EP1066367A1 - Application of lipase in brewing - Google Patents

Application of lipase in brewing

Info

Publication number
EP1066367A1
EP1066367A1 EP99914541A EP99914541A EP1066367A1 EP 1066367 A1 EP1066367 A1 EP 1066367A1 EP 99914541 A EP99914541 A EP 99914541A EP 99914541 A EP99914541 A EP 99914541A EP 1066367 A1 EP1066367 A1 EP 1066367A1
Authority
EP
European Patent Office
Prior art keywords
lipolytic activity
cereal material
lipase
wort
process according
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP99914541A
Other languages
German (de)
French (fr)
Inventor
Jerome Souppe
Marie-Paule Laroye
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
DSM IP Assets BV
Original Assignee
DSM NV
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by DSM NV filed Critical DSM NV
Priority to EP99914541A priority Critical patent/EP1066367A1/en
Publication of EP1066367A1 publication Critical patent/EP1066367A1/en
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
    • C12C5/00—Other raw materials for the preparation of beer
    • C12C5/004—Enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
    • C12C7/00—Preparation of wort
    • C12C7/04—Preparation or treatment of the mash
    • C12C7/047—Preparation or treatment of the mash part of the mash being unmalted cereal mash
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
    • C12C7/00—Preparation of wort
    • C12C7/14—Lautering, i.e. clarifying wort
    • C12C7/16—Lautering, i.e. clarifying wort by straining

Definitions

  • the present invention consists in making wort from unmalted cereals like barley with improved filterabiiity.
  • the invention also relates to the use of lipase in brewing in combination with routinely used mashing enzymes.
  • Barley is the most important grain in use for making beer all over the world. Total lipid content in barley ranges from 3 to 5 % of grains'dry matter. The distribution of lipids in barley is:
  • trigiycerides 65-78% glycolipids 7-13% phospholipids 15-26%
  • Lipase is produced during germination of barley.
  • the malt lipase thus obtained can hydrolyse most trigiycerides into glycerol and free fatty acids. Consequently, barley grains contain higher amounts of lipids than malt grains. These lipids are able to build inclusion complexes with starch which makes starch degradation more difficult during mashing when barley is used as crude adjunct. Moreover lipids in the mash create an emulsion like medium leading to low filtrate rates. After wort filtration these lipids are found mainly in the spent grains causing rancidity, off-flavours, and others, which render cattlefeed application less acceptable. A minor amount (5-1 0%) is still found in the filtered wort with bad consequences on organoleptic properties of the final beer: no foam stability, staling, hazing.
  • the present invention enables to produce worts with high levels of crude adjuncts by combining the action of exogenous lipases with other mashing enzymes (e.g. amylase, protease, b-glucanase, endoxylanase) . Also the invention enables to produce beer and any other alcoholic beverage, wherein a wort is obtained according to the invention. Also the invention includes the use of a lipase having 1 ,3-specificity on trigiycerides, e.g. lipase from Rhizopus oryzae.
  • the invention comprises in one aspect the use of exogenous lipolytic activity in a process for making wort.
  • the wort is preferably made from cereal material, more preferably crude cereal material. Preferred is crude barley.
  • the lipolytic activity is a lipase or a phospholipase, preferably in conjunction with one or more mashing enzymes.
  • the mashing enzyme may be one or more of the group consisting of: amylase, protease, beta-glucanase, (endo)xylanase and exopeptidase.
  • a preferred lipolytic activity has 1 ,3 specificity.
  • the lipolytic activity may be of fungal origin, preferably it originates from Rhizopus oryzae.
  • 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 sacchariflcation an enzyme having lipolytic activity is present.
  • the cereal material may comprise crude cereal material, such as crude barley.
  • the lipolytic activity may be a lipase or a phospholipase.
  • Mashing enzymes if present are selected from one or more of the group consisting of: amylase, protease, beta-glucanase, (endo)xylanase and exopeptidase.
  • a process for making an alcoholic beverage comprising the steps of making wort according to the invention and fermenting the said wort, optionally further treating the fermentate, to obtain the alcoholic beverage.
  • a preferred alcoholic beverage is beer.
  • an exogenous lipolytic enzyme for the treatment of cereal material.
  • the instant invention comprises a process of making wort and/or alcoholic beverages, characterised by the presence of lipase during one of the stages of its production.
  • the lipase is characterised as being exogenous, by which is meant that the lipase is not naturally present in the wort or alcoholic beverage making process.
  • Wort is usually made from cereal material, which may accidentally have some lipase activity. If present at all, the lipase activity in cereal material is insufficient, as the Examples of the specification point out, to bring about the advantages of the invention. Therefore a lipase may be added from another source, such as a microbial or non-cereal vegetable source, which is one way of being exogenous, or from the same source as the cereal material from which the wort is made, but in higher amounts than usually present.
  • Such higher amounts may be brought about by over-expressing genes coding for the lipolytic activity, or by adding a lipolytic activity of any source, including from a cereal source, which may be the same or different, at some stage in the process.
  • the enzyme is already present prior to the onset of the process, such 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 prior to the onset of the process.
  • the lipolytic activity may be a lipase or a phospholipase.
  • a preferred lipolytic activity is one having 1 ,3-specificity (1 and 3 referring to the C-atoms in the alcohol backbone, usually glycerol, of the (phospho-)lipid, as these lipases have shown to have a very advantageous effect on filtration rates of the wort.
  • Outstanding performance has been observed when lipolytic activity from Rhizopus oryzae or phospholipase C is used.
  • the cereal material may be any cereal material, although the advantages are most outspoken when at least part of the cereal material is "crude” . With crude is meant non-malted. Those of skill in the art are aware of the fact, that malting is a complex and expensive process. In some countries, the import of malt is prohibited. To save money, or to circumvent any legislative hurdles, it is preferred to make beer, or any kind of alcoholic beverage which relies on malting for yield, taste and/or other organoleptic properties, the use of the process according to the invention is very advantageous. Not only the filtration rates are improved, also the properties of the beer, or any other alcoholic beverage is greatly improved by the use of lipases. Thus the invention will find use in making beer of crude cereals, or mixed brews (crude cereals mixed with malt). Preferred cereal material is barley, wheat, sorghum, oat or mixtures thereof.
  • Liquefying and saccharifying enzymes comprise amylases (including alpha- and beta-amylases, hemicellulases (notably xylanoiytic enzymes), (endo-)glucanases, proteases, exopeptidases, and the like.
  • lipase will advantageously be accompanied by the use of commonly used mashing enzymes.
  • the conditions for liquefaction and saccharification are well known to those of skill in the art. Reference is made to handbooks of brewing. The following examples illustrate the advantages of lipase in a process of making fermentable wort, and in making beer. However, the use of lipase in other alcoholic beverages, such as whisky and bourbon and other spirits is contemplated.
  • 1 -Lipase Lipase activity is assayed by pH-stat monitoring the production of free fatty acids from olive oil.
  • 1 PLi unit is the amount of enzyme needed to produce 1 ⁇ moi free acid per minute at pH 7.5 and 37°C for a neutral olive oil/water emulsion.
  • Lipase from Rhizopus oryzae is commercially available from Gist-brocades under the trademark Lipase 80000 having an activity of 80000 PLi/g.
  • b-glucanase is from Bacillus amyloliquefaciens, commercially available from Gist-brocades under the trademark Filtrase L3000( + ) .
  • protease is from Bacillus amyloliquefaciens , commercially available from Gist-brocades under the trademark Brewers' Protease ( + ) .
  • saccharifying amylase is from Aspergillus oryzae, commercially available from Gist-brocades under the trademark Brewers Fermex.
  • endoxyianase is from Aspergillus niger, commercially available from Gist-brocades under the trademark Lyxasan Forte.
  • Example 1 Wort was prepared from crude barley grains, variety PLAISANT.
  • Barley grains were ground with the EBC MIAG mill in order to make filter press type barley flour.
  • 57g barley flour are added in 300ml water or aqueous solution of enzymes at 50°C. This temperature is maintained for 1 h ; it is then heated up to 63°C (1 °C/min) and maintained at that temperature during 30 minutes. The medium is then heated up to 90°C
  • Mashing enzymes were the same as in Example 1 . No endoxylanase was used in the series. Lipase addition was varied as shown below:
  • Worts were prepared from barley as described in Example 1 . Mashing enzymes, were the same as in Example 3. Lipolytic enzymes were varied as given below:
  • Lipase 80000 from Rhizopus oryzae has an absolute 1 ,3 specificity and preferably yields 1 ,2-diglycerides.
  • Phospholipase C which yields 1 ,2-diglycerides too but from phospholipids, seems to perform nearly as well as Lipase 8000.
  • malt lipase is given as non regiospecific lipase and malt phospholipase is given as being of B-type.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Food Science & Technology (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Distillation Of Fermentation Liquor, Processing Of Alcohols, Vinegar And Beer (AREA)

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 BREWING
Field of the invention
The present invention consists in making wort from unmalted cereals like barley with improved filterabiiity. The invention also relates to the use of lipase in brewing in combination with routinely used mashing enzymes.
Background of the invention
Barley is the most important grain in use for making beer all over the world. Total lipid content in barley ranges from 3 to 5 % of grains'dry matter. The distribution of lipids in barley is:
trigiycerides 65-78% glycolipids 7-13% phospholipids 15-26%
Lipase is produced during germination of barley. The malt lipase thus obtained can hydrolyse most trigiycerides into glycerol and free fatty acids. Consequently, barley grains contain higher amounts of lipids than malt grains. These lipids are able to build inclusion complexes with starch which makes starch degradation more difficult during mashing when barley is used as crude adjunct. Moreover lipids in the mash create an emulsion like medium leading to low filtrate rates. After wort filtration these lipids are found mainly in the spent grains causing rancidity, off-flavours, and others, which render cattlefeed application less acceptable. A minor amount (5-1 0%) is still found in the filtered wort with bad consequences on organoleptic properties of the final beer: no foam stability, staling, hazing.
Although much is known about the bad effects of lipids present when crude adjuncts are used in brewing, no solution has ever been proposed to reduce these effects. Conversely much has been proposed to solve problems linked to the absence of endogenous enzymes when brewing with unmalted cereals.
Summary of the invention
The present invention enables to produce worts with high levels of crude adjuncts by combining the action of exogenous lipases with other mashing enzymes (e.g. amylase, protease, b-glucanase, endoxylanase) . Also the invention enables to produce beer and any other alcoholic beverage, wherein a wort is obtained according to the invention. Also the invention includes the use of a lipase having 1 ,3-specificity on trigiycerides, e.g. lipase from Rhizopus oryzae.
Accordingly, the invention comprises in one aspect the use of exogenous lipolytic activity in a process for making wort. The wort is preferably made from cereal material, more preferably crude cereal material. Preferred is crude barley.
According to another aspect of the invention, the lipolytic activity is a lipase or a phospholipase, preferably in conjunction with one or more mashing enzymes. The mashing enzyme may be one or more of the group consisting of: amylase, protease, beta-glucanase, (endo)xylanase and exopeptidase. A preferred lipolytic activity has 1 ,3 specificity. The lipolytic activity may be of fungal origin, preferably it originates from Rhizopus oryzae.
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 sacchariflcation an enzyme having lipolytic activity is present. The cereal material may comprise crude cereal material, such as crude barley. •The lipolytic activity may be a lipase or a phospholipase. Mashing 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 a process is provided for making an alcoholic beverage, comprising the steps of making wort according to the invention and fermenting the said wort, optionally further treating the fermentate, to obtain the alcoholic beverage. A preferred alcoholic beverage is beer.
In another aspect the use is provided of an exogenous lipolytic enzyme for the treatment of cereal material.
Detailed Description of the Invention
The instant invention comprises a process of making wort and/or alcoholic beverages, characterised by the presence of lipase during one of the stages of its production.
The lipase is characterised as being exogenous, by which is meant that the lipase is not naturally present in the wort or alcoholic beverage making process. Wort is usually made from cereal material, which may accidentally have some lipase activity. If present at all, the lipase activity in cereal material is insufficient, as the Examples of the specification point out, to bring about the advantages of the invention. Therefore a lipase may be added from another source, such as a microbial or non-cereal vegetable source, which is one way of being exogenous, or from the same source as the cereal material from which the wort is made, but in higher amounts than usually present. Such higher amounts may be brought about by over-expressing genes coding for the lipolytic activity, or by adding a lipolytic activity of any source, including from a cereal source, which may be the same or different, at some stage in the process. Alternatively, the enzyme is already present prior to the onset of the process, such 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 prior to the onset of the process.
The lipolytic activity may be a lipase or a phospholipase. A preferred lipolytic activity is one having 1 ,3-specificity (1 and 3 referring to the C-atoms in the alcohol backbone, usually glycerol, of the (phospho-)lipid, as these lipases have shown to have a very advantageous effect on filtration rates of the wort. Outstanding performance has been observed when lipolytic activity from Rhizopus oryzae or phospholipase C is used.
The cereal material may be any cereal material, although the advantages are most outspoken when at least part of the cereal material is "crude" . With crude is meant non-malted. Those of skill in the art are aware of the fact, that malting is a complex and expensive process. In some countries, the import of malt is prohibited. To save money, or to circumvent any legislative hurdles, it is preferred to make beer, or any kind of alcoholic beverage which relies on malting for yield, taste and/or other organoleptic properties, the use of the process according to the invention is very advantageous. Not only the filtration rates are improved, also the properties of the beer, or any other alcoholic beverage is greatly improved by the use of lipases. Thus the invention will find use in making beer of crude cereals, or mixed brews (crude cereals mixed with malt). Preferred cereal material is barley, wheat, sorghum, oat or mixtures thereof.
Those of skill in the art know that in order to ferment cereal material, certain pretreatments are required in order to increase the content of glucose, or other fermentable sugars, freely available nitrogen. Whereas the malting process usually causes the content of fermentable nutrients to rise, with crude cereals, or mixed brews, use is made of exogenous enzymes. Liquefying and saccharifying enzymes, usually collectively referred to as mashing enzymes, comprise amylases (including alpha- and beta-amylases, hemicellulases (notably xylanoiytic enzymes), (endo-)glucanases, proteases, exopeptidases, and the like. It will be clear, that the use of lipase will advantageously be accompanied by the use of commonly used mashing enzymes. The conditions for liquefaction and saccharification are well known to those of skill in the art. Reference is made to handbooks of brewing. The following examples illustrate the advantages of lipase in a process of making fermentable wort, and in making beer. However, the use of lipase in other alcoholic beverages, such as whisky and bourbon and other spirits is contemplated.
Examples
Experimental part
1 -Lipase Lipase activity is assayed by pH-stat monitoring the production of free fatty acids from olive oil. 1 PLi unit is the amount of enzyme needed to produce 1μmoi free acid per minute at pH 7.5 and 37°C for a neutral olive oil/water emulsion.
Lipase from Rhizopus oryzae is commercially available from Gist-brocades under the trademark Lipase 80000 having an activity of 80000 PLi/g.
2-other mashing enzymes
* liquefying amylase is from Bacillus licheniformis, commercially available from Gist-brocades under the trademark B.A.T.S.
* b-glucanase is from Bacillus amyloliquefaciens, commercially available from Gist-brocades under the trademark Filtrase L3000( + ) .
* protease is from Bacillus amyloliquefaciens , commercially available from Gist-brocades under the trademark Brewers' Protease ( + ) . * saccharifying amylase is from Aspergillus oryzae, commercially available from Gist-brocades under the trademark Brewers Fermex. *' endoxyianase is from Aspergillus niger, commercially available from Gist-brocades under the trademark Lyxasan Forte.
Example 1 Wort was prepared from crude barley grains, variety PLAISANT.
Barley grains were ground with the EBC MIAG mill in order to make filter press type barley flour. 57g barley flour are added in 300ml water or aqueous solution of enzymes at 50°C. This temperature is maintained for 1 h ; it is then heated up to 63°C (1 °C/min) and maintained at that temperature during 30 minutes. The medium is then heated up to 90°C
(1 °C/min) and maintained at that temperature during 20 minutes. Water is added to compensate for water evaporation. The mash is then poured into a funnel containing Schleicher and Schull paper filter.
Mashing enzymes were applied in each brew at the following dosage (in g per T barley) :
- B.A.T.S. : 300
- Filtrase L3000( + ) : 1 7
- Brewers' protease ( + ) : 1 10
- Brewers Fermex : 2000 whereas Lipase 80000 and Lyxasan Forte were used at varied levels:
Results clearly show the influence of lipase on the filtration rate, which is stronger than the well known effect of endoxylanase.
Example 2
In this series wort was prepared from various malts using the same brewing diagram as in Example 1 . However, mashing enzymes were different:
- assay 1 no enzyme added
- assay 2 300g Filtrase Br (commercially available from
Gist-brocades) per T malt
- assay 3 25g Lipase 80000 per T malt
- assay 4 300g Filtrase Br + 25g Lipase 80000 per T malt.
Malt from Korea
Malt from China
Results clearly indicate the poor effect of lipase in malt brews whereas a filtration enzyme (mix of amylase, protease and hemicellulases) usually applied show normal efficiency. Moreover no synergistic effect between lipase and filtration enzyme can be demonstrated.
Example 3
In this series we determine the optimal dosage of lipase to be applied. Wort was prepared from barley as in Example 1 .
Mashing enzymes were the same as in Example 1 . No endoxylanase was used in the series. Lipase addition was varied as shown below:
Results indicate that high performance is already obtained as from 1 2g/T whereas overdosing ( > 250g/T) has detrimental effect. Example 4
In view of correlating lipase's specificity to brewing performance, we have compared different lipase and phospholipase commercially available.
Worts were prepared from barley as described in Example 1 . Mashing enzymes, were the same as in Example 3. Lipolytic enzymes were varied as given below:
Lipase 80000 from Rhizopus oryzae has an absolute 1 ,3 specificity and preferably yields 1 ,2-diglycerides. Phospholipase C which yields 1 ,2-diglycerides too but from phospholipids, seems to perform nearly as well as Lipase 8000.
These results are rather surprising since malt lipase is given as non regiospecific lipase and malt phospholipase is given as being of B-type.

Claims

Claims
1 . Use of exogenous lipolytic activity in a process for making wort.
2. Use according to claim 1 , wherein the wort is made from cereal material.
3. Use according to claim 2, wherein the cereal material comprises crude cereal material.
4. Use according to claim 3, wherein the crude cereal material comprises crude barley.
5. Use according to any one of claim 1 to 4, wherein the lipolytic activity is a lipase or a phospholipase.
6. Use according to any one of claim 1 to 5, wherein in addition to the lipolytic activity one or more mashing enzymes are added.
7. Use according to claim 6, wherein the mashing enzyme is selected from one or more of the group consisting of: amylase, protease, beta-glucanase, (endo)xylanase and exopeptidase.
8. Use according to any one of claim 1 to 7, wherein the lipolytic activity has 1 ,3 specificity.
9. Use according to any one of claim 1 to 8, wherein the lipolytic activity is of fungal origin. O. Use according to any one of claim 1 to 9, wherein the lipolytic activity originates from Rhizopus oryzae.
1 . 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 and saccharifying enzymes, optionally subjecting the so-treated cereal material to filtration to obtain a fermentable wort, characterized in that during liquefaction and saccharification an enzyme having lipolytic activity is present.
2. A process according to claim 1 1 , wherein the cereal material comprises crude cereal material.
3. A process according to claim 1 2, wherein the crude cereal material comprises crude barley.
4. A process according to any one of claim 1 1 to 1 3, wherein the lipolytic activity is a lipase or a phospholipase.
5. A process according to any one of claim 1 1 to 14, wherein in addition to the lipolytic activity one or more mashing enzymes are added.
6. A process according to claim 1 5, wherein the mashing enzyme are selected from one or more of the group consisting of: amylase, protease, beta-glucanase, (endo)xyianase and exopeptidase.
7. A process according to any one of claim 1 1 to 1 6, wherein the lipolytic activity has 1 ,3 specificity. 1'8. A process according to any one of claim 1 1 to 1 7, wherein the lipolytic activity is of fungal origin.
1 9. A process according to claim 1 8, wherein the lipolytic activity originates from Rhizopus oryzae.
20. A process making an alcoholic beverage, comprising the steps of making wort using a process according to any one of claims 1 1 to 1 9 and fermenting the said wort to obtain the alcoholic beverage.
21 . A process according to claim 20, wherein the alcoholic beverage is beer.
22. Use of an exogenous lipolytic enzyme for the treatment of cereal material.
EP99914541A 1998-03-24 1999-03-24 Application of lipase in brewing Withdrawn EP1066367A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP99914541A EP1066367A1 (en) 1998-03-24 1999-03-24 Application of lipase in brewing

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
EP98200916 1998-03-24
EP98200916 1998-03-24
PCT/EP1999/002047 WO1999049011A1 (en) 1998-03-24 1999-03-24 Application of lipase in brewing
EP99914541A EP1066367A1 (en) 1998-03-24 1999-03-24 Application of lipase in brewing

Publications (1)

Publication Number Publication Date
EP1066367A1 true EP1066367A1 (en) 2001-01-10

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Country Status (4)

Country Link
EP (1) EP1066367A1 (en)
AP (1) AP2000001891A0 (en)
BR (1) BR9908966A (en)
WO (1) WO1999049011A1 (en)

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7312062B2 (en) 1998-11-27 2007-12-25 Novozymes A/S Lipolytic enzyme variants
EP1905821B1 (en) * 2002-09-26 2016-08-10 Novozymes North America, Inc. Fermentation methods and compositions
US20040063184A1 (en) * 2002-09-26 2004-04-01 Novozymes North America, Inc. Fermentation processes and compositions
US8765199B2 (en) 2006-06-15 2014-07-01 Novozymes A/S Mashing process
DK2222831T3 (en) * 2007-12-12 2015-08-10 Novozymes As A method for brewing
WO2009074650A2 (en) 2007-12-12 2009-06-18 Novozymes A/S Brewing process
BR112021000369A2 (en) * 2018-07-11 2021-04-13 Novozymes A/S PROCESSES FOR PRODUCTION OF FERMENTATION PRODUCTS

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS6030488B2 (en) * 1982-11-10 1985-07-17 協和醗酵工業株式会社 Fabric improvers and fabrics containing them
JPS61254178A (en) * 1985-05-04 1986-11-11 Hirofumi Akano Production of food vinegar
JPS6255069A (en) * 1985-09-03 1987-03-10 Aoto Shoten:Kk Production of sake using corn grits as raw material
JP2858875B2 (en) * 1990-05-23 1999-02-17 サントリー株式会社 Alcohol production method
JPH08214863A (en) * 1995-02-14 1996-08-27 Musashino Kagaku Kenkyusho:Kk Production of sake by using liquefied rice
JPH08266213A (en) * 1995-03-31 1996-10-15 Kyowa Hakko Kogyo Co Ltd Production of bread dough

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9949011A1 *

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AP2000001891A0 (en) 2000-09-30
WO1999049011A8 (en) 1999-11-04
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BR9908966A (en) 2000-11-28

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