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Driving the nitrogen/carbon ratio of the culture medium determines Propionibacterium freudenreichii survival during spray-drying.

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HAL Id: hal-01908153

https://hal.archives-ouvertes.fr/hal-01908153

Submitted on 29 Oct 2018

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Driving the nitrogen/carbon ratio of the culture medium determines Propionibacterium freudenreichii

survival during spray-drying.

Floriane Gaucher, Houem Rabah, Koffigan Kponouglo, Sylvie Bonnassié, Philippe Blanc, Romain Jeantet, Gwénaël Jan

To cite this version:

Floriane Gaucher, Houem Rabah, Koffigan Kponouglo, Sylvie Bonnassié, Philippe Blanc, et al.. Driv- ing the nitrogen/carbon ratio of the culture medium determines Propionibacterium freudenreichii survival during spray-drying.. IDF WDS 2018 Dairy for the next generation, Sep 2018, Daejeon, South Korea. 2018. �hal-01908153�

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labellisé

P. freudenreichii is used both as Swiss-type cheese starter and as probiotic. For these purposes, it should be converted into a powdered form to be stabilized, without significant loss of viability. Spray-drying is a stressful process, with simultaneous thermal, oxidative and osmotic stresses. To tolerate to these latter, bacteria should adapt. We investigate whether growth media could constitute a means to adapt bacteria in the aim to increase their survival during spray drying.

Milk Ultrafiltrate (MU) Sweet Whey (SW)

Yeast Extract Lactate (YEL)

Yeast Extract Lactate + Lactose (YEL+L)

Oxidative challenge 1,15mM H

2

O

2

, 1h

Heat challenge 60°C, 10 min

Spray drying Osmoprotectants

quantification

+ Salt - Salt

P. freudenrichii CIRM- BIA 129 growth

0 20 40 60 80 100 120

0 2 4 6 8 10

Survialafterheatchallange (%)

Trehalose/Glycine Betaine

-10 0 10 20 30 40 50 60

0 2 4 6 8 10

Survialafteroxidative challange(%)

Trehalose/Glycine betaine

0,00 20,00 40,00 60,00 80,00 100,00 120,00

YEL YEL + NaCl YEL + L YEL+ L + NaCl Surivivalafterheat chllange(%)

0,00 20,00 40,00 60,00 80,00 100,00

YEL YEL + NaCl YEL + L YEL+ L + NaCl Survivalafteroxidative challange(%)

0,00 20,00 40,00 60,00 80,00 100,00

SW SW+ NaCl MU MU + NaCl

Survivalafterheat challange(%)

0 10 20 30 40 50 60

SW MU

Concentration (g.L-1 )

Lactose Lactate Nitrogen

0 20 40 60 80 100 120

YEL YEL+NaCl YEL+L+NaCl YEL+L+NaCl

Survivalaftrerspary- drying

0 100 200 300 400 500 600 700 800 900

SW SW + NaCl MU MU+NaCL

Osmoprotectantsconcenctration (relative)

Tréhalose Glutamate GB

0 200 400 600 800 1000 1200 1400 1600 1800

YEL YEL + NaCl YEL+ L YEL + L + NaCl Osmoprotectantsconcentration (relative)

Trehalose Glutamate GB

0 10 20 30 40 50 60 70 80

YEL YEL+L

Concentration (g.L-1 )

Lactose Lactate Nitrogen

Carbon/nitrogen composition of the culture medium

The trehalose/glycine betaine ratio is linked to P. freudenreichii surivival during lethal challenges

The nitrogen/carbon ratio of the growth medium influence P. feudenreichii viability during spray drying

Osmoprotection protects P. freudenreichii during lethal challenges

P. freundeichii was cultivated in the different growth media and collected during exponential phase.

Cytoplasmic molecules were extracted and the osmprotectants were identified and quantified by C and H NMR

P. freundeichii was cultivated in the different growth medium. Both lethal challenges were applied at the

beginning of the stationary phase. P. freudenreichii quantification was made by CFU numeration before and after challenges

P. freundeichii was cultivated in the different growth media, and spray-dried at the beginning of the stationary phase. P. freudenreichii quantification was made by numeration before and after spray-drying

Osmoprotectants accumulations is determined by the medium composition

1

2

3

5

4

The relation between the trehalose/glycine betaine ratio and P. feudenreichii survival with the experiments 2 and 3

0,01 0,1 1 10

0 20 40 60 80

We demonstrate here that driving the nitrogen/carbon ratio of the culture medium modulates the trehalose/glycine betaine ratio which, in turn, determines P. freudenreichii survival during spray-draying. Spray-drying needs less energy than freeze-

drying. This work thus opens new perspectives for sustainable drying of beneficial bacteria

YEL and YEL+LactoseSweetwheyMilk ultrafiltrate

0,00 0,05 0,10 0,15 0,20 0,25 0,30 0,35

SW SW+ NaCl MU MU + NaCl

Survivalafteroxidative challange(%)

YEL+NaCl

YEL+L+NaCl

SW+NaCl

MU+NaCl

YEL+NaCl YEL+L+NaCl

SW+NaCl MU+NaCl

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