Abstract

This study was undertaken to investigate the impact of culture pH (4.5–6.5) and temperature (32–37 °C) on the stress resilience of Lactobacillus reuteri DSM 17938 during freeze-drying and post freeze-drying exposure to low pH (pH 2) and bile salts. Response-surface methodology analysis revealed that freeze-drying survival rates left( {frac{Ncells;after;drying}{Ncells ;before;drying};100} right) were linearly related to pH with the highest survival rate of 80% when cells were cultured at pH 6.5 and the lowest was 40% when cells were cultured at pH 4.5. The analysis further revealed that within the chosen temperature range the culture temperature did not significantly affect the freeze-drying survival rate. However, fermentation at pH 4.5 led to better survival rates when rehydrated cells were exposed to low pH shock or bile salts. Thus, the effect of pH on freeze-drying survival was in contrast to effects on low pH and bile salts stress tolerance. The rationale behind this irreconcilability is based on the responses being dissimilar and are not tuned to each other. Culturing strain DSM 17938 at pH values higher than 5.5 could be a useful option to improve the survivability and increase viable cell numbers in the final freeze-dried product. However, the dissimilar responses for the process- and application parameters tested here suggest that an optimal compromise has to be found in order to obtain the most functional probiotic product possible.

Highlights

  • As stated by Rosenstiel and Stange (2010), probiotics are living microorganisms that—if taken in appropriate dosage—may result in a health benefit for the host

  • The results shown here will be of relevance for future development of stable desiccated formulations of L. reuteri DSM 17938 with high levels of probiotic activity

  • Fermentation behavior at 1 L scale The initial number of cells is an important variable to be considered when bacterial cells are preserved by desiccation methods (Morgan et al 2006; García 2011), and most often the aim is to maximize the yield at the end of the fermentation (Alonso 2016)

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Summary

Introduction

As stated by Rosenstiel and Stange (2010), probiotics are living microorganisms that—if taken in appropriate dosage—may result in a health benefit for the host. In 2015, it has exceeded 35 billion USD and is expected to reach revenues of 74 billion USD in 2024 (Grand View Research Inc. 2016). To help realize this expectation, it is essential to better understand how production processes influence product quality. Cell stress tolerance is an important issue in the development of stable probiotic products based on freezedried formulations of Lactic Acid Bacteria (LAB). Among the different culture variables, temperature, pH, and dissolved oxygen concentration, have previously been shown to be important for the stabilization of freeze-dried bacterial cells of the genera Lactobacillus (Schoug et al 2008; Liu et al 2014; Béal and Fonseca 2015) and Bifidobacterium (Mozzetti et al 2013)

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