Techno-economic analysis for probiotics preparation production using optimized corn flour medium and spray-drying protective blends
Graphical abstract
Introduction
Probiotics, as defined by Food and Agriculture Organization of the United Nations (FAO) and World Health Organization (WHO), are “live micro-organisms which when administered in adequate amounts confer a health benefit on the host” (FAO/WHO, 2001). New probiotics formulations and functional foods are the major driving factors for continuous growth in the food and feed sector. It has been estimated that the total market value of both human and animal probiotics will gain revenues of 52 (2020) and 4.71 billion dollars (2021), respectively (Zoumpopoulou et al., 2018). Amongst different strains used as the probiotics, those belonging to Lactobacillus and Bifidobacterium genera have gained the greatest importance (Vinderola et al., 2017). While many technologies of the probiotics production are well established on the market, research in this field is further continued. Recent scientific efforts have been focused on characterization of probiotic properties of new strains, and formulation of the final products as solid and dehydrated preparations, which assure stability during long-term storage. Newly obtained Lactobacillus plantarum strains are considered particularly suitable for the production of desiccated probiotics (Bustamante et al., 2017; Kalita et al., 2018).
To meet the demands of the growing food and feed sector, innovative production solutions are required. Highly-efficient and cost-effective methods for the probiotic bacteria preservation are currently intensively sought (Tan et al., 2018). Spray-drying, being the most high-throughput and high-capacity unit operation of converting liquid probiotics into a solid and dehydrated form, is gaining significant attention. High capacity of this process, reaching over 20 tons of powder per hour, is highly desired in the probiotics production (Fu et al., 2018). Nevertheless, in the last few years, freeze-drying was the preferred method of the probiotics preservation, especially for non-thermotolerant strains, due to milder process conditions than in the case of spray-drying. Indeed, lower processing temperature can positively affect the number of living cell counts in the final preparation. However, freeze-drying is more expensive due to high specific energy consumption and much lower capacity of this process, when compared to spray-drying (Huang et al., 2017b). On the other hand, viability of the microbial cells during spray-drying can be increased by a number of different strategies, making this process highly competitive and attractive (Golowczyc et al., 2010). It has been evidenced, that the main reasons for inactivation of the microbial cells during spray-drying are dehydration (causing osmotic stress) and high temperature (Perdana et al., 2013). In contrast to the other groups of microorganisms, lactic acid bacteria (LAB) are not capable to synthesize osmo-protecting compounds that minimize the harmful effects of dehydration. Thus, adding specific protectants, either during growth of LAB or directly prior to the spray-drying process, is recommended to compensate the difference in the osmotic pressure between the cell interior and the environment (Morgan et al., 2006). The composition of the applied protectants, together with the drying conditions (the inlet and especially, the outlet temperature) are the key factors affecting bacterial cells viability during spray-drying. The other factors impacting the cells survival during the drying process are: the phase of bacterial growth, pre-heating adaptation and, above all, characteristics of the strain (Fu and Chen, 2011).
In order to reduce the overall the probiotics production costs, cultivation of the probiotic bacteria can be carried out using renewable sources of carbon and nitrogen, such as wastes from the food industry, animal-free agro-residues, and starchy crops or wastes (Coghetto et al., 2016; Hofvendahl and Hahn-Hägerdal, 2000). Flour-rich waste streams and by-products from bakery, confectionery and wheat milling plants were indicated as sustainable streams of raw materials for microbial media formulations to be used in large scale production (Tsakona et al., 2014). Corn is the third most cultivated cereal, but only 5% of the corn grain, either cooked or processed into flour, are directly consumed by human. The remaining 95% of the corn grain is transformed into fodder or ethanol (Yang et al., 2017). It was suggested that it is crucial to find further ways to improve the applicability of corn (Yang et al., 2017). Corn flour hydrolysate was successfully used as an economical substrate for the production of simple organic compounds, such as ethanol and d-lactic acid (Szymanowska-Powałowska et al., 2014; Zhao et al., 2014). Using starchy crops or wastes is one of the solutions which may contribute to lowering the cost of nutrients, necessary for the probiotics bacteria growth. On the other hand, exploitation of starchy substrates for some of the probiotic strains may be problematic, due to the lack of appropriate enzymatic machinery decomposing starch, which consequently necessitates the addition of hydrolysing enzymes to secure the supply of simple carbohydrates for microbial growth (Mazzoli et al., 2014). On the other hand, some of the LABs, like L. plantarum, are known for their efficient amylolytic activity (Zielińska et al., 2000). Furthermore, limited ability of LAB to synthesize B-vitamins and amino-acids, makes the bacteria able to grow only in complete, rich media. For this reason, starch-based media require supplementation with complex nutrients that increases the overall costs (Hofvendahl and Hahn-Hägerdal, 2000). Nevertheless, in spite of applying these pre-treatments, such starch-based production still remains profitable at industrial scale, when compared to the production based on synthetic media.
Another aspect in the production of probiotic preparations is the possibility of combining them with prebiotics. Prebiotic is defined as a substrate that is selectively utilised by host microorganisms conferring a health benefit (Gibson et al., 2017). Inulin, fructo-oligosaccharides (FOS) and galactooligosaccharides (GOS) are the most commonly used prebiotic agents (Pineiro et al., 2008). The combination of the prebiotic and the probiotic is termed a synbiotic. The synbiotics are known to confer a synergistic health benefit, which results from specific stimulation of the probiotic component by the prebiotic agent (de Vrese and Schrezenmeir, 2008). This mechanism does not rely only on improving the cells survival during transit through gastrointestinal tract, but also on stimulating their growth in the intestine (Pandey et al., 2015).
The ultimate aim of this study was to optimize a process of probiotics production by adopting a multidirectional strategy. Exploitation of an inexpensive substrate for the probiotic strain growth, a reduced number of unit operations, and optimization of the spray-drying procedure, were all implemented in this strategy. Inexpensive, starch-based medium was double used for the strain growth and for the preservation of the probiotics during spray-drying, which allowed to reduce the number of unit operations. The drying procedure was preceded by the addition of an optimized blend of protectants. Experimental data, were further used to conduct simulation of a synbiotic preparation production at industrial scale. The simulated synbiotic was composed of the here developed probiotic preparation and inulin, used as the prebiotic and standardization agent. Techno-economic evaluation of the process model and sensitivity analysis of the key parameters was conducted.
Section snippets
Microorganism
Lactobacillus plantarum LOCK 0860 was used in this study as the model probiotic strain. The strain was obtained from Lodz University of Technology as cryobank beads (Mast Diagnostica, Germany) and stored at −20 °C. Its probiotic properties were previously documented (Śliżewska et al., 2012).
Phenotype MicroArrays tests
Phenotype MicroArrays (PMs) tests were conducted using Biolog technology (Biolog, USA) and PM1-2 plates. The PM1-2 are separate sets of 96 C-source substrates. L. plantarum LOCK 0860 was grown overnight at
Phenotype profiling of L. plantarum LOCK 0860 - C-source utilization profile
Cell-based phenotypic testing technologies, as the here used Phenotype MicroArrays, greatly assist medium optimization efforts and allow to gain deeper understanding of a given microorganism performances in a specific growth medium (Greetham, 2014). Here conducted phenotype profiling, evidenced that L. plantarum LOCK 0860 is able to assimilate the following C-sources: l-arabinose, N-acetyl-d-glucosamine, d-galactose, d-trehalose, d-mannose, d-gluconic acid, d-xylose, d-mannitol, d-ribose, d
Conclusions
The competitive production processes of stable probiotics and synbiotics preparations is currently an urgent need of the food and feed industry. Amongst different strategies of increasing the market attractiveness of a particular process, development of inexpensive, sustainable growth medium, reduction of the number of unit operations, and exploitation of cost-effective preservation method, are the most promising. All these issues were addressed in the present study.
In the current research we
Declaration of interests
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Declaration of Competing Interest
The authors report no declarations of interest.
Acknowledgements
This study was financially supported by the National Centre for Research and Development (NCBR) under the project PBS3/A8/32/2015.
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