MXPA00009321A - Application of lipase in brewing - Google Patents

Abstract

The invention provides for a process for the production of a fermentable wort comprising the steps of:liquefying and saccharifying cereal material in the presence of one or more mashing enzymes, optionally subjecting the so-treated cereal material to filtration to obtain a fermentable wort, characterized in that during liquefaction and/or saccharification an enzyme having lipolytic activity is present. The cereal material may comprise crude cereal material, such as crude barley.

Description

APPLICATION OF LIPASE IN THE PREPARATION OF BEER Field of the Invention The present invention consists in the production of beer wort from malt-free cereals such as barley with improved filtering capacity. The invention also relates to the use of lipase in brewing in combination with the pastes enzymes routinely used.

Background of the Invention Barley is the most important grain in the use for the manufacture of beer in the whole world. The total lipid content in the barley is in the range of 3 to 5% of the grain dehydrated material. The distribution of lipids in barley is: Triglycerides 65 - 7 8% Glucolipids 7 - 13% Phospholipids 1 5 - 2 6% Ref: 123109 Lipase is produced during the germination of barley. The malt lipase obtained in this way can hydrolyse most of the triglycerides in glycerol and free fatty acids. Consequently, barley grains contain higher amounts of lipids than malt grains. These lipids are capable of building inclusion complexes with the starch, which makes starch degradation more difficult during pasting when the barley is used as a raw adjunct. In addition, the lipids in the paste create an emulsion as the medium that leads to low filtration rates. After the filtration of the wort these lipids are found mainly in the spent grains, causing rancidity, bad tastes and others, which makes the application to cattle feed less acceptable. A smaller amount (5-10%) is still found in the filtered wort with bad consequences on the organoleptic properties of the final beer: lack of foam stability, stripped and turbidity. Although much is known about the bad effects of lipids present when raw adjuncts are used in beer brewing, no solution has yet been proposed to reduce these effects. Conversely, much less has been proposed to solve the problems associated with the absence of endogenous enzymes when brewing beer with no malt.

Brief Description of the Invention The present invention makes it possible to produce musts with high levels of raw adjuncts by combining the action of exogenous lipases with other passaging enzymes (for example amylase, protease, β-glucanase, endoxylanase). The invention also makes it possible to produce beer and any other alcoholic beverage, where a wort is obtained according to the invention. The invention also includes the use of a lipase having 1,3 specificity on triglycerides, for example the lipase from Rhi zopus oryza e. Accordingly, the invention comprises in one aspect the use of exogenous lipolytic activity in a process for making wort. The must is preferably made from cereal material, more preferably raw cereal material. Crude barley is preferred.

According to yet another aspect of the invention, the lipolytic activity is a lipase or a phospholipase, preferably in conjunction with one or more plating or bracketing enzymes. The brace enzyme may be one or more of the group consisting of: amylase, protease, β-glucanase, (endo) ylanase and exopept idase. A preferred lipolytic activity has 1,3 specificity. The lipolytic activity can be of fungal origin, preferably it originates from Rhi zopus oryza e. The invention provides a process for the production of a fermentable must comprising the steps of: liquefaction and saccharification of the material in the presence of one or more brachial enzymes, optionally subjecting the treated cereal material to filtration to obtain a fermentable must , characterized in that during liquefaction and / or saccharification, an enzyme having lipolytic activity is present. The cereal material may comprise the raw cereal material, such as raw barley.

The lipolytic activity can be a lipase or a phospholipase. Braceage enzymes if present are selected from one or more of the group consisting of: amylase, protease, beta-glucanase (endo) xylanase and exopeptidase. According to a further aspect of the invention, there is provided a process for the preparation of an alcoholic beverage, comprising the steps of making the must according to the invention and fermenting said must, optionally treating the fermented one, to obtain the beverage alcoholic A favorite alcoholic beverage is beer. In still another aspect, the use of an exogenous lipolytic enzyme is provided for the treatment of the cereal material.

Detailed description of the invention The present invention comprises a process for the preparation of must and / or alcoholic beverages, characterized by the presence of lipase during one of the stages of its production. Lipase is characterized by being exogenous, which is why it is understood that lipase is not naturally present in the must or in the process of making the alcoholic beverage. The must is usually made from the cereal material, which may accidentally have some lipase activity. If it is present at all, the lipase activity in the cereal material is insufficient, as indicated by the examples of the specification, to explain the advantages of the invention. Therefore, a lipase can be added from another source, such as a microbial or non-cereal vegetable source, which is a way of being exogenous, or from the same source as the cereal material from which the must is made , but in higher quantities than those usually present. Such higher amounts may be caused by the over-expression genes encoding the lipolytic activity, or by the addition of a lipolytic activity from any source, including from a cereal source, which may be the same or different, in some stage in the process. Alternatively, the enzyme is already present before the start of the process, as will be the case when the lipolytic activity is present as a consequence of its production in the cereal, by genetic modification of the cereal, or by adding it before the beginning of the process . The lipolytic activity can be a lipase or a phospholipase. A preferred lipolytic activity is one that has specificity 1.3 (1 and 3 refer to the carbon atoms in the backbone of the alcohol, usually glycerol, of the (phospho-) lipid, since these lipases have been shown to have a very advantageous over wort filtration rates A surprising performance has been observed when the lipolytic activity of Rhi zopus oryza eo phospholipase C is used. The cereal material can be any cereal material, although the advantages are more open when at least Part of the cereal material is "raw." By crude it is meant unfermented or germinated.Those of experience in the technique are aware of the fact that malting is a complex and expensive process.In some countries, importation of the product is prohibited. malt or fermented barley To save money, or to circumvent any legislative obstacles, it is preferred to brew beer, or any type of alcoholic beverage that relies on malting for performance, taste and / or other organoleptic properties, the use of the process according to the invention is very advantageous. Not only are filtration rates improved, but also the properties of beer, or any other alcoholic beverage, which is greatly enhanced by the use of lipases. In this way, the invention will find use in the preparation of raw cereal beers, or mixed boilers (raw cereals mixed with malt). The preferred cereal material is barley, wheat, sorghum, oats or mixtures thereof. Those of skill in the art know that in order to ferment the cereal material certain pretreatments are required in order to increase the glucose content, or other fermentable sugars, freely available nitrogen. While the process of malting usually causes the content of fermentable nutrients to rise, with raw cereals, or mixed boilers, exogenous enzymes are used. Liquefaction and saccharification enzymes, usually referred to collectively as brace enzyme, comprise amylases (including alpha- and beta-amylases, hemicellulases (mainly xylanolytic enzymes), (endo) glucanases, proteases, exopeptidases, and the like. of the lipase will be advantageously accompanied by the use of commonly used brace enzymes The conditions for liquefaction and saccharification are well known to those skilled in the art Reference is made to the brewing manuals The following examples illustrate the advantages of lipase in a process for the manufacture of fermentable must, and in the manufacture of beer, however, the use of lipase in other alcoholic beverages, such as whiskey and bourbon and other alcohols, is also contemplated.

Eg emplos Experimental part 1. Lipase Lipase activity is evaluated by periodically verifying by pH the production of free fatty acids from olive oil. The lPli unit is the amount of enzyme needed to produce 1 μmol of free acid per minute at pH 7.5 and at 37 ° C, for a neutral emulsion of olive oil / water. Rhizopus oryzae lipase is commercially available from Gist-brocades _ under the trade name Lipase 80000 which has an activity of 80000 PLi / g. 2. Other Braceage Enzymes * The liquaction amylase is from Bacillus licheniformis, commercially available from Gist-brocades under the trade name B.A.T.S. * the β-glucanase is from Bacillus amyloliquefaciens, commercially available from Gist-brocades under the trade name Filtrase L3000 (+). * The protease is from Bacillus amyloliquefaciens, commercially available from Gist-brocades under the trade name Brewers' Protease (+). * the saccharification amylase is. of Aspergillus oryzae, commercially available from Gist-brocades under the trade name Brewers Fermex.

* The endoxylanase is from Aspergi l l us niger, commercially available from Gist-brocades under the trade name Lyxasan Forte.

Example 1 The must was prepared from raw barley grains, variety PLAISANT. The barley grains were crushed with the MIAG EBC mill in order to make the barley flour filter press type. 57 g of barley flour are added to 300 ml of water or to an aqueous solution of enzymes at 50 ° C. This temperature is maintained for 1 hour; This is then raised to 63 ° C (1 ° C / minute) and maintained at that temperature for 30 minutes. The medium is then heated to 90 ° C (1 ° C / minute) and maintained at that temperature for 20 minutes. Water is added to compensate for water evaporation. The dough or paste is then emptied into a funnel containing Schleicher and Schull filter paper. The braceage enzymes were applied in each boil at the following doses (in grams per ton of barley). - B.A.T.S. : 300 - Filter L3000 (+): 17 Brewers Protease (+): 110 Fermex Brewers: 2000 while Lipase 80000 and Lyxasan Forte were used at varying levels: The results clearly show that the influence of lipase on filtration rate, which is stronger than the well-known effect of endoxylanase.

Example 2 In this series, the must was prepared from various malts using the same brewing diagram as in Example 1. However, the brachial enzymes were different: assay 1: no added enzyme - assay 2: 300 g of Filtrase Br (commercially available from Gistbrocades) per ton of malt - test 3: 25 g of lipase 80000 per ton of malt test 4: 300 g of Filtrase Br + 25 g of Lipase 80000 per ton of malt.

Malta of Korea The results clearly indicate the poor effect of lipase in malt boilers, while a filtration enzyme (amylase, protease and hemicellulase mixture) usually applied shows normal efficiency. In addition, no synergistic effect between the lipase and the filtration enzyme can be demonstrated.

Example 3 In this series, the optimal dose of the lipase to be applied was determined. The must was prepared from barley as in Example 1. Braceage enzymes were the same as in Example 1. Endoxylanase was not used in the series. The addition of the lipase was varied as shown below: The results indicate that the high yield is already obtained from 12 g / T while the overdose (> 250 g / T) has a harmful effect.

Example 4 In view of the correlation of the specificity of lipase to the performance of brewing, different commercially available lipases and phospholipases have been compared. The musts were prepared from barley as described in Example 1. The braceage enzymes were the same as in Example 3. The lipolytic enzymes were varied as follows: Lipase 80000 from Rhi zopus oryza e has an absolute 1,3 specificity and preferably produces 1,2 diglycerides. The phospholipase C that produces 1,2-diglycerides also but from phospholipids, seems to work almost also as the Lipase 8000.

These results are rather surprising, since the malt lipase is given as non-regiospecific lipase and the malt phospholipase is given as type B.

It is noted that in relation to this, date the best method known by the applicant to carry out the said invention, is that which is clear from the present description of the invention.

Claims (22)

CLAIMS Having described the invention as above, the content of the following claims is claimed as property:

1. The use of exogenous lipolytic activity in a process for the manufacture of must.

2. The use according to claim 1, characterized in that the must is made from cereal material.

3. The use according to claim 2, characterized in that the cereal material comprises raw cereal material.

4. The use according to claim 3, characterized in that the raw cereal material comprises raw barley.

5. The use according to any of claims 1 to 4, characterized in that the lipolytic activity is a lipase or a phospholipase.

6. The use according to any of claims 1 to 5, characterized in that one or more brachial enzymes are added to the lipolytic activity.

7. The use according to claim 6, characterized in that the brackish or paste enzyme is selected from one or more of the group consisting of: amylase, protease, beta-glucanase, (endo) -i-lane and exopeptidase.

8. The use according to any of claims 1 to 7, characterized in that the lipolytic activity has a specificity of 1.3.

9. The use according to any of claims 1 to 8, characterized in that the lipolytic activity is of fungal origin.

10. The use according to any of claims 1 to 9, characterized in that the lipolytic activity originates from Rhi zopus oryza e.

11. A process for the production of a fermentable must, which comprises the steps of: liquefaction and saccharification of the cereal material in the presence of one or more brachial and saccharification enzymes, optionally subjecting the treated cereal material thus to filtration, to obtain a fermentable must, characterized in that an enzyme having lipolytic activity is present during liquefaction and saccharification.

12. A process according to claim 11, characterized in that the cereal material comprises raw cereal material.

13. A process according to claim 12, characterized in that the raw cereal material comprises raw barley.

14. A process according to any of claims 11 to 13, characterized in that the lipolytic activity is a lipase or a phospholipase.

15. A process according to any of claims 11 to 14, characterized in that, in addition to the lipolytic activity, one or more brachial enzymes are added.

16. A process according to claim 15, characterized in that the brace enzyme is selected from one or more of the group consisting of: amylase, protease, beta-glucanase, fenddxi-lane, and exopeptidase.

17. A process according to any of claims 11 to 16, characterized in that the lipolytic activity has 1,3 specificity.

18. A process according to any of claims 11 to 17, characterized in that the lipolytic activity is of fungal origin.

19. A process according to claim 18, characterized in that the lipolytic activity originates from Rhi zopus oryza e.

20. A process for the preparation of an alcoholic beverage, characterized in that it comprises the steps of making the must using a process according to any of claims 11 to 19, and fermenting said must to obtain the alcoholic beverage.

21. A process according to claim 20, characterized in that the alcoholic beverage is beer.

22. The use of an exogenous lipolytic enzyme for the treatment of cereal material.