Maximilian Schmacht

A process development from a traditional grain-based fermentation to a defined water kefir fermentation using a co-culture of one lactic acid bacterium and one yeast was elaborated as a prerequisite for an industrially scalable, controllable, and reproducible process. Further, to meet a healthy lifestyle, a low ethanol-containing product was aimed for. Five microbial strains—Hanseniaspora valbyensis, Dekkera bruxellensis, Saccharomyces cerevisiae, Liquorilactobacillus nagelii, and Leuconostoc mesenteroides—were used in pairs in order to examine their influence on the fermentation progress and the properties of the resulting water kefir products against grains as a control. Thereby, the combination of H. valbyensis and L. mesenteroides provided the best-rated water kefir beverage in terms of taste and low ethanol concentrations at the same time. As a further contribution to harmonization and reduction of complexity, the usage of dried figs in the medium was replaced by fig syrup, which could have been proven as an adequate substitute. However, nutritional limitations were faced afterward, and thus, an appropriate supplementation strategy for yeast extract was established. Finally, comparative trials in 5-L scale applying grains as well as a defined microbial consortium showed both water kefir beverages characterized by a pH of 3.14, and lactic acid and aromatic sensory properties. The product resulting from co-culturing outperformed the grain-based one, as the ethanol level was considerably lower in favor of an increased amount of lactic acid. The possibility of achieving a water kefir product by using only two species shows high potential for further detailed research of microbial interactions and thus functionality of water kefir.

Manufacturers of starter cultures and probiotics aim to provide preparations with the highest possible amount of living cells and assurance of long-term storage stability. Thereby the industrial economy and thus an efficient outcome of the processes is of utmost importance. Earlier research has shown that the sterilization procedure of the microbial culture medium tremendously impacts growth performance of heating product-sensitive Lactobacillus strains. Thus, three different strains, i.e., L. acidophilus NCFM, L. johnsonii La-2801 and L. delbrueckii subsp. delbrueckii La-0704, were investigated for the influence of media heat pretreatment on cell morphology and stability during fermentation and further freeze drying and storage. The data indicate a relationship between the heating time of the culture medium, which is associated with an increase in browning reactions, and the cultural characteristics of the three strains. The resulting characteristic cell sizes of the cultures could be a major reason for the different stability properties during processing and storage that were observed. Besides the obvious relevance of the results for the production of starter cultures and probiotics, the pleomorphic phenomenon described here could also be a subject for other biotechnological processes, where heat-mediated media conversions, and thereby related cellular effects, could be a topic. Future studies have to show if further functional properties are influenced by the cell morphology and which cellular mechanisms lead to the observed pleomorphism.

Lactic acid bacteria (LAB) are widely applied microorganisms in food, feed, and beverage applications, where they can provide essential functionality for product modification, increase product shelf life, or act as beneficial organisms after consumption. Among these, strains of the genus Lactobacillus are often used as starters, probiotics, or biopreservatives. For all these types of bacterial preparations, a transportable shelf-stable form of concentrated bacteria, preserving their intrinsic properties, is essential for commercial distribution. Former studies revealed a relationship between the culture medium, cellular morphology, and the robustness of Lactobacillus acidophilus NCFM (name derived from North Carolina Food Microbiology) cultures. Due to these insights, a multitude of Lactobacillus strains representative of the genus were screened regarding their sensitivity to thermal medium pretreatment possibly accompanied by the alteration of their chemical composition, such as the formation of Maillard reaction products (MRPs). This study reveals a quite diverse and different growth behavior of those strains in the form of altered or non-altered cell concentrations and the size distributions of the populations, whereby five strains of the L. delbrueckii group in particular showed increased cell concentrations combined with decreased mean cell volumes. The results are of both scientific and industrial relevance, as they highlight the necessity to consider and understand the effects of media sterilization for the applied production strain.

Glutathione (GSH) is a non-coding tripeptide thiol with several important regulative and protective functions in eukaryotes and in most prokaryotes. The primary function of GSH is to maintain the redox potential of the cell, which is directly connected to GSH concentration, and to prevent cellular damages caused by reactive oxygen species or toxic heavy metals. Due to its antioxidant character, it is widely used in pharmaceutical, cosmetic and food industry. There have been different strategies to optimize GSH yield and productivity in bacteria and yeasts by means of metabolic and evolutionary engineering, media optimization and bioprocess engineering. The fed-batch procedure with yeasts of the genera Saccharomyces and Candida is still common method for industrial production. However, for an economic bioprocess production of GSH key factors like media costs, strain performance and process scalability are essential. Beside the extraction and purification of GSH as bulk product, GSH-enriched yeast cells are used for food and beverage applications, as well. This review outlines current applications of microbially produced GSH and illustrates current developments and strategies for its production.