MXPA99010037A - INHIBITORS OF CELLULOLYTIC, XYLANOLYTIC AND&bgr;-GLUCANOLYTIC ENZYMES - Google Patents
INHIBITORS OF CELLULOLYTIC, XYLANOLYTIC AND&bgr;-GLUCANOLYTIC ENZYMESInfo
- Publication number
- MXPA99010037A MXPA99010037A MXPA/A/1999/010037A MX9910037A MXPA99010037A MX PA99010037 A MXPA99010037 A MX PA99010037A MX 9910037 A MX9910037 A MX 9910037A MX PA99010037 A MXPA99010037 A MX PA99010037A
- Authority
- MX
- Mexico
- Prior art keywords
- spp
- malted
- cereals
- spores
- barley
- Prior art date
Links
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Abstract
The present invention concerns an inhibitor of xylanolytic and/or&bgr;-glucanolytic enzymes, method for obtaining the inhibitor, said inhibitor and processes for obtaining micro-organism, plant or plant material wherein the activity of the inhibitor according to the invention is increased or reduced and to the use of the inhibitor, the cited micro-organism, plant or plant material in a variety of processes and applications.
Description
PROCEDURE FOR THE PREPARATION OF MALTEED CEREALS
SCOPE OF THE INVENTION
The present invention relates to an improved process for the preparation of malted cereals, improved malted cereals obtained and their use, especially in biotechnological processes for the preparation of beverages, or in feed / feed applications, laundry and food delivery systems. detergents and paper and pulp technology, as well as bleaching applications.
TECHNOLOGICAL BACKGROUND OF THE INVENTION
Cereals such as barley, wheat, rye, corn, oats, rice, millet and sorghum are used for the production of beverages. In most cases, they have undergone a malting procedure to take advantage of their enhanced enzymatic potential. In traditional malting processes, the moisture content of the cereals is increased, either by immersion (or dipping) and / or spraying (s) and the cereal with a high moisture content is allowed to germinate. After reaching the appropriate physiological conditions, it is preferably subjected to a phase (or phases) of desiccation. In the following, the term "soak" refers to the increase in the moisture level, while the term "germination" is used in the way it is explained in plant physiology. Drying operations refer to oven drying and the malting term involves all operations necessary to convert barley (or other cereals) to malted barley (or other malted cereals). The quality of the malt obtained is largely determined by the presence of endogenous enzymes of the plant generated during the malting process. For example, with cereals such as barley used as raw material for the production of malt, the variety, the composition of the microbial flora and environmental factors, such as agricultural practice, influence the quality of the malt. During cultivation and storage, cereals are contaminated with bacteria and fungi. In the malt factory, neither the air, nor the water or the equipment are sterile, and the conditions of humidity, pH and temperature favor the growth of microbial populations. The variable quality of the cereals and the lack of means to supply deficiencies during the malting process lead to variations in the quality of the malt. In many cases, this has to do with an imbalance of the specific enzymatic potential and the insufficient degradation of the cell wall. Apart from this, problems with microbial safety can occur. As a consequence of the defects in the malt, problems of quality in the production of beer take place, such as a low leaching of the must.
STATE OF THE ART
During the malting of the barley, the microflora develops and the quality of the drinks and of the malt is influenced by the activity of the endogenous microorganisms. Analogously to other biotechnological procedures, attempts have been made to optimize the aspects of the quality of the malt, adding initiation cultures during the malting process (Boivin, P & alanda, M., M., influence of Starter Cultures in Malting on the Microflora Development and Malt Quality, EBC, Meetings of the 24th Congress, pp. 95-102 (1993), Haikara, A. et al., Lactic Starter Cultures in Malting - A.Novel Solution to Gushing Problems, EBC, Meetings of the 24th Congress, pp. 163-172 (1993)). The addition of spores of Geotrichum candidum to the steeping water leads to the inhibition of the development of undesirable microorganisms and to a decrease in the time of filtration of the must produced from the obtained malt. The treatment with Geotrichum candidum also inhibits the formation of mycotoxins by Fusarium spp. The influence of Lactobacillus plantarum and Pediococcus pentosaceus has been tested on the microflora during malting and it has been found that these crops act as natural preservatives because they restrict the growth of Fusarium and prevent spillage. International patent application WO94 / 29430 describes a process for improving the properties of malted cereals in which starter cultures comprising molds, yeasts or bacteria are added before and / or during the malting of such cereals. Preferred bacteria used are lactic acid producing bacteria such as several Lactobacilli, for example, Lactobacillus casei, Lactobacillus casei var rhamnosus, Lactobacillus fermentum, Lactobacillus plantarum and Lactobacillus brevis, and bacteria of the genus Pediococcus, for example, Pediococcus acidilactici. Preferred fungi are fungi of the genus Aspergillus and Geotrichum, such as Geotrichum candidum. International patent application WO94 / 16053 describes a process for treating cereals to inhibit the growth of unwanted microbial species by inoculating the cereals during the germination process with a preparation of lactic acid bacteria or a preparation produced by the lactic acid bacteria. The preferred bacteria are lactic acid bacteria belonging to the genus Lactococcus, Leuconostoc, Pediococcus or Lactobacillus. British patent application GB-1211779 provides a method for the automatic control and regulation of a malting process. It allows to determine the necessary parameters for a malting procedure regulated and controlled automatically. At the Meetings of the European Beer Factory Convention, volume 16, 1977, pages 245 to 254, the influence of some fungi on the quality of the malt is described, more specifically, the contamination of the barley malt with mushrooms that have led to spillage and other qualitative changes in beer. Reference is also made to the spores of such fungi. German patent application DE-3028360 discloses a method for preparing malt from corn. However, the malt prepared according to the present invention is of better quality than that prepared according to any of the aforementioned documents. This is exemplified by the more intense ß-glucanase and xylanase activities, the lower ß-glucan content in the malt and must, and the better analytical data from the European Beer Factory Convention.
OBJECTIVES OF THE INVENTION
The present invention aims to provide an improved preparation process for malted cereals and improved malted cereals. A principal objective of the present invention is to provide an improved preparation process for malted cereals and improved malted cereals in terms of brewing performance, especially of maize cereals having an improved quality in terms of enzyme potential and microbial safety. Another objective is to provide a process and improved malted cereals whose quality varies to a lesser extent with the raw material used. A further objective of the present invention is to obtain malted cereals that improve the process of biotechnological production of beverages and can improve the properties of said beverages obtained. Another objective of the present invention is to use malted cereals with improved properties in food technology, such as in the baking industry as a bread additive, in feed technology, for the production of high-efficiency animal feed, in the technology of pulp and paper, as a bleaching agent, or in laundry and detergent systems such as laundry liquids, laundry powders, liquids and dishwashing powders, softeners, cleansers and soap bars as a source of enzymatic cleaning agents.
SUMMARY OF THE INVENTION
The present invention relates more specifically to a process for the preparation of malted cereals, in which the soaking phase includes one or more humidification states at a temperature between 5 and 30 ° C, preferably between 10 and 20 ° C , until the material has a moisture content of between 20 and 60% by weight, preferably between 38 and 47%, in which after a germination period of between 2 and 7 days, preferably between 3 to 6 days at a temperature between 10 and 30 ° C, preferably between 14 and 18 ° C, the soaked and sprouted cereals are preferably dried in the oven increasing the temperature to values between 40 and 150 ° C, preferably between 45 and 85 ° C, until the material has a moisture content of between 2 and 15% by weight, preferably between 4 and 7%, and in which one or more microbial cultures selected from the group consisting of one or more bacteria and / or one or more fungi, one is added or more times, either before or during or after the malting process of said cereals, and in which at least one of said microbial cultures is inoculated by activated spores, which are significantly more swollen than the inactive size, increasing the size of the spores by a factor of between 1, 2 and 10 preferably relative to the inactive type and / or having one or more germ tubes per spore. The term "fungi" as used in the present application includes both fungi and yeasts. This procedure, in this way, allows a wide flexibility in the conditions of malting. Preferably, for the preparation of the malted barley, said bacteria are selected from the group comprising Micrococcus spp., Streptococcus spp., Leuconostoc spp., Pediococcus spp., Preferably Pediococcus halophilus, Pediococcus cerevisiae, Pediococcus damnosus, Pediococcus hemophilus, Pediococcus parvulus, Pediococcus soyae, Lactococcus spp., Lactobacillus spp. preferably Lactobacillus acidophilus, Lactobacillus amylovorus, Lactobacillus bavaricus, Lactobacillus bifermentans, Lactobacillus brevis var lindneri, Lactobacillus casei var casei,
Lactobacillus delbrueckii, Lactobacillus delbrueckii var lactis, Lactobacillus delbrueckii var bulgaricus, Lactobacillus fermenti, Lactobacillus gasserii,
Lactobacillus helveticus, Lactobacillus hilgardii, Lacbotacillus renterii,
Lacbotabacillus sake, Lactobacillus sativorius, Lactobaciilus cremoris,
Lactobacillus kefir, Lactobacilius pentoceticus, Lactobacillus cellobiosus,
Lactobacillus bruxellensis, Lactobacillus buchnerii, Lacboacillus coryneformis, Lactobacillus confusus, Lactobacillus florentinus, Lactobacillus viridescens, Corynebacterium spp., Propionibacterium spp., Bifidobacterium spp., Streptomyces spp., Bacillus spp., Sporolactobacillus spp., Acetobacter spp., Agrobacterium spp., Alcaligenes spp., Pseudomonas spp. preferably Pseudomonas amylophilia, Pseudomonas aeruginosa, Pseudomonas cocovenenans, Pseudomonas mexicana, Pseudomonas pseudomallei, Gluconobacter spp., Enterobacter spp., Erwinia spp., Klebsiella spp., Proteus spp. Preferably, for the preparation of the malted barley, the fungi are selected from the group comprising the genera (genera described by the Ainsworth and Bisby Dictionary of Mushrooms, 8th Edition, 1995, edited by DL Hawksworth, PM Kirk, BC Sutton and DN Pegler (632 pp) Cab International) comprising Ascomycota preferably Dothideales preferably Mycosphaereláceas preferably Mycosphaerella spp., Venturiaceae preferably Venturia spp; Eurotiales preferably Monascaceae preferably Monascus spp., Trichocomaceae preferably Emericilla spp., Euroteum spp., Eupenicillium spp .; .. Neosartorya spp, Talaromyces spp, Hypocreales preferably Hypocreceae preferably Hypocrea spp Saccharomycetales preferentially Dipodascaceae preferentially Dipodascus spp, Galactomyces spp, Endomycetaceae preferentially Endomyces spp, Metschnikowiaceae preferentially Guilliermondella spp, Saccharomycetaceae preferentially Debaryomyces spp, Dekkera spp, Pichia...... spp., Kluyveromyces spp., Saccharomyces spp., Torulaspora spp., Zygosaccharomyces spp., Saccharomycodaceae preferably Hanseniaspora spp.
Schizosaccharomycetales preferentially Schizosaccharomycetaceae preferentially Schizosaccharomyces spp., Sordariales preferentially Chaetomiaceae preferentially Chaetomium spp., Sordariaceae preferentially Neurospora spp., Zygomycota preferentially Mucorales preferentially Mucoraceae prefentemente Absidia spp., Amylomyces spp., Rhizomucor spp., Actinomucor spp., Thermomucor spp., Chlamydomucor spp ., Mucor spp., Preferably Mucor circinelloides, Mucor grisecyanus, Mucor hiemaiis, Mucor indicus, Mucor, Mucor piriformis, Mucor plumbeus, Mucor praini, Mucor pusillus, Mucor silvaticus, Mucor javanicus, Mucor racemosus, Mucor rouxianus, Mucor rouxii, Mucor aromaticus, Mucor flavus, Mucor miehei, Rhizopus spp., preferably Rhizopus arrhizus, Rhizopus oligosporus, Rhizopus oryzae preferably strains ATCC 4858, ATCC 9363, NRRL 1891, NRRL 1472, Rhizopus stolonifer, Rhizopus thailandensis,
Rhizopus formosaensis, Rhizopus chinensis, Rhizopus cohnii, Rhizopus japonicus, Rhizopus nodosus, Rhizopus delemar, Rhizopus acetorinus,
Rhizopus chlamydosporus, Rhizopus circinans, Rhizopus javanicus, Rhizopus peka, Rhizopus saito, Rhizopus tritici, Rhizopus niveus, Rhizopus microsporus; Mitosporic fungi preferably Aureobasidium spp., Acremonium spp.,
Cercospora spp., Epicoccum spp., Moniiia spp., Preferably Monilia candida, Moniiia sitophila, Mycoderma spp., Candida spp., Preferably
Candida diddensiae, Candida edax, Candida etchelysii, Candida kefir, Candida krisei, Candida lactose, Candida lambica, Candida melinii, Candida utilis, Candida milleri, Candida mycoderma, Candida parapsilosis, Candida cosex, Candida tropicalis, Candida valid, Candida versatilis, Candida guilliermondii , Rhodotoruia spp., Torulopsis spp., Geotrichum spp., Preferably Geotrichum amycelium, Geotrichum armillariae, Geotrichum asteroides, Geotrichum bipunctatum, Geotrichum dulcitum, Geotrichum eriense, Geotrichum fici, Geotrichum flavo-brunneum, Geotrichum fragans, Geotrichum gracile, Geotrichum heritum, Geotrichum klebaknii, Geotrichum penicillatum, Geotrichum hirtum, Geotrichum pseudocandidum, Geotrichum rectangulatum, Geotrichum suaveolens, Geotrichum vanryiae, Geotrichum loubieri, Geotrichum microsporum, Cladosporium spp., Trichoderma spp., preferably Trichoderma hamatum, Trichoderma harzianum, Trichoderma koningii, Trichoderma pseudokoningii, Trichoderma reesei, Trichoderma vi rgatum, Trichoderma viride, Oidium spp., Alternaria spp., preferably Alternaria alternata, Alternaria tenuis, Helminthosporium spp., preferably
Helminthosporium gramineum, Helminthosporium sativum, Helminthosporium teres, Aspergillus spp. as described by R.A: Samson (1994) in the Biotechnology Manual, volume 7: Aspergillus, edited by Smith, J.E. (273 pp), Plenum Press), preferably the Aspergillus ochraseus Group (Thom &
Church), the Aspergillus nidulans Group (Thom &Church), the Group of
Aspergillus versicolor (Thom &Church), the Aspergillus group wentii (Thom &
Raper), Aspergillus candidus Group (Thom &Raper), Aspergillus flavus Group (Raper &Feneil), Black Aspergillus Group (Thom &Church), Penicillum spp. preferably Penicillium aculeatum, Penicillum citrinum, Penicillum claviforme, Penicillum funiculosum, Penicillum italicum, Pnicillum lanoso-viride, Penicillum emersonii, Penicillum lilacinum, Penicillum expansum. Preferably, for the preparation of malted cereals other than barley, especially for the preparation of malted wheat, rye, corn, oats, rice, millet and sorghum, said bacteria are selected from the group consisting of Micrococcus spp., Streptococcus spp. , Leuconostoc spp., Pediococcus spp., Lactococcus spp., Lactobacillus spp., Corynebacterium spp., Propionibacterium spp., Bifidobacterium spp., Streptomyces spp., Bacillus spp., Sporolactobacillus spp., Acetobacter spp., Agrobacterium spp., Alcaligenes spp., Pseudomonas spp., Gluconobacter spp., Enterobacter spp., Erwinia spp., Klebsiella spp., Proteus spp. or its mixture; and said fungi are fungi selected from the group consisting of Ascomycota preferably Dothideales preferably Mycosphaereláceas preferably Mycosphaerella spp., Venturiaceae preferably Venturia spp; Eurotiales preferably Monascaceae preferably Monascus spp., Trichocomaceae preferably Emericiila spp., Euroteum spp.,
Eupenicillium spp; Neosartorya spp., Talaromyces spp., Hypocreales preferably Hypocreceae preferably Hypocrea spp.:
Saccharomycetales preferably Dipodascaceae preferably
Dipodascus spp., Galactomyces spp., Endomycetaceae preferably
Endomyces spp., Metschnikowiaceae preferably Guilliermondella spp.,
Saccharomycetaceae preferably Debaryomyces spp., Dekkera spp., Pichia spp., Kluyveromyces spp., Saccharomyces spp., Torulaspora spp., Zygosaccharomyces spp., Saccharomycodaceae preferably Hanseniaspora spp. Schizosaccharomycetals preferably
Schizosaccharomycetaceae preferably Schizosaccharomyces spp., Sordariales preferably Chaetomiaceae preferably Chaetomium spp., Sordariaceae preferably Neurospora spp., Zygomycota preferably Mucorales preferably Mucoraceae preferably Absidia spp., Amylomyces spp., Rhizomucor spp., Actinomucor spp., Thermomucor spp., Clamydomucor spp., Mucor spp., Rhizopus spp., Mitosporic fungi preferably Aureobasidium spp., Acremonium spp., Cercospora spp., Epicoccum spp., Monilia spp., Mycoderma spp., Candida spp., Rhodotorula spp., Torulopsis spp., Geotrichum spp. , Cladosporium spp., Trichoderma spp., Oidium spp., Alternaria spp., Helminthosporium spp., Aspergillus spp., Penicillium spp.
According to a preferred embodiment, the process for preparing malted cereals according to the present invention comprises the following steps: the soaking step includes one or several stages of humidification or the total time of immersion in water during soaking for physiological reasons does not exceed 30 hours (preferably 10 to 25 hours) or the oven drying step includes more than two temperature ranges and the microbial cultures that are added are preferably selected from the group consisting of Rhizopus spp., Preferably from
Rhizopus oryzae, such as strain ATCC 9363 of Rhizopus oryzae and / or Pseudomonas spp., Preferably Pseudomonas herbicola, or Aspergillus spp., Preferably Aspergillus oryzae such as strain ATCC 14156 of Aspergillus oryzae. According to the present inventionMalted cereals are selected from the group consisting of barley, wheat, rye, corn, oats, rice, millet and sorghum. In the process according to the present invention, identical or different microbial cultures are added in the presence of once or several activated spores. The microbial cultures used are preferably fungal cultures. The use of activated spores greatly enhances their contribution to improved malt quality, most likely because of more vigorous growth. The activated spores possess one of the following properties: the treated spores are more swollen than the inactive ones, more particularly, the size of the spores increases by a factor preferably between 1, 2 and 10 with respect to their inactive size and / or form one or more germ tubes per spore. The activated spores are prepared by subjecting them to environmental changes, preferably by means of at least one or a combination of the following treatments: (a) humidification and / or drying cycles, (b) addition of nutritional provisions (such as a nitrogen source, preferably mono or disaccharides) or addition of spore elements, (c) exposure to changes in temperature, preferably of the order of 0 to 80 ° C, (d) exposure to changes in pH, preferably of the order of 2.0 to 8, 0, more preferably between 3.0 and 6.0. The person skilled in the art can easily select precise treatment steps to obtain either the swelling of the spores or the germ tubes, as mentioned above. The present invention also relates to the malted cereals obtained, which present improved analytical results according to the European Convention of Beer Factories. Said improvements may be related to modifications and / or increased hydrolytic enzymatic activities. At the same time, a decreased level of toxins, increased microbial safety for example, non-competitive non-competitive microbial flora such as Fusarium and / or increased acceptability compared to malted cereals can be observed, according to the state of the art. For example, malted cereals according to the invention may have a lower β-glucan content or a higher xylase or β-glucuronase activity (represented in the following examples and figures) than malted cereals according to the state of the art. This allows a better processing of the malt in the must and beer production, as exemplified by the increased filtration rates. Another objective of the present invention relates to the use of the malted cereals according to the invention for the preparation of beverages. The present invention also relates to these improved beverages. The improved malted cereals according to the invention can also be used advantageously during the manufacture of beer without alcohol or with low content of alcohol or light beer, since the greater enzymatic activity will enhance the elimination of alcohol from beer. The improved malt cereals according to the invention could also be used in other biotechnological processes well known to those skilled in the art, in which in most cases their improved quality is utilized. Another object of the present invention relates to the use of malted cereals with improved properties in food technology such as the baking industry as bread additives, in feed technology for the production of animal feed with superior conversion characteristics , in the technology of paper and pulp; as bleaching agents, or in detergent-based compositions. The present invention will be further described, in several examples, in view of the following drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 represents the β-glucanase activity of the malted barley obtained according to the preparation procedure of example 1 (legend: see example 1) Figure 2 represents the xylanase activity of the malted barley obtained according to the preparation procedure of example 1 ( legend: see example 1) Figure 3 represents the β-glucanase activity of the malted barley obtained according to the preparation procedure of example 3 (legend: see example 3). Figure 4 represents the xylanase activity of the malted barley obtained according to the preparation procedure of example 3 (legend: see example 3). Figure 5 represents the relative magnification factor (RIF) for the bacterial populations (see text, evaluation of the malt, example 2) (legend: see example 2).
EXAMPLE 1
1. Preparation of microbial cultures
Strain S46: Rhizopus oryzae ATCC 9363
Preparation of the spore suspension - The strain was grown on PDA (Dextrose Agar potato, Oxoid) for approximately 10 days at 28 ° C; - The spores were collected by flooding the culture with sterile physiological saline (0.9% NaCl) and gently rubbing the sporulated mycelium with a sterile spatula; - The spore suspension was washed twice with sterile physiological saline (0.9% NaCl) by centrifugation (5500 rpm, Sorvall centrifuge type SS-34 ° for 15 minutes) and resuspended in sterile physiological saline (NaCl 0.9%); - The density of the spores was determined microscopically using a Thoma counting chamber.
Activation of spore suspension. - 107 spores were transferred to 20 ml of sterile acidified TSB (Tryptic soybean broth, Oxoid), pH = 4.0 and incubated in an aqueous bath with stirring for 5 to 6 hours at ± 42 ° C; - Activated spores were collected by centrifugation
(3500 rpm, Sorvall SS-34 ° centrifuge for 15 minutes), washed once with sterile physiological saline (0.9% NaCl) by centrifugation (3500 rpm, Sorvall SS-34 ° centrifuge for 15 minutes) and resuspended in sterile physiological saline (0.9% NaCl).
2. Cebada Plaisant - French Harvest 1994;
3. Process
Processing The malts were obtained by four different malting procedures: A1. Traditional malting (without inoculation of any spore suspension) B1. Malting procedure with inoculation of non-activated spores (inoculation of barley soaked with a suspension of non-activated spores of Rhizopus oryzae ATCC 9363) - C1. Malting process according to the invention (inoculation of the barley soaked with a suspension of activated spores of Rhizopus oryzae ATCC 9363) - D1. Malting process according to the invention (inoculation of the soaked barley during the first stage of humidification with a suspension of activated spores of Rhizopus oryzae ATCC 9363).
Soaking - Soaking was carried out on a 2 kg base with a ratio of total water (tap water) to air dried barley of 1: 5: 1; - Two termendores (Bioflo III,
New Brunswick Scientific), in which perforated plates were placed; - The temperature was only controlled during the humidification phases; during the phases of resting in the air, the system was allowed to reach room temperature (± 20 ° C); - During the entire soaking period, the barley was aerated (4 liters of sterile air per minute); - The soaking was carried out by immersion using the following scheme:
Addition of microbial cultures - ± 460 g of soaked barley were immersed in 0.5 I of tap water containing no spores (A1), non-activated spores of Rhizopus oryzae ATCC 9363 (B1) or activated spores of Rhizopus oryzae ATCC 9363 (C1 according to the invention): for B1 and C1, the soaked barley was inoculated with 10 spores per gram of air-dried barley; - During the soaking, 104 activated spores were inoculated per gram of air-dried barley in the water of the first humidification phase (D1); - The liquid was removed by drainage.
Germination - The germination was carried out in a cylindrical container with perforated lids at a temperature of 16-18 ° C for 4 days; - The air was supplied by natural diffusion; - The containers were turned slowly in an electronically controlled rotation system (Cellroll®, Tecnorama), that is, every two hours the containers were turned for 15 minutes at 1 rpm.
Baked Drying - Oven drying was carried out in a malting unit Joe White (Australia)
4. Methods of analysis and results. Methods for the determination and units of humidity, extracts, difference of extracts, color, total protein content, content of soluble proteins, Kolbach index, pH, diastatic power, according to the
European Analytical Convention of Beer Factories (4th edition, 1987,
Brauerei and Getránke-Rundschau). Methods for the determination and units of turbidity, friability, homogeneity, whole grain seeds, β-glucan content, according to the European Analytical Convention of Beer Factories (4th Edition, 1987, Brauerei and Getránke-Rundschau, supplement published in 1989 ). The post-coloration of the must is determined after boiling under reflux at 108 ° C for 2 hours.
The must viscosity is determined with the Delta viscometer. For the determination of the volume of filtration, the must is filtered on a Schleicher filter and Schuell 597 folded 1/2. The volume (in me) that is obtained after one hour of filtration is the volume of filtration of the must. The modification is determined with the Calcofluor device
(Haffmans) according to the Carsiberg method (European Analytical Convention of Beer Factories, 4th edition, 1987, Brauerei and Getránke-Rundschau). The xylanic and ß-gluconase activities are determined with the ß-glucazym method (Megazyme (Austr.) Pty Ltd (April, 1993)] and the xylazym method (Megazyme (Austr) .Pty Ltd (September, 1995) ), respectively.
ND: Not determined Figures 1 and 2 represent the β-glucanase and xylanase activity, respectively, of the malted barley obtained (A1, B1, C1, D1). The ß-glucanase activity was determined by the ß-glucazyme method [(Megazyme (Austr.) Pty Ltd (April, 1993)]. Therefore, the ß-glucanase activity of the malt (U / kg) was calculated as 380 × E (590 nm) + 20. The xylanase activity was determined with the endo 1-4-xylazyme method [(Megazyme (Austr.) Pty Ltd (September 1995)]. Therefore, the xianic activity of the malt (U / kg) ) was calculated as (46.8 x E (590 nm) + 0.9) x 5.
EXAMPLE 2
1. Preparation of microbial cultures
Strain - S46: Rhizopus oryzae ATCC 9363
Preparation of the spore suspension - as described in example 1
Activation of the spore suspension - as described in example 1
2. Barley - Stander- North American Harvest 1995
3. Process
Processing Malts were obtained by six different malting procedures: - A2. Traditional malting (without inoculation of any spore suspension) - B2. Malting procedure with inoculation of non-activated spores (inoculation of barley soaked with a suspension of non-activated spores of Rhizopus oryzae ATCC 9363) - C2. Malting process according to the invention (inoculation of the soaked barley during the first humidification phase with a suspension of activated spores of Rhizopus oryzae ATCC 9363) - D2. Malting process according to the invention (inoculation of the soaked barley during the second stage of humidification with a suspension of activated spores of Rhizopus oryzae ATCC 9363). - E2. Malting process according to the invention (inoculation of the soaked barley during the third stage of humidification with a suspension of activated spores of Rhizopus oryzae ATCC 9363). - F2. Malting process according to the invention (inoculation of the barley soaked with a suspension of activated spores of Rhizopus oryzae ATCC 9363).
Soaking and addition of microbial cultures. The soaking was carried out on a base of 300 g with a ratio of total water (tap water) to air-dried barley of 5: 3; 2000 ml flasks were used for the soaking; A temperature of 18 ° C was maintained during the humidification phases and during the phases of resting in the air; During the period of total soaking the barley was aerated by compressed air; The soaking was carried out by immersion using the following scheme:
During the soaking, 10 activated spores were inoculated per gram of air-dried barley in the water of the first humidification phase (C2), of the second humidification phase (D2) or of the third humidification phase (E2) before immersion of barley; The soaked barley was immersed in 0.5 liters of tap water that did not contain spores (A2, C2, D2, E2), non-activated spores (B2) or activated spores (F2);
4 For B2 and F2, the soaked barley was inoculated with 10 spores per gram of air-dried barley; The liquid was removed by drainage.
Germination - as described in example 1
Baked in the oven - as described in example 1
EVALUATION OF THE MALTES
Determination of the increase of the bacterial population. To judge the evolution of the bacterial population during the malting process, a relative increase factor (R.I.F) was determined by dividing the total bacterial count present in the green malt by the total bacterial count in the barley. The total bacterial count was determined after appropriate dilutions of an extract of the pips in tryptic soy agar (Oxoid) supplemented with 100 ppm of pimaricin and after incubation at 28 ° C for 3 days. Figure 5 shows the increase of the bacterial population during the malting according to the preparation procedure of example 2.
EXAMPLE 3
1. Preparation of microbial cultures.
Strain - S46: Rhizopus oryzae ATCC 9363
Preparation of the spore suspension - as described in example 1
Activation of Is spore suspension - as described in example 1
2. Cebada Plaisant - French Harvest 1994;
3. Process
Processing The malts were obtained by three different malting procedures: - A3 Traditional malting (no inoculation of any spore suspension) B3 Malting procedure with non-activated spore inoculation (inoculation of the barley soaked with a suspension of non-activated Rhizopus spores) oryzae ATCC 9363) - C3 Malting process according to the invention (inoculation of barley soaked with a suspension of activated spores of Rhizopus oryzae ATCC 9363)
Soaking - Soaking was carried out on an air-dried barley of 2 kg base with a ratio of total water (tap water) to air-dried barley of 1: 5: 1; the pH of the soaking water was controlled to pH = 5.5 by adding lactic acid and NaOH; - A fermentor was used for the soaking (Bioflo III,
New Brunswick Scientific), in which a perforated plate was placed; The temperature was only controlled during the humidification phases; during the phases of rest in the air, the system was allowed to reach room temperature (ca. 20 ° C); - During the entire soaking period the barley was aerated (4 liters of sterile air per minute); The soaking was carried out by immersion using the following scheme:
Addition of microbial cultures 460 g of soaked barley were immersed in 0.5 I of tap water containing no spores (A3), non-activated spores of Rhizopus oryzae ATCC 9363) (B3) or activated spores of Rhizopus oryzae ATCC 9363
(C3 according to the invention): for B3 and C3, the soaked barley was inoculated with
1. 10 spores per gram of air-dried barley; The liquid was removed by drainage.
Germination as described in example 1
Baked in the oven - as described in example 1
4. Methods of analysis and results. These were as described in example 1 (4, Methods of analysis and results). See box on the next page. In this frame:
A1 / 3: Traditional malting process B1 / 3: Malting process with inoculation of non-activated spores C1 / 3: Malting process according to the invention
? ?
Figure 3 represents the β-glucanase activity, measured according to the β-Glucazyme method [(Megazyme (AUSTR) Pty, Ltd)], of the malted cereals A3, B3 and C3. The ß-glucanase activity of the malt (U / kg) was calculated as described in Example 1. A3 was obtained by the traditional malting method with pH control of the steeping water (pH =
.5). B3 came from the malting process according to the invention with the inoculation of the barley soaked with a suspension of non-activated spores of Rhizopus oryzae ATCC 9363 and with pH control of the soaking water (pH = 5.5). C3 was obtained by the malting process according to the invention with the inoculation of the barley soaked with a suspension of activated spores of Rhizopus oryzae ATCC 9363 and with pH control of the soaking water (pH = 5.5). These results show the increased β-glucanase activity when the pH of the steeping water is maintained at around 5.5. Figure 4 gives the corresponding results for the xylanase activity. These were measured according to the method Xylazyme, Megazyme [(AUSTR), Pty. Ltd. (September 1995))]. The xylanase activity of the malt was calculated as described in example 1.
COMPARISON OF THE ACTIVITY OF ß-GLUCANASE OBTAINED
ACCORDING TO EXAMPLES 1 AND 3 WITH THE ACTIVITY OF ß-GLUCANASE
ACCORDING TO THE STATE OF THE TECHNOLOGY AS DESCRIBED IN THE
PATENT DOCUMENT WO94 / 29430
In order to compare the improved results with respect to the activity of the β-glucanase by the present invention, we defined the factor μ as follows: Activity of the β-glucanase of the treated malt μ = Activity of the β-glucanase of the control malt This factor was calculated for the control malt and the malt treated with Rhizopus oryzae ATCC 9363 as described in examples 1 and 3 of the present invention. It was also calculated for the data described in patent document WO94 / 29430 (example 1) in which Geotrichum candidum was used. As described in both documents, WO94 / 29430, and in the present application, the activity of β-glucanase was determined with the beta-glucazyme method [(Megazyme (Austr) Pty. Ltd. (April 1993)] Therefore, the activity of the β-glucanase (U / kg) was calculated as 380 x E (590 nm) + 20 and one unit of activity was defined as the amount of the enzyme needed to release a micromole of sugar equivalents reducer per minute under the conditions defined above.
Comparison of results:
Ge: Geotrihum candidum The results clearly show that the present invention provides a more significant increase in β-glucanase activity than that described above (WO 94/29430). It is observed, therefore, that it is possible to obtain malted cereals having an increased β-glucanase activity by at least a factor of 4 compared to the conventional malting process in which the addition of microbial culture is omitted. From Figures 2 and 4 it is also apparent that it is possible to obtain malted cereals possessing a xylanase activity increased by at least a factor 4, compared to the conventional malting process in which the addition of the microbial culture is omitted.
EXAMPLE 4
1. Preparation of microbial cultures
Strain - S40: Aspergillus oryzae ATCC 14156
Preparation of Is spore suspension The strain was grown on PDA (Dextrose Agar potato, Oxoid) for about 7 days at 28 ° C; The spores were collected by flooding the culture with sterile physiological saline (0.9% NaCl) and gently rubbing the sporulated mycelium with a sterile spatula; The spore suspension was washed once with sterile physiological saline (0.9% NaCl) by centrifugation (5500 rpm, Sorvall centrifuge type SS-34 ° for 15 minutes) and resuspended in sterile physiological saline solution. The density of the spores was determined microscopically using a Thoma counting chamber.
Activation of spore suspension. 5,107 spores were transferred to 20 ml of sterile acidified TSB (Tryptic soybean broth, Oxoid), pH = 5.0 and incubated in a shaking aqueous bath for 3 hours (1) or 1 hour (2) to 35 minutes. ° C;
2. Barley Cereal Clarine- French harvest 1995
3. Process
Processing Malts were obtained by two different malting procedures A4 Traditionally (without inoculation of any spore suspension) E4 Method of maiteating according to the invention (inoculation of the soaked barley during the first and third humidification period with a suspension of activated spores of Apergillus orizae ATCC 14156)
Soaking - as described in example 1
Addition of microbial cultures During the soaking, 5,103 activated spores (1) were inoculated per gram of air-dried barley in the water of the first humidification period and 1,104 activated spores (2) per gram of air-dried barley were inoculated into the water from the third humidification period (E4);
Germination The germination of ± 460 g of a soaked barley was carried out in cylindrical containers with perforated lids at a temperature of 16-18 ° C for 4 days; The air was supplied by natural diffusion; The containers rotated slowly in an electronically controlled rotation system (Cellroll®, Tecnorama), that is, every two hours the containers were rotated for 15 minutes at 1 rpm.
Baked in the oven - as described in example 1
4. Methods of analysis and results. These were as those described in example 1 (4. methods of analysis and results). The lengths of the desoiled plumules of the seeds were determined by sorting the pips into 6 types, that is, those with pips that did not have seeds developed from the seeds (O) and those that had a length of the plumules developed from the seeds. seeds from 0 to 25% (0-1 / 4), 25 to 50% (1 / 4-1 / 2), from 50% to 75% (3 / 4-1) and > 100% (> 1) of the length of the nugget.
It was reported that the use of activated spores of Aspergillus oryzae ATCC 14156 improved the analytical specifications of the malt (see below). Furthermore, it was unexpectedly found that during the malting process of the barley, the lengths of the plumules of the seeds were significantly longer when the method according to the invention was used instead of the traditional method.
EXAMPLE 5
1. Preparation of microbial cultures
Strains S40: Aspergillus oryzae ATCC 14156 S46: Rhizopus oryzae ATCC 9363
Preparation of spore suspensions. - as described in example 4 Activation of S 40 spore suspensions: 5,107 spores were transferred to 20 ml of sterile acidified TSB (Tryptic soy broth, Oxoid), pH = 5.0 and incubated in an aqueous bath stirring for one hour at 35 ° C; The activated spores were harvested by centrifugation (3500 rpm, Sorvall centrifuge of SS-34 ° type for 15 minutes) and resuspended in sterile physiological saline (0.9% NaCl). S 46: 5,107 spores were transferred to 20 ml of sterile acidified TSB (Tryptic soy broth, Oxoid), pH = 4.0 and incubated in a water bath for 5 hours at 42 ° C; The activated spores were harvested by centrifugation (3500 rpm, Sorvall centrifuge of SS-34 ° type for 15 minutes) and resuspended in sterile physiological saline (0.9% NaCl).
2. Clarine Barley Cereal - French vintage 1995 3. Procedure
Processing Malts were obtained by two different malting procedures: A5. Traditional malting (without inoculation of any spore suspension) F5. Malting process according to the invention
(inoculation of the soaked barley during the first period of humidification with a suspension of activated spores of Apergillus oryzae
ATCC 14156 and following soaking with a suspension of activated spores of Rhizopus oryzae ATCC 9363)
Soaking - as described in example 1
Addition of microbial cultures During the soaking, 1,104 activated spores of Apergillus oryzae ATCC 14156 were inoculated per gram of air-dried barley in the water of the first humidification period (F5, according to the invention); ± 460 g of soaked barley were immersed in 0.5 I of tap water that did not contain spores (A5) or activated spores of Rhizopus oryzae ATCC 9363 (F5 according to the invention); for F5 the soaked barley was inoculated with 1,104 activated spores per gram of air-dried barley; The liquid was removed by drainage:
Germination - As described in example 4
Baked in the oven As described in example 1
4. Methods of analysis and results. These were as those described in example 1 (4. methods of analysis and results). The method for determining the length of the plumules of the seeds as in example 4.
EXAMPLE 6
1. Preparation of microbial cultures.
Strain S46: Rhizopus oryzae ATCC 9363
Preparation of spore suspensions. - as described in example 4
Activation of spore suspensions 5,107 spores were transferred to 20 ml of sterile acidified TSB (tryptic soybean broth, Oxoid), pH = 4.0 and incubated in an aqueous shaking bath for five hours at 42 ° C; The activated spores were harvested by centrifugation (3500 rpm, Sorvall centrifuge of the SS-34 ° type for 15 minutes) and resuspended in sterile physiological saline (0.9% NaCl).
2. Wheat cereal: Belgian vegetable harvest 1996 3. Procedure
Processing Malts were obtained by two different malting procedures: A6. Traditional malting (without inoculation of any spore suspension) D6. Malting process according to the invention (inoculation of the soaked wheat during the first humidification period with a suspension of activated spores of Rhizopus oryzae ATCC 9363)
Soaking Soaking was carried out on a 2 kg base with a ratio of total water (tap water) to air of 1: 5: 1; - Two termendores were used for the soaking
(Bioflo III, New Brunswick Scientific), in which a perforated plate was placed; The temperature was only controlled during the humidification phases; during the phases of rest in the air, the system was allowed to reach room temperature (± 20 ° C); - During the period of total soaking the wheat was aerated
(4 liters of sterile air per minute); - The soaking was carried out by immersion using the following scheme:
Addition of microbial cultures ± during the soaking, 1,104 activated spores were inoculated per gram of wheat dried in the air in the water of the first humidification phase (D6);
Germination as described in example 4
Baked in the oven as described in example 1
4. Methods of analysis and results These were as described in example 1 (4, Methods of analysis and results).
EXAMPLE 7
1. Preparation of microbial cultures
Strain - S46: Rhizopus oryzae ATCC 9363 Preparation of the spore suspension The strain was grown on PDA (Dextrose Agar potato, Oxoid) for approximately 7 days at 28 ° C; The spores were collected by flooding the culture with sterile physiological saline (0.9% NaCl) and gently rubbing the sporulated mycelium with a sterile spatula; The spore suspension was washed once with sterile physiological saline (0.9% NaCl) by centrifugation (3500 rpm, Jouan C312 centrifuge, for 15 minutes) and resuspended in sterile physiological saline (NaCl 0, 9%); The density of the spores was determined microscopically using a Thoma counting chamber.
Activation of spore suspension. - 5,107 spores were transferred to 20 ml of sterile acidified TSB (Tryptic soybean broth, Oxoid), pH = 4.0 and incubated in an aqueous shaking bath for 5 hours (1) at 42 ° C;
2. Sorghum cereal (S14) 3. Procedure
Processing Malts were obtained by two different malting procedures: A7. Traditional malting (without inoculation of any spore suspension) D7. Malting process according to the invention (inoculation of sorghum during the first period of humidification with a suspension of activated spores of Rhizopus oryzae ATCC 9363)
Cleaning Sorghum washing is done using 6 liters of tap water per kilogram of sorghum and removing excess water
Soaking Soaking was carried out on a 2 kg base with a ratio of total water (tap water) to air of 1: 5: 1; Two soils were used for the soaking (Bioflo III, New Brunswick Scientific), in which a perforated plate was placed; The temperature was only controlled during the humidification phases; during the phases of resting in the air, the system was allowed to reach room temperature (± 20 ° C);
During the period of total soaking, the barley was aerated (2 liters of sterile air per minute); - The soaking was carried out by immersion using the following scheme:
Addition of microbial cultures ± During the soaking, 1.10 activated spores (1) were inoculated per gram of air-dried sorghum in the water of the first humidification phase (D7);
Germination The germination of ± 460 g of a soaked sorghum was carried out in cylindrical containers with perforated lids at a temperature of 28 ° C for 4 days; The air was supplied by natural diffusion; The containers were rotated slowly in an electronically controlled rotation system (Cellroil®, Tecnorama); that is, every two hours the containers were turned for 15 minutes at 1 rpm.
Baked in the oven - as described in example 1
4. Methods of analysis and results. These were as those described in example 1 (4. methods of analysis and results).
EXAMPLE 8 Bread making
The execution of the wheat malts described in example 6 (A6: traditional malting process, D6: malting process according to the invention) were compared in a 100 g procedure described by Finney, KF, An optimized straight-dought breadmaking method after 44 years, cereal Chemistry, 61, pp 20-27 (1984). The recipe used commercial wheat flour, 6.0% sugar, Crisco 3.0% (Crisco, Procter and Gamble, Cincinatti, OH, USA), salt at 1.5% and yeast at 2.5 % (Bruggeman, Belgium). The malts were tested in a concentration range of 0 to 0.25% and replaced an equal weight of flour.
Method of analysis and results The specific volumes of the bread (ie the volume in ce by weight in g of bread) were determined using the substitution of rapeseed seeds, evaluating bread crumb. It was clearly observed that the malt according to the invention was a much more potent agent increasing the bread volume than the malt obtained by the traditional malting process. At the same time, no significant differences were found in the structure of the crumb of the breads prepared with the malt according to the invention and the malt of the conventional processes.
Therefore, the present invention also includes the process for making bread that demonstrates an increase in bread volume of 3% compared to bread made from known malt.
REFERENCES
Thom, C. and Church, M.B., 1926, The Aspergilli, Williams and Wilkins, Baltimore. Thom, C. and Raper, K.B., 1945, A Manual of the Aspergilli, Williams and Wilkins, Baltimore. Raper, K.B. and Fennel, D.I., 1965, The Genus Aspergillus, Williams and Wilkins, Baltimore. Haffmans B.V., Marinus Dammeweg 30, Postbus 3150 5902 RD Venlo Holland, The Netherlands.
LIST OF SEQUENCES (1) GENERAL INFORMATION:
(i) APPLICANT: (A) NAME: LEUVEN RESEARCH & DEVELOPMENT (B) STREET. Groat Begijnhof 59 (C) CITY: Leuven (E) COUNTRY: BELGIUM < • (F) POSTAL CODE. B-3000
(A) NOM8RE: DEBYSER Winok (B) STREET: illem Coosemansstraat 23 (C) CITY: Kessel-Lo (E) COUNTRY: BELGIUM (F) POSTAL CODE: B-3010
(A) NAME: DELCOR Jan (B) STREET: Kastapjelaap, 14 (C) CITY: Leuven (E) COUNTRY. BELGIUM (F) POSTAL CODE: B-3001 (¡i) TITLE OF THE INVENTION- INHIBITORS OF CELLULOLITIC ENZYMES, XYLANOLITICS AND BETA-GLUCANOLITICS (iii) NUMBER OF SEQUENCES: 2 (iv) LEGIBLE FORM OF THE COMPUTER: (A) TYPE OF MEDIUM: soft disk (B) COMPUTER: IBM PC Compatible (C) OPERATING SYSTEM: PC-DOS / MS-DOS (D) SOFTWARE: Patentlp Relay # 1.0. Version # 1.30 (EPO) (2) INFORMATION FOR SEQ ID NO: 1 (I) CHARACTERISTICS OF THE SEQUENCE "(A) LENGTH: 14 amino acids (B) TYPE: Amyloacid (C) TYPE OF FILAMENT: simple (D) TOPOLOGY : linear
(ii) TYPE OF MOLECULE: peptide (ix) CHARACTERISTICS: (A) NAME / KEY: modified sITy (B) LOCALl2AClON: 12 (D) OTHER INFORMATION: / product? = "Asp preferably"
(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO. Lys Gly Leu Pro Val Leu Pro Wing Val Thr Lys Xaa Thr Wing 1 5 10 (2) INFORMATION FOR S'EC ID NO.2 (I) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 17 amino acids (B) TYPE: amino acid (C) TYPE OF FILAMENT: simple (D) TOPOLOGY: linear
(p) TYPE OF MOLECULA. peptide (1X) CHARACTERISTICS: (A) NAME / KEY, modified site (B) LOCATION: 1 (D) OTHER INFORMATION: / product ^ "Ser, Phe or Gly" (ix) CHARACTERISTICS: ( A) NAME / KEY: modified site (B) LOCATION: 16 (D) OTHER INFORMATION: / product = "unidentified" (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 2 Xaa Ala Pro Val Ala Lys Met Val Leu Pro Val Ala Met Lys Glu Xaa
1 5 10 15
Val
Claims (5)
- NOVELTY OF THE INVENTION CLAIMS 1. - Process for the preparation of malted cereals, comprising one or more phases of humidification at a temperature of between 5 and 30 ° C, until the material reaches a moisture content of between 20 and 60% by weight, in which after From a period of germination between 2 and 7 days at a temperature between 10 and 30 ° C, the dampened and sprouted cereals are preferably dried in the oven increasing the temperature to values between 40 and 150 ° C until the material possesses a moisture content of between 2 and 15% by weight, and in which one or more microbial cultures selected from the group consisting of one or more bacteria and / or one or more fungi is added once or several, characterized in that less one of said microbial cultures is inoculated by activated spores, whose size is increased by a factor preferably between 1.2 and 10 with respect to the inactive size and / or having one or more germ tubes per spore.
- 2. Method according to claim 1, characterized in that the activation of the spores comprises at least one or a combination of the following treatments: (a) cycles of humidification and / or drying; (b) addition of nutritional provisions or addition of spore elements; (c) exposure to changes in temperature, preferably of the order of 0 to 80 ° C; (d) exposure to changes in pH, preferably of the order of 2.0 to 8. 0, more preferably between 3.0 and 6.0.
- 3. Method according to claim 1 or 2, for the preparation of malted barley, characterized in that the bacteria are selected from the group formed by Micrococcus spp., Streptococcus spp., Leuconostoc spp., Pediococcus spp., Lactococcus spp., Lactobacillus spp., Corynebacterium spp., Propionibacterium spp., Bifidobacterium spp., Streptomyces spp., Bacillus spp., Sporolactobacillus spp., Acetobacter spp., Agrobacterium spp., Alcaligenes spp., Pseudomonas spp., Gluconobacter spp., Enterobacter spp., Erwinia spp., Klebsiella spp., Proteus spp.
- 4. Method according to claim 1 or 2 for the preparation of malted barley in which the fungi are selected from the group (genera described by the Ainsworth and Bisby dictionary of fungi, 8th edition, 1995, edited by DL Hawksworth, PM Kirk , BC Sutton, and DN Pegler (632 pp) Cab International) comprising Ascomycota preferably Dothideales preferably Mycosphaereláceas preferably Mycosphaerella spp., Venturiaceae preferably Venturia spp; Eurotiales preferably Monascaceae preferably Monascus spp., Trichocomaceae preferably Emericilla spp., Euroteum spp., Eupenicillium spp .; Neosartorya spp., Talaromyces spp., Hypocreales preferably Hypocreceae preferably Hypocrea spp: Saccharomycetales preferably Dipodascaceae preferably Dipodascus spp., Galactomyces spp., Endomycetaceae preferably Endomyces spp., Metschnikowiaceae preferably Guilliermondella spp., Saccharomycetaceae preferably Debaryomyces spp., Dekkera spp., Pichia spp., Kluyveromyces spp., Saccharomyces spp., Torulaspora spp., Zygosaccharomyces spp., Saccharomycodaceae preferably Hanseniaspora spp.; Schizosaccharomycetals preferably Schizosaccharomycetaceae preferably Schizosaccharomyces spp., Sordariales preferably Chaetomiaceae preferably Chaetomium spp., Sordariaceae preferably Neurospora spp., Zygomycota preferably Mucorales preferably Mucoraceae preferentially Absidia spp., Amylomyces spp., Rhizomucor spp., Actinomucor spp., Thermomucor spp., Chlamydomucor spp., Mucor spp., Rhizopus spp. ., Mitosporic fungi preferably Aureobasidium spp., Acremonium spp., Cercospora spp., Epicoccum spp., Monilia spp., Mycoderma spp., Candida spp., Rhodotorula spp., Torulopsis spp., Geotrichum spp., Cladosporium spp., Trichoderma spp., Oidium spp., Alternaria spp., Helminthosporium spp., Apergillus spp. as described by R.A: Samson [(1994) in Biotechnology Handbook, volume 7: Aspergillus, edited by Smith, J.E. (273 pp), Plenum Press)], Penicillum spp.
- 5. Process according to claim 1 or 2 for the preparation of malted cereals other than malted barley, in which the bacteria are selected from the group consisting of Micrococcus spp., Streptococcus spp., Leuconostoc spp., Pediococcus spp. , Lactococcus spp., Lactobacillus spp., Corynebacterium spp., Propionibacterium spp., Bifidobacterium spp., Streptomyces spp., Bacillus spp., Sporolactobacillus spp., Acetobacter spp., Agrobacterium spp., Alcaligenes spp., Pseudomonas spp., Gluconobacter spp., Enterobacter spp., Erwinia spp., Klebsiella spp., Proteus spp. Method according to claim 1 or 2 for the preparation of malted cereals other than malted barley, in which the fungi are selected from the group consisting of: Ascomycota preferably Dothideales preferably Mycosphaereláceas preferably Mycosphaerella spp., Venturiaceae preferably Venturia spp; Eurotiales preferably Monascaceae preferably Monascus spp., Trichocomaceae preferably Emericilla spp., Euroteum spp., Eupenicillium spp .; Neosartorya spp., Talaromyces spp., Hypocreales preferably Hypocreaceae preferably Hypocrea spp.: Saccharomycetales preferably Dipodascaceae preferably Dipodascus spp., Galactomyces spp., Endomycetaceae preferably Endomyces spp., Metschnikowiaceae preferably Guilliermondella spp., Saccharomycetaceae preferentially Debaryomyces spp., Dekkera spp., Pichia spp., Kluyveromyces spp., Sacharomyces spp., Torulaspora spp., Zygosaccharomyces spp., Saccharomycodaceae preferably Hanseniaspora spp. Schizosaccharomycetals preferably Schizosaccharomycetaceae preferably Schizosaccharomyces spp., Sordariales preferably Chaetomiaceae preferably Chaetomium spp., Sordariaceae preferably Neurospora spp., Zygomycota preferably Mucorales preferably Mucoraceae preferentially Absidia spp., Amylomyces spp., Rhizomucor spp., Actinomucor spp., Thermomucor spp., Chlamydomucor spp., Mucor spp., Rhizopus spp., Fungi Mitosporic preferentially Aureobasidium spp., Acremonium spp., Cercospora spp., Epicoccum spp., Monilia spp., Mycoderma spp., Candida spp., Rhodotorula spp., Torulopsis spp., Geotrichum spp., Cladosporium spp., Trichoderma spp., Oidium spp., Alternaria spp., Helminthosporium spp., Apergillus spp., Penicillium spp. 7 .- Method according to any of the preceding claims, wherein the humidification phase is a soaking phase and the total time of aqueous immersion during the same does not exceed 30 hours, preferably takes 10 to 25 hours, or that the oven drying includes more than two temperatures and in which the microbial culture comprises Rhizopus spp., Pseudomonas spp. and / or Aspergillus spp. 8. The method according to claim 7, wherein Rhizopus spp. is a Rhizopus oryzae such as an ATCC 9363 strain of Rhizopus oryzae. 9. Process according to claim 7, wherein Aspergillus spp. is an Aspergillus oryzae such as an ATCC strain 14156 of Aspergillus oryzae. 10. Process according to any of the preceding claims, wherein said cereals are disinfected. 11. Malted cereal characterized by an increased β-glucanase activity by at least a factor 4 and / or increased xylase activity by at least a factor 4, compared to the corresponding malted cereal obtained with the conventional malting method without spores added 12. Malted barley, in which the β-glucanase activity is higher than 700 U / kg, and / or the xylanase activity is greater than 250 U / kg. 13. Malted cereal according to claim 11 or 12 obtained by the method according to any of claims 1 to 10. 14. Malted cereal according to any of claims 11 to 13, characterized in that it has an improved modification or an increased enzymatic activity, for example, an increased hydrolytic enzymatic activity and / or a lower level of toxins and / or increased microbial safety and / or acceptability, compared to the corresponding maize cereal obtained by the conventional malting process without the addition of spores. 15. Malted cereal according to any of claims 1 to 14, which can show a significantly longer seed plumule length compared to that of the corresponding malted cereal obtained by the conventional malting process without the addition of spores. 16.- Combination of cereals and at least one activated spore. 17. Use of malted cereals according to any of claims 11 to 14 obtainable by the method according to any of claims 1 to 10, for the preparation of drinks. 18. Use of malted cereals according to any of claims 11 to 14 that can be obtained by the process according to any of claims 1 to 10, in a detergent composition. 19. Use of malted cereals according to any of claims 11 to 14, which can be obtained by the process according to any of claims 1 to 10, as a bread additive. 20. Use of the malted cereals according to any of claims 11 to 14, which can be obtained by the process according to any of claims 1 to 10, in animal feed compositions. 21. Use of the malted cereals according to any of claims 11 to 14, which can be obtained by the process according to any of claims 1 to 10, in bleaching technology. 22. Use of the malted cereals according to any of claims 11 to 14, which can be obtained by the method according to any of claims 1 to 10, in the technology of pulp and paper.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP97870060.7 | 1997-04-30 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| MXPA99010037A true MXPA99010037A (en) | 2000-12-06 |
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