HAL Id: hal-02961841
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Submitted on 8 Oct 2020
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Exploring the potentialities of Metschnikowia species during sequential wine fermentation
Pauline Seguinot, Anne Ortiz-Julien, Pascale Brial, Carole Camarasa
To cite this version:
Pauline Seguinot, Anne Ortiz-Julien, Pascale Brial, Carole Camarasa. Exploring the potentialities of Metschnikowia species during sequential wine fermentation. 7. conference on Yeasts and Filamentous Fungi, Oct 2019, Milan, Italy. �hal-02961841�
Exploring the potentialities of Metschnikowia species during sequential wine fermentation
PAULINE SEGUINOT1, 2, ANNE ORTIZ-JULIEN2, PASCALE BRIAL1, CAROLE CAMARASA1
1 INRA
2 place Viala
F-34080, MONTPELLIER www.montpellier.inra.fr/
fermentation performances
Currently, in the winemaking sector, there is a renewed interest of the use of non-Saccharomyces species during the fermentation process. Long regarded as spoilage microorganisms, the potential of these yeasts, including Metschnikowia pulcherrima species, to improve the sensory quality of wines is now recognized. Due to their inability to complete the fermentation, M. pulcherrima strains must be used in combination with Saccharomyces cerevisiae, added in sequential or co- inoculation. However, the limited knowledge on their traits prevents to more widely gain the benefits and potentialities of M. pulcherrima during wine fermentation. To fill this shortcoming, this study investigated the specificities and the contribution of M. pulcherrima during sequential fermentation with S. cerevisiae.
Compared with S. cerevisiae pure culture, sequential fermentation has a negative impact on:
the implantation of S. cerevisiae, due to the important
consumption of glutamate, glutamine and branched amino acids of M. pucherrima
the fermentation rate, resulting in an increased fermentation duration
the production of medium chain fatty acids and their esters derivatives
production of medium chain fatty acids and ethyl esters
1 2 3 4 5
Octanoïc acid (mg/L)
M. pulcherrima pure culture
sequential fermentation
S. cerevisiae pure culture
0.25 0.50 0.75
M. pulcherrima pure culture
sequential fermentation
S. cerevisiae pure culture
Ethyl Octanoate (mg/L)
0,0E+00 4,0E+07 8,0E+07 1,2E+08 1,6E+08
S. cerevisiae pure sequential fermentation
S. cerevisiae final population (cells/mL)
0 20 40 60
Ala Arg NH4 Asp Gln Glu Gly His Ile Leu Lys Met Phe Ser Thr Trp Tyr Val
ConsumedN-source after48 h (mg N/L)
S. cerevisiae Must M. pucherrima
Nitrogen sources consumption
0 0,2 0,4 0,6 0,8
0 100 200 300
tim e (h) 1,0
1,2
dCO2/dT(g/l/h)
Fermentation rate
Compared with S. cerevisiae pure culture, sequential fermentation has a positive impact on:
the production of acetate esters (and some higher alcohols, depending on lipids, sugars and nitrogen availability)
the production of acetate (decrease) and glycerol ( increase)
the production of positive thiols, which increases by a factor up to 6
production of higher alcohols and acetate esters
production of thiols
●
200 400 600 800
3MH (ng/L)
S. cerevisiae pure culture sequential
fermentation
100 200 300
4MMP (ng/L)
S. cerevisiae pure culture sequential
fermentation
Thiols production in MP/SC sequential and SC pure fermentations
Incidence of nutrients availability on thiols production in MP/SC fermentation
production of CCM compounds
Example of octanoïc acid and ethyl octanoate.
Similar profile observed with hexanoïc, decanoïc and dodecanoïc acids and their ester derivatives
SM with 200 mg N/L, 240 g/L sugars and 5 mg/L lipids
No production of 3-MHA in S.cerevisiae pure culture
Range of production of 3-MHA in sequential fermentation: 0-30 ng/L
Nitrogen ( mg/L)
100
150 200 250 300
Lipi ds
(m g/L)
2 3 4 5 6 7 8
3M HA (ng/
L)
0 10 20 30
Slice at sucre = 220
Nitrogen ( mg/L)
100
150 200 250 300
Lipi ds (m
g/L)
2 3 4 5 6 7 8
4M MP (ng/
L)
50 100 150 200 250 300
Slice at sucre = 220 Nitrogen (
mg/L)
100 150
200 250 300
Lipi ds
(m g/L)
2 3 4 5 6 7 8
3M H ( ng/
L)
200 400 600 800
Slice at sucre = 220 Nitrogen (
mg/L)
100
150 200 250 300
Sugar
(g/
L)
180 200 220 240 260
Pheny lethanol
(m g/L)
80 90 100 110 120 130
Slice at lipide = 5
Sugar (g/L)
180 200
220
240 260
Lipi ds
(m g/L)
2 3 4 5 6 7 8
Phe nyl eth anol (m
g/L) 80
90 100 110 120 130
Slice at nitrogen = 190
Nitrogen ( mg/L)
100 150 200 250 300
Lipi ds
(m g/L) 2
3 4 5 6 7 8
Iso but anol (m g/L)
120 130 140 150 160
Slice at sugar = 220
Nitrogen ( mg/L)
100
150 200 250 300
Lipi ds
(m
g/L) 2
3 4 5 6 7 8
Iso but yl ac
etat e ( mg/
l) 0
1000 2000 3000
Slice at sugar = 220
Nitrogen ( mg/L)
100
150 200 250 300
Sugar
(g/
L) 180
200 220 240 260
Phe nyl eth anol (m g/L)
60 80 100 120
Slice at lipide = 5
Sugar (g/L)
180 200
220
240 260
Lipi ds
(m
g/L) 2
3 4 5 6 7 8
Phe ny lethanol
(m
g/L) 60
80 100 120
Slice at nitrogen= 190
Nitrogen ( mg/L)
100
150 200 250 300
Lipi ds
(m
g/L) 2
3 4 5 6 7 8
Iso but yl ac
etat e ( mg/
L)
200 400 600 800
Slice at sugar = 220 Nitrogen (
mg/L)
100
150 200 250 300
Lipi ds (
mg/
L) 2
3 4 5 6 7 8
Iso but anol (m
g/l) 30
40 50 60
Slice at sugar= 220
The comparison between sequential inoculation and pure cultures pointed out the benefits but also the shortcomings of the use of M. pulcherrima in fermentation. Thanks to the Box-Behnken model, the impact and interaction of the parameters were assessed, showing that nitrogen was the most influencing nutrient on the sensory and organoleptic profile of wines.
These results pave the way for the design of an improved management of sequential inoculation during winemaking to better exploit the potentialities of M. pulcherrima strains.
MP/SC sequential fermentation
SC pure
fermentation
Nitrogen ( mg/L)
100
150 200 250 300
Sugar
(g/
L) 180
200 220 240 260
ac etat
e ( g/L)
0.0 0.1 0.2 0.3 0.4
Slice at lipid = 5
Nitrogen ( mg/L)
100 150 200 250 300
Sugar
(g/
L) 180
200 220 240 260
glycer ol (
g/L) 9
10 11
Slice at lipid = 5
MP/SC sequential fermentation
Nitrogen ( mg/L)
150 200 250 300
Sugar
(g/
L) 200
220 240 260
glycer ol ( g/L)
5.0 5.5 6.0 6.5 7.0 7.5 8.0
Slice at lipid = 5 Nitrogen (
mg/L)
100
150 200 250
Sugar
(g/
L)
180 200 220 240 260
ac etat e ( g/L)
0.3 0.4 0.5 0.6
Slice at lipid = 5
SC pure
fermentation
phenylethanol Sugar: negativeC
Lipid: quadratic Nitrogen: no impact Sugar: no impactC
Lipid: no impact Nitrogen: quadratic
isobutanol Lipid: quadratic C
Nitrogen: quadratic Nitrogen: quadraticC
isobutyl acetate Lipid: + low NC
- high N
Lipid: no impact low NC
+ high N
acetate Sugar: no impactC
Nitrogen: negative Sugar: positiveC
Nitrogen: quadratic
glycerol Sugar: postiveC
Nitrogen: + low sugar - high sugar Sugar: positiveC
Nitrogen: negative
Similar response of 3MH (3-
mercaptohexan-1-ol) and 4MMP (4-mercapto-4-méthylpentan-2- one) as branched higher alcohols Similar profile of 3MHA (3-
mercaptohexyl acetate)l as acetate esters
The response to changes in nutrient availability may
differ between MP/SC fermentation and SC pure culture
strenght of the response opposite variations
Interactions between factors depending on the compound
2 Lallemand SAS
F-31700, BLAGNAC
