Growth conditions have been frequently studied in optimizing polyhydroxybutyrate (PHB) production, while few studies were performed to unravel the dynamic mixed microbial consortia (MMCs) in the process. In this study, the relationship between growth conditions (C/N ratios and pH) and the corresponding key-microbes were identified and monitored during PHB accumulation. The highest PHB level (70 wt% of dry cell mass) was obtained at pH 9, C/N 40, and acetic acid 10 g/L. Linking the dominant genera with the highest point of PHB accumulation, Thauera was the most prevalent species in all MMCs of pH 9, except when a C/N ratio of 1 was applied. Notably, dominant bacteria shifted at pH 7 (C/N 10) from Thauera (0 h) to Paracoccus, and subsequently to Alcaligenes following the process of PHB accumulation and consumption. Further understanding of the relationship between the structure of the microbial community and the performance will be beneficial for regulating and obtaining high PHB accumulation within an MMC. Our study illustrates the impact of C/N ratios and pH on microbial prevalence and PHB production levels using a mixed microbial starter culture. This knowledge will broaden industrial perspectives for regulating high PHB production and timely harvesting.
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This study aimed to evaluate technological (acidification, proteolysis, lipolysis, resistance to low pH, NaCl, and bile salts) and biopreservation (antimicrobial activity against foodborne pathogens) features of 1002 LAB by high throughput screening (HTS) methods. The LAB was isolated from 11 types of Brazilian artisanal cheeses (BAC) marketed in the main 5 producing regions. Remarkable intra-species variability in acidification rates have been found, which was most pronounced between isolates from Mina's artisanal cheeses, Caipira and Coalho cheeses. Lacticaseibacillus paracasei and Levilactobacillus brevis showed the fastest acidification rate; however, all isolates showed slower acidification rates than a lactococcal control strain (4.3 × lower). When testing inhibitory effects, > 75% of LAB isolates could inhibit the growth of Staphylococcus aureus ATCC 19095 and Listeria monocytogenes ATCC 7644. Two of these isolates, identified as Lactiplantibacillus plantarum and Lentilactobacillus buchneri, the sterile and neutral supernatants alone, were sufficient to inhibit L. monocytogenes growth. Principal component analysis (PCA) allowed the identification of functional groups based on proteolytic and lipolytic activity, osmotic stress resistance, and inhibition of L. monocytogenes. The type of cheese the isolates were recovered from influenced properties such as anti-listerial compounds and lipolytic enzyme production. The use of HTS and multivariate statistics allowed insights into a diverse set of LAB technological and biopreservation properties. These findings allow a profound knowledge of the heterogeneity of a large set of isolates, which can be further used to design starter cultures with varied and combined properties, such as biopreservation and technological features. Besides that, HTS makes it possible to analyze a vast panel of LAB strains, reducing costs and time within laboratory analysis, while avoiding the loss of information once all LAB are tested at the same time (differently from the traditional labor-intensive approach, in which a few numbers of strains is tested per time).
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In the production of fermented foods, microbes play an important role. Optimization of fermentation processes or starter culture production traditionally was a trial-and-error approach inspired by expert knowledge of the fermentation process. Current developments in high-throughput 'omics' technologies allow developing more rational approaches to improve fermentation processes both from the food functionality as well as from the food safety perspective. Here, the authors thematically review typical bioinformatics techniques and approaches to improve various aspects of the microbial production of fermented food products and food safety.
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