Discovery of two novel bioactive algicidal substances from Brevibacillus sp. via metabolomics profiling and back-validation.

Identifying potent bacterial algicidal agents is essential for the development of effective, safe, and economically viable algaecides. Challenges in isolating and purifying these substances from complex secretions have impeded progress in this field. Metabolomics profiling, an efficient strategy for identifying metabolites, was pioneered in identifying bacterial algicidal substances in this study. Extracellular secretions from different generations of the algicidal bacterium Brevibacillus sp. were isolated for comprehensive analysis. Specifically, a higher algicidal efficacy was observed in the secretion from Generation 3 (G3) of Brevibacillus sp. compared to Generation 1 (G1). Subsequent metabolomics profiling comparing G3 and 1 revealed 83 significantly up-regulated metabolites, of which 9 were identified as potential algicidal candidates. Back-validation highlighted the potency of 4-acetamidobutanoic acid (4-ABC) and 8-hydroxyquinoline (8-HQL), which exhibited robust algicidal activity with 3d-EC50 values of 6.40 mg/L and 92.90 µg/L, respectively. These substances disrupted photosynthetic activity in M. aeruginosa by ceasing electron transfer in PSⅡ, like the impact exerted by Brevibacillus sp. secretion. These findings confirmed that 4-ABC and 8-HQL were the main algicidal components derived from Brevibacillus sp.. Thus, this study presents a streamlined strategy for identifying bacterial algicidal substances and unveils two novel and highly active algicidal substances. ENVIRONMENTAL IMPLICATION: Harmful cyanobacterial blooms (HCBs) pose significant environmental problems and health effects to humans and other organisms. The increasing frequency of HCBs has emerged as a pressing global concern. Bacterial-derived algicidal substances are expected to serve as effective, safe, and economically viable algaecides against HCBs. This study presents a streamlined strategy for identifying bacterial algicidal substances and unveils two novel substances (4-ABC and 8-HQL). These two substances demonstrate remarkable algicidal activity and disrupt the photosynthetic system in M. aeruginosa. They hold potential as prospective algaecides for addressing HCBs.

Fruit peel crude enzymes for enhancement of biohydrogen production from synthetic swine wastewater by improving biohydrogen-formation processes of dark fermentation.

Biohydrogen is a promising clean fuel but with a low yield. This study aims to enhance biohydrogen production from synthetic swine wastewater by employing crude enzymes obtained from different fruit peels (orange, mandarin, and banana) to improve the biohydrogen-formation processes of dark fermentation. Results indicated that dosing with crude enzymes affected volatile fatty acids (VFAs) and biogas composition insignificantly, while increased biohydrogen yield from 1.62 ± 0.00 (blank) to 1.90 ± 0.08 (orange peel), 2.01 ± 0.00 (mandarin peel), and 1.96 ± 0.01 (banana peel) mol H2/mol glucose, respectively. Banana peel crude enzyme was the most effective additive, with 1 g/L protein improving 97.41 ± 3.72 % of biohydrogen yield. The crude enzymes wielded less influence on acetic acid and butyric acid pathways but enhanced other biohydrogen production pathways. These observations demonstrated that fruit peel-based crude enzymes as additives are advantageous to improving biohydrogen yield towards higher biohydrogen production.

Wastewater-derived biohydrogen: Critical analysis of related enzymatic processes at the research and large scales.

Organic-rich wastewater is a feasible feedstock for biohydrogen production. Numerous review on the performance of microorganisms and the diversity of their communities during a biohydrogen process were published. However, there is still no in-depth overview of enzymes for biohydrogen production from wastewater and their scale-up applications. This review aims at providing an insightful exploration of critical discussion in terms of: (i) the roles and applications of enzymes in wastewater-based biohydrogen fermentation; (ii) systematical introduction to the enzymatic processes of photo fermentation and dark fermentation; (iii) parameters that affect enzymatic performances and measures for enzyme activity/ability enhancement; (iv) biohydrogen production bioreactors; as well as (v) enzymatic biohydrogen production systems and their larger scales application. Furthermore, to assess the best applications of enzymes in biohydrogen production from wastewater, existing problems and feasible future studies on the development of low-cost enzyme production methods and immobilized enzymes, the construction of multiple enzyme cooperation systems, the study of biohydrogen production mechanisms, more effective bioreactor exploration, larger scales enzymatic biohydrogen production, and the enhancement of enzyme activity or ability are also addressed.

Volatile fatty acids production from waste streams by anaerobic digestion: A critical review of the roles and application of enzymes.

Volatile fatty acids (VFAs) produced from organic-rich wastewater by anaerobic digestion attract attention due to the increasing volatile fatty acids market, sustainability and environmentally friendly characteristics. This review aims to give an overview of the roles and applications of enzymes, a biocatalyst which plays a significant role in anaerobic digestion, to enhance volatile fatty acids production. This paper systematically overviewed: (i) the enzymatic pathways of VFAs formation, competition, and consumption; (ii) the applications of enzymes in VFAs production; and (iii) feasible measures to boost the enzymatic processes. Furthermore, this review presents a critical evaluation on the major obstacles and feasible future research directions for the better applications of enzymatic processes to promote VFAs production from wastewater.

Treatment of Synthetic Ammonium Sulfate Wastewater by Mixed Culture of Chlorella pyrenoidosa and Enriched Nitrobacteria.

Ammonium sulfate wastewater can cause eutrophication and black odor of water body. Although ammonia nitrogen can be used as nutrient of microalgae, high ammonia nitrogen levels could inhibit the growth of microalgae. Nitrobacteria can transform ammonia nitrogen into nitrate nitrogen. In this study, mono Chlorella pyrenoidosa culture (mono-C.py), synchronous mixed culture (mixed-a), and asynchronous mixed culture (mixed-b) systems were examined for their ability to treat ammonium sulfate wastewater. Nitrogen removal rate of mixed-b at the end of culture (52.96%) was higher than that of the mono-C.py (46.37%) and the mixed-a (39.11%). Higher total suspended solid concentration (2.40 g/L), crude protein yield (0.76 g/L), and heating value yield (35.73 kJ/L) were obtained in mixed-b, meanwhile with excellent settlement performance (91.43 ± 0.51%). Mechanism analysis of settlement showed that the relative abundance of floc-forming-related bacteria Sphingopyxis and Acidovorax were increased generally, while nitrification/denitrifying members were decreased in mixed-b along with the culture proceeding.

Roles and applications of enzymes for resistant pollutants removal in wastewater treatment.

Resistant pollutants like oil, grease, pharmaceuticals, pesticides, and plastics in wastewater are difficult to be degraded by traditional activated sludge methods. These pollutants are prevalent, posing a great threat to aquatic environments and organisms since they are toxic, resistant to natural biodegradation, and create other serious problems. As a high-efficiency biocatalyst, enzymes are proposed for the treatment of these resistant pollutants. This review focused on the roles and applications of enzymes in wastewater treatment. It discusses the influence of enzyme types and their sources, enzymatic processes in resistant pollutants remediation, identification and ecotoxicity assay of enzymatic transformation products, and typically employed enzymatic wastewater treatment systems. Perspectives on the major challenges and feasible future research directions of enzyme-based wastewater treatment are also proposed.

The joint effect of ammonium and pH on the growth of Chlorella vulgaris and ammonium removal in artificial liquid digestate.

Although ammonium containing digestate is an ideal alternative medium for microalgae cultivation, high ammonium or unfavorable pH may inhibit microalgal growth. In this study, the joint effect of ammonium and pH on the growth of C. vulgaris and nutrient removal in artificial digestate was investigated. Our results show that ammonium and pH both affected algal growth, but free ammonia (FA) was the main actual inhibitory factor. Algal specific growth rate presented a negative correlation with FA and their relationship was well fitted by a linear regression model. Microalgal growth was little affected below 36.8 mg L-1 FA, while the obvious inhibition occurred at 184 mg L-1 FA (EC50), indicating a high tolerance to FA. Ammonium removal was well described by a first-order kinetics model. FA stress stimulated the production of extracellular organic matters (EOMs), which was good for microalgae adaptation but adverse to pollutant removal.

Synthesis and biological evaluation of novel pyrazoline derivatives containing indole skeleton as anti-cancer agents targeting topoisomerase II.

In order to develop potent anticaner agents, a novel series of 3-(1H-indol-3-yl)-2,3,3a,4-tetrahydrothiochromeno[4,3-c]pyrazole derivatives were synthesized. Structures of all compounds were confirmed. MTT assay has been employed to study antiproliferative activity of these compounds with four human cancer cell lines (MGC-803, Hela, MCF-7 and Bel-7404) and a normal cell line L929. Most of these compounds showed potential anticancer activity and low cytotoxicity on normal cell in vitro. 7d and 7f showed the best anticancer activity, whose IC50 value is 15.43 µM and 20.54 µM towards MGC-803, respectively. Most of them exhibited topoisomerase II selective inhibitory. Cleavage reaction assay and DNA unwinding assay showed that 7f was a nonintercalative Topo II catalytic inhibitor, which was consistent with the docking results. Laser scanning confocal microscopy system tracks the location of representative compounds 7d and 7f which can be abundantly entering the nucleus. In particular, the most potent compounds 7d and 7f were shown to be able to induce G2/M cell cycle arrest and apoptosis in MGC-803 cells.

Performance of a microalgal-bacterial consortium system for the treatment of dairy-derived liquid digestate and biomass production.

To enhance the treatment performance of dairy-derived liquid digestate (DLD) using microalgal-bacterial consortium system composed of Chlorella vulgaris and indigenous bacteria (CV), activated sludge was introduced to form a new microalgal-bacterial consortium system (Co-culture). The activated sludge shortened the lag phase and increased the specific growth rate of C. vulgaris (0.56 d-1). The biomass yield in the Co-culture was 2.72 g L-1, which was lower than that in the CV (3.24 g L-1), but the Co-culture had an improved COD (chemical oxygen demand) removal (25.26%) compared to the CV (13.59%). Quantitative PCR and metagenomic analyses demonstrated that microalgae also promoted bacterial growth, but influenced differently on the bacterial communities of indigenous bacteria and activated sludge. Compared with indigenous bacteria, activated sludge was more prone to forming a favorable symbiosis with C. vulgaris. These findings contribute to the construction of efficient microalgal-bacterial consortium system in wastewater treatment.

Cultivation of Chlorella vulgaris on unsterilized dairy-derived liquid digestate for simultaneous biofuels feedstock production and pollutant removal.

In order to assess viability of microalgae cultivation using unsterilized dairy-derived liquid digestate (DLD) for simultaneous biofuels feedstock production and contaminant removal, four DLD concentrations (25%, 50%, 75% and 100%) were used to grow Chlorella vulgaris in batch photobioreactors (PBRs). The 25% DLD was an ideal alternative medium in that high growth rate (0.69 d-1), high lipid productivity (112.9 mg L-1 d-1) as well as high nutrient removal were attained. The high DLD concentration caused inhibition of microalgal growth, where COD was more inhibitive than ammonium. The presence of bacteria did not influence microalgae production because of limited growth. Microalgal growth reduced the richness and diversity of bacterial community. Furthermore, the species of Bacteroidetes, Candidatus Saccharibacteria, and Chlamydiae rather than Proteobacteria benefited microalgal-bacterial symbiosis. These findings contribute to better application of microalgal-bacterial system for large-scale microalgae cultivation as well as environmental sustainability.

Synthesis of Novel Pyrazole Derivatives as Promising DNA-Binding Agents and Evaluation of Antitumor and Antitopoisomerases I/II Activities.

Molecules bearing pyrazole nucleus present diverse biological properties such as antitumor and anti-inflammatory activities that can be associated with DNA interactions. This study aimed to the synthesis of new pyrazol derivatives and evaluated their ability to interact with the DNA and antitumor and topoisomerase inhibition activities. All derivatives were successfully synthesized, and their structures were elucidated by 1H-NMR and high resolution (HR)-MS (electrospray ionization positive mode (ESI+)). Antiproliferative inhibition assays, UV titration assays, fluorescence titration assays, circular dichroism (CD) assays, KI quenching studies, topoisomerase inhibitory activity assays and molecular docking were evaluated for these compounds. Especially, compounds 5e and 5q showed higher antitumor activity with IC50 values <13 µM for the tested cell lines. However, compounds 5e and 5q did not inhibit the topoisomerase activity evaluated by relaxation assay. These results show that the pyrazole nucleus contributes to the incorporation of molecules into the DNA. Moreover, it was highlighted that positive charges are relevant for the design of promising antitumor and DNA binding compounds.