High-Throughput rRNA Gene Sequencing Reveals Highand Complex Bacterial Diversity Associated withBrazilian Coffee Bean Fermentation.

Coffee bean fermentation is a spontaneous, on-farm process involving the action of different microbial groups, including bacteria and fungi. In this study, high-throughput sequencing approach was employed to study the diversity and dynamics of bacteria associated with Brazilian coffee bean fermentation. The total DNA from fermenting coffee samples was extracted at different time points, and the 16S rRNA gene with segments around the V4 variable region was sequenced by Illumina high-throughput platform. Using this approach, the presence of over eighty bacterial genera was determined, many of which have been detected for the first time during coffee bean fermentation, including Fructobacillus, Pseudonocardia, Pedobacter, Sphingomonas and Hymenobacter. The presence of Fructobacillus suggests an influence of these bacteria on fructose metabolism during coffee fermentation. Temporal analysis showed a strong dominance of lactic acid bacteria with over 97% of read sequences at the end of fermentation, mainly represented by the Leuconostoc and Lactococcus. Metabolism of lactic acid bacteria was associated with the high formation of lactic acid during fermentation, as determined by HPLC analysis. The results reported in this study confirm the underestimation of bacterial diversity associated with coffee fermentation. New microbial groups reported in this study may be explored as functional starter cultures for on-farm coffee processing.

Whole-cell synthesis of 2,5-furandicarboxylic acid from pineapple waste under various fermentation strategies.

2,5-Furandicarboxylic acid (FDCA) is one of the platform chemicals and monomers used in plastic industries, currently synthesized by carcinogenic and toxic chemical processes with high pressure and temperature. The aim of this study was to develop a bioprocess for the production of FDCA. 5-(Hydroxymethyl)furfural (HMF) was synthesized (22.67 ± 1.36 g/l/h) from pineapple peel using chromium(III) chloride (CrCl3) at 100 °C. After optimization, approximately 3 mg/l/h FDCA was produced by Aspergillus flavus APLS-1 from HMF in a 2.5 L fermenter in a batch strategy. Parallel and immobilized packed bad bioreactors showed less production of FDCA. A fed-batch strategy produced 3.5 ± 0.3 mg/l/h of FDCA in shake flasks. Also, approximately 0.55 mg/l/h of FDCA was produced from pineapple waste derived HMF. However, these bioprocesses may be improved to increase the yield of renewable FDCA, in the future. This is the first report on FDCA production from pineapple waste.

Bioprospecting lipid-producing microorganisms: From metagenomic-assisted isolation techniques to industrial application and innovations.

Traditionally, lipid-producing microorganisms have been obtained via conventional bioprospecting based on isolation and screening techniques, demanding time and effort. Thus, high-throughput sequencing combined with conventional microbiological approaches has emerged as an advanced and rapid strategy for recovering novel oleaginous microorganisms from target environments. This review highlights recent developments in lipid-producing microorganism bioprospecting, following (i) from traditional cultivation techniques to state-of-the-art metagenomics approaches; (ii) related topics on workflow, next-generation sequencing platforms, and knowledge bioinformatics; and (iii) biotechnological potential of the production of docosahexaenoic acid (DHA) by Aurantiochytrium limacinum, arachidonic acid (ARA) by Mortierella alpina and biodiesel by Rhodosporidium toruloides. These three species have been shown to be highly promising and studied in research articles, patents and commercialized products. Trends, innovations and future perspectives of these microorganisms are also addressed. Thus, these microbial lipids allow the development of food, feed and biofuels as alternative solutions to animal and vegetable oils.

Integrating metagenetics and high-throughput screening for bioprospecting marine thraustochytrids producers of long-chain polyunsaturated fatty acids.

Omega-3 produced by marine thraustochytrids has appeared as an alternative to fish oil and an eco-friendly solution to overfishing. Herein, an integrative analysis of metagenetics and high-throughput screening was used for bioprospecting marine thraustochytrids from southern Brazil mangrove and coastal seawater. All sampled environments showed biodiversity and abundance of SAR clade. Environmental samples detected with potential lipid-accumulating labyrinthulomycetes were further processed for direct plating and pollen baiting isolation. Microtiter plate system and fluorescence spectroscopy were combined for high-throughput screening of 319 isolates to accumulate lipids. Twenty isolates were selected for submerged cultivation and lipid characterization. Among them, B36 isolate, identified as Aurantiochytrium sp. by 18s rRNA sequencing, achieved the highest biomass (25.60 g/l CDW) and lipids (17.12 g/l CDW). This lipid content had a high biological value with 44.37% LC-PUFAs and 34.6% DHA, which can be used as a sustainable source in vegan, seafood-free and animal feed diets.

Monascus: a Reality on the Production and Application of Microbial Pigments.

Monascus species can produce yellow, orange, and red pigments, depending on the employed cultivation conditions. They are classified as natural pigments and can be applied for coloration of meat, fishes, cheese, beer, and pates, besides their use in inks for printer and dyes for textile, cosmetic, and pharmaceutical industries. These natural pigments also present antimicrobial activity on pathogenic microorganisms and other beneficial effects to the health as antioxidant and anticholesterol activities. Depending on the substrates, the operational conditions (temperature, pH, dissolved oxygen), and fermentation mode (state solid fermentation or submerged fermentation), the production can be directed for one specific color dye. This review has a main objective to present an approach of Monascus pigments as a reality to obtaining and application of natural pigments by microorganisms, as to highlight properties that makes this pigment as promising for worldwide industrial applications.