ALCOHOLIC BEVERAGES

▪ Sugar converted into ethanol by yeasts

▪ Fermentable carbohydrates: honey, cereals, root crops, palm saps, fruits
(grapes, apples, pears, plums...)
▪ Ethanol concentration limited by sugar concentration and ethanol tolerance
of yeast: 14% v/v
▪ Can be increased by distillation: whisky, vodka, brandy, calvados, cognac,
armagnag...
▪ Addition of partially purified ethanol: fortified wine – porto, sherry, madeira
BEER

Definition:
Beer is an alcoholic beverage obtained by fermentation made
from water, malt (germinated and roasted barley and/or
other cereals) and hops. Germany: 100% malt
France: at least 70% malt
Beer is made in every country in the world!

Composition of beer
+ Water: (80-90 %)absolutely very good quality. It takes 6 to 8 liters for a liter of
beer made
+ Malt: barley in general, sometimes sprouted and dried wheat, or rice
 + Hops: for bitterness, aroma, aseptization and digestibility + Yeasts: allow
alcoholic fermentation but also give taste and fragrance;
+ Raw grains (possibly): corn, rice, rye ...
+ Substitutes (possibly): glucose, sucrose, maltodextrin, dextrose.

Water
• Water is one of the most important components in beer making. The brewers
pay particular attention to the quality of the water.
• It must, above all, meet the standards set by the World Health Organization
(WHO)
• Ex: “Bia hơi Hà Nội”: at main siege (Hoang Hoa Tham, Hanoi) great water makes
an unique taste! At other branches of Hanoi Beer (Hai Duong, Nam Dinh, Hung
Yen): other water sources => Quality reduced!!!!
The composition of the water is variable, with great diversity of ions in different proportions.
The density of some of these ions can affect more or less substantially the progress of certain
stages of the manufacture of beer.
Ex:
• NH3, NaHCO3, HNO2: not allow
• NO3-ions: harmful to the work of yeasts.

• Some ions also influence the final taste of the product (Na+, SO4 2-, , etc.)
Water: Recommended values of some ions

 Ca2+, Mg2+ ions are essential but at high level => hard water => need treatment
 Fe: high level => reaction with tannin in houblon => decrease in color and final taste
 Standard water for beer making:
 Permanent hardness: 0.26-0.72 mg/L equivalent
 Temporary hardness: 0.72 mg/L equivalent
 Total microflora: < 100/L
 Total E.coli: < 3/L
 pH: 6/8-7.2

Methods for water treatment:

 Neutralization by acid
 Addition of sulfate or chlorure (Cl-, SO4 )
 Decarbonation (addition of lime, decantation, filtration or precipitation of carbonates by heating);
 Bacteriological treatment (activated carbon filter);
 Reverse osmosis (R.O)
 Electrolysis

Hops
Hops (Humulus lupulus) are a vineclimbing perennial of the Cannabinaceae family whose inflorescences of
female plants produce a yellowish resin, called lupulin, which gives bitterness to beer
Hops was again mainly used for its conservative virtues (antibacterial)
(Indian Pale Ale-IPA)
 Precipiation of proteins (albumin)
The most famous hops come from the Czech Republic and western Slovenia.
The main variety: Saaz.
There are also other good quality varieties in Germany (Hallertau, Spalt, Tettnang) and in Croatia (Styria).
Main hops components
• Resins (14%), tannins (4%)
 • essential oils (0.5%)
• alpha-acids (humulone)
beta-acids (lupulone)

MALT
▪ Principal cereal: barley, 160 g for 1 L of beer
▪ Occasionally wheat, rice, corn
▪ Africa: regional variants from sorghum, millet – mixed lactic/ethanolic fermentation
▪ Low gelatinization temperature of malted barely starch of 52-59°C – solubilization without inactivation of
alpha-amylase
▪ Presence of beta-amylase: rapid conversion starch and dextrins to maltose
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Malt : provide natural enzymes

▪ Saccharomyces cerevisiae unable to ferment starch
▪ Conversion of starch to fermentable sugars
▪ Oriental Technique: mould enzyme preparations (koji)
▪ Western technique: endogenous enzymes during malting
▪ South America: salivary amylase
BEER
Production processmalting; saccharification; boiling called 'hopping'); fermentation ; guard
(secondary fermentation); conditioning
Malting
The principle of malting is to reproduce the natural development of barley grain so that it
produces certain enzymes necessary for the transformation of starch into sugars
(saccharification), and therefore, to the manufacture of alcohol during fermentation.
Malting takes place in 4 stages: steeping, germination, kilning and
degerming.
 Steeping: barley in water with an ‘air rest’ period in between, total of 2-3 days:
 cleaning and activation of enzyme formation
 6-16 h in water (33-37%)
 12-24 h air rest: plant embryo can take up oxygen
 then 10-20 h in water (43- 46%)
 Germination: formulation and acccumulation of enzymes; partial hydrolysis of
protein, starch, beta glucan…Lasts ~ 50 hours, optimal water content 40-43%, 14-18
° C , depending on the barley, and gives rise to what is called the "green malt".
 Kilning consists of drying the green malt in an air oven at 40 °C for 30 hours.
 Reduction of water content from >40% to 1.5-2% (dark malt) or 3.5-4% (light
malt)
 At this stage, the malt maker performs a "shot" where the green malt is heated to
85 °C (light malts)-105 °C (dark malts) for 1 to 4 hours. This step is crucial because
it is the duration of the "shot" that will determine the color of the beer dress.
 Maillard reaction at T > 100°C between amino acids and sugars; formation of
colour and flavour
 Sulfur compounds are formed during kilning, e.g., dimethyl sulfide DMS, which
gives lager beer a typical flavour, but is undesired in higher concentrations. DMS is
formed from S-methylmethionine SMM, which is synthesized during germination, at
higher temperatures, but evaporates in contrast to SMM
 Typical concentration in beer is 10-150 µg/L
 Degerming consists of ridding the malt of its rootlets. At the end of this stage the "malt"
can be kept for almost a year
Enzyme formed during malting
– -(1,4)-glucanases (cellulases), -(1,3)glucanases

– Pentosanases: xylanase, xylosidases, arabinosidases,

– Proteases

– Amylases: -amylases, amylases

– Lipases
Malts
Lager/Pils malt
Pale Ale malt
Carapils malt

– 1 h at 100-120°C

Crystal malt

– < 2 h at 135°C

Chocolate malt

– 75 bis 150°C in 1 h, then increase to220° C

Black malt

– Analogous but with stronger roasting

Saccharification
Saccharification involves diluting the malt sugars in heated water to activate the
enzymes. The malt is crushed roughly before moisturizing with water which forms the
 "mash". This mixture will then be heated, there are three main techniques: Decoction,
stepwise infusion and Simple infusion
Mashing and wort production
▪ Grinding of malt and other ingredients
▪ Mashing and boiling results in the preparation of an extract, wort, mainly
containing fermentable sugars.
▪ Separation of the aqueous extract from insolubles by lautering or filtration
▪ Boiling of cleared wort with hops
▪ Cooling of the wort
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Decoction: part of the brew is removed and boiled before being added to
the mash, called “tempers".
 First steeping of the grist in cold water, then 1/3 the mash is heated at 100 °
C for 10-20 min and pumped back , mashing at 35-37°C (for β-Glucanase)
 Again heating, mashing at 50-53°C; increased enzyme activities (protease)
 Again heating, mashing at 62-67°C, for β-Amylase (=> maltose)
 Again heating, mashing at 71-72°C, for a-Amylase (=> dextrin)
 Total duration of 5-6 h
Single, double or triple-decoction mashing, produced good
quality wort but is expensive; boiling results in Maillard reactions
Stepwise infusion: The water is heated with the mash, or hot water is added
at regular intervals. It is a very flexible and precise method using a heating jacket
rather than decoction
Simple infusion: water is heated and mixed with malt.
The "primitive must" is then extracted from the mash by filtering and percolating
with hot water (as for coffee) in a special vat called "filter vat". The solid residue is
called "duff" and is recycled to feed the livestock.
Flavoring or hopping
It is at this stage that we incorporate hops and sometimes spices. The mixture is
brought to a boil, in 1-2h. Boiling is conducive to the appearance of bitter flavors.
The bitterness comes mainly from a yellowish resin produced by the female cones
of hops: lupulin.
Objective:
• Destroy the enzymes whose role is then complete.
• Sterilize the must before fermentation
• Promote the formation of proteintannin complexes
• Concentrate must
• Extracting the aroma of hops. Remove of undesired flavor component (DMS).
Isomerization of alpha acid to iso alpha acid => improve solubility

Primary fermentation
• This step involves sowing the must with a certain amount of yeast so that
these yeasts convert the present sugars into alcohol and CO2.
• Use of starter cultures, yeast from previous fermentations
can be reused (up to 15 times) “pitching yeast‘, addition of yeast
▪ 500 mL of yeast slurry per 100 L 1 2 ° wort, equivalent to 15 x 106 yeast cells
temperature of wort of 5-6 ° C
 ▪ Initial aeration of the wort to provide oxygen
▪ In typical fermentation yeast population increases by factor of ≈8, 3 generations
▪ Duration of fermentation of 1 2 ° wort: 6 to10 days

Bottom fermentation using Saccharomyces uvarum

• Beers have a taste of hops and malt.
• Beers less fruity and less alcoholic than beers top fermentation, but more
loaded with CO2. Fresh, between 4 and 7 ° C => protect beer against
bacteria and fungi.
• During the fermentation the yeast migrates to the bottom of the barrel =>
“bottom". This lasts 7 to 10 days between 4 and 12 ° C.
• Shelf life > top fermentation
• Lower alcohol production than top (yeast is held back by the alcohol it
produce).
The name Lager (from lagern, store in German).
Top fermentation using Saccharomyces cerevisiae.
• 3 to 8 days at 15 to 25 ° C.
• When the yeast has exhausted the glucose, it rises to the surface of the beer,
=> top fermentation.
• The most common brewing method before the invention of refrigerator, the
high temperature does not protect the beer from bacteria or fungi
• Conservation: shorter than that of bottom fermentation.
• Obtaining high levels of alcohol (yeast is not slowed down in the presence of
alcohol) as well as complex aromas.
• Beers produced less loaded with CO2 than low fermentation beers.
 • Consume at higher temperatures (between 6 and 12 ° C).
• => Ale : beer from bottom fermentation

Spontaneous fermentation does not require the addition of yeast to the must:
exposed to the open air, it is inoculated with wild yeasts.
This was the way of producing beer before yeast culture, mastered during the Middle
Ages. Today it is little more used than in Belgian beers lambic type and gives an acid taste
due to lactic acid and acetic acid.
Mixed fermentation combines bottom fermentation and spontaneous
fermentation.

Yeast S. Cerevisiae Ale S. carlsbergensis Lager

Characteristic

Temperature 14 – 25oC 4 – 12oC

Substrates Monosaccharides Both mono-, di- and tri –

(glucose, fructose), saccharides: glucose, maltose,
disaccharides galactose,
(saccharose, maltose), low fructose, saccharose, mannose,
effect with melibiose and raffinose, melibiose.
trisaccharides (raffinose).
Fermentation Top Bottom
type
Fermentation Faster Longer
time
 Beer flavor Robust, hearty and fruity Smooth, elegant, crips, clean

Fermentation Yeast(s) Temperature (°C) Appellation(s)

Bottom Saccharomyces 4-12 lager, pils,
uvarum Schwarzbier, Dunkles, Export
Bier, Helles

Top Saccharomyces 15-25 ale, stout, weizen, etc.

cerevisiae

Spontaneous Brettanomyces NC lambic, gueuze, faro, kriek

bruxellensis,
Brettanomyces
lambicus
Mixed Brettanomyces NC Red beer
bruxellensis,
Brettanomyces
lambicus,
Saccharomyces
cerevisiae

Secondary fermentation
• It is carried out for several weeks around 1 ° C
• Decisive changes on the organoleptic level and therefore no significant
biochemical transformation:
 • Disappearance of substances with an unpleasant flavor such as acetaldehyde,
acetolactate, diacetyl ...
• Appearance of alcohols and esters, byproducts of nitrogen metabolism
contributing positively to the flavor of beer
• Precipitation of protein-tannin complexes to ensure the subsequent stability of beer
Conditioning
• Once the final product is reached, it is essential to avoid any contact with
pathogens and with oxygen
• Beer is stored in refrigerated tanks with a counter pressure of CO2. After a certain
period of "storage", the beer is packaged in barrels (pressure), bottles or aluminum
can, under vacuum. To increase shelf life, many brewers use pasteurization in
bottle.
• Craft beers or microbreweries, beer is called refermented. Indeed, the gasification
("carbonation") of the beer is done naturally with active yeasts in the bottle, process
that does not allow pasteurization. The inactive yeasts are deposited in the bottom
of the bottle forming a binding.

CONCLUSION

▪ Microbiologically robust process

▪ Low nutrient status, ethanol content, low pH
▪ Klebsiella, Enterobacter spp.: sensitive to low pH and ethanol but can grow in
wort – produce dimethyl sulfide, nitrosamines
▪ Acetobacter, Gluconobacter spp.: associated with cask-conditioned beer
▪ Lactobacillus, Pediococcus: lactic acid, diacetyl, polysaccharides
Composition

Flavors
▪ Bitterness, 5-50 mg/L iso-α-acids, rather labile and decompose in light and with oxygen
▪ Esters, produced by yeasts; ethyl acetate (20-40 mg/L), isoamyl acetate (2-5 mg/L)
(bananas), ethyl butyrate (pinapple, mango), ethyl hexanoate (apple)
▪ Aldehydes (from oxidation of alcohols or from lipids) , acetaldehyde, 510 mg/L
▪ Higher alcohols, 2-methylpropanol, 2-methylbutanol, 3-methylbutanol, 2-phenylethanol
(from yeasts)
▪ 4-Vinylguaiacol, phenolic or clove-like flavor (mùi gia vị), from ferulic acid through thermal or
enzymatic decarboxylation
▪ Vicinal diketones, 2,3-butanedione (diacetyl), 2,3-pentanedione, formed from α-
acetolactate, degrade by yeast, threshold value of diacetyl in Pilsner beer 0.03 mg/L
▪ Sulfur components, dimethyl sulfide DMS
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