of the genus Pediococcus E-65, E-67 and E-68 were grown aerobically at 25 ° C and other yielding strains anaerobically at 30 ° C with a growing time of 2-5 days. The cells were then centrifuged and the supernatant was sterile filtered.

4. Breeding of test strains:

Spore suspensions were produced from Fusarium mold broth by growing the mold strain in CMC solution (carboxymethylcellulose) at 25 ° C for 5-6 days as a shaking culture, breaking the spore formations in TWEEN solution, filtering the suspension and collecting the resulting filtrate.

A spore suspension of Asperqillus mold was produced directly on PD agar (25 ° C, 3 days) (Potato dextrose, 25 Difco).

· Gram-negative bacteria were grown aerobically in NB broth (Nutrient broth, Difco) for 1 day, Enterobacter strain at 30 ° C and Flavobacterium and Pseudomonas strains at 25 ° C.

30 Lactic acid bacteria were grown in section 3.

as set out in

, 5. Investigation of the microbicidal effect of lactic acid bacterial strains:

The microbicidal activity in the growth medium was determined by the disc method or by turbidometry.

5.1. Disc method for the determination of microbicidal activity: · Sterile filtered growth solution or dilution was pipetted into a filter paper disc (diameter 12.7 mm) 100 μΐ. The discs were placed on Plate Count agar plates plated with 0.3 ml dilution of test organism, 10-2. The samples were grown for 24 h at 30 ° C, after which the diameter of the blocking ring formed was expressed in millimeters.

5.2. Turbidometric turbidity measurement method for the determination of mirobicidal activity:

An automatic turbo-10 meter (Bioscreen, Labsystems) was used in the method.

The sample contained 10% by volume of the test organism and 10% by volume of the sterile-filtered preparation obtained from the growth of the yield stock, as well as the culture medium. In the controls, sterile water was adjusted in place of the sterile-15 supplement preparation, the pH of which was adjusted with lactic acid to the same level as the pH of the sterile-filtered preparation.

For each test strain, the same medium as for the test strain was used as the culture medium.

In Fusarium molds and Aspergillus strain, the growing conditions were 5 days, 25 ° C and vigorous shaking. For Gram-negative bacteria, the culture conditions were: Enterobacter strain 30 ° C, others 25 ° C, 2 25 days and shaking. For lactic acid bacteria, the growth conditions were 3 days, 30 ° C and shaking.

The apparatus determined the absorbance of the samples at visible wavelengths of 420 to 580 nm. After culturing, a kas-30 flow curve of each sample could be generated and the area of the growth curve calculated.

The microbicidal effect of the yield strain on the growth of the test strain was expressed as the percentage inhibition obtained by comparing the size of the growth area obtained with the control and the sterile-filtered preparation of the yield strain.

35 6. Investigation of the fungicidal effect of certain lactic acid bacteria:

The fungicidal activity of the six lactic acid bacterial strains, E-76, E-98, 12,94875 E-315, E-317, E-414 and E-415, was studied separately for all Fusarium molds used as test strains. The experiment visually examined the formation of turbidity in mold cultures to which 5 sterile-filtered preparations from each lactic acid bacterial culture had been added at different dilutions as defined in Table 3.

Sterilized Milli-Q water and Milli-Q water adjusted to pH 3.6 with lactic acid were used as controls instead of the growth solution.

The culture was performed as a test tube culture in CMC broth, at 25 ° C, for 5 days. The results were read visually.

7. Results: 15 Tables 1 and 2 and Figure 1 show the microbicidal activity of different lactic acid bacterial strains determined by the disc method and the turbidometric method, respectively.

Table 3 shows the visually determined fungicidal activity.

TABLE 1 . Microbicidal activity of lactic acid bacterial growth solutions by the disc method.

j25 Lactic acid Cell count Growth medium Blocking ring PMY / ml pH diameter, mm E-76 3.2x108 3.70 19 E-98 1.2x108 3.72 17 E-315 1.6x108 3.90 16 * 30 E-317 1 .0x108 3.86 16 E-390 1.9x108 3.92 15 |

Test organism E-396 cell count 4.0x107 PMY / ml; on a plate 1,2x105 PMY / ml 1 3 ip '00000 £ 2 2 2 2 £ £ 22222 Γ) λ O * 7 r I f I ii 5 5 0 4 ° 0 <5 <5 0 <£ 0 0 5 0 94875 • 5 C. N e N Μ e o 'n' - «-Ι -r * · - vo cm cm - irt _ _ ΓΤ TTTT N» 2 “> <? a - g; ^ r- »Ό f> rp cn α ^ v v.

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- no growth

D-141 LACTIC ACID- GROWTH SOLUTION

bacterial strain dilution __L6__IA__3j6__4 ^ 6__5A__6: 6 E-76 __ + __ + __-__; __-__- E-98 __ + __ + __; __-__-__- E-31S __ * __ + __ + __; __-__- E -317 __ + __ + __-__-__-__ · E-414 __ + __ + __ + __ + __-__- E-4IS __ + 1 + 1 + 1-1-1 control_____ + jH-jcontroU ^^^^^ ^^^ —j ^ BSSssa = s ^ ssB = SBSsanB ^ aas ^ BSSsssassaBssasssssss

D-147 LACTIC ACID- GROWTH SOLUTION

bakteenkanu dilution __L6__2 ^ 6__3k6__4 * __ 5t6__6: 6 E-76 __ + __ + __; __; __-__- E-98 __ + __ + __; __; __-__- E-315 __ + __ + __ + __; __; __ - E-317 __ + __ + __ + __-__-__- E-414 __ + __ + __ + __ + __-__- E-415 + __ + __ + __; __; __- control + pH4comroHi _ ^ _ ^ _

0-148 LACTIC ACID- GROWTH SOLUTION

bakteenkanu dilution __L6__2A__3 ^ 6__4 ^ 6__5 * __ 6: 6 E-76_ + __; __; __; __; __- E-98 __ + __ + __-__-__; __- E-315__i __ + __; __; __; __ - E-317 __ + __ + __; __; __; __- E-tl4 __ + __ + __ + __-__; __-

E-415 __ + I + 1 - I - I - I

control + I jyi-control ^^^^^^

D 149 LACTIC ACID - GROWTH SOLUTION

bakteenkanu dilution __L6__2A__3 ^ 6__4JS__5j6__6: 6 E-76 __ + __ + __; __; __; __- E-98 __ + __ + __; __; __; __- E-315 __ + __ + __ + __; __; __- E -317 __ + __ + __ + __-__-__- E-414 __ + __ + __ + __; __; __-

E-415 __ + 1 + I + 1 - 1 - I

control + __nH control ^ _ 15 94875

The results show that said lactic acid bacterial strains inhibit the growth of harmful Fusarium molds and other harmful microbes present in malting without substantially affecting the beneficial-5 microbes. The results demonstrate the utility of lactic acid bacteria in the method of the present invention.

Example 2: MICRO-10 BICIDIC EFFECT OF CERTAIN LACTIC ACID BACTERIAL STEPS ON FOOD PATHOGENS AND FOOD HARMFUL MICROBES

The experiment investigated the micro-15-biocidal effect of Pediococcus pentosaceus VTT-E-90390 (E-390) and Lactobacillus plantarum VTT-E-78076 (E-76) on food pathogens and food-harmful microbes. As a test organism, Bacillus was used. Yersinia. Listeria. Pseudomonas. Strains belonging to the genera Salmonella and Staphylococcus.

A sterilized growth solution of lactic acid bacteria prepared as described in Example 1 was used as a preparation. All other test strains were grown for 16-18 h in Iso-sensitest broth (Oxoid) except Listeria strain. grown in tryptose phosphate broth. The growth temperature was 30 ° C, except Salmonella-. For Listeria and Staphylococcus strains 37 ° C.

Microbicidal activity was determined by the turbidimetric method described in Example 30 1. The experimental conditions in the turbidometric assay were as described above for medium and temperature. The rearing time was 24 h, except for Bacillus and

For Yersinia strains 48 h.

The results are shown in Table 4, from which it can be seen that the addition of the lactic acid bacterial preparation according to the invention inhibits the growth of food pathogens and food-harmful microbes.

94875 TABLE 4. Growth inhibition by P. pentosaceus E-390 and L. plantarum E-76.

Yield stocks E-390 E-76

Test organisms Decrease in growth area (%)

Bacillus cereus 93 80 ATCC9139 _ *) ___

Yersinia enterolitica 85 54 ELI351_2___

Listeria monocytogenes 41 49 KTL4126 '*) ___

Pseudomonas fluorescens 89 97 ELI97___

Pseudomonas fragi 84 93 ATCC4973___

Salmonella infantis 90 98 ELI5_2___

Staphylococcus aureus 73 86 ELI200 -) *) Foodborne pathogens ATCC; American Type Culture Collection ELI; VTT, Food Laboratory KTL; Public Health Laboratory: «94875 1 7

Example 3; EFFECT OF LACTIC ACID BACTERIAL PREPARATIONS AND LACTIC ACID BACTERIAL PREPARATIONS ON THE MICROBIFLORE AND MALT QUALITY OF MAL

1. Strains used: 5 The bacterial strain Pediococcus pentosaceus VTT-E-90390 (E-390) was used in the experiment.

MRS broth was used as the growth solution. The cultures were fresh, rejuvenated twice. At rejuvenation, bacteria were grown in 10 ml of MRS broth 10 anaerobically, at a temperature of 30 ° C. The inoculum volume was 1% of the volume of the growth solution.

2. Barley:

The barley used was 1990 barley, which accounted for 55% of the grains contaminated with Fusarium molds.

3. Malting process: 1000 g batches of barley were immersed in a 12 ° C water bath for 1 h for rinsing. The rinsing water was changed to the first wastewater, which in turn was changed to the second wastewater after 20 5 h. The dry menu was started after 16 h. The purpose of the dryer was to remove water from the surface of the grains. The dry menu lasted 8 h. The barley reached 44% moisture during the dirt. Barley was aerated throughout the soaking.

25 Liko was followed by germination. Barley was germinated in a germination cabinet for 6 days at 14 ° C. To keep the moisture level, 44% of the barley batches were moistened and turned daily.

The resulting green malt was dried for 21 h with a temperature program. At the beginning of drying, the temperature was 50 ° C for 4.5 h. During the next 4.5 h, the temperature was raised to 60 ° C, where it was kept for 4 h. The temperature was further raised steadily over 5 h to 85 ° C, where it was kept for the remaining 3 h. The final humidity in Malta was about 4 %. Finally, the root germs were removed from the malt mechanically.

Malting without any addition was used as a control experiment.

18 94875 4. Lactic acid bacterial preparation and preparation produced by lactic acid bacteria:

Lactic acid bacterial cells isolated from the culture medium and growth solutions containing microbicidal compounds were used together and separately. The medium with the cells was added to 120 ml / kg of barley or the cells were separated from 120 ml of the medium. Separation was performed from the growth solution by centrifugation and the cells were suspended in water. When the growth solution was added, the solution 10 was used as it was. The number of cells of the preparations to be added was of the order of about 10® -10® cfu / ml.

5. Lactic acid bacterial supplements:

Lactic acid bacterial supplementations were performed on either barley, I Lion top, I and II Lion top 15, or germination start.

6. Analyzes performed: Samples were taken from each malting step.

6.1. Molds were determined as follows: The percentage of grains contaminated with Fusarium molds was determined on Fusarium molds with selective CZAPEK IPRODION DICLORAL agar (CZID agar) and a moist blotting paper support (IP, EBC-Analytica Microbiologica, Part II, 1987). Fusarium molds were identified based on typical colony and spore morphology and red color.

Aspergillus and Penicillium molds were determined on selective malt salt agar (EBC-Analytica Microbiologica, Part II, 1987). The other most common 30 mold families were determined on a damp blotting paper tray.

6.2. Lactic acid bacteria were determined by MRS-aga-. in the case of both crops to be added and malting samples.

6.3. Total bacterial concentrations were determined on 35 Plate Count agar (Difco).

6.4. The chemical properties of malt were determined by methods known from the malting process (EBC-Analytica, 1987, 4th edition).

7. Results:

Table 5 shows the concentrations of Fusarium molds and lactic acid bacteria at different stages of malting with the addition of E-390 growth medium.

Figures 2 and 3 show the total bacterial concentrations at different stages of malting for E-390 growths and E-390 cells.

Table 6 shows the results of mal-10 glass analysis when E-390 growth medium or E-390 cells have been added to the malting.

t # i 20 94875 TABLE 5. Effect of P. peniosaceus E-390 growth medium added to different stages of digestion on the Fi / vine M fungal content of malting and the number of lactic acid bacteria.

__Fujarium mold (percentage of contaminated grains.%) _

Addition Stage Barley Liko Germination Malt_

Control 55 72 96 25__ 120 ml of growth solution for barley 3 6 77 3 120 ml of growth solution for 1st grade 55 62 98 14 120 ml of growth solution for 1st + 2nd grade 55 42 90 9___ 120 ml of growth solution for germination 55 78 91 3 __Lactic acid bacteria (cell density, cfu / g) _

Control__6,0x10 * _ 3,4x1ο2 2,2x1ο4 3,0x10 * _ 120 ml of growth solution of barley__1,2x10 * _ l.lxltf 2,2x10 * 1,8x10 * _ 120 ml of growth solution 1st grade 6,0x10 '3, lxl07 2 , 8xl07 l, 4xl07 120 ml of growth solution for the 1st + 2nd lion 6.0x10 '7.4xl07 8.3xl07 5.3xl07__ 120 ml of growth solution for germination 6.0x10' 1.5x10 * 2.7x10 * 2.6xltf 21

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From the results obtained, it can be seen that the treatments according to the invention reduce in particular the amount of Fusarium mold and the total amount of bacteria at different stages of malting.

5 The addition of the lactic acid bacteria preparation did not adversely affect the quality of the malt. On the contrary, the treatment according to the invention improved the filterability of the mash obtained from the wort and reduced the B-glucan content of the malt.

10

Example 4: MANUFACTURE OF PREPARATIONS PRODUCED BY LACTIC ACID BACTERIA

1. Preparation of concentrated growth solution:

The yield strains, Pediococcus pentosaceus VTT-E-90390 (E-390) and Lactobacillus plantarum VTT-E-78076 (E-76), were grown in a fermentor in a volume of 15 l.

The inoculum volume was 6% to 7%, the culture was performed in MRS medium, at 30 ° C, for 2 days under microaerophilic conditions. The growth solutions were concentrated 10- and 20-fold by the lyophilization and evaporation method. The microbicidal activity of the concentrates was determined by the disc method.

2. Preparation of a purified solution containing microbicidal compounds: The lactic acid bacteria growth solution was purified by gel chromatography by fractionation according to molecular size. Fractions found to be microbicidally active by the disk method and turbidometrically were collected and run again through a gel column.

30

Example 5: LACTIC ACID BACTERIAL PREPARATIONS AND LACTIC *; EFFECT OF PREPARATIONS PRODUCED BY ACID BACTERIA

FOR MICROBIFLORA AND MALT QUALITY OF CHARGING

1. Strains used and preparations prepared from the strains: Bacterial strains were used:

Lactobacillus plantarum VTT-E-78076 (E-76)

Il i II I ll | i I <liri 23 94875

Pediococcus pentosaceus VTT-E-90390 (E-390)

The strains were obtained from the stock collection of VTT's biotechnology laboratory.

Preparations were made as described in Example 1, Section 3, Example 5, Sections 1 and 4, and Example 4, Sections 1 and 2.

2. Barley: The barley used was ten barley from the 1991 harvest.

10 3. The malting process:

The malting was performed as described in Example 3, but the germination phase lasted 8 days. The final humidity in Malta was less than 5%.

4. Malting: 15 Two pilot maltings were performed. Malting without additions was used as a control experiment.

4.1. First malting:

In the experimental malting, 10-fold concentrated growth solutions without cells 20 and untreated growth solution with cells were used as preparations. The preparation was added at the beginning of Lion I or at the beginning of I and II Lions. Mal loading tests 1-8, were performed as follows:

Experiment No. 1: Control Ten Barley 1991;

Experiment No. 2: At the beginning of Lions I and II, 120 ml of E-76 2b medium with cells are added; ‘Experiment No. 3: At the beginning of the Lion, 120 ml of 10-fold concentrated E-76 medium is added;

Experiment No. 4: At the beginning of Lions I and II, 120 ml of 10-fold concentrated E-76 medium is added; Experiment No. 5: At the beginning of Lions I and II, 120 ml of E-390 medium with cells are added; *, Experiment No. 6: At the beginning of the I Lion, 120 ml of 10-fold concentrated E-390 medium is added.

Experiment No. 7: At the beginning of Lions I and II, 120 ml of 10-fold concentrated E-390 growth medium is added;

Experiment No. 8: At the beginning of Lions I and II, 60 ml of E-76 medium with cells and 60 ml of E-390 medium with 24,94875 cells are added.

4.2. Second malting:

In the experimental malting, 20-fold concentrated growth solutions without cells, 5 purified microbicidal fractions without cells and untreated growth solution with cells were used as a preparation. The pH of the wastewater was adjusted. The preparation was added at the beginning of I and II Lions. Malting experiments 9-16 were performed as follows: 10 Experiment No. 9: Control Ten Barley 1991;

Experiment No. 10: At the beginning of Lions I and II, 120 ml of water, pH 3.8, is added;

Experiment No. 11: At the beginning of Lions I and II, 120 ml of E-76 medium with cells are added; Experiment No. 12: At the beginning of Lions I and II, 120 ml of E-76 fractionated concentrate, pH 3.8 is added;

Experiment No. 13: At the beginning of Lions I and II, 120 ml of 20-fold concentrated E-76 medium is added;

Experiment No. 14: At the beginning of Lions I and II, 120 ml of E-390 20 growth medium with cells are added;

Experiment No. 15: At the beginning of Lions I and II, 120 ml of E-390 fractionated concentrate, pH 3.8, is added;

Experiment No. 16: At the beginning of Lions I and II, 120 ml of 20-fold concentrated E-390 medium is added.

25 5. Analyzes performed: Samples were taken from each malting step.

Molds, lactic acid bacteria, total bacterial concentrations and physical and chemical quality characteristics of malt were determined as described in Example 3.

6. Results:

Figures 4 and 5 show the total bacterial concentrations at different stages of malting using lactic acid bacterial growth solutions with cells or concentrated or fractionated growth solutions without cells in leachate.

25 94875

Tables 7, 8, 9 and 10 show the concentrations of Fusarium molds and other molds as well as lactic acid bacteria at different stages of malting in both maltings.

5 Tables 11 and 12 show the malt analysis results from both maltings.

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Sample numbering in Example 5.

Sample Soaking__Idätvs__Mallas 1 __5.5x10 * __ 3.3x10 * __ 2.5x10 * 2 __7.3x10 * __ 1.2xl07__5.5x10 * 3 __1.4x10 * __ 4.4x10 * __ 1.1x10 * 4 __3.2x10 * __ 1.4x10 * __ 9, 5x10 * 5 __1.3xl07__7.5x10 * __ 2.6x10 «6 __4.8x10 * __ 5.0x10 * __ 4.0x10« 7 __7.3x10 * __ 2.4x10 * __ 2.6x10 * 8 l, 2xl07 l, 4xl07 2.3x10 «

The lactic acid bacterial content of barley is 7.5x10 * CFU / g.

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Sample Soaking__Idätvs Malt 9 __8.0x10 * __ 4.2x10 * __ 8.0x10 * 10 __8.0X102__5.0x10 * __ 5.6x10 * 11 __2.7xl07__2, lxl07__8.5x10 * 12 __2.2x10 * __ 3.4x10 * __ 2.5x10 * 13 __1.2x10 * __ 1.4xl07__2.3x10 * 14 __4. lxlO7__3.Sxl07__1.1x10 * 15 __4,8x10 * __ 4.8x10 * __ 3.9x10 * 16 3.5x10 * 2.1x10 * 1.7x10 *

The lactic acid bacterial content of barley is 7.5x10 * CFU / g. 1 30 94875 O in «N 2 _ V / -J _ (Μ,« 00 ®i _- r- * -: «-e £ £" 3 $ - £ 3 - 5 .2 n "" "Ϊ - S ". * h · 4> E _________________.-f

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From the results obtained, it can be seen that the treatments according to the invention reduce in particular the amount of Fusarium mold and the total amount of bacteria at different stages of malting. Furthermore, it can be seen that in addition to the growth solutions, especially concentrated and fractionated growth solutions have a beneficial effect over e.g. Fusarium molds.

From the malt analyzes it can be concluded that the additions of the growth solutions of strains E-76 and E-390 had an improving effect on the filterability of the wort made from 10 malts, the β-glucan concentrations are also lower in the corresponding malts than in the control malt.

EXAMPLE 6: LACTIC ACID ADDED IN MALT—

15 EFFECT OF PREPARATIONS PRODUCED BY BLADES ON MASK FILTERS

Figure 6 illustrates the effect of the treatment according to the invention (120 ml of culture medium / kg of barley) with preparations prepared from different strains of lactic acid bacteria on the filterability of the mash produced from the treated malt.

The strains mentioned in Example 1 were used in the experiment: E-390, E-416, E-98, E-317, E-390, E-76 and E-315. The experiment was performed using the Tepral filtration method (BIOS. 19, 1988, Grandclerc, J. et al., "Simplification of the method of filtration by Brass tepral description of the method", pp. 88-92).

It can be seen from the results that the treatment according to the invention improves the filterability of the mash.

30

Claims (6)

Process for treating industrial germinated seed material for use in food, characterized in that, in connection with the germination process, a lactic acid bacterial preparation or a preparation of a lactic acid bacterium is added to the seed material, which has the growth of a microbe. preventive activity during the germination process. 2. A method according to claim 1, characterized in that the lactic acid bacterium belongs to the Lactococcus, Leuconostoc, Pediococcus or Lactobacillus genus. 3. A method according to claim 1 or 2, characterized in that the lactic acid bacterial preparation or the preparation produced by lactic acid bacteria is obtained from Lactococcus lactis, Leuconostoc mesenteroides, Pediococcus damnosus, Pediococcus parvulus, Lactococcus parvulus, Pediococcus The Lactobacillus plantarum species or a mixture thereof, advantageously the Lactobacillus plantarum or Pediococcus pentosaceus species or a mixture thereof. 4. A process according to any one of claims 1 to 3, characterized in that, in the grain process, in addition to the digestion process, a lactic acid bacterial preparation or a preproduction produced by lactic acid bacteria is added, which has the growth-preventing activity of Fusarum mold. 5. A process according to claim 4, characterized in that the lactic acid bacterial preparation or the preparation produced by lactic acid bacteria is added at the soaking or germination stage. Process according to any of claims 1-5, characterized in that the lactic acid bacterial preparation or the preparation produced by the lactic acid bacteria is added to seed material which is to produce sprouts intended as food.