dry hopping - université catholique de louvain | … yeast by dry hopping improves the beer value...

Dry HoppingMyths versus Reality

1

Major volatile compounds

after main fermentation

(at start of dry hopping !)

2

Compounds present in beer after main fermentation

IR CPSil5 Compounds Flavour Samples

T 0 Day

C (ppm)

bièreFD

730 isobutylacetate banana, fruity 0.037 128

774 ethylbutyrate banana, fruity 0.058 512

812 3-methylbutanoic acid cheese 0.101 512

852 isoamylacetate Banana, apple 0.843 128

921 4-methylpentanoic acid cheese 0.009 <128

957 hexanoic acidDry leaves, goaty,

fatty acids0.094 512

974 2-methyl-2-pentanoic acid grassy, acid 0.056 <128

982 β-myrcene lemon, sweet 0.014 <128

1001 hexylacetate + sulfitic grassy, hop 0.002 512

1018 phenylacetaldehyde sweet, hony 0.010 <128

1088 linalool lemon 0.009 <128

1101 β-phenylethanol rose 6.672 512

1226 citronellol lemon 0.037 <128

1260 4-ethylguaiacol spicy 0.011 ND

1294 4-vinylguaiacol cloves 0.70 <128

YEAST

HOP

GLYCOSILES PRECURSORS

CYSTEINES BY PRODUCTSACTION ON FATTY ACIDS

BA

CK

GR

OU

ND

IN

FOR

MA

TIP

ON

3

IR CPSil5 Compounds Flavour Samples

T 0J

C (ppm)bière

FD

808 3-methyl-2-buten-1-thiol (MBT) hop, cheese ND 512

850 3-mercaptopropanol potatoes ND 512

890 2-mercaptoethylacetate grilled meat ND 512

905 4-mercapto-4-methyl-2-pentanone catty ND <128

1374 beta -damascenone cooked apple ND <128

1432 gamma-decalactone peach ND <128

1465 delta-decalactone coconut ND <128

1441 4-vinylsyringol Cloves ND <128

1578 ethyldodecalactone grassy ND <128

1687 delta-dodecalactone fruity, olive ND <128

4

YEAST

HOP

GLYCOSILES PRECURSORS

CYSTEINES BY PRODUCTSACTION ON FATTY ACIDS

Compounds present in beer after main fermentation

Analysis performed

at the lab of Prof Sonia Collin

UCL

5EBC HOP SYMPOSIUM GDX/JMR/SDK/2009

6

ALCOHOLS

ALDEHYDES

LACTONES

TERPENES

Monoterpenes

Sesquiterpenes

MCF-ACIDS (Medium chain fatty acids)

LCF-ACIDS

GLUCOSIDE OF TERPENES

Monoterpenes

Sesquiterpenes

GLUCOSIDE OF PHENOLS

Phenolic acids

Polyphenols

7

ACIDS

ALCOHOLS

TERPENES

Monoterpenes

Sesquiterpenes

VOLATILE PHENOLS

CYSTEINE DERIVATIVES (THIOLS)

MCF-ACIDS

LACTONES

GLUCOSIDE OF TERPENES

Monoterpenes

Sesquiterpenes

GLUCOSIDE OF PHENOLS

Phenolic acids

Polyphenols

Fate of major beer volatiles

by dry hopping (maturation) in presence of S.cerevisiae

8

MONOTERPENES

MAJOR BEER VOLATILES

CITRUS FLAVORS

9

IR CPSil5 Compounds Flavour Samples

TERPENEST 0J T 14J T 21J

C (ppm)bière

FD C (ppm)bière

FD C (ppm)bière

FD

982 β-myrcene Citrus, sweet 0.014 <128 0.014 <128 0.040 <128

1088 linalool Citrus 0.009 <128 0.05 >128 0.06 >1024

1226 citronellol Citrus, fresh 0.037 <128 0.020 <128 0.118 512

YEAST

HOP

GLYCOSILES PRECURSORS

10

CITRUS FLAVORS

IR CPSil5 Compounds Flavour Samples

T 0J T 14J T 21J

C (ppm)bière

FD C (ppm)bière

FDC (ppm)

bièreFD

982 β-myrcene Citrus, sweet 0.014 <128 0.014 <128 0.040 <128

1088 linalool Citrus 0.01 <128 0.05 >128 0.06 >1024= dil 1024x

1226 citronellol Citrus, fresh 0.037 <128 0.020 <128 0.118 512

TERPENES

11

DRY HOPPING: evolution of linalool

Influenced by :

- Time

- Temperature

- Hop variety (oil content)

- Hop quantity added

- Hop dispersion methods

(static /dynamic)- Beer type and yeast type

12

Linalool

CITRUS FLAVOURS

IR CPSil5 Compounds Flavour Samples

TERPENEST 0J T 14J T 21J

C (ppm)bière

FD C (ppm)bière

FD C (ppm)bière

FD

982 β-myrcene Citrus, sweet 0.014 <128 0.014 <128 0.040 <128

1088 linalool Citrus 0.009 <128 0.034 >128 1.192 >1024

1226 citronellol Citrus, fresh 0.037 <128 0.02 <128 0.12 512= dil 512x

13

DRY HOPPING: evolution of citronellol

Influenced by :

- Time

- Temperature

- Hop variety

- Hop quantity

- Hop dispersion methods

(static /dynamic)

- Beer type and yeast type

14

15

METABOLISM CASCADE OF MONOTERPENE ALCOHOLS BY LAGER

AND ALE YEASTS PROPOSED BY KING AND DICKENSON. THE ASTERIC

INDICATES A CHIRAL CENTER.

PRESENCE OF CITRONELLOL

16

IN LATE HOPPED WORT

(10 min before end of boiling)

Citation :

HAPPOSHU

17

G

Fate of the other volatile compounds

as the …

• Acids

• Esters

• Alcohols

• Aldehydes

• Lactones

• Thiols

• Others

18

CHEESY FLAVOURS

IR CPSil5

Compounds Flavour Samples

SHORT CHAIN FATTY ACIDST 0J T 14J T 21J

C (ppm) bière FD C (ppm) bière FD C (ppm) bière FD

812

3-methylbutanoic

acidcheesy

0.101 512 0.102 512 0.179 >1024

921

4-methylpentanoic

acidcheesy

0.009 <128 0.003 <128 0.018 512

957 hexanoic acid mild cheesy0.094 512 0.102 512 0.179 >1024

974

2-methyl-2-

pentanoic acidgrassy, acid

0.056 <128 0.037 <128 0.160 512

YEAST

HOP

19

HOP CONDITIONNING AND STORAGE CONDITIONS

CHEESY : linked to bad storage of hop bales and/or pellets and to the use of aged hop (presence of degradation products as 2-methyl-propionic acid/ isobutyric acid and of 2-methyl-butyric acid/ isovaleric acid)

20

FRUITY, FLORAL FLAVOURS

IR CPSil5 Compounds Flavour Samples

ESTERST 0J T 14J T 21J

C (ppm)bière

FD C (ppm)bière

FD C (ppm)bière

FD

730 isobutylacetate banana, fruity 0.037 128 0.065 512 0.096 >1024

774 ethylbutyratebanana,

fruity, flowery0.058 512 0.113 >1024 0.156 >1024

852 isoamylacetatebanana,

sweet0.843 128 1.086 128 1.821 >1024

1001 hexylacetate grassy, hop 0.002 512 0.002 512 0.011 512

YEAST

HOP

21

FRUITY, FLORAL FLAVOURS

YEAST

HOP

GLYCOSILES PRECURSORS

CYSTEINES BY PRODUCTS

ACTION ON FATTY ACIDS

IR CPSil5 Compouds Flavour Samples

HIGHER ALCOHOLST 0J T 14J T 21J

C (ppm)bière

FD C (ppm)bière

FD C (ppm)bière

FD

1101 ββββ-phenylethanol rose 6.672 512 6.532 512 11.685 >1024

22

IR CPSil5 Compouds Flavour Samples

ALDEHYDEST 0J T 14J T 21J

C (ppm)bière

FD C (ppm)bière

FD C (ppm)bière

FD

1018phenylacetaldehyde sweetened,

honey0.010 <128 0.015 <128 0.022 512

23

IR CPSil5 Compounds Flavour Samples

LACTONEST 0J T 14J T 21J

C (ppm)bière

FD C (ppm)bière

FD C (ppm)bière

FD

1432gamma-

decalactonepeach ND <128 ND <128 ND 512

1465delta-

decalactonecoconut ND <128 ND <128 ND 512

1578Ethyl-

dodecalactonegrassy ND <128 ND <128 ND 512

1687delta-

dodecalactone

fruity,

oliveND <128 ND <128 ND 512

FRUITY, FLORAL FLAVOURS

YEAST

HOP

GLYCOSILES PRECURSORS

CYSTEINES BY PRODUCTS

ACTION ON FATTY ACIDS

24

FRUITY FLAVORS

WHY LACTONES ?

25

FRUITY FLAVORS

YEAST PEROXISOME

BIOSYNTHESIS

26

FRUITY FLAVORS

YEAST PEROXISOME

BIOSYNTHESIS

27

IR CPSil5 Compounds Flavour Samples

T 0J T 14J T 21J

C (ppm)bière

FD C (ppm)bière

FD C (ppm)bière

FD

8083-methyl-2-buten-1-thiol

(MBT)

hop,

cheeseND 512 ND >1024 ND >1024

850 3-mercapto-propanol potatoes ND 512 ND >1024 ND >1024

890 2-mercapto-ethylacetategrilled

meatND 512 ND >1024 ND >1024

9054-mercapto-4-methyl-

2-pentanonecatty ND <128 ND <128 ND 512

YEAST

CYSTEINES BY PRODUCTS

SULFITIC FLAVOURS

28

Concentration of

major non-volatile compounds

after maturation in presence of raw hops• Polyphenols

• Glucosides

29

Non volatile compounds extractedPOLYPHENOLS

30

Year 2008

POLYPHENOLS

MOST INTERESTING PHENOLIC COMPOUNDS

• Phenolic acids

• Flavonoids

Flavanoids

Proanthocyanidin = anthocyanogen

• Resveratrol

31

IR CPSil5 Compounds Flavour Samples

PHENOLST 0J T 14J T 21J

C (ppm)bière

FD C (ppm)bière

FD C (ppm)bière

FD

1260 4-ethylguaiacol spicy 0.011 ND 0.000 ND 0.067 >1024

1294 4-vinylguaiacol Cloves, dentist 0.70 <128 1.32 <128 3.36 >1024

CLOVES AND SPICY FLAVORS

YEAST

ACTION ON PHENOLIC ACIDS

PRESENCE OF “POF or PAD YEASTS”

(Phenolic acid decarboxylase activity)

Several TOP fermenting yeasts

ONLY IF PRESENCE OF

BRETTANOMYCES sp

(Vinyl reductase activity)

POLYPHENOLS PHENOLIC ACIDS

HOP

32

FERULIC ACID = Phenolic acid

(Barley, Hops)

Formation of vinylguaiacol vinylphenol and styrene

from phenylalanine by S. cerevisiae !

33

POLYPHENOLS

FLAVANOIDS

WHY ARE “FRESH” FLAVANOIDS INTERESTING !

34

BECAUSE….

Non volatile compounds extracted

- POLYPHENOLS

- GLUCOSIDES35

NON VOLATILE COMPOUNDS

EXTRACTED

POLYPHENOLS

PHENOLIC ACIDS

FLAVANOIDS

GLUCOSIDES

GLYCOSIDES

• IMPORTANCE OF THE HOPS

• INFLUENCE OF THE YEAST STRAINS

• CULTURE YEASTS

• WILD YEASTS

Brettanomyces sp

36

- GLUCOSIDES

A few theory…

Glucosidic bound can be hydrolysed by :

- High temperature

- Enzymatic activity

- endogenous presence (yeast by

maturation, bottle refermentation)

- (addition of exogenous beta-

glucosidase during the process) –

No

n v

ola

tile

co

mp

ou

nd

s e

xtr

act

ed

37

No

n v

ola

tile

co

mp

ou

nd

s e

xtr

act

ed- GLyCOSIDES

In beer, the aglycon part is liberated by two

different types of activities

- by ββββ -1-3-glucanase activity in case of S. cerevisiae strains

- by β β β β -glucosidase activity in case of Brettanomyces sp. strains

38

Glycosidic bounds hydrolysed by endogenous

presence (by yeast during maturation and/or during bottle fermentation)

Preferential links detected in hopsTerpenes

• TERPENIC SUB – UNIT + SUGAR : FLAVOUR PRECURSOR

linalool - β-D- glucoside

α – terpineol – β – D - glucoside

1- octen-3-ol- β – D – glucoside

3 – hydroxy – β – damascenone – D – glucoside

• PHENOLIC SUB – UNIT + SUGAR : SHELF – LIFE PROTECTOR PRECURSOR

Flavonoids

Kaempferol and Quercetin – 3 – glycosides

glycosidic sub-unit : mono or di- glycosides – glucose , galactose, rhamnose

Flavanoids

As yet, flavanol dimers or multimers glucoside aren’t detected nor described

No

n v

ola

tile

co

mp

ou

nd

s e

xtr

act

ed

39

Glucosidic bound hydrolyses by endogenous presence (yeast by maturation, bottle refermentation)

No

n v

ola

tile

co

mp

ou

nd

s e

xtr

act

ed

• Importance of the yeast strain and

yeast species

40

41

LEGEND :SC : S.CEREVISIAE (ALE YEAST)

BB : BRETTANOMYCES BRUXELLENSIS

BC : BRETTANOMYCES CUSTERSII

*SC + D : S. CEREVISIAE + CARBOHYDRATE

SOURCE (GLUCOSE)

AR : AR 2000 GLYCOSIDASIC CONCENTRATE OF

ASPERGILLUS NIGER (COMMERCIALLY AVAILABLE)

TYPICAL YEASTS ββββ-glucanase and ββββ glucosidase activities

of different yeast strains and yeast species

Influence of hop quantity and of yeast strain

Glucosidic bound hydrolyses by endogenous presence (yeast by maturation, bottle refermentation)

• Importance of the aglycon moiety

(glucoside of higher alcohols)

1-octen-3-ol

No

n v

ola

tile

co

mp

ou

nd

s e

xtr

act

ed

cis-3-hexen-1-ol

42

YEAST

HOP

GLYCOSILES PRECURSORS

1374beta -

damascenone

cookedapple, rose

ND <128 ND <128 ND >1024

IR CPSil5 Compounds Flavour Samples

OTHERST 0J T 14J T 21J

C (ppm)bière

FD C (ppm)bière

FD C (ppm)bière

FD

43

FRUITY, FLORAL FLAVOURS

BEER DRY HOPPING IS A CONFLICTUAL ART

WHEN RAW HOP EXTRACTION TAKES PLACE IN PRESENCE OF ACTIVE YEAST

ACTIVE YEAST BY DRY HOPPING IMPROVES THE BEER VALUE BY :

- PROTECTING BEER FROM OXIDATION BY CAPTURING MOLECULAR OXYGEN

- PROTECTING BEER BY EXERTING A REFRESHING EFFECT

- IMPROVING (IN A FIRST STEP) THE BEER FLAVOUR BY :

• increasing the content of citronellol

• synthesizing nice flavours as lactones

• liberating aglycons from glucosides :

- Linalool, Geraniol,…

44

BEER DRY HOPPING IS A CONFLICTUAL ART

WHEN RAW HOP EXTRACTION TAKES PLACE IN PRESENCE

OF ACTIVE YEAST

ACTIVE YEAST by dry hopping

DECREASES THE BEER VALUE BY :

- LIBERATING ESTERASES THAT DECREASE THE AROMA

- METABOLIZING H+ ACCEPTORS AS SOME ALDEHYDES

AND OTHER POTENT SUBTILE FLAVOURS AS LACTONES

45

46

BEER DRY HOPPING IS A CONFLICTUAL ART

WHEN BOTTLING TAKES PLACE IN PRESENCE OF ACTIVE YEASTs

Sierra Nevada

Orval

47

TRADITIONNAL EUROPEAN METHODS

Methods by batch dry hoppingA

B

C

D

E

A : Parts of 4 KG of dried hop cones are compacted under inert gas (N2) in onion bags that are sealed in aluminium packets by the hop supplier. of 4 KG. These are stored in the fridge of the brewery at + 2 /+ 4°C. B : Just before use, the aluminium packets are opened and the onion bags containing the hop cones are decompressed.C : Before introduction in the empty tank,the hop cones are decompacted tooD : In order to improve the contact between the beer and the hop bells, the hop bag can be attached to the tank. E : Hop plugs of 5 Oz ( 140 Gr) are also available by hop suppliers and can be added to beer casks directly.

48

Pneumatic Method

= dry methodOut- Indoor hop

cellar canoon

Lagunitas Brewery

Methods by continuousextraction in separated vessels

1.2. by hop pelletsStone Brewery (designed plant)

New Belgium Brewery

1.1. by hop conesSierra Nevada Brewery

NEW AMERICAN METHODS

With authorization of Steven Pauwels, CBC Dry hopping subcommittee

HOP and BEER

49