BEER PRODUCTION

Submitted by: Group 5 3A Biochemistry Golez, Karla Gonzales, Erika Juliano, Karla Kongsun Ching, Laine Mamerto, Therriz Submitted to: Dr. Librado Santiago

INTRODUCTION Brewing is almost certainly the most ancient manufacturing art known to man, and is probably as old as agriculture. Beer is also as old as bread - in fact it is probable that either beer or bread may have been a by-product of the other. According to archaeologists, 'beerbread' was known in many eras. The word beer is derived from the Anglo-Saxon baere, meaning barley. Thus beer is traditionally a product of yeast fermentations of barley grains. The first evidence of beer manufacture has been traced to ancient Babylonia, possibly dating as far back as 500 to 700 B.C. Some 18 varieties of beer, called bousa, were aid to have been prepared in Babylonia as long ago as 220 B.C. by 100 B.C., the Egyptians had produced hopped beer and spliced beers were prepared. The brewing beer and the baking of bread were co-associated and were womens work. The sediment from brewing was consumed by the poorer classes. Certainly, the solid sediment was nutritious. Barley was the predominant cereal used, and not always malted. At first, malting was apparently practiced in baking and brewing. At some time, barley was moistened and when germination began, it was crushed with a pestle, roughly ground, and made into loaves with sour dough or leaven. They were baked sufficiently to form a crust without cooking the interior. When beer was required, the loaves were broken, mixed with water, and allowed to ferment. The liquid was pressed, separated from the dough, and when fermentation was completed, the resulting acid-alcoholic beverage was called boozah or bousa. One can appreciate the fact that the solids given to the poor were nutritive. Beer was an important beverage in the daily life of the ancient Egyptians. An Egyptian been known as hek or heqa was brewed from malt barley and later from malted barley and fruit. The Egyptians may have learned the art from the Babylonians. Beer also had status - a keg of beer was considered the only proper gift to be offered to the Pharaoh by a suitor seeking the hand of a royal princess. It is recorded that a similar amount was also offered to appease the gods when they became angry. Beer was used as a medicine in both Babylon and Egypt. The art of brewing was passed on to the Greeks and Romans and later to the Celts, Germans and Scandinavians. The ancient Hebrews prepared a bread beer called machmetzth from stale bread, malt flour and water. The Chinese might have been the first peoples to prepare beer-like beverage from cereal grain. Samshu was brewed from rice. A beer-like beverage, kin, was supposedly produced about 2300 B.C. the Chinese may also have been the first to produce distilled liquors. Santchoo was prepared by distilling the rice and millet beer, tchoo, Hung-chu, called a red wine, is a beer colored with ang-kak. Germanic tribes made their bior or peor with germinated and steeped barley, and it was usually in sour taste. Even today, some of the beers have a slightly sour taste, indicating lactic acid bacterial fermentation. The Celts fermented wheat and honey. Honey and grain beverages were made in the British Isles in the 4th century. Columbus reputably was presented by friendly Indians with a beer made from corn. Beer was made in Virginia by its settlers as early as 1548 and later in New England. A brewery was erected in

Manhattan in 1612 by Dutch settlers and in Pennsylvania in 1683 by William Penn. Other notable figures in early history of America include Washington, Jefferson, Samuel Adams, Madison, Henry, and others. The Germans and Dutch however, apparently exerted the greater influence on brewing in later years. The application of scientific instruments in breweries occurred comparatively late. It was not until about 1760 that brewers appreciated the value of a thermometer and 1785 before the first saccharometer was used. Fermentation was formerly a mystery. Several theories were offered before 1800 to explain the changes, some of which sound almost fantastic today. For example, vinous fermentation, acetous fermentation, and putrefaction were considered revelations of the same process. Malting was regarded as a vegetable degree of fermentation. According to Hind (1940), Dubrunfant in 1830 converted starch to sugar by means of an extract of malt. Three years later, Pagen and Persoz precipitated and dried an active substance from similar extracts and called it diastase. Thnard in 1803 (Hind 1940) stated that yeasts were the cause of fermentation but believed the yeasts were of animal origin. In 1863 Cagnard de la Tour and Schwann and Ktzing independently declared that yeasts were of vegetative origin. This started the dispute between those lead by Liebig and supported by Berzelius and Whler who believe fermentation was strictly a chemical process and those who believed it was a biological process. Liebigs theory, stated in 1839, was that fermentation waas brought about by an unstable body called a ferment, and waas essentially a chemical process. A new era opened when Pasteur published the results of his study of yeasts. He found hat sugar was not decomposed exactly according to Gay-Lussacs equation but that glycerol and succinic acid were formed at the same time. The fundamental outcome of this was the doctrine that fermentation was coincident with the life of the microorganism. This was a universal doctrine for all fermentations, whether by yeasts, lactic acid, or acetic acid bacteria. Pateurs Etudes sur le Bire published in 1876 had a far-reaching influence on brewery practice. When in 1883, Emil Christian Hansen introduced his pure culture of yeasts at the Carlsberg brewery, a system of fermentation control was set up that proved to be of inestimable value. Hansen had previously shown that brewery yeasts were a mixture of several races of yeasts and that defects were caused by wild yeasts. The result of Hansens pure yeast culture were so successful in Carlsberg brewery in Copenhagen that the method was adopted permanently in 1886. Pure cultures maintain regularity in larger breweries. It was not until 1897 that Buchner demonstrated the connecting link between the ideas of Liebeg and those of Pasteur. He extracted the enzymes from yeasts and demonstrated that fermentation was possible in the absence of the living cell. This gave impetus to the vast amount of scientific investigation that followed that also has been of such great value in many other branches of knowledge, e.g. medicine, nutrition, chemistry and other sciences. The brewing industry is developed step-by-step as a result of observation and research. It is sometimes stated that this began with the notable research of Emil Christian Hansen. One cannot, however, discount the careful observations of numerous other individuals

throughout history who were responsible for earlier developments. The advances not only involve microbiology, but also chemistry, physics, engineering, agronomy, and other sciences. INGREDIENTS USED FOR BREWING 1. Water Water is a very substantial ingredient of beer. Beer is composed of 90-95 percent water. It is utilized in almost every step of the brewing process. The chemistry of the water can influence not just the taste but also the brewing efficiency. Therefore, it is essential that the water supply used is adequate for the job. Chlorine is added to water to eliminate bacteria or other contaminants. However, excessive chlorine can greatly affect other beer ingredients, resulting in a harsh aroma and flavor. Chlorine can easily be removed by boiling or filtration. Also, Calcium is a trace element found in water and is necessary for the complete yeast life cycle. Bacteria is a major concern in water quality. The presence of bacteria can spoil the wort and cause a souring of the beer. Boiling is again effective in the removal of contaminants. 2. Malted Barley Malted barley is essentially a barley that has been allowed to germinate to a degree and then dried. Barley is a seed of the barley plant. During the seed's initial stage of selfsufficient growth, germination occurs. The seed contains its own storage of starch for food. The germination and drying stages capture fermentable sugars, soluble starch, and the diastase enzymes for beer brewing. Malted barley is the eventual source of the fermentable sugar consumed by the yeast. 3. Hops Hops are the cones (flowers) of the female hop vine. The cones contain a soft resin called alpha acid that gives beer its bitterness flavor. Hops also help to inhibit the growth of certain beer spoiling bacteria, to maintain flavor stability, and to retain the head of the beer. There are many different varieties of hops. They are selected from the rest for their unique bitterness in flavor or aroma. Hops are broadly classified into two groups. Noble hops are low in bitterness and have a pleasant aroma and flavor. High-alpha hops have high bitterness but a less pleasant aroma. The high-alpha hops can be used more economically due to the less amount required. However, the flavor of the beer is slightly sacrificed. . 4. Yeast Yeast is the most important ingredient in beer brewing. It is a single-celled organism belonging to the fungus family. Yeast is a living creature, metabolizing, reproducing, and living off the ingredients in the beer. It is responsible for the converting of sugar to alcohol and carbon dioxide in the fermentation stage. Yeast is also the final component

that determines the flavor of the beer. The two main varieties of yeast used for beer brewing are top-fermenting yeast (Saccharomyces cerevisiae) and bottom-fermenting yeast (Saccharomyces uvarum). The top-fermenting yeast is similar to the yeast for baking bread. It is applied for making ales and stouts. The bottom-fermenting yeast is utilized for production of lagers and steam beer BREWING SCHEMATICS

1. Mash Tun In the mash tun, ingredients for wort are mixed according to the recipe. The product which leaves this unit is called mash. The mash produced is sent to the kettle where it is treated with water. 2. Kettle The brew kettle is the apparatus where the hot water and mash are combined and heated. This unit is strategically placed below the Mash Tun so that the wort can flow by gravity to the vessel. It is made from stainless steel. From here the slurry moves to the heat exchanger. 3. Heat Exchanger A heat exchanger is necessary to cool the mixture to 70 degrees Celsius before it enters the fermenters. Yeast can not function if the temperature becomes too high. A countercurrent heat exchanger works best for maximizing the amount of heat transfer that occurs. 4. The Primary Fermenter

The fermenter is where the most important step takes place the production of ethanol. Usually it takes approximately 8-10 hours for fermentation to start. A hydrometer is used daily to measure the specific gravity of the fermenting solution (which leads to a measure of ethanol content). Once the desired ethanol is reached, the beer is chilled by means of a cooling plate within the fermenter. The beer is then sent to the ageing tank where it matures. The fermenter is then cleaned for one-half hour using a 180 degree Celsius caustic solution. 5. Ageing Tank The ageing tank allows the beer to age at cool temperatures (4-15 degrees Celsius) for a few weeks to months, thereby allowing the beer to develop more carbonation and mellow. In addition, "harsh flavor notes" subside and yeast fall to the bottom of the tank. An Evolution of brewing* B.C. 300,000 15,000 11,000 4000 1500 500 500 A.D. 500 500 1100 1200 1680 1800 1835 1870 1883 1903-1930 1935 1940 Formalized mashing and mash clarification Size of fermentation vats and brewing vessels increased Use of hops in brewing Develpment of big European brewing centers Microscope invented Modern lrge-scale brewing facilities developed Brewingscience initiated Pasteurization invented Pure culture developed for brewer's yeast Scientists elucidate biochemistry of yeast fermentation Beer can developed Stainless steel, automation, unitized cleaning,etc. starts new generation of breweries (still evolving) Alcoholic fermentation of plant sap and fruit juices Alcoholic/lactic fermentation of grains Soaking grain to make malt for beer and bread Specialized malting for beer Use of copper vessels in brewing Construction of separate integrated brewing facility Formalized malting in a malting facility

BREWING Brewing is the production of malt beverages. Beer, ale, porter and stout are examples of these beverages. A malt beverage is prepared from infused grains which have undergone sprouting (malting) and by the fermentation of the sugary solution (wort) by yeasts. A

portion of the carbohydrate is changed into ethyl alcohol and carbon dioxide. A normal beer contains dextrin, maltose, glucose, peptones, amino acids, resins, essential oils, and tannic acids. The fundamental basis of brewing is the conversion of sugars to ethyl alcohol and carbon dioxide by the action of enzymes. The process of conversion of cereal starches to alcohol is not a direct one and involves many steps that are capable of modification. Furthermore, the stimulations conversions of non sugar comments may have a marked effect upon the ultimate product. In addition, non enzymatic chemical changes, such as those involved in aging, occur and affect quality. The significant difference between the beer and whiskey industries and the wine industries is the composition of the raw material. Beer and ale are malt beverages made with malt, malt adjuncts, hops, yeasts and water. Malt is prepared from barley grains, germinated and dried after removal of the sprouts. Hops are the dried flowers of the hop plant. Malt adjuncts are starch or sugar containing materials, such as corn and corn products, rice, wheat, barley, sorghum grain, soybeans, cassava, potatoes, sugars and syrups. Overview of Brewing Process:

Starting material: Malt extract, usually with extracts of other grains (wheat, rice, corn) added Organism: Saccharomyces cerevisiae The reasons for the use of Saccharomyces in beer and wine production are relatively straightforward. Both beer mash and grape juice are mixtures characterized by a high concentration of sugar and low pH. During fermentation these sugars lead to the production of ethanol and carbon dioxide. It is important that the yeast strain used is able to survive the higher ethanol concentrations produced. For beer, these concentrations generally range from 3-9%, while for wine they are substantially higher at 11-15%. Saccharomyces strains are fairly resistant to high ethanol concentrations. Even when present as a very small percentage of the initial yeast population, these strains tend to be the dominant strain at the end of natural fermentation due to their ethanol tolerance [4]. Saccharomyces strains are also chosen based on their ability tolerate other compounds, such as sulphite. In modern winemaking, the native yeast strains are not killed by heat as they are in beer-making, because the act of heating alters the final taste of the wine. Instead, the grape juice is treated with sulphite to remove the native strains, thus making it necessary for the Saccharomyces strains to survive the addition of these compounds. In addition to these practical reasons for choosing Saccharomyces strains, vintners may also want to impart a particular characteristic to the final wine produced. For this, the winemaker may select strains that have been characterized in the past as having a particular taste. Thus, there is a great deal of trial and error in selecting yeast strains. It is also not yet entirely clear what genes are responsible for giving a particular yeast strain a particular flavor in the final product. A partial list of desired characteristics in wine is given in the table below. Conditions: Anaerobic; temperature, yeast strain, and time of incubation vary with type of beer. Malting Malting refers to the process of activating germinating and roasting barley Malting is a process now generally performed by large breweries or more commonly specialised malting companies Barley is cleaned, sized, viability measured and nitrogen content determined prior to malting rodent dropping and insect infestations removed Barley sizing is important to ensure even germination Different varieties malted separately First step in malting is the activation of enzymes within the barley Process called steeping Addition of cool water for 2-3 days, which is changed regularly (removes microbes, DO) Removal of microbes essential for malt stability

Respiration of barley increases therefore O2 demand rises (anaerobiosis -> alcohol -> toxin) Post steeping - germination Barley spread thinly in a germination box and turned regularly to dissipate CO2, keep temperature constant, and prevent roots from matting Embryo activated - catylases dissolve wall of endosperm Enzymes secreted by the developing embryo result in modification malted barley can be referred to as under, well or over modified Germination is completed once barley root is approximately 2/3 length of husk The processes of steeping and germination can be combined and a continuous process evolved Dormancy Profound dormancy (embryos temporarily unable to germinate) Water sensitivity - barley germinates only if not submerged Biochemistry of Malting Numerous proteases are present (breakdown proteins -> AAs) Starch the most important material Occurs in two forms, amylose and amylopectin Glucose polymers consisting of 1000-4000 Glc units During germination barley starch degraded into polyglucose molecules Enzymes involved include glucosidase, amylase, amylase and debranching enzymes After kilning only amylase, amylase activity remains Two most important enzymes during malting amylase Cleaves starch randomly (except near ends or branch points) Produces mainly dextrin molecules Requires calcium for optimal activity Conditions, pH 5.5, temp 700C Not present in mature barley, forms during germination More thermostable than amylase amylase Cuts off maltose from non-reducing ends of molecules Produces maltose (major sugar in wort) Conditions, pH 5.2, temp 600C Present in mature barley however increases during germination Other compounds liberated during Malting hemicelluloses and gums (10% weight of barley from endosperm cell walls i.e. glucans (glucose polymers) are undesirable in large quantities Precipitate during fermentation --> jelly Must be degraded to products soluble during brewing Fats (3.5%) Unsaturated fatty acids most important --> cell wall synthesis of yeast during fermentation

Phosphates (1%) phytic acid - a hexaphosphate of inositol (Vit B) Degraded by the yeast to produce myoinositol and phosphoric acid Germination is arrested by drying kilning the barley reduces moisture content (45 -> 5%) Imparts colour to malt - higher temperatures used to produce dark coloured beers Malt is then graded and sold to brewery All these process result in a malt that can be readily converted into the fermentation medium required by the brewer. The following steps are thus typically carried out on site to produce the wort (sugar medium) used by the yeast to produce beer

Milling Process in which malt is ground into meal or grist, or gritty flour Important to keep barley husk relatively intact as they are needed during wort recovery Endosperm fragments are however required to be small to ensure rapid degradation Two types of mills dry mills wet mills - addition of water prior to milling wet mills inflict less damage on the malt Grist then pumped / gravity fed into mash-tun Mashing Mashing is the process in which the ground malt and adjuncts of the corn, rice and other cereals are mixed with water and extracted to obtain the fermentable liquid known as wort or sweet wort. Mashing is a process which digests the starch to fermentable sugars. Different blends of barley malt, roasted barley, oat malt, caramel, sugar, and other materials are used for distinctive types of beer or ale. Grist sprayed into mash-tun along with water (doughing-in) Heat applied to mash (65-670C) amylase and amylase continue to degrade starch molecules into more readily fermentable sugars and dextrins Mash conditions favour one or the other enzyme Control of mashing conditions through using profiles enables brewer to produce vastly different worts Two types of mashing Infusion mashing heating of mash is relatively constant throughout the process generally used for top fermenting worts Decoction mashing

heating of mash is achieved by removing portion of mash to separate vessel, heating and returning Results in stepwise increase in temp Decoction results in optimum tempertures being reached for proteolysis (40-540C) starch hydrolysis (54 - 650C) Wort separation (730C) Some breweries use a mixture of the systems Changing conditions of mash as mentioned = large changes in wort Wort is then extracted from the mash-tun (or through a separate larger SA vessel called a lauter tun) through filtering through the husks of the barley Wort produced 1.060 - 1.100 SG However this is diluted due to sparging of water through the husks Spent grains sent for animal feed Solid adjuncts added at this stage of brewing Double mashing Process as described but with an additional cereal cooker which is used to hydrolyse maize or rice grists Mashing process begins as usual with the subsequent addition of adjunct later in mashing cycle when temperature is highest Used in production of American worts Temperature programming of mash also used Important Considerations in Mashing: a. The malt should be ground, but not too fine b. The starch should be gelatinized c. The water should contain 200-300 ppm calcium. d. The pH should be 5.0 5.2 e. Temperature of 600C produces nitrogen compounds of high molecular weight. A high temperature favors alpha-amylase activity more than betaamylase. f. Of course, the time used for enzyme hydrolysis of substrates as well as the concentration of enzyme and substrate are important.

Wort Boiling Sweet wort brought to the boil in large kettles (or coppers) Principle effects arrest enzyme activity sterilise wort (at least partially) coagulate proteins and tannins (removed as trub) precipitation of calcium phosphate, pH distillation of volatile compounds evaporation of H2O, concentrate wort colour production, caramelisation, oxidation of tannins (colour of final product controlled at this stage) if a very dark beer is required caramel can be added at this stage

extraction of bitter residues from hops essential oils infused iso acids improve beer foam (from hops) Dissolved matter within the wort is called extract (and expressed in oPlato) hot break - hop residues removed via strainer cold break - <600C protein precipitate - trub Addition of finings during kettle boiling enhances trub removal (haze) Further trub removal occurs though either filtration or centrifugation (whirlpool) Trub produced during wort boiling - 30-60 g/hl Kettle boiling isomerises the humulones to isohumulones and allo-isohumulones humulinic acid has little bitter taste Wort is then cooled (plate heat exchangers) and aerated before transfer to fermentation vessels Oxygen is required as a growth factor during anaerobic fermentation Used to synthesise unsaturated fatty acids and sterols for the membrane Yeast added to fermentation from propagation facility

Fermentation Fermentation rooms are special rooms that can be kept clean and are at the constant and uniform temperature and humidity. Completely enclosed tanks or tanks with domed lids that can be lowered into the tank are employed because of the need of Carbon dioxide. Larger beers are generally bottom fermentations conducted at a relatively low temperature by strains of Saccharomyces carlbergensis which are said to have stronger fermentive capacity but weaker respiratory activity than Saccharomyces cerevisiae. The lower temperature fermentation is usually thought to yield beers of finer quality and fuller flavor and supposedly retain their character over a longer period. Pale beers are fermented at lower temperature and dark beers are produced from more vigorous fermentation. The most active fermentation proceeds while yeasts are remained suspended in the wort. Clarified, aerated wort is pitched (inoculated) with approximately 10 million cells per ml Metabolism of substrates occurs via pathways First fermentation stage is termed primary fermentation this stage continues until SG levels off (or in brewing terms the beer has attenuated) 24-48 hours after pitching clumps of foam (krasen) appear Yeast will double in mass during ale production and quadruple in mass in lager Process time 3-5 days for ale and 8-10 days for lager Once beer has attenuated the yeast is removed (lager) by cone cooling (00C) and the beer left or pumped to maturation tanks Beer at the end of this stage is referred to as green beer Continuous fermentation A process developed here in New Auckland in the late 50s and early 60s through collaboration between Lion and DB breweries (Morton Coutts) Almost all NZ beer was at one stage produced by this method

 DB breweries currently one of the only breweries world-wide to be producing beer by this method Two major types of continuous process Cascade Method - A series of stirred tanks into which wort is introduced and flows up and out into the next - Degree of attenuation is a function of temperature, dwell time, strength of the wort R - Recycling of yeast enables higher flow rates - Dwell time 25-30 hours Plug flow reactor (tower fermenters) - Use of strongly bottom flocculant yeast - Wort pumped up through the yeast - Flow rate adjusted such that wort reaching top of vessel is correctly attenuated - Now used primarily in vinegar production Operating costs significant reduced Process however susceptible to infection (lactobacillus) and brewery limited to production of one beer although beer is generally very consistent Equilibrium time 2-3 weeks Possible to run fermenter for over 9 months Fermentation is carried out in breweries in two distinct ways: Top fermentation yeasts rise to the surface Bottom fermentation yeasts sink to the bottom Total fermentation is better defined as three phase: Adaptation or Lagtime phase, Primary or Attenuative phase and Secondary or Conditioning phase. Lagtime or Adaptation Phase At the beginning of the adaptation phase, the yeast take stock of the sugars, The yeast use their own glycogen reserves, oxygen, and wort lipids to synthesize sterols to build up their cell membranes. Once the cell walls are permeable, the yeast can start metabolizing the amino nitrogen and sugars in the wort for food. Like every animal, the goal of life for the yeast cell is to reproduce. Yeast reproduce asexually by "budding". Primary or Attenuative Phase The primary or attenuative phase is marked by a time of vigorous fermentation when the gravity of the beer drops by 2/3-3/4 of the original gravity (OG). The majority of the attenuation occurs during the primary phase, and can last anywhere from 2-6 days for ales, or 4-10 days for lagers, depending on conditions. Secondary or Conditioning Phase The reactions that take place during the conditioning phase are primarily a function of the yeast. When the vigorous primary stage is over, the majority of the wort sugars have been

converted to alcohol, and a lot of the yeast cells are going dormant - but some are still active. The secondary phase allows for the slow reduction of the remaining fermentables. The yeast have eaten most all of the easily fermentable sugars and now start to turn their attention elsewhere. The yeast start to work on the heavier sugars like maltotriose. Maturation Aging and Finishing The fermented beer is aged two weeks to several months at 0 0C. During this time, unstable proteins and resins precipitate. Esters are formed, giving the beer its mellow flavor. Secondary fermentation - yeast that did not settle during cone cooling converts harsh flavours For example diacetyl --> 2,3 butanediol (burnt butterscotch flavour to a flavourless compound) Aging of beer at 00C helps with formation of large protein-tannin complexes which impact on beer stability As these form at low temperatures they can effectively be removed by filtration If secondary fermentation not required the beer is filtered (diatomaceous earth) and clarified Carbonated with CO2 recovered during fermentation Additional flavours, caramels added to produce the desired product hop extracts added to raise bitterness levels caramels to enhance colour antioxidants (oxidation reduces shelf life of beers) Finished beer then sent to packaging line

Defects in Beer Turbidities in beer may be caused by a number of factors: unstable protein, protein-tannin complexes, starch, resin, and microorganisms. Gluten or albumin turbidity occurs at a low temperature and when the malt has been improperly dried in the kiln or when a barley with very high protein content used. Warming the beer causes the turbidity to disappear. Oxidation turbidity (haze) is due in part to protein-tannin compounds. The presence of oxygen; the shaking of the beer during transportation; the collisions of beer bottles, which impart supersonic and high pitch sound vibrations; and sunlight affect the formation oxidation haze. Saturation of the beer with carbon dioxide does much prevent this turbidity. Tannin-protein hazes appear also at low temperatures. In order to produce stable, chillproof beers, which will not become hazy or turbid when cold, the use of a small amount of a proteolytic enzyme preparation is advantageous. . Starch turbidities develop as a result of the improper conversion of starch during mashing. Lack of proper digestion at this time may due to the use of a malt in which the

diastase has been destroyed during the kilning. Sparging with water at a temperature much higher than 80*C may also result in the production of turbidity. Amylases may be added to the storage vats to remove starch turbidity. The presence of resin oil containing pitch may, rarely cause turbidity, as may calcium oxalate. Proper filtration will prevent bad types of turbidities. Yeasts turbidity may be due to lack of proper clarification during secondary fermentation, which in turn is caused by an unsatisfactory wort. The use of chips, or krausening is usually effective in correcting this type of turbidity. Wild yeasts especially of the Saccharomyces pastorianum species produce turbidity. By excluding air and keeping the concentration of fermentable sugars in the beer low, growth of yeasts is initiated. A low pH, 12 or below, will also inhibit the growth of yeast. Faulty beer may be the result of the use of a low-grade raw material in the mash; the use of hops of poor quality, too much hops, or prolonged boiling of the hops; contact of the beer with iron, causing an inky taste; contact with tin; an unsuitrable brewing water, young or green beer; or carbon dioxide deficiency; etc. Beer Infection The term "beer infection" or "beer disease" is commonly used in describing the undesirable condition that occasionally exists in beers as the results of the presence of microorganisms, chief among which are the bacteria. The microbial agents causing the symptoms may be designated as beer-infection microorganisms or beer disease microorganisms. The infection bacteria described in literature are inlcuded in five families: the Pseudomoadaceae, Bacillaceae, Bacteriaceae, Micrococacceae, and Lactobacteriaceae, are of particular significance to the brewing industry. The actual number of genera involved is also small, as illustrated by Tosic, who reviewed the subject of beer-infection organisms in 1945 and who reported that beer infections are caused by species of the following genera: Acetobacter, Lactobacillus, Streptococcus, Flarobacterium, and Ahcromobacter. There appears to be some doubt as to whether bacteria of the genera Bacillus and Micrococcus cause infections of malt baverages. The mere fact that spore formers exists in the raw materials used in brewing is no indication that they are producers of infection. Spoilage in Beer A wide variety of microorganisms are capable of growing in liquid substances. In beers, hops will inhibit many species of bacteria but hops cannot solely be relied on. Hansen has used the term wild yeast to denote any yeast that may cause trouble in brewing or produce defects in beer. Five examples of wild yeasts: Saccharomyces ellipsoideus I and II (S. cerevisiae var ellipsoideus, S. cerevisiae var turbidans); Saccharomyces pastorianus I,II, and III (S. pastorianus var intermedius and var validus). Two most common bacterial species attributed to beer spoilage:

1. Pediococcus cerevisiae 2. Lactobacilus pastorianus Two types of bacteria that may cause spoilage in beer: 1. Acid tolerant Acetobacter species (microaerophilic and can grow in beer) intolerant to hops 2. Acid intolerant species of genera Achromobacter, Flavobacterium, Bacillus, and others. (aerobic and cannot grow in acid beer) spoilage is confined to nonacid mashes and worts Acetobacter sp. May be detected in equipment that has been exposed to beer and subsequently to air, such as returned empty barrels. The alcohol is converted to acetic acid. Pasteur suggested that yeasts contaminated with bacteria can be purified by growing in a slightly acidic solution of sugars. Although the lactics wont be killed their growth will be attenuated. Pasteur also observed that the cocci associated with the long rod baceria were the so-called causes of beer sickness. These organisms were referred to as sarcina by Hansen.

REFERENCES Atlas, R., (1995) Principles of Microbiology. Missouri: Mosby-Yearbook, inc. Alcamo, I., (1997) Fundamentals of Microbiology 5th ed. USA: Addison Wesley Longman, Inc. Anderson, Nester, Pearsall, Roberts. (2001) Microbiology: A human Perspective 3rd ed. USA: McGraw-Hill Book Company. Pederson, Carl. (1971) Microbiology of Food Fermentation. Westport, Connecticut: The AVI Publishing Company, Inc. Prescott, S., Dunn, C., (1949) Industrial Microbiology 2nd ed. New York: McGraw-Hill Book Company. Brock, Thomas. (1970) Biology of Microorganisms. New Jersey: Prentice-Hall, Inc.