Effects of Inhibitory Compounds Present in Lignocellulosic Biomass Hydrolysates on the Growth of Bacillus subtilis

This study evaluated the individual and combined effects of inhibitory compounds formed during pretreatment of lignocellulosic biomass on the growth of Bacillus subtilis. Ten inhibitory compounds commonly present in lignocellulosic hydrolysates were evaluated, which included sugar degradation products (furfural and 5-hydroxymethylfurfural), acetic acid, and seven phenolic compounds derived from lignin (benzoic acid, vanillin, vanillic acid, ferulic acid, p-coumaric acid, 4-hydroxybenzoic acid, and syringaldehyde). For the individual inhibitors, syringaldehyde showed the most toxic effect, completely inhibiting the strain growth at 0.1 g/L. In the sequence, assays using mixtures of the inhibitory compounds at a concentration of 12.5% of their IC50 value were performed to evaluate the combined effect of the inhibitors on the strain growth. These experiments were planned according to a Plackett–Burman experimental design. Statistical analysis of the results revealed that in a mixture, benzoic acid and furfural were the most potent inhibitors affecting the growth of B. subtilis. These results contribute to a better understanding of the individual and combined effects of inhibitory compounds present in biomass hydrolysates on the microbial performance of B. subtilis. Such knowledge is important to advance the development of sustainable biomanufacturing processes using this strain cultivated in complex media produced from lignocellulosic biomass, supporting the development of efficient bio-based processes using B. subtilis.

Sugar Recovery from Bakery Leftovers through Enzymatic Hydrolysis: Effect of Process Conditions and Product Characterization

This study evaluates the process conditions, (enzyme concentration (120-1200 U/g substrate), temperature (30-60 °C), and pH (3-9)) of enzymatic hydrolysis (EH) for sugar recovery from leftover croissants (LC) and leftover doughnut (LD), and characterizing its residue and hydrolysate. The highest sugar yield recovered from LC was 574.21 ± 0.74 mg/g (840 U/g substrate, 49 °C and pH 3) and for LD was 460.53 ± 0.74 mg/g (1176 U/g substrate, 47 °C and pH 3). The highest fructose and glucose yield for LC and LD were 14.47±0.73 mg/g and 11.84±0.21 mg/g, and 13.26±0.63 mg/g and 10.34±0.11 mg/g, respectively. Morphology analysis (SEM) showed that the structure of LC and LD had changes in its starch granules that indicates hydrolysis process occurrence. The presence of monosaccharides and oligosaccharides were detected from FTIR. HMF was also detected from sugar degradation due to EH, (0.043 ± 0.0334 mg/g for LC) and (0.023 ± 0.0124 mg/g for LD).

Genome-centric metagenomics reveals insights into the evolution and metabolism of a new free-living group in Rhizobiales

Background The Rhizobiales (Proteobacteria) order is an abundant and diverse group of microorganisms, being extensively studied for its lifestyle based on the association with plants, animals, and humans. New studies have demonstrated that the last common ancestor (LCA) of Rhizobiales had a free-living lifestyle, but the phylogenetic and metabolism characterization of basal lineages remains unclear. Here, we used a high-resolution phylogenomic approach to test the monophyly of the Aestuariivirgaceae family, a new taxonomic group of Rhizobiales. Furthermore, a deep metabolic investigation provided an overview of the main functional traits that can be associated with its lifestyle. We hypothesized that the presence of pathways (e.g., Glycolysis/Gluconeogenesis) and the absence of pathogenic genes would be associated with a free-living lifestyle in Aestuariivirgaceae. Results Using high-resolution phylogenomics approaches, our results revealed a clear separation of Aestuariivirgaceae into a distinct clade of other Rhizobiales family, suggesting a basal split early group and corroborate the monophyly of this group. A deep functional annotation indicated a metabolic versatility, which includes putative genes related to sugar degradation and aerobic respiration. Furthermore, many of these traits could reflect a basal metabolism and adaptations of Rhizobiales, as such the presence of Glycolysis/Gluconeogenesis pathway and the absence of pathogenicity genes, suggesting a free-living lifestyle in the Aestuariivirgaceae members. Conclusions Aestuariivirgaceae (Rhizobiales) family is a monophyletic taxon of the Rhizobiales with a free-living lifestyle and a versatile metabolism that allows these microorganisms to survive in the most diverse microbiomes, demonstrating their adaptability to living in systems with different conditions, such as extremely cold environments to tropical rivers.

Physiological Changes of Bamboo (Fargesia yunnanensis) Shoots During Storage and the Related Cold Storage Mechanisms

The mechanisms for physiological senescence in bamboo shoots after harvest remain unclear. This study investigated physiological changes in Fargesia yunnanensis shoots during storage at different temperatures. The relationship between morphological and physiological changes in bamboo shoots during storage was also analyzed. The results show that cold storage can reduce weight loss, browning, respiration rates, and sugar degradation in bamboo shoots; decrease related enzymatic activities; and inhibit the increase in lignin and cellulose content. The quality of bamboo shoots declines more during the first 3d after harvesting than it does during subsequent periods. The increase in the degree of lignification and fibrosis is the main reason for senescence and for the decline in quality of bamboo shoots after harvest. The bamboo shoots under RT conditions began browning during the third 3d of storage, with a browning rate of 688gkg−1 even in the upper parts; the increase in shoot browning degrees significantly decreased the quality. Low temperatures had better inhibitory effects on browning than they did on lignification and fibrosis. Nonstructural carbohydrates in bamboo shoots are degraded and flow into sheath and shoot respiration, phenols, and shoot fibrosis and lignification at room temperature, but only flow into sheath respiration, shoot fibrosis, and lignification at cold temperature. Soluble protein and free amino acids are primarily distributed into shoot and sheath respiration and into phenols at room temperature, but that process is well inhibited at cold temperature. Bamboo shoots, once removed from cold storage, should be consumed rapidly because enzyme activity recovers quickly. This research provides new theoretical information on the preservation of bamboo shoots.

Thermococcus bergensis sp. nov., a Novel Hyperthermophilic Starch-Degrading Archaeon

A novel hyperthermophilic archaeon, termed strain T7324T, was isolated from a mixed sulfate-reducing consortium recovered from hot water produced from a deep North Sea oil reservoir. The isolate is a strict anaerobic chemo-organotroph able to utilize yeast extract or starch as a carbon source. The genes for a number of sugar degradation enzymes and glutamate dehydrogenase previously attributed to the sulfate reducing strain of the consortium (Archaeoglobus fulgidus strain 7324) were identified in the nearly completed genome sequence. Sequence analysis of the 16S rRNA gene placed the strain in the Thermococcus genus, but with an average nucleotide identity that is less than 90% to its closest relatives. Phylogenomic treeing reconstructions placed the strain on a distinct lineage clearly separated from other Thermococcus spp. The results indicate that the strain T7324T represents a novel species, for which the name Thermococcus bergensis sp. nov. is proposed. The type strain is T7324T (=DSM 27149T = KCTC 15808T).

Natural Medicines for the Treatment of Epilepsy: Bioactive Components, Pharmacology and Mechanism

Epilepsy is a chronic disease that can cause temporary brain dysfunction as a result of sudden abnormal discharge of the brain neurons. The seizure mechanism of epilepsy is closely related to the neurotransmitter imbalance, synaptic recombination, and glial cell proliferation. In addition, epileptic seizures can lead to mitochondrial damage, oxidative stress, and the disorder of sugar degradation. Although the mechanism of epilepsy research has reached up to the genetic level, the presently available treatment and recovery records of epilepsy does not seem promising. Recently, natural medicines have attracted more researches owing to their low toxicity and side-effects as well as the excellent efficacy, especially in chronic diseases. In this study, the antiepileptic mechanism of the bioactive components of natural drugs was reviewed so as to provide a reference for the development of potential antiepileptic drugs. Based on the different treatment mechanisms of natural drugs considered in this review, it is possible to select drugs clinically. Improving the accuracy of medication and the cure rate is expected to compensate for the shortage of the conventional epilepsy treatment drugs.

Development of a Cofactor Balanced, Multi Enzymatic Cascade Reaction for the Simultaneous Production of L-Alanine and L-Serine from 2-Keto-3-deoxy-gluconate

Enzymatic reaction cascades represent a powerful tool to convert biogenic resources into valuable chemicals for fuel and commodity markets. Sugars and their breakdown products constitute a significant group of possible substrates for such biocatalytic conversion strategies to value-added products. However, one major drawback of sugar cascades is the need for cofactor recycling without using additional enzymes and/or creating unwanted by-products. Here, we describe a novel, multi-enzymatic reaction cascade for the one-pot simultaneous synthesis of L-alanine and L-serine, using the sugar degradation product 2-keto-3-deoxygluconate and ammonium as precursors. To pursue this aim, we used four different, thermostable enzymes, while the necessary cofactor NADH is recycled entirely self-sufficiently. Buffer and pH optimisation in combination with an enzyme titration study yielded an optimised production of 21.3 +/− 1.0 mM L-alanine and 8.9 +/− 0.4 mM L-serine in one pot after 21 h.

Artificial consortium demonstrates emergent properties of enhanced cellulosic-sugar degradation and biofuel synthesis

Planktonic cultures, of a rationally designed consortium, demonstrated emergent properties that exceeded the sums of monoculture properties, including a >200% increase in cellobiose catabolism, a >100% increase in glycerol catabolism, a >800% increase in ethanol production, and a >120% increase in biomass productivity. The consortium was designed to have a primary and secondary-resource specialist that used crossfeeding with a positive feedback mechanism, division of labor, and nutrient and energy transfer via necromass catabolism. The primary resource specialist was Clostridium phytofermentans (a.k.a. Lachnoclostridium phytofermentans), a cellulolytic, obligate anaerobe. The secondary-resource specialist was Escherichia coli, a versatile, facultative anaerobe, which can ferment glycerol and byproducts of cellobiose catabolism. The consortium also demonstrated emergent properties of enhanced biomass accumulation when grown as biofilms, which created high cell density communities with gradients of species along the vertical axis. Consortium biofilms were robust to oxic perturbations with E. coli consuming O2, creating an anoxic environment for C. phytofermentans. Anoxic/oxic cycling further enhanced biomass productivity of the biofilm consortium, increasing biomass accumulation ~250% over the sum of the monoculture biofilms. Consortium emergent properties were credited to several synergistic mechanisms. E. coli consumed inhibitory byproducts from cellobiose catabolism, driving higher C. phytofermentans growth and higher cellulolytic enzyme production, which in turn provided more substrate for E. coli. E. coli necromass enhanced C. phytofermentans growth while C. phytofermentans necromass aided E. coli growth via the release of peptides and amino acids, respectively. In aggregate, temporal cycling of necromass constituents increased flux of cellulose-derived resources through the consortium. The study establishes a consortia-based, bioprocessing strategy built on naturally occurring interactions for improved conversion of cellulose-derived sugars into bioproducts.