Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12H—PASTEURISATION, STERILISATION, PRESERVATION, PURIFICATION, CLARIFICATION OR AGEING OF ALCOHOLIC BEVERAGES; METHODS FOR ALTERING THE ALCOHOL CONTENT OF FERMENTED SOLUTIONS OR ALCOHOLIC BEVERAGES
  • C12H1/00—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages
  • C12H1/003—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages by a biochemical process

Definitions

• the present invention relates to the use of microbial laccase in order to produce a storage stable beer.
• Storage life of a beer depends on many factors, e.g. temperature, haze forming potential and oxygen content.
• PVPP insoluble polyvinylpolypyrrolidone
• a storage stable beer may be produced by adding laccase to the fermented beer.
• the present invention relates to a beer- making process, comprising a) fermenting wort into beer, and b) adding a laccase to the fermented beer so as to improve the storage stability of the beer.
• the microbial laccase may be derived from bacteria or fungi (including filamentous fungi and yeasts) .
• the microbial laccase is preferably obtained from a fungus.
• Some preferred fungi include strains belonging to the subdivision Basidiomvcotina and to the subdivision Ascomvco- tina. Suitable examples include a laccase derivable from a strain of Aspergillus, Neurospora, e.g. , N. crassa, Podospora, Botrytis, Collybia, Fomes, Lentinus, Pleurotus, Trametes, e.g. , T. villosa and T. versicolor, Rhizoctonia, e.g., R. solani, Coprinus, e.g., C. plicatilis and C. cinereus, Psatyrella, Myceliophthora, e.g., M.
• thermophila Schytalidium, e.g., S.thermophilum
• Polyporus e.g., P. pinsitus
• Phlebia e.g., P. radita
• Coriolus e.g., C.hirsutus (JP 2- 238885)
• Hygrophoropsis Agaricus, Vascellum, Crucibulum, Myrothecium, or Sporormiella.
• laccases derivable from T. villosa, T. ersicolor or M. thermophila are preferred.
• the laccase may furthermore be one which is producible by a method comprising cultivating a host cell transformed with a recombinant DNA vector which carries a DNA sequence encoding said laccase as well as DNA sequences encoding functions permitting the expression of the DNA sequence encoding the laccase in a culture medium under conditions permitting the expression of the laccase and recovering the laccase from the culture.
• LACU Laccase Activity
• Laccase activity is determined from the oxidation of
• LACU 1 laccase unit
• the starting material is barley which is malted (i.e. dampened, germinated and subsequently dried) .
• the malt is grounded and mixed with water, heated and stirred. This mixture is then filtered. After addition of hops, the so called beer wort is boiled. Hereby a precipitation of some of the poly ⁇ phenols will take place. After removing the precipitates by filtration or other means of separation, the finished beer wort is aerated to typically 8-10 ppm oxygen and yeast is added.
• VPP polyvinylpoly- pyrrolidone
• laccase is added at the end of the process, because oxygen is unwanted in the finished beer, so addition of laccase may remove any excess oxygen whereby storage life of the beer is enhanced, and at the same time the laccase will remove some of the polyphenols that may still remain in the beer.
• a suitable amount of laccase is in the range of from 0.1-1000 LACU per liter of fermented beer, preferably in the range of from 1-50 LACU per liter of fer ⁇ mented beer.
• the polyphenol complexes formed by laccase may be removed by filtration or other means of separation.
• polyvinyl- polypyrrolidone may also be added to the fermented beer.
• Storage stability tests may be performed in many different ways; a quick way of predicting shelflife may be performed as follows (for reference see Analytica-EBC 1987, 9.17, Prediction of Shelflife) : For determining initial haze the turbidity of the beer is measured at 0°C. The sample is then stored at 60°C for 48 hours whereafter the sample is coo- led and kept at 0°C overnight. For determining final haze the turbidity is measured at 0°C. The results are reported in EBC formazin units .
• Beer Pilsner type beer stabilized normally using PVPP, 10.7%P, 4.6% v/v alcohol.
• Laccase SP 710 available from Novo Nordisk A/S, product former called Laccase SP 504; a Trametes villosa laccase
• Bottle turner 16 turns pr. minute.
• Oxygen Meter DIGOX EC-401.
• Laccase was diluted with oxygen-free water to a concentration of 25 x 2/3 and 50 x 2/3 LACU/ml, respectively.
• the beers were cooled to 0°C, opened, and ⁇ ml of the enzyme solutions or oxygen-free water was added with a syringe resulting in beers with 0, 25 and 50 LACU/liter beer, respect- ively.
• the beer bottles were knocked at until a foam plug had developed, then air was added using a syringe, in the amounts of 0, 3 and 6 ml, respectively.
• the bottles were crowned, turned for 10 minutes and stored for 1 hour or 24 hours at 0°C.
• the beer bottles were turned by hand 10 times before placed in the sampling device where the crown was penetrated and the beer pressed through the oxygen meter by C0 2 at a flow rate of approx. 9 liter/hour.
• the oxygen content in mg/liter was measured using the DIGOX EC-401. For all treatments triple determinations were carried out.
• an oxygen content greater than 1.0 mg/liter is believed to form unwanted flavour in beer.
• the oxygen content can be reduced to below the acceptable level of 0.2 mg/liter in 1 hour by the Laccase treatment, thereby reducing detrimen ⁇ tal chemical changes of the beer during storage.
• Laccase treatment has no significant influence on the formation of haze particles, which might reduce the shelflife of the beer, when measured using the quick shelflife test as described herein.

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• Chemical & Material Sciences (AREA)
• Organic Chemistry (AREA)
• Engineering & Computer Science (AREA)
• Health & Medical Sciences (AREA)
• Physiology (AREA)
• Genetics & Genomics (AREA)
• Life Sciences & Earth Sciences (AREA)
• Biochemistry (AREA)
• Bioinformatics & Cheminformatics (AREA)
• General Engineering & Computer Science (AREA)
• General Chemical & Material Sciences (AREA)
• Food Science & Technology (AREA)
• General Health & Medical Sciences (AREA)
• Wood Science & Technology (AREA)
• Zoology (AREA)
• Distillation Of Fermentation Liquor, Processing Of Alcohols, Vinegar And Beer (AREA)
• Detergent Compositions (AREA)
• Cosmetics (AREA)
• Enzymes And Modification Thereof (AREA)

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Citations (7)

• 1995
  • 1995-01-26 AU AU15324/95A patent/AU680721B2/en not_active Ceased
  • 1995-01-26 MX MX9602923A patent/MX9602923A/es not_active IP Right Cessation
  • 1995-01-26 EP EP95906907A patent/EP0741780A1/en not_active Withdrawn
  • 1995-01-26 CN CN 95191175 patent/CN1138348A/zh active Pending
  • 1995-01-26 CA CA 2181338 patent/CA2181338A1/en not_active Abandoned
  • 1995-01-26 WO PCT/DK1995/000037 patent/WO1995021240A2/en not_active Application Discontinuation
• 1996
  • 1996-07-26 FI FI962975A patent/FI962975A0/fi unknown

Patent Citations (7)

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