TWISTAROMA D.Steyer
1
Sesquiterpenol Monoterpenol
Oxygenated Sesquiterpene
Ester Ketone
Aldehyde
Monoterpene
Compounds
Description
Threshold (beer)
Linalool1)
floral
8-20-80 g/L
Geraniol
rose-like, ‘citrus’ odour,
5g/L
Hop ether
typical
5g/L
What are the influences of yeast on hop volatile compounds ?
1) ” Linalool—A Key Contributor to Hop Aroma” S.Hanke
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1) what are the hop volatile compounds influenced by yeast ? 2) what kind of metabolic pathway(s) is (are) involved
4
Yeasts Choices
UCD915 (Ale)
CLIB267 (Lager) CLIB279 (Lager)
Yeast strains from the CLIB collection (Institut Pasteur)
Nguyen, Legras et al, Plos one 2011
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Strisselspalt
Aramis
Bouclier
Triskel
old variety
new variety
new variety
Linalool
7-8mg/g
14 mg/g
6 mg/g
new variety 10 mg/g
α-acids
3,4%
8,1%
9,5%
8,7% 6
Wort (13°P)
10min before end of ebulltion
Strisselspalt
Aramis
Bouclier
2,1g/L
Triskel
yeast
Wort
Stir bar (not SBSE)
CO2
• Volatile compounds analysed by SBSE-GC-MS
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Compounds Ethyl hexanoate Ethyl octanoate Ethyl heptanoate Nerol Heptanoic acid Isoamyl alcool 3-methylpentanol 1-nonanal Decanoic acid 9-decenoic acid Ethyl nonanoate a-Humulene Isoamyl acetate Ethyl pentanoate Styrene Hexyl acetate Decanal 2-Undecanone 1-Octanol Ethyl decanoate Isoamyl octanoate Ethyl dodecanoate 2-Phenyl Ethanol 1-Butanol 1-Hexanol Heptyl acetate octyl acetate 1-Heptanol 2-Nonanol Linalool Ethyl cis-dec-4-enoate Ethyl 9-decenoate 2-undecanol Citronellol 2-Phenethyl acetate Geraniol Octanoic acid isobutyl butanoate 3-penten-2-ol 4-methyl-1-pentanol 2-Ethyl hexanol Isovaleric acid
KI Validation Odor 1255 MS,RI fruit, green apple, sweet 1432 MS,RI fruit, sweet, floral 1336 MS,RI fruit, wine 1800 MS,RI floral, sweet 1781 MS,RI unpleasant, sweaty 1230 MS,RI alcoholic, malty, fusel 1347 MS,RI vinous, herbaceous 1395 MS,RI floral, fruit 2235 MS,RI rancid, soapy 2315 MS,RI fatty, wax 1535 MS,RI fruit, rose, floral 1690 MS,RI wood 1186 MS,RI banana, fruit, sweet 1193 MS,RI fruit, apple, floral 1277 MS,RI sweet 1282 MS,RI fruit, floral, pear 1499 MS,RI green, tallow 1602 MS,RI floral, Orange 1551 MS,RI green, floral, rose 1642 MS,RI fruit, grape fruit, pleasant 1665 MS,RI fruit, sweet 1851 MS,RI fruit, sweet, floral 1908 MS,RI rose, floral, honey 1191 MS,RI medicinal, alcoholic, fruit 1347 MS,RI green, floral, resinous 1372 MS,RI Herbal, wine 1472 MS,RI fruit, pear 1445 MS,RI green, fruit 1511 MS,RI green, watermelon 1538 MS,RI floral, citrus, fruit 1670 MS,RI unknown, 1695 MS,RI fruit, unknown 1720 MS,RI fruit, 1764 MS,RI rose, green 1819 MS,RI floral, rose, honey 1845 MS,RI rose, floral, sweet 2057 MS,RI fatty, rancid 1172 MS,RI fruit, 1206 MS,RI burnt, vinyl 1347 MS,RI toasted 1480 MS,RI green, floral, rose 1690 MS,RI rancid, cheesy, acidic
Compounds 2-Octanol 1-Nonanol
KI Validation 1511 MS,RI 1658 MS,RI
Odor mushroom roseorange
cis-3-Hexenol
1324
MS,RI
green, grassy, fresh
Geranyl acetone 1-Decanol
1860 1758
MS,RI MS,RI
magnolia, floral floral, Orange, fatty
alpha-Terpineol Hexanoic acid Hexyl formate Geranyl acetate b-Myrcene Ethyl 3-hexenoate Nonanoic acid Citronellyl acetate 2-Butanol 2,7-Dimethyl-2,7-octadiene 3-methyl-2-Buten-1-ol
1702 1871 1346 1759 1209 1309 2183 1664 1170 1175 1312
MS,RI MS,RI MS,RI MS,RI MS,RI MS,RI MS,RI MS,RI MS,RI MS,RI MS,RI
floral, lilac, oily cheesy, rancid, sweaty wood rose, sweet spicy
Isoamyl propanoate 1-Dodecene 2-Hexanol Methyl 4-Methyl-2-hexenoate Hop ether
1225 1259 1312 1340 1382
MS,RI MS,RI MS,RI MS,RI MS,RI
pineapplenotes,
hexyl butanoate 3-(methylthio)propyl acetate Decyl acetate Methyl geranate 2-tridecanol 2-Phenylethyl butanoate Caryophyllene oxide Humulene epoxide II spathulenol pentadecanol
1458 1629 1685 1699 1920 1966 2002 2053 2223 2150
MS,RI MS,RI MS,RI MS,RI MS,RI MS,RI MS,RI MS,RI MS,RI MS,RI
fruit, floral, pine apple
2-Phenethyl hexanoate a-Cadinol
2154 2158
MS,RI MS,RI
fruit, unknown, wood
4-Vinylguaiacol Sesquiterpene1 a-Eudesmol b-Eudesmol 7-epi-amiteol Juniper Camphor Farnesol 2,3-dihydro-benzofuran 2-Phenylethyl octanoate dodecanoic acid
2163 2166 2204 2212 2231 2268 2302 2321 2333 2426
MS,RI MS MS,RI MS,RI MS,RI MS,RI MS,RI MS,RI MS,RI MS,RI
spicy, clove, smoky
fatty, rancid rose, fruit, wine herbaceous,
floral, green musty, wood Herbal, unknown,
green, wood
unknown, metallic, wax
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Terpenyl acetate; 2 Monoterpenol; 5
Phenol; 2
Ester; 22
Terpene; 2 Ketone; 1 Autre; 27
Higher Alcohol; 2
Alcohol; 15 Aldehyde; 2
Acetate Ester; 5 Sesquiterpene; 6
Acid; 8
High Isoamyl acetate + 4VG content Triskel and Bouclier +Sesquiterpenol (spicy and wood) Yeast volatile compounds
Strisselspalt and Aramis +Monoterpenol (floral)
Hop volatile compounds 11
Linalool 400
µg/L rel
300
200
1 00
0 Yeast
Hop
2 IB L C
77
2 IB L C
79
am Ar
U
5 91 D C
7 9 5 27 27 91 B B D I I C CL CL U
is Bo
lie uc
2 IB L C
77
r St
2 IB L C
79
t al sp l se ris
U
5 91 D C
7 9 5 27 27 91 B B D I I C CL CL U el ir s k T
12
Geraniol 35 30
µg/L rel
25 20 15 10 5 0 Yeast
Hop
77 79 15 2 2 9 IB IB CD CL CL U am Ar
77 79 15 2 2 9 IB IB CD CL CL U
is Bo
77 79 15 2 2 9 IB IB CD CL CL U
r lie uc St
t al sl p se ris
77 79 15 2 2 9 IB IB CD CL CL U el k s i Tr
13
By
UCD915 (Ale)
CLIB267 (Lager) CLIB279 (Lager)
14
+1mg/L geraniol Yeast strain
Stir bar CO2
Synthetic must
Fermented medium
Strain : Bywt
∑ Citronellol Geraniol Linalool
µg/L
Nerol
Geranylacetate Citronellylacetate Nerylacetate 16
Geranyl acetate 40
µg/L rel
30
20
10
0 Yeast
Hop
2 IB L C
77
2 IB L C
79
am Ar
U
6 91 D C
7 9 6 27 27 91 B B D I I C CL CL U
is Bo
lie uc
2 IB L C
77
2 IB L C
r St
sp el s r is
79
t al
U
6 91 D C
7 9 6 27 27 91 B B D I I C CL CL U is Tr
l ke
Des écarts types individuels ont été utilisés pour calculer les intervalles.
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Acetic acid
+
Geraniol
=
Geranylacetate
Acetic acid
+
Nerol
=
Nerylacetate
Acetic acid
+
Citronellol
=
Acetic acid
+
Isoamylalcool
Citronellylacetate
= Isoamylacetate
banana
ATF1/ATF2
18
Bywt + 1 mg/L geraniol
ByΔATF1 + 1 mg/L geraniol
[Geranyl+citronellyl+neryl acetate]
wt
ΔATF1 19
Synthetic must
Fermented medium
Strain : Bywt
∑ Citronellol Geraniol Linalool
µg/L
Nerol
Geranylacetate Citronellylacetate Nerylacetate 20
Citronellol 60
µg/L rel
50 40 30 20 10 0 Yeast
Hop
5 79 77 91 B2 B2 D I I C CL CL U am Ar
5 79 77 91 B2 B2 D I I C CL CL U
is B
7 5 9 27 91 27 D IL B IL B C C C U
r l ie c ou St
t al p s el s s ri
2 IB L C
77
9 5 27 91 D IL B C C U el k s i Tr
21
Fermented wort
Wort
Géraniol
reduction Géraniol Citronellol
22
NADPH,H+
NADP+
OYE21 OH
OH
3-methyl -2-buten-1-ol
3 methyl butanol
OH
OH
Geraniol
Citronellol
1Chou,
Howard H.Keasling, Jay D, US Patent Application 20100205855 (2009) 23
Bywt + 1 mg/L geraniol
ByΔOYE2 + 1mg/L geraniol wt ΔOYE2 ΔOYE3
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In our conditions ◦ Linalool concentration is controlled only by Hop varieties ◦ To protect Monoterpenol content in beer yeast has to be carrefully chosen to have low acetylsynthase (ATF1) activity ◦ Geraniol can be preservered by choosing yeast with low OYE2 activity
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In our conditions
◦ Sesquiterpenes, hop esters, ketone, are also influenced by yeast strains ◦ Yeast volatile compounds are also influenced by hop varieties
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Pauline Tristram Student-Engineer
Céline Clayeux PhD Chief Research and Development Officer
Comptoir Agricoles’ team
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[email protected]
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