Influence of fermentation time, cryoprotectant and neutralization of cell concentrate on freeze‐drying survival, storage stability, and acid and bile exposure of Bifidobacterium animalis ssp. lactis cells produced without milk‐based ingredients

Maria Saarela (Corresponding Author), Ilkka Virkajärvi, Hanna-Leena Alakomi, Tiina Mattila-Sandholm, Anu Vaari, T. Suomalainen, Jaana Mättö

Research output: Contribution to journalArticleScientificpeer-review

69 Citations (Scopus)

Abstract

Aims: To investigate the stability of Bifidobacterium animalis ssp. lactis VTT E‐012010 (=Bb‐12) during freeze‐drying, storage and acid and bile exposure. The effect of harvesting time and composition and pH of the cryoprotectant on the survival was evaluated. The procedure was performed by using a milk‐free culture medium and cryoprotectants to produce cells for nonmilk‐based applications.

Methods and Results: Bifidobacterial cells were grown in fermenters in general edible medium for 15 or 22 h. The cell mass was freeze‐dried either as non‐neutralized or neutralized using sucrose, betaine or reconstituted skim milk (control) as cryoprotectants. For stability studies freeze‐dried powders were stored at 37, 5 and −20°C for 2–6 months. In addition, acid and bile tolerance of the powders was tested. Sucrose‐formulated B. animalis ssp. lactis preparations had an excellent stability during storage at refrigerated and frozen temperatures for 5–6 months. They also had a good survival during storage at 37°C for 2 months as well as during exposure to pH 3 and 1% bile acids. No difference was observed between 15 and 22 h grown cells or between non‐neutralized and neutralized cells. Betaine proved to be a poor cryoprotectant compared with sucrose.

Conclusions: Fermentation time and neutralization of cell concentrate before freeze‐drying had no impact on the storage stability and bile and acid tolerance of freeze‐dried bifidobacterial cells. The nonmilk‐based production protocol using sucrose as a cryoprotectant yielded powdery preparations with excellent stability in adverse conditions (storage at elevated temperatures and during acid and bile exposure).

Significance and Impact of the Study: The results indicate that it is feasible to develop nonmilk‐based production technologies for probiotic cultures. This provides new possibilities for the development of nondairy‐based probiotic products.

Original languageEnglish
Pages (from-to)1330 - 1339
Number of pages10
JournalJournal of Applied Microbiology
Volume99
Issue number6
DOIs
Publication statusPublished - 2005
MoE publication typeA1 Journal article-refereed

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Bile Acids and Salts
Fermentation
Sucrose
Betaine
Probiotics
Powders
Temperature
Bifidobacterium animalis
Culture Media
Milk
Technology

Keywords

  • acid tolerance
  • Bb-12
  • Bifidobacterium
  • fermentation
  • freeze-drying

Cite this

@article{065b4b39f3594696895ec879b17a7de2,
title = "Influence of fermentation time, cryoprotectant and neutralization of cell concentrate on freeze‐drying survival, storage stability, and acid and bile exposure of Bifidobacterium animalis ssp. lactis cells produced without milk‐based ingredients",
abstract = "Aims: To investigate the stability of Bifidobacterium animalis ssp. lactis VTT E‐012010 (=Bb‐12) during freeze‐drying, storage and acid and bile exposure. The effect of harvesting time and composition and pH of the cryoprotectant on the survival was evaluated. The procedure was performed by using a milk‐free culture medium and cryoprotectants to produce cells for nonmilk‐based applications. Methods and Results: Bifidobacterial cells were grown in fermenters in general edible medium for 15 or 22 h. The cell mass was freeze‐dried either as non‐neutralized or neutralized using sucrose, betaine or reconstituted skim milk (control) as cryoprotectants. For stability studies freeze‐dried powders were stored at 37, 5 and −20°C for 2–6 months. In addition, acid and bile tolerance of the powders was tested. Sucrose‐formulated B. animalis ssp. lactis preparations had an excellent stability during storage at refrigerated and frozen temperatures for 5–6 months. They also had a good survival during storage at 37°C for 2 months as well as during exposure to pH 3 and 1{\%} bile acids. No difference was observed between 15 and 22 h grown cells or between non‐neutralized and neutralized cells. Betaine proved to be a poor cryoprotectant compared with sucrose. Conclusions: Fermentation time and neutralization of cell concentrate before freeze‐drying had no impact on the storage stability and bile and acid tolerance of freeze‐dried bifidobacterial cells. The nonmilk‐based production protocol using sucrose as a cryoprotectant yielded powdery preparations with excellent stability in adverse conditions (storage at elevated temperatures and during acid and bile exposure). Significance and Impact of the Study: The results indicate that it is feasible to develop nonmilk‐based production technologies for probiotic cultures. This provides new possibilities for the development of nondairy‐based probiotic products.",
keywords = "acid tolerance, Bb-12, Bifidobacterium, fermentation, freeze-drying",
author = "Maria Saarela and Ilkka Virkaj{\"a}rvi and Hanna-Leena Alakomi and Tiina Mattila-Sandholm and Anu Vaari and T. Suomalainen and Jaana M{\"a}tt{\"o}",
year = "2005",
doi = "10.1111/j.1365-2672.2005.02742.x",
language = "English",
volume = "99",
pages = "1330 -- 1339",
journal = "Journal of Applied Microbiology",
issn = "1364-5072",
publisher = "Wiley-Blackwell",
number = "6",

}

Influence of fermentation time, cryoprotectant and neutralization of cell concentrate on freeze‐drying survival, storage stability, and acid and bile exposure of Bifidobacterium animalis ssp. lactis cells produced without milk‐based ingredients. / Saarela, Maria (Corresponding Author); Virkajärvi, Ilkka; Alakomi, Hanna-Leena; Mattila-Sandholm, Tiina; Vaari, Anu; Suomalainen, T.; Mättö, Jaana.

In: Journal of Applied Microbiology, Vol. 99, No. 6, 2005, p. 1330 - 1339.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Influence of fermentation time, cryoprotectant and neutralization of cell concentrate on freeze‐drying survival, storage stability, and acid and bile exposure of Bifidobacterium animalis ssp. lactis cells produced without milk‐based ingredients

AU - Saarela, Maria

AU - Virkajärvi, Ilkka

AU - Alakomi, Hanna-Leena

AU - Mattila-Sandholm, Tiina

AU - Vaari, Anu

AU - Suomalainen, T.

AU - Mättö, Jaana

PY - 2005

Y1 - 2005

N2 - Aims: To investigate the stability of Bifidobacterium animalis ssp. lactis VTT E‐012010 (=Bb‐12) during freeze‐drying, storage and acid and bile exposure. The effect of harvesting time and composition and pH of the cryoprotectant on the survival was evaluated. The procedure was performed by using a milk‐free culture medium and cryoprotectants to produce cells for nonmilk‐based applications. Methods and Results: Bifidobacterial cells were grown in fermenters in general edible medium for 15 or 22 h. The cell mass was freeze‐dried either as non‐neutralized or neutralized using sucrose, betaine or reconstituted skim milk (control) as cryoprotectants. For stability studies freeze‐dried powders were stored at 37, 5 and −20°C for 2–6 months. In addition, acid and bile tolerance of the powders was tested. Sucrose‐formulated B. animalis ssp. lactis preparations had an excellent stability during storage at refrigerated and frozen temperatures for 5–6 months. They also had a good survival during storage at 37°C for 2 months as well as during exposure to pH 3 and 1% bile acids. No difference was observed between 15 and 22 h grown cells or between non‐neutralized and neutralized cells. Betaine proved to be a poor cryoprotectant compared with sucrose. Conclusions: Fermentation time and neutralization of cell concentrate before freeze‐drying had no impact on the storage stability and bile and acid tolerance of freeze‐dried bifidobacterial cells. The nonmilk‐based production protocol using sucrose as a cryoprotectant yielded powdery preparations with excellent stability in adverse conditions (storage at elevated temperatures and during acid and bile exposure). Significance and Impact of the Study: The results indicate that it is feasible to develop nonmilk‐based production technologies for probiotic cultures. This provides new possibilities for the development of nondairy‐based probiotic products.

AB - Aims: To investigate the stability of Bifidobacterium animalis ssp. lactis VTT E‐012010 (=Bb‐12) during freeze‐drying, storage and acid and bile exposure. The effect of harvesting time and composition and pH of the cryoprotectant on the survival was evaluated. The procedure was performed by using a milk‐free culture medium and cryoprotectants to produce cells for nonmilk‐based applications. Methods and Results: Bifidobacterial cells were grown in fermenters in general edible medium for 15 or 22 h. The cell mass was freeze‐dried either as non‐neutralized or neutralized using sucrose, betaine or reconstituted skim milk (control) as cryoprotectants. For stability studies freeze‐dried powders were stored at 37, 5 and −20°C for 2–6 months. In addition, acid and bile tolerance of the powders was tested. Sucrose‐formulated B. animalis ssp. lactis preparations had an excellent stability during storage at refrigerated and frozen temperatures for 5–6 months. They also had a good survival during storage at 37°C for 2 months as well as during exposure to pH 3 and 1% bile acids. No difference was observed between 15 and 22 h grown cells or between non‐neutralized and neutralized cells. Betaine proved to be a poor cryoprotectant compared with sucrose. Conclusions: Fermentation time and neutralization of cell concentrate before freeze‐drying had no impact on the storage stability and bile and acid tolerance of freeze‐dried bifidobacterial cells. The nonmilk‐based production protocol using sucrose as a cryoprotectant yielded powdery preparations with excellent stability in adverse conditions (storage at elevated temperatures and during acid and bile exposure). Significance and Impact of the Study: The results indicate that it is feasible to develop nonmilk‐based production technologies for probiotic cultures. This provides new possibilities for the development of nondairy‐based probiotic products.

KW - acid tolerance

KW - Bb-12

KW - Bifidobacterium

KW - fermentation

KW - freeze-drying

U2 - 10.1111/j.1365-2672.2005.02742.x

DO - 10.1111/j.1365-2672.2005.02742.x

M3 - Article

VL - 99

SP - 1330

EP - 1339

JO - Journal of Applied Microbiology

JF - Journal of Applied Microbiology

SN - 1364-5072

IS - 6

ER -