BELGIAN ACIDIC BEERS

DAILY WELCOME REMINISCENCES OF THE PAST

Hubert Verachtert University of Leuven

 Acidic beers

From (1) historic aspects

Through (2) biochemical aspects
To (3) technical aspects
 Acidic beers

Historical aspects
 6000 years BC ago, independent of regio’s,it was discovered that

different tasteless milky suspensions of sugary material in water

changed to enjoyable beverages on standing.The magic tool was
spontaneous fermentation.Several types of ”Beer”were invented.Soon
however it was prescribed to consume it rapidly as a vinegar sourness
already known from the souring of grape juice,started to dominate.
Acidic beers were invented.The souring must have been very
common as known from texts such as “..the dead pharaos were
promised bread that doesn’t crumble and beer that doesn’t turn sour”.
 (acidic) beers from 6000 BC to 2012
From reeds straws to drink the milky (acidic) beers
avoiding the deposits to cups,bowls,pots and…glasses
!!
Slide 6
Peasant dancing and enjoying
more acidic beer

Slide 7
Medieval brewing

Slide 8
Slide 9
" A green wreath indicated
that the beer was fresh"

Slide 10
Slide 11
ACIDIC BEERS AROUND 1560
FERMENTATIONS QUASI SPONTANEOUS OR
WITH added FOAM "INOCULA" FROM PREVIOUS BATCHES

LOW ATTENUATIONS and ALCOHOL CONTENT

THE BEERS ARE OPAQUE
UNAVOIDABLE BACTERIAL INFECTIONS FORM ACIDS
- during wort cooling (lactic acid )
- during mashing - in (lactic acid)
- during storage (lactic + acetic acid)
INFECTIONS INTRODUCED FROM THE AIR, RAW
MATERIALS,EQUIPMENT and INOCULA

Depending on region use of barley + wheat. Rye leads to “beer vinegar”

HOPS BETTER THAN HERBS AGAINST HIGH ACIDIFICATIONS

Slide 12
 The famous beers of Louvain
the Peterman and
the Bière de Louvain (18 -20 century)
th th

Peterman (yellow-brown)
Bière de Louvain (white)
barley + wheat
Acidity fresh beer 600 ppm (as lactic acid)
Old beer up to 4000 ppm
Alcohol :around 3 %
 STATEMENTS CONCERNING
BEER ACIDIFICATION AND WORT ATTENUATIONS
(1895-1905)
" HIGH ATTENUATION LEADS TO HIGH ALCOHOL LEVELS WHICH KILL THE YEASTS ,
NEEDED DURING STORAGE TO FAVORABLY COMPETE WITH BACTERIAL INFECTIONS "

" HIGH ATTENUATED WORT CONTAINS RESIDUAL SUBSTRATES WHICH CAN ONLY BE
USED BY NON-YEAST INFECTIONS "

CONSEQUENTLY KEEP THE ATTENUATION LOW :

" TURN TO BARLEY KILNING OR RE-KILNING TO REDUCE

ITS DIASTASIC POWER AND INCREASE THE FERMENTABLES
CONTENT OF THE WORT "

" USE THE TURBID MASH BREWING METHOD "

" INCREASE WORT AERATION TO PRODUCE MORE YEAST AND

REDUCE NITROGENOUS COMPOUNDS "

" USE MORE NON - MALTED GRAINS "

Slide 14
Optimal lactic acid acidity of acidic beers advized by
H.Verlinden (brewmaster in Brasschaat) around 1930
calculated from NaOH titration values

White beers of Louvain: 1800 ppm

Antwerp,Brabant,Limburg: 2700 ppm
Flanders beers: 4500 ppm
Lambics: 9000 ppm

Main Problem: how to control the concentration of

acetic acid (bacteria)!!!
For lambics remain under the 1000 ppm limit.
Slide 15
 More old acidic beers in
Het Hageland
Hoegaarden,Aarschot,Tienen,
Diest…
All involve a type of spontaneous fermentation in casks

Folklore : yearly « Grietmuyl » celebration in Tienen

« Grietmuyl »: Refers to the awful mouth of brill fish

(=griet=kind of turbot).Sometimes…
a drinker at a first sip of a Hoegaarden showed
a sour face… It indicated that the beer was
turning to vinegar,meaning the beer
was an acidic beer which
according
to indications had to be drunk before
summertime.

Slide 16
 SPARKLING ACIDIC BEERS

AT THEIR BEST...

MEETING POINTS BEWEEN BEER AND WINE

AT THEIR WORST ...

A TASTE OF HISTORY

17
Slide 17
 Acidic beers

Biochemical aspects
ACETIC ACID PRODUCTION from ethanol BY ACETOBACTER
energy for growth=oxidation of ethanol with O2 to acetic acid!!

CH3-CH2OH + NAD+ > CH3-CHO + NADH + H+ with membrane dehydrogenase !

CH -CHO+ NADP+ >CH -COOH + NADPH + H+ with membrane dehyrogenase!

3 3
Transport of electrons and protons from reduced co-enzymes to oxygen:

NADH >>electrons >>>> O >> H O + trans membrane proton gradient >>ATP synthesis
2 2
 Growth on ethanol by Acetobacter
Growth = from ethanol through acetate and
acetyl-CoA (c-2) and through malate (c-4) and the
glyoxylate cycle
2 acetyl-CoA (2 C-2) >> malate (C-4) + 2 CoA
then:malate>>oxalacetate>>pyruvate>>sugars
 Unraveling the lactic acid acidification

Centuries ago:appreciated in milk,not in beer!

1780 Scheele: lactic acid isolated from milk
1873 Wislicenus:acid=CH3-CHOH-COOH
1876 Pasteur:acid=biological origin (rods+cocci)
1878 Lister:name Bacterium lactis
1892 Van Laer:Saccharobacillus
1901Beijerinck:Lactobacillus
1812 Goodsir:Cocci Sarcina (latin=bundles)
1884 Balcke:Pediococcus (latin cells in a plane)
 Acetification by Brettanomyces
Brettanomyces:fermentation=stimulated by O2
but leads to acetic acid !!(Scheffers,1966)
glucose+NAD++NAD+ ==)) CH3-CHO+NADH++H++CO2
==)) CH3-CHO+NADH++H++CO2
CH3-CHO+NADH+H+=))CH3-CH2OH+NAD+ (alcohol dehydrogenase)
CH3-CHO+NAD+=))CH3-COOH+NADH+H+ (aldehyde dehydrogenase)
Glucose+2 NAD+=))CH3-CH2OH+CH3-COOH
+2 NADH + 2 H+ + 2 CO2
Excess NADH stops further glucose metabolism
Oxygen regenerates needed NAD+
!!!!!Fermentation resumes!!!!!
 Types of Belgian beers
 Trappist ales (strong, with yeast deposit, brewed in abbeys)

 Abbey ales
(strong, with /without yeast deposit, not brewed in abbeys)

 Ales and strong ales (different colours, with/without yeast deposit)

 Pilsner types

 White beers (barley+wheat+spices,turbid)

 ACID ALES and FRUIT BEERS

Slide 24
 Acidic beers
technical aspects

Belgian lambics
and gueuzes
 Belgian acidic beers anno 2012
-2 main types-
1.spontaneous fermentation:lambic
and gueuze
2.yeast-bacterial mixed culture
fermentation
sparkling acidic
beers
At their best...
Meeting points between
beer and wine

At their worst…
A tast of history
Where lambic and gueuze are produced
(more recent :lambic types in West
Vlaanderen:Ingelmunster and Bellegem)

Slide 28
LAMBIC : (end 18 th century) from alambics
found in breweries also making spirits
GUEUZE : gueuze = some old lambic sold in
bottles
>>>with time bubbles and foam production observed
>>>cheap used champagne bottles available (begin 19th
century)
>>>Champenoise method known and competition from
sparkling new pilsner beers
>>>improve lambic sparkling by blending old lambic
with young lambic
>>>good bottled lambic first sold in 1844 as gueuze-
lambic by some breweries.

what’s in a name?
Gueuze not mentioned in standard work of G.Lacambre in
1851
Slide 29
 CHARACTERISTICS
OF TRADITIONAL LAMBIC BREWING

 RAW MATERIALS : BARLEY + WHEAT

 HOP : AGED HOP CONES / HIGH DOSIS

 BREWING: TURBID MASH

 FERMENTATION : SPONTANEOUS

 PRODUCTION : COLD SEASON

SELECTED REGIONS

Slide 30
Filling of open shallow trays

Slide 31
Cooling

Slide 32
 Turbid mash method
(slijmmethode)
Mash at 38°C (10 min)

Proteinrest at 50°C (15 min)

take slime
warm up until 63°C heat separately
hot water addition to until 80-90 °C
warm up until 72°C
α-amylaserest at 72°C until iodine normality
direct the hot slime
to the mash
filtration and sparging at 78°C

go to the kettle
Slide 33
The so called madammen
(madams)
A typical device of rotating
perforated discs used by lambic
brewers to filter the mash

Slide 34
Traditional gueuze brewing
from bags of grain to glass
Wooden casks (300-8000 L)
cask filling

Slide 36
Wooden casks (300-8000 L)

Slide 37
Wooden casks (8000 L)

Slide 38
Slide 39
Slide 40
Large metal tanks

Slide 41
 Main lambic yeasts
-1. Saccharomyces and typical Brettanomyces sp.

-2. Discovered in old English stock beer and named

Brettanomyces in 1903 by Claussen (Carlsberg
Laboratories)
-3. Discovered in lambic in 1921 and named
Br.bruxellensis by Kufferath and Van Laer

-4. Gives typical Brett flavour consisting of

e.g.ethylphenol,ethylguaiacol,isovaleric acid,acetic
acid,ethylacetate ,2-acetyl 1,4,5,6 tetrahydropyridine and
ethyllactate

-5. Ferments higher oligo-saccharides ! Slide 42

 Brettanomyces present in all
acidic beers ?
Prefer acidic conditions established by lactic acid
(and acetic ?)bacteria
Actually 5 Species
Br.bruxellensis,intermedia,intermedium,lambicus,custersii
=Brettanomyces (Dekkera)bruxellensis
Br.abstinens,claussenii,anomalus
=Brettanomyces(Dekkera) anomala
Brettanomyces naardenensis
Brettanomyces custersianus
Brettanomyces nanus
The famous Brettanomyces
and Pediococcus

Slide 44
Lambic fermentation microbiological profile
 Comparison of
different brewing and fermentation
conditions

METAL TANKS versus WOODEN CASKS

INFUSION versus TURBID MASH BREWING

FRESH versus OLD HOP / TURBID MASH

FRESH versus OLD HOP / INFUSION

DIFFERENT WORT AT SAME LOCATION

SAME WORT AT 2 DIFFERENT LOCATIONS

(data from G.Aerts/Kuleuven/Kaho)

Slide 46
Lambic characteristics determined during
18 months lasting fermentations
 ENTEROBACTERIACEAE (normally found /first weeks)
 ACTIDIONE-RESISTANT YEASTS ( Brettanomyces)
 NON ACT.-RES. YEASTS (e.g. Saccharomyces)
 LACTIC ACID BACTERIA

 ATTENUATION
 ETHANOL
 LACTIC ACID
 ACETIC ACID
 pH
 ETHYL LACTATE
 ETHYL ACETATE

Slide 47
WHATEVER CONDITIONS EXAMINED :

Microbial Sequence in lambic fermentation

Enterobacteriaceae sp

Saccharomyces sp + LAB sp

Brettanomyces sp + LAB sp

Slide 48
 METAL TANKS versus WOODEN CASKS

Lactate ( LACT ) + ethyllactate ( E.LACT ) Acetate ( ACET ) + ethylacetate ( E.ACET )

PPM
PPM
ETHYLACTATE –- PPM

ETHYLACETATE –- PPM
7450 700 1500 140
– PPM

PPM
LACTATE- PPM

ACETATE -–PPM
6450 600 1250 120

ETHYLLACTATE
5450 500 100

ETHYLACETATE
1000
4450 400 80

ACETATE
LACTATE

750
3450 300 60
2450 200 500 40
1450 100 250 20
450 0 0 0
0 2 4 6 8 10 12 14 16 18 20 0 2 4 6 8 10 12 14 16 18 20
MONTHS MONTHS
MONTHS
WOOD –- LACT.
WOOD LACT. METAL
METAL –- LACT.
LACT. WOOD –- ACET.
WOOD ACET. METAL -– ACET.
METAL ACET.

WOOD –- E.LACT.
WOOD E.LACT. METAL –- E.LACT.
METAL E.LACT. WOOD
WOOD –- E. ACET.
E.ACET. METAL
METAL -– E. ACET.
E.ACET.

Turbid mash method / old hop / comparison between metal tank and wooden cask

WOOD METAL

Original extract °P 9.8 9.8

Final attenuation (app) % 93 . 1 97 . 2

Final attenuation (real) % 74 . 4 78 . 6

pH 3.3 3.1
Slide 49
Lambic fermentation microbiological profile
Lambic fermentation:Brettanomyces type yeasts,actidione resistant
tanks:wood vs metal hops:fresh vs old
 METAL TANKS versus WOODEN CASKS
Enterobacteriaceae ( ENTERO’S ) + Actidione resistant yeasts ( A. Res. Y. ) + Non
Lactic acid bacteria ( LAB ) actidione resistant yeasts ( Non A. Res. Y. )
8 8

LOG N c.f.u./mL
c.f.u./mL
N c.f.u./mL

7
c.f.u./mL

7
6 6
5 5
4 4
LOGN

3 3

LOG
LOG

2 2
1 1
0 0
0 2 4 6 8 10 12 14 16 18 20 0 2 4 6 8 10 12 14 16 18 20
MONTHS
MONTHS MONTHS
MONTHS
WOOD
WOOD –- ENTERO'S
ENTERO’S METAL
METAL -– ENTERO'S
ENTERO’S
WOOD -– A.
WOOD A. RES.
RES. Y.
Y. METAL –- A. RES.
METAL RES. Y.
Y.
WOOD –- LAB.
WOOD LAB METAL –- LAB.
METAL LAB WOOD -– NON
WOOD NON A.
A. RES.
RES.Y.Y. METAL -– NON
METAL NON A.
A. RES.
RES.Y.Y.
Eapp ( °P ) + ethanol ( ETH ) + pH Lactate ( LACT ) + Lactic acid bacteria ( LAB )
20 6 8 8000
pH

c.f.u./mL

PPM
v/v) and pH

LACTATE -–PPM
LOGNNc.f.u./mL
5 7 7000
15 6 6000
4
5 5000

LACTATE
(% v/v)
°P
°P

10 3 4 4000
3 3000
ETH(%

2
LOG
5 2 2000
ETH

1
1 1000
0 0 0 0
0 2 4 6 8 10 12 14 16 18 20 0 2 4 6 8 10 12 14 16 18 20
MONTHS MONTHS
MONTHS
WOOD –- °P
WOOD METAL -– °P
METAL °P WOOD –-LAB.
WOOD LAB METAL – -LAB.
METAL LAB
WOOD –-ETH
WOOD ETH METAL –
METAL -ETHETH
WOOD –- pH
WOOD METAL -–pH
METAL pH WOOD –-LACT.
WOOD LACT METAL – -LACT.
METAL LACT

Slide 52
Under proper conditions

• Lambic can be
produced in metal
tanks
Under proper conditions

•
• Fresh hops
compared to aged
hops were not
detrimental to
lambic brewing
 Under proper conditions

• Infusion
brewing
compared to
turbid mash
brewing is
possible
Under proper conditions

• Infected lambic
worts from site
A or B give a
good similar
cask fermenting
lambic at site C
 Under proper conditions

•
• Seeded lambic
wort at site A
shows a similar
behavior when
cask fermented at
different sites a
or b
 Spontaneous infected lambic wort
from a brewery A is transfered to
brewery a and b where it is only
cask- and bottle fermented.
Or:the infected worts from brewery A
and B is transfered to brewery a
OBSERVATIONS:
The cask environment has more final
effects than the open trays
environment
The steker gueuze has its own
characteristics.
Slide 59
 Bottle refermentation scheme

CLASSIC BOTTLE TRADITIONAL LAMBIC

REFERMENTATION BOTTLE REFERMENTATION

YOUNG FINISHED BEER OLD LAMBIC

(enzymes + micro-organisms)
MIXED WITH YOUNGER
LAMBIC (substrates)
ADDITION OF SUGAR + YEASTS

BOTTLING BOTTLING

INCUBATION INCUBATION

REFERMENTED BEER GUEUZE (« FOND GUEUZE »)

Slide 60
Slide 61
 Experimental conditions
to study bottle refermentations
4 MOTHER LAMBICS

A - 20 months (0.95°P) D - 20 months (2.28°P)

B - 12 months (3.10°P) F - 20 months (1.99°P)
AND
10 DIFFERENT MICROBIAL INOCULATIONS
A B C D E F G H I J
Brettanomyces bruxellensis x x x
Brettanomyces lambicus x x x
Saccharomyces cerevisiae x
Saccharomyces bayanus x x x
Pediococcus (2 strains) x x x
Brettanomyces (isolated, 2 strains) x x

Slide 62
Other gueuze beers
1.Filtered gueuzes: lambic
pasteurized,not-
refermented,carbonated (sweetened)

2.Fruit beers: As above but

mixed (diluted) with fruit juices.(Not
always with lambic)

3.Kriek gueuze: cask-fermented

with added sour cherries and
refermented:The real thing!)
Slide 63
 FARO
much preferred sweetened version of lambic
Faro from latin far:a type of wheat
Faro is a blend of old strong lambic
and a less strong version.The blending
is traditionally done by the inn-keeper
or later by the brewer or
a »préparateur »,an intermediate
between brewer and inn-keeper
Anyhow the blend is sweetened by
the inn-keeper by adding dark
concentrated lambic wort syrup or later
candy sugar.The préparateur in
general may clarify,colour and sweeten
the faro blends…. Slide 64
 Preferential sweetening of acidic beers
Faro is a lambic locally sweetened by pub holders
Sweetening can also be an individual preference.An
individu is then provided with sugar and a crusher
(“stoemper”)This practice was also common for other
acidic beers such as Aarschotse bruine etc.
 2.Acidification by blending an ale with lambic or gueuze

JACK-OP !!!

A succesful acidic beer around

1920 ,brewed in Werchter ,
called the students beer, is a BLEND of a top
fermented ale and a lambic or gueuze.

A similar process was used for Aarschotse bruine

which has recently been re-introduced.
 Are white beers acidic ?
Verlinden in 1944 described our white beers to
contain around 2000 ppm acid (as lactic acid)NOW in
contrast to the German Berliner Weisse(+mixed
cultures)
the Belgian white beers(+pure cultures) are not
really sour
(although drinkers may appreciate sourness by
adding a slice of lemon!)
A so called white lambic on the contrary can be
appreciated by its charming low acidity
Lambic wort+spices--spontaneous fermentation--
young lambic--carbonatation--bottling--
pasteurisation
 THE FUTURE OF
LAMBIC AND GUEUZE

SPONTANEOUS FERMENTATION CAN SURVIVE

BY USING TIME SAVING PROCESS ADAPTATIONS
WHICH DO NOT LARGELY AFFECT
THE REQUIRED CHARACTERISTICS OF THE FINAL REAL PRODUCT

FOR RELIABLE ADAPTATIONS HOWEVER

A MINIMUM OF SCIENTIFIC KNOWLEDGE
BECOMES A NECESSITY

Slide 68
 Acidic beers

Belgian red and brown

sour ales
Non gueuze TYPES OF ACID BEERS
ANNO 2012
The regional Flandres acid ales

type Rodenbach….

Slide 70
Rodenbach,Liefmans,Petrus,Bacchus,Bourgogne
des Flandres,Duchesse de Bourgogne…
Red-brown Flandres acidic beers fermented in barrels or
metal tanks.
1. Flanders' red-brown beer : Rodenbach

Large wooden casks

Slide 72
Rodenbach beer fermentation steps
WORT

“yeast”
7 d 16 - 21°C
CYLINDRO CONICAL PRIMARY FERMENTATION

TANK
a
SECONDARY FERMENTATION
EPOXY COATED TANK 4 - 5 W 15 - 21°C

CASK
TERTIARY FERMENTATION
young beer
18 – 24 M 15 - 21°C
▼
aged beer
►
on-line blending
to GRAND CRU
to RODENBACH
◄◄◄◄◄◄ ◄◄◄◄◄◄

cooling 0°C ► centrifugation ► carbonatation ► pasteurization

Slide 73
 Evolution of microbial populations
of Rodenbach fermentation
: total yeasts 9,00
8,00

Log (CFU / mL)

7,00
: actidione-resistant 6,00
5,00
yeasts
4,00
: lactic acid bacteria 3,00
2,00
1,00
: acetic acid bacteria 0,00
0 0,5 1 1,5 2 2,5 3 3,5 4 4,5 5 5,5 6 6,5
9,00 Weeks
8,00
7,00
Log (CFU/mL)

6,00
5,00
4,00
3,00
2,00
1,00
0,00
0 10 20 30 40 50 60 70 80 90 100 110
Weeks Slide 74
 Evolution of lactic acid during
Rodenbach fermentation
900
750

Lactic acid (ppm)

: total lactic acid 600
450
: D lactic acid
300
: L lactic acid 150
0
0 0,5 1 1,5 2 2,5 3 3,5 4 4,5 5 5,5 6 6,5 7

6000 Weeks

5000
Lactic acid (ppm)

4000
3000
2000
1000
0
0 10 20 30 40 50 60 70 80 90 100 110
Weeks
Slide 75
 Evolution of acetic acid during
Rodenbach fermentation
1200

1000

Acetic acid (ppm)

800
600
400

200
0
0 0,5 1 1,5 2 2,5 3 3,5 4 4,5 5 5,5 6 6,5
Weeks
1600
1400
Acetic acid (ppm)

1200
1000
800
600
400
200
0
0 10 20 30 40 50 60 70 80 90 100 110
Weeks
Slide 76
 CONCLUSIONS
The renewed appreciation for high quality
gueuzes and sour ales proves that
traditional processes
CAN survive modern times provided that
the brewer is prepared for
HARD WORK , incomparable enthusiasm
for the final product and awareness and
understandig of possible scientific progress.
Slide 77
TYPES OF ACID BEERS ANNO 2009

BERLINER WEISSBIER (Germany)

Slide 78
 Goslar/Leipzig Gose
beer
Spontaneous fermented acidic
beer around 1740
Cask fermentation achieved in
long neck bottles
Bottles closed by yeast
crop,not corked !

Slide 79