Miller Brewing Company

Etzer Chicoye

William F Line

Henry Goldstein

Robert J Mizerak

Edward J Kot

Patrick L Ting

Michael C Barney

Vinod K Chaudhary

Charles L Cronan

John M Cowles

George R Duncombe

Walter H Fly

J Raymond Helbert

James F Rice

Lance T Lusk

Matthew L Tripp

Donald H Westermann

Nick J Huige

Patricia A Bower

Geoffrey H Bertkau

Thomas Rhodes

Charles E Lowery

Chris E Lindberg

Darrell R Hoffmann

Janna K Jilek

Yeast is genetically engineered by transformation with an expression vector containing a natural yeast secretion signal sequence combined appropriately with a chemically synthesized gene encoding antifreeze protein resulting in the expression, proper processing, and secretion of antifreeze protein which is heterologous to yeast in recoverable amounts. Disclosed are DNA sequences comprising structural genes encoding peptides having amino acid sequences with the biochemical or physiochemical properties of antifreeze protein and a method of combining the antifreeze protein gene sequences with appropriate expression vectors.

Method of expressing antifreeze proteins in yeast

Oral care compositions containing hop acids or their salts are effective in inhibiting Gram positive bacteria which can cause plaque or periodontal disease. A representative composition is a toothpaste containing tetrahydroisohumulone.

Oral care compositions containing hop acids and method

Beer spoilage microorganisms are detected with a culture medium that is advantageous for detecting Lactobacillus spp., Pediococcus spp. and wild yeast, and which allows detection to be carried out in four days. The medium contains tomato juice, peptones, cysteine hydrochloride, sorbitan nonooleate, mono and disaccharides, yeast extract, beef extract, potassium acetate, malic acid, mono and dibasic potassium phosphate, magnesium sulfate, maganese sulfate, sodium chloride, ferrous sulfate, beer, water and agar.

Culture medium for detection of beer spoilage microorganisms

A process for enhancing yeast growth during the fermentation of beer which comprises adding to the wort ashed trub. The ashed trub is obtained by heating whole trub until all organic combustibles have been removed or destroyed. The ashed trub contains valuable trace elements, including zinc.

Process for brewing beer while enhancing yeast growth

A hopping material useful as a kettle additive in preparing anactinic malt beverages is prepared by a method which includes first forming an aqueous suspension of crude hops, containing alpha acids and cellulosic material and alkali hydroxide and an alkali borohydride. The suspension, which has a pH above about 10.5, is heated to isomerize and reduce the alpha acids initially present in the hops. The pH is lowered to about 2 to convert the reduced iso-alpha acids to their free acid forms so that they are adsorbed on cellulosic hop material in the reaction mixture. The cellulosic material which now contains the free isomerized and reduced alpha acids is isolated and is useful as an anactinic kettle additive which can be stored for prolonged periods with no decomposition. If desired, the cellulosic material thus obtained can be further treated by adjusting the pH with an alkaline solution to obtain an aqueous solution containing only reduced iso-alpha acids, which is useful as an anactinic post-kettle additive, and a cellulosic material, which is useful as an anactinic kettle additive.

Anactinic hopping materials and method of preparation

A sugar syrup is prepared by saccharifying a liquefied starch hydrolyzate at a pH of about 4 to about 5.5 and a temperature above 55.degree. C. with a heat stable, aciduric, pullulanase obtained from rice and an .alpha.-1,4 carbohydrase. The pullulanase employed is substantially free of maltase and transglucosidase activity. In one embodiment, a dextrose syrup is prepared by saccharifying a thinned starch hydrolyzate with glucoamylase and the rice pullulanase. Maltose syrup is prepared using the rice pullulanase and a maltogenic enzyme.

Production of dextrose and maltose syrups using an enzyme derived from

A superattenuated low calorie beer is prepared from a wort containing as a liquid adjunct a starch-derived syrup which contains dextrins by adding to the wort during fermentation a glucoamylase type debranching enzyme obtained from Schwanniomyces castellii.

Preparation of low calorie beer

Tetrahydroisohumulones are prepared from beta-acids without using objectionable organic solvents by treating the beta-acids to form desoxytetrahydrohumulones which are dissolved in an aqueous alcoholic caustic solution and then oxidized and isomerized with an oxygen-containing gas in the presence of metallic ion to form the desired tetrahydroisohumulones.

Preparation of tetrahydroisohumulones

A traditional flavored fermented alcoholic beverage of low alcohol content is prepared by reverse osmosis with a thin layer composite membrane which has a support layer of polysulfone, a barrier layer of polymer prepared from a polyamine and a polyacyl, and a fiber backing. The membrane has a molecular weight cut-off of less than about 100 for organics and rejects the volatile components which contribute to flavor and aroma and permits about 25 to about 30% alcohol in the beverage to pass.

Preparation of low alcohol beverages by reverse osmosis

Beta-acids are separated from alpha-acids in a hop extract without using organic solvents by bubbling CO.sub.2 through an aqueous hop extract having a pH of about 9.6 to about 13 to lower the pH to about 8.5 to about 9.5 to precipitate the beta-acids and leave the alpha-acids in the extract.

Process for separation of beta-acids from extract containing alpha-acids

An aqueous extract containing .alpha.-1,6 and .alpha.-1,4 carbohydrases obtained from diastatic malt is treated with an adsorbent to remove yeast lethal factors and can be used to produce a superattenuated low calorie beer. The enzyme-containing extract preferably is introduced into a pitched aerated wort during fermentation. Crude aqueous extracts of diastatic malt cannot be used because they contain factors lethal to yeast which adversely affects the yeast viability throughout the fermentation.

Preparation of low calorie beer with malt extract free of yeast lethal

A superattenuated low caloric beer is produced by passing fermenting beer through an immobilized glucoamylase reactor having active glucoamylase immobilized on a ceramic monolith. The ceramic monolith has open ended passages ranging from 50 to 3,000 microns in diameter through which the beer passes, and glucoamylase is covalently bonded to internal surfaces of the passages. The glucoamylase is preferably covalently bonded by reacting an aldehydic derivative of glucoamylase with an aminofunctional group on the internal surfaces of the passages to form an aldimine which is reduced to a secondary amine. The reactor can sustain high flow rates of beer containing yeast and other particulate matter without plugging or fouling, and it is operationally stable under fermentation conditions over a long period of time.

Immobilized glucoamylase reactor for preparing a low calorie beer

Low calorie beer is prepared by introducing into the brewing process a debranching enzyme (pullulanase) obtained from rice, a traditional brewing material. The debranching enzyme reduces the real extract of the beer by cleaving alpha 1,6 linkages of unfermentable limit dextrins to form alpha 1,4 dextrins which can be converted by alpha 1,4 carbohydrases to sugars that can be fermented by brewer's yeast. The enzyme may be introduced into the brewing process by adding rice or the enzyme extracted from rice to the mash or to the wort before or during fermentation. The debranching enzyme may be obtained from polished dry milled rice by extraction with an aqueous buffer solution. When malted rice is used as the enzyme source a particularly useful mixture of the debranching enzyme and alpha 1,4 carbohydrases is obtained.

Method of preparing a low calorie beer

A debranching enzyme (pullulanese) useful in the preparation of a low calorie beer may be obtained from rice by extraction of the rice with an aqueous buffer system having a pH of about 6. A preferred buffer system is 0.1 M potassium phosphate-0.2 M-sodium chloride. Extraction is preferably carried out at a temperature of about 50.degree. C. for about 3 hours. When malted rice is used as the enzyme source a particularly useful mixture of the debranching enzyme and alpha 1,4 carbohydrases is obtained.

Preparation of debranching enzyme from rice for producing a low calorie

Hop extracts useful in the preparation of anactinic or light stable malt beverages are obtained by a method which does not use organic solvents. The extracts are obtained by first treating a crude hop extract containing humulone with an aqueous reducing solution, heating the resulting mixture to form reduced isohumulone, acidifying the reaction mixture to form an aqueous phase and an organic phase, and then adjusting the temperature to facilitate the separation of the two phases. The organic phase which is obtained contains the desired isomerized and reduced humulone and it can be used per se as an anactinic brew kettle flavoring additive for beer or ale. In a preferred embodiment, the organic phase is further treated with an amount of an aqueous KOH solution sufficient to neutralize the reduced isohumulone and to form both an aqueous phase and a solid phase. The aqueous phase which contains reduced isohumulone of about 90+% purity can be used as an anactinic post-kettle additive and the solid phase which contains the remainder of the reduced isohumulone and other hop constituents can be used as an anactinic brew kettle additive.

Growing community of inventors

Milwaukee, WI, United States of America

Etzer Chicoye