Optimum pH Research Articles

Preparation and performance of MOF-808 (Zr-MOF) for the efficient adsorption of phenoxyacetic acid pesticides

The organic herbicides of chlorophenoxyacetic acids, including 2,4-dichlorophenoxyacetic acid (2,4-D) and 2-methyl-4-chlorophenoxyacetic acid (MCPA), have posed serious threat to aquatic ecosystems and human health due to extensive use, high solubility and degradation-resistant. In this study, the efficient adsorbent (MOF-808) was synthesized through solvothermal synthesis method and further evaluated for the adsorption performances of 2,4-D and MCPA. Results showed that MOF-808 exhibited the Type I nitrogen adsorption–desorption isotherm with a specific surface area of 2,155 m2·g−1, pore volume of 0.59 cm3·g−1 and pore size of 1.41 nm. It was suggested that the pseudo-second-order and Langmuir model were much better to illustrate the adsorption performances of MOF-808 for both 2,4-D and MCPA, and the maximum adsorption capacities of 793 and 775 mg·g−1 were achieved, respectively; besides, the adsorption processes were spontaneous and primarily driven by chemical adsorption with contributions from physical-chemical adsorption. Additionally, the adsorption capacities for 2,4-D and MCPA were decreased with the increasing of pH and cations concentrations; the optimal pH was 3.0 for 2,4-D and 4.0 for MCPA, and the effects of four cations on pesticides absorption followed the order of Ca2+ > Mg2+ > Na+ > K+. Reuse experiments demonstrated that MOF-808 could retain a high removal rate of 95.9 % (2,4-D) and 95.8 % (MCPA) at the fifth cycle. The spectral analysis revealed that the adsorption mechanisms of 2,4-D and MCPA by MOF-808 involved electrostatic interactions, π-π interactions, hydrogen bonding, and complexation. In conclusion, the well-defined physicochemical properties and outstanding adsorption performance of the synthesized MOF-808 may provide a potential application for the removal of chlorophenoxyacetic acid herbicides from real wastewater.

Study on the selective separation mechanism of tannin acid in chalcopyrite and talc: Insights from adsorption morphology and electrochemical analysis

A key strategy for achieving the selective separation of talc and chalcopyrite is the development of chemical depressants with high solubility and strong selectivity. Tannic acid (TA), a polyphenol found in green plants and has good solubility, contains abundant phenolic hydroxyl groups. Moreover, the adsorption mechanism of TA with chalcopyrite and talc was examined after TA was chosen as the separation depressant for chalcopyrite and talc. Single-mineral flotation test results found that at optimal pH and TA and sodium butyl xanthate (SBX) dosages, the flotation recoveries of talc and chalcopyrite were 9% and 92.17%, respectively, and the difference in recovery between them was 83.17%. Results from real ores and artificially mixed minerals tests also confirmed that TA can achieve selective separation of chalcopyrite and talc. Since chalcopyrite has distinct electrochemical properties, the results of atomic force microscope and electrochemical tests demonstrated that there are significant differences in the adsorption behavior of TA on the surface of chalcopyrite and talc. And the adsorption selectivity of SBX on the chalcopyrite surface is better than that of TA on the chalcopyrite surface, which is an important reason for the selective separation of chalcopyrite and talc.

L-arabinose isomerase from Lactobacillus fermentum C6: Enzymatic characteristics and its recombinant Bacillus subtilis whole cells achieving a significantly increased production of D-tagatose

L-arabinose isomerase (L-AI) is a functional enzyme for the isomerizing of D-galactose to produce D-tagatose. In this study, L-AI-C6-encoding gene from the probiotic Lactobacillus fermentum C6 was cloned and expressed in Bacillus subtilis WB600 for investigating enzymatic characteristics and bioconverting D-tagatose by means of whole-cell catalysis. Results showed that the engineered B. subtilis WB600-pMA5-LAI achieved a maximum specific activity of L-AI-C6 (232.65 ± 15.54 U/mg protein) under cultivation in LB medium at 28 °C for 40 h. The recombinant L-AI-C6 was purified, and enzymatic characteristics test showed its optimum reaction temperature and pH at 60 °C and 8.0, respectively. In addition, L-AI-C6 exhibited good stability within the pH range of 5.5–9.0. By using B. subtilis WB600-pMA5-LAI cells as whole-cell catalyst, the highest D-tagatose yield reached 42.91 ± 0.28 % with D-galactose as substrate, which was 2.41 times that of L. fermentum C6 (17.79 ± 0.11 %). This suggested that the cloning and heterologous expression of L-AI-C6 was an effective strategy for improving D-tagatose conversion by whole-cell catalysis. In brief, the present study demonstrated that the reaction temperature, pH, and stability of L-AI-C6 from L. fermentum C6 meet the demands of industrial application, and the constructed B. subtilis WB600-pMA5-LAI shows promising potential for the whole-cell biotransformation of D-tagatose.

Exploring of new Poly(Thiourea-Amide) as a prospective bismarck Brown Y dye adsorbent: Synthesis via ultrasound irradiation, isotherms, kinetic, thermodynamic, and DFT studies

In this work, a facial ultrasound technique was employed to synthesize poly (N1-((6-aminopyridin-2-yl)carbamothioyl)-N3-carbamothioylisophthalamide) (PTUA) as a new polymer from a condensation reaction of isophthaloyl diisothiocyanate and 2, 6-diaminopyridine. Different techniques were utilized to identify the chemical structure of the synthesized polymer including FTIR spectra, 1HNMR spectra, thermal gravimetric analysis (TGA), Brunauer-Emmett-Teller (BET), and field emission scanning electron microscopy (FESEM). The capability of the synthesized polymer to adsorb Bismarck brown Y (BBY) dye from an aqueous solution was analytically evaluated. Several effective parameters on the adsorption process including contact time, pH, dye concentration, adsorbent dose, and temperature were extensively investigated. The adsorption process was found to be completed with a contact time of 45 min. The results indicated that the optimal adsorption pH was 8 with removal efficiency of 85.331 % and the adsorbed amount of BBY dye increased when the temperature was increased. The adsorption isotherms analysis revealed that the Langmuir model with a correlation coefficient (R2 = 0.9982) was the most relevant to describe the adsorption process compared with the Freundlich model (R2 = 0.9863). By applying the Langmuir isotherm, the maximum adsorption capacity was obtained with a value of 132.39 mg/g. The study of adsorption kinetics showed that the adsorption model of BBY dye onto the polymer surface significantly correlated with a pseudo-second-order model. Based on the thermodynamic studies, the calculated ΔG was negative, ΔH was endothermic, and ΔS was positive, verifying the adsorption process is spontaneous. The desorption percentage of BBY dye was calculated with a value of 81.574 % utilizing methanol as an optimal solvent. The density functional theory (DFT) approach was applied to acquire good understanding for the adsorption process.

The individual and combined effect of DHA and high-fat diet on flesh quality, antioxidant capacity and myofiber characteristics of grass carp (Ctenopharyngodon idellus)

The impact of a high-fat diet (HFD) on the meat quality of farmed fish is a subject of considerable interest among researchers. While docosahexaenoic acid (DHA) is recognized for enhancing growth performance and muscle mass in fish, its potential to mitigate the adverse effects of HFD on meat quality remains uncertain. This study examined the nutritional value, textural properties, and muscle characteristics of grass carp (Ctenopharyngodon idellus) fed varying levels of DHA (0.0%, 0.5%, and 1.0%) under either a regular fat diet (5.0% lipid) or an HFD (10.0% lipid) for a period of eight weeks. Results indicated that the addition of DHA to both the normal fat diet and HFD significantly improved the nutritional and sensory qualities of fish fillets, increasing crude protein, polyunsaturated fatty acid (PUFA), and collagen content while preserving optimal muscle hardness and pH levels. The HFD group exhibited diminished antioxidant capacity, reduced sarcomere length, and mitochondrial structural damage, all effectively ameliorated through DHA supplementation. Moreover, dietary DHA supplementation at both lipid levels upregulated the expression of genes associated with myogenesis and protein synthesis. A two-way ANOVA revealed significant interactions between dietary DHA and lipids, influencing muscle pH, fatty acid composition, myofiber characteristics, antioxidant capacity, and gene expression related to muscle growth and protein synthesis. In conclusion, DHA supplementation can effectively mitigate the deterioration of fish flesh quality induced by HFD.

Direct evolution of an alkaline fungal laccase to degrade tetracyclines

A fungal laccase-mediator system capable of high effectively oxidizing tetracyclines under a wide pH range will benefit environmental protection. This study reported a directed evolution of a laccase PIE5 to improve its performance on tetracyclines oxidization at alkaline conditions. Two mutants, namely MutA (D229N/A244V) and MutB (N123A/D229N/A244V) were obtained. Although they shared a similar optimum pH and temperature as PIE5, the two mutants displayed approximately 2- and 5-fold higher specific activity toward the mediators ABTS and guaiacol at pHs 4.0 to 6.5, respectively. Simultaneously, their catalytic efficiency increased by 8.0- and 6.4-fold compared to PIE5. At a pH range of 5–8 and 28 °C, MutA or MutB at a final concentration of 2.5 U·mL−1 degraded 77 % and 81 % of 100 mg·L−1 tetracycline within 10 min, higher than PIE5 (45 %). Furthermore, 0.1 U·mL−1 MutA or MutB completely degraded 100 mg·L−1 chlortetracycline within 6 min in the presence of 0.1 mM ABTS. At pH 8.0, MutA degraded tetracycline and chlortetracycline following the routine pathways were reported previously based on LC-MS analysis.

Insights into the environmental benefits of using apple pomace for biosorption of lead from contaminated water

The apple processing industry generates large quantities of organic waste, presenting a major source of organic contamination. Consequently, finding an effective solution for valorizing this waste has become a pressing issue. This study aims to address two key concerns: (i) solving an agricultural problem by efficiently using agri-food residue, and (ii) removing lead, an extremely toxic element, from contaminated waters to mitigate environmental pollution. Two biosorbents were tested: raw apple waste (RA), obtained from a mixture of apple varieties, and the same material after extracting valuable bioactive and reusable components, extracted apple (EA). The study evaluated the influence of pH, initial biosorbent mass, adsorption kinetics, and equilibrium isotherms. The results are very promising, showing a lead removal efficiency of 82% for RA and 100% for EA at a low initial concentration of the solution of 20 mg Pb2⁺/L and an optimal pH of 5 ± 0.5. The Langmuir model predicted a maximum adsorption capacity of 44.6 mg/g for RA and 48.6 mg/g for EA. These findings demonstrate that apple waste, even after selective extraction of valuable bioactive components, can be effectively used for environmental remediation on a practical scale.

Advanced electrochemical strategies for simultaneous removal of Microcystis aeruginosa and microcystin-LR: On-demand optimization and mechanistic insights

The frequent occurrence of harmful cyanobacterial blooms and toxins poses significant threats to aquatic life and drinking water safety. This work reports the removal mechanisms of M. aeruginosa and microcystin-LR (MC-LR) using the Pt-Graphite felt (GF) electrochemical system, which builds upon prior research to investigate diverse processes driven by distinct mechanisms within a single reactor, primarily by pH regulation. Factors affecting removal were assessed through single-factor trials and Box-Behnken design, identifying optimal conditions for successful algae removal, finally a 96% removal efficiency of algae cells and an 83% elimination efficiency of MC-LR within a 60-minute timeframe. Particularly at non-optimal pH 7 conditions, the Pt-GF system showcases synergistic effects, enhancing electro-flocculation and weakening electro-Fenton reactions. Conversely, at an optimal pH of 3, the electro-Fenton process is favorably enhanced, leading to improved removal efficiency through escalated reactive oxygen species generation. Additionally, the system exhibits consistent repeatability in its performance. Notably, its effectiveness is substantiated across varying nitrogen and phosphorus levels and in simulated water samples containing multiple algal species. In essence, this subtly adaptable reactor tuning integrates electro-Fenton, electro-flocculation, electro-adsorption, and electrochemical direct oxidation effects for the first time, potentially providing a more effective and manageable solution for addressing harmful algae in water.

Studies of hybrid ceria polypyrrole CPP-2 composite for effective defluoridation from potable water

Drinking water sources should eliminate fluoride ions, as they pose a serious threat to human health. By using in situ oxidative polymerization and Pechini’s method of CeO2 (CO), a new hybrid composite, CPP-2, was designed. Several physicochemical techniques were used to establish the properties of the adsorbent in its as synthesized state. The F- ions were removed from effluent using the hybrid composite adsorbent. The adsorption isotherm model was best represented by the Langmuir adsorption isotherm model. It yielded a maximum adsorption capacity of 351.8 mg/g at an optimal pH of 4 ± 0.2 at room temperature. The pseudo second order model best describes the adsorption process according to kinetic results. To get a detailed understanding of the fluoride adsorption mechanism, FTIR and XPS spectra are analyzed. Anionic exchange of the hydroxyl groups with F- ions and electrostatic attractions of the protonated hydroxyl on the surface of the adsorbent, as well as the nitrogen atoms of the amino groups from the polypyrrole (PP) moiety with the F- ions, were responsible for the process. The enthalpy change (ΔH° = +6.32 KJ/mol) and Gibbs free energy (ΔG° = -8.95 KJ/mol) were obtained from the thermodynamic data, which demonstrated the spontaneity, feasibility, and endothermic nature of adsorption. It had a great potential for defluoridation from groundwater at a pH level below the thresholds specified by the WHO and could withstand up to five cycles of adsorption and desorption. The overall results exhibited that CPP-2 has an enormous promise for defluoridation.

Methylomonas rivi sp. nov., Methylomonas rosea sp. nov., Methylomonas aurea sp. nov. and Methylomonas subterranea sp. nov., type I methane-oxidizing bacteria isolated from a freshwater creek and the deep terrestrial subsurface.

Four methane-oxidizing bacteria, designated as strains WSC-6T, WSC-7T, SURF-1T, and SURF-2T, were isolated from Saddle Mountain Creek in southwestern Oklahoma, USA, and the Sanford Underground Research Facility (SURF) in Lead, South Dakota, USA. The strains were Gram-negative, motile, short rods that possessed intracytoplasmic membranes characteristic of type I methanotrophs. All four strains were oxidase-negative and weakly catalase-positive. Colonies ranged from pale pink to orange in colour. Methane and methanol were the only compounds that could serve as carbon and energy sources for growth. Strains WSC-6T and WSC-7T grew optimally at lower temperatures (25 and 20 °C, respectively) compared to strains SURF-1T and SURF-2T (40 °C). Strains WSC-6T and SURF-2T were neutrophilic (optimal pH of 7.5 and 7.3, respectively), while strains WSC-7T and SURF-1T were slightly alkaliphilic, with an optimal pH of 8.8. The strains grew best in media amended with ≤0.5% NaCl. The major cellular fatty acids were C14 : 0, C16 : 1 ω8c, C16 : 1 ω7c, and C16 : 1 ω5c. The DNA G+C content ranged from 51.5 to 56.0 mol%. Phylogenetic analyses indicated that the strains belonged to the genus Methylomonas, with each exhibiting 98.6-99.6% 16S rRNA gene sequence similarity to closely related strains. Genome-wide estimates of relatedness (84.5-88.4% average nucleotide identity, 85.8-92.4% average amino acid identity and 27.4-35.0% digital DNA-DNA hybridization) fell below established thresholds for species delineation. Based on these combined results, we propose to classify these strains as representing novel species of the genus Methylomonas, for which the names Methylomonas rivi (type strain WSC-6T=ATCC TSD-251T=DSM 112293T), Methylomonas rosea (type strain WSC-7T=ATCC TSD-252T=DSM 112281T), Methylomonas aurea (type strain SURF-1T=ATCC TSD-253T=DSM 112282T), and Methylomonas subterranea (type strain SURF-2T=ATCC TSD-254T=DSM 112283T) are proposed.

Innovative potassium hexacyanoferrate intercalated into layered double hydroxide adsorbent for efficient cesium removal from seawater

Effective radioactive wastewater treatment technology is essential to ensure the sustainable development of nuclear energy. Here, the potassium ferrocyanide (HCF) intercalating with layered double hydroxide (LDH) (HCF@LDH) was successfully synthesized by coprecipitation method for selective cesium (Cs) removal from seawater. Characteristics of the adsorbent, and adsorption performance under different environmental conditions including Cs+ ion selectivity, and underlying adsorption mechanism were systematically investigated. The experimental results demonstrated that the adsorption capacity of HCF@LDH was 52.08 mg/g and it effectively removed 100 % of Cs+ from up to 200 mg/L Cs solution within 1 h under an optimal pH of 6.0–10.0. Moreover, the adsorbent displayed high selectivity towards Cs+ even in seawater and more than 90 % of Cs+ was removed from seawater and the highest cesium removal capacity from seawater was 44.64 mg/g. Both the Langmuir isotherm model (R2 = 0.997) and the pseudo-second-order kinetic model (R2 = 0.999) provided a good fit for the adsorption data signifying the adsorption process is homogeneous and formed monolayer through chemisorption. Furthermore, the reusability analysis exhibited this adsorbent was recycled and reused for at least 5 cycles without losing its original adsorption performance. The physicochemical analysis of the HCF@LDH proved that K4Fe (CN)6 successfully intercalated onto LDH and cation exchange between K+ and Cs+ played an important role in effective Cs adsorption. More specifically, the interstitial spaces of the hexacyanoferrate crystals facilitate the cation exchange process. The findings of this study demonstrate that HCF@LDH is a unique, highly effective, and selective adsorbent for Cs+ from complex seawater.

Applying the experiment design approach to minimize pollutants in olive mill wastewater using aluminum sulfate and ferric chloride as coagulants. West Bank—occupied Palestinian territories

A significant environmental contaminant in the West Bank and the Mediterranean Basin is olive mill wastewater (OMWW). OMWW was released between mid-September and mid-November, when harvesting was taking place. This led to environmental issues with soil, water resources, and the ecology. The optimization of the removal of organic compounds as total suspended solids (TSS), chemical oxygen demand (COD), and total polyphenols group (TP) pollutants is the main goal of this study. Both aluminum sulfate (AS) and ferric chloride (FC) are utilized as coagulants. The ideal dosages, coagulant types, and pH values to reduce the concentration of these pollutants were determined using the Design of Expert program (DoE). For the AS and FC, the optimal pH range is between 4 and 5, which reduces the TSS by roughly 93 % and 80 %, respectively. By using AS and FC, respectively, COD was removed at rates of 68 % and 58 %, and 0.5 g/l was the ideal coagulant concentration dose. The most important factor affecting the elimination procedure was the pH level. The outcomes show that those three contaminants can be successfully eliminated by the coagulation process. TSS ranges between 90 % and 96 %, COD between 53 % and 73 %, and TP between 11 % and 37 % when the two coagulate salts are used

Design and synthesis of a macro-porous chitosan-sodium alginate aerogel adsorbent for efficient adsorption of SDBS from aqueous solutions

The treatment of wastewater containing surfactants is an increasingly serious issue that requires immediate remediation. Adsorption is an effective way to treat wastewater containing surfactants. However, for the adsorption of macromolecular substances like surfactants, adsorbents typically require a macro-porous structure to ensure efficient adsorption of surfactants. In this study, a macro-porous aerogel adsorbent was successfully fabricated with renewable biopolymers of chitosan (CS) and sodium alginate (SA) by a novel internal gelation strategy with a ternary solvent system of acetic acid/water/tetrahydrofuran (THF). The as-prepared adsorbent was systematically characterized by XRD, FT-IR, SEM, and XPS. Equilibrium adsorption experiments were conducted to investigate the effects of pH, adsorbent dosage, contact time, initial concentration, and temperature on the Sodium dodecylbenzene sulfonate (SDBS) adsorption performance. At the optimal pH of 3.0, the maximum adsorption capacity reached 254.83 mg/g at 308 K with 50 mg/L initial concentration and 0.20 g/L adsorbent dosage. The macro-porous structure of the adsorbent facilitated the diffusion of SDBS and exposed more adsorption sites, which enabled the adsorption of SDBS in aqueous solutions. The adsorption process conformed to the pseudo-second-order kinetic model and the Freundlich isotherm model. Thermodynamic results indicated that the adsorption process was spontaneous and endothermic. Moreover, the synthesized adsorbent demonstrated excellent reusability after five cycles. The fixed-bed adsorption experiments further showed the potential of the synthesized adsorbent as an efficient adsorbent for continuous removal of SDBS. This study offers a feasible strategy for the development of macro-porous aerogel materials and demonstrates the as-prepared adsorbent for SDBS removal.

Rhodamine-benzothiazole-thiophene: A triangular molecular tool for simultaneous detection of Hg2+ and Cu2+

A novel rhodamine-benzothiazole fluorescent chemosensor, bridged with a thiophene unit, is designed for the turn-on detection of Hg2+ and Cu2+. Noteworthy findings include impressively low limits of detection (0.007 μM for Hg2+ and 0.013 μM for Cu2+), a determined sensing mechanism involving spirolactam ring opening and hydrolytic cleavage, consistent 1:1 binding stoichiometry for both metal ions and an optimal pH of around 6. The sensor’s practical applicability is demonstrated through successful implementation in HeLa cells and zebrafish, along with effective analyte recovery in various water samples. This work advances the field by providing a highly sensitive and versatile chemosensor for environmental and biological applications.

Evaluation of turbidity removal efficiency from Karun river water using polyaluminum chloride and chitosan

This study was conducted with the aim of evaluation of turbidity removal efficiency from Karun river water using polyaluminum chloride and chitosan. Chitosan is a linear cationic polyelectrolyte with high positive electron charge (one positive charge per glucose amine unit). Chitosan is used in water purification systems as coagulant along with common coagulants (alum, chlorferric). This biopolymer is also used as a primary coagulant. Chitosan has a higher ability to coagulate colloidal particles than synthetic polyelectrolytes, most of which lack the property of biological degradation in the short term, and some of them cause carcinogenesis and genetic mutation in humans. Therefore, the use of chitosan is preferable to artificial polyelectrolytes from an environmental point of view. This research was conducted on the laboratory scale using a jar test device at the water treatment plant in Ahvaz. In this study, various parameters such as pH, polyaluminum chloride concentration and chitosan concentration were present as variables. After preparing the samples and just before starting jar tests, in the rapid mixing cycle of 200 rounds per minute, the optimal pH to remove the turbidity obtained 8. Also, the optimal dosages of polyaluminum chloride and chitosan were selected as 4 ppm and 0.04 ppm, respectively. Also the main mechanism for destabilizing clots and removing turbidity was bridging between the particles. In this condition, the removal of turbidity showed the highest efficiency. On the other hand, chitosan created clots with a higher weight and, as a result, better settling ability. Therefore, chitosan was a suitable choice to remove turbidity from water.

Ecofriendly synthesized Zeolite 4A for the treatment of a multi-cationic contaminant-based effluent: Central composite design (CCD) statistical approach

One of the key aspects of futureproofing the sustainability of life on earth lies in the protection of the hydrosphere, particularly from soluble heavy metal ion pollutants. In the current study, the central composite design and optimization of the ion-exchange process have been carried out for the simultaneous removal of selected cations; Cd2+, Cu2+, and Zn2+ cations using synthesized zeolite 4A. X-ray diffraction analysis confirmed the formation of zeolite 4A. The Brunauer-Emmett-Teller (BET) surface area of the synthesized zeolite was 32 m2/g. Results mainly indicate that there is a strong relationship between the experimental data and central composite design-based models of ion removal efficiency with R2 > 0.9 and the lack of fit less than 0.1 %. All the selected ion exchange parameters (time, dosage, pH, and temperature) were found to be statistically significant, with a p-value less than 0.05.For the complete simultaneous removal of selected cations, the optimal zeolite dosage, pH, and contact time are 1.2 g/100 cm3, 6, and 3 h. The optimal temperature ranges from 25 to 27 °C. The initial concentration of each selected cation is 450 mg/L. The ion exchange is in good agreement with the Freundlich and Langmuir isotherm models. Based on the Langmuir isotherm model, the maximum Cd2+, Cu2+, and Zn2+ uptake capacity values of zeolite are 103, 99.89, and 82.08 mg/g, respectively. In this study, it has been mainly inferred that CCD can be considered a useful tool for the modeling and optimization of zeolite ion exchange applications.

Cloning Cellulase Genes from Victoria Falls Rainforest Decaying Logs Metagenome.

The Victoria Falls rainforest is a protected site whose forest floors harbor a host of cellulolytic microorganisms involved in biomass degradation. This study collected decaying logs and soil from the rainforest for bioprospecting cellulases from their metagenomes. Metagenomic DNA was isolated from the compound sample. Degenerate cellulase primers were used to amplify cellulase genes in the metagenome. The resulting amplicons cloned into Z-competent Escherichia coli DH5α were analyzed by functional screening for the production of cellulase extracellularly. Functional screening of the clones resulted in one clone (Clone-i) testing positive for extracellular cellulase production. Submerged fermentation of Clone-i was carried out for cellulase production. The cellulases were characterized to determine their activity's optimum pH and temperature. The diversity of the cellulases produced by Clone-i was determined. Clone-i's optimum enzyme activity was observed after 72 hours of incubation at 50°C and pH 5. Clone-i produced 80% more exoglucanases as compared to endoglucanases. The cellulolytic Clone-i' isolate shows Victoria Falls rainforest's potential as an enzyme bioprospecting site, reflecting that metagenomics is a valuable tool in microbial ecology.

Biological Characteristics, Artificial Domestication Conditions Optimization, and Bioactive Components of Beauveria caledonica.

In this study, one strain of Beauveria caledonica was isolated from wild fruiting bodies collected from Guizhou Province, China, and its species identification, biological characteristics, domestication, and cultivation methods were studied along with polysaccharide and adenosine content analysis. The mycelia were identified by ITS sequencing, and the fruiting bodies of B. caledonica were domestically cultivated for the first time using wheat and rice as basic cultivation media. The carbon sources, nitrogen sources, cultivation temperatures, and pH for mycelial growth were optimized through single-factor experiments and response surface methodology (RSM) experiments. The polysaccharide content was detected by the phenol-sulfuric acid method, and the adenosine content was measured by high-performance liquid chromatography (HPLC). The results confirmed that the identified mycelia were B. caledonica. The optimum medium for solid culture was 25.8 g/L glycerol, 10.9 g/L yeast extract, 1 g/L MgSO4·7H2O, 1 g/L KH2PO4, 10 mg/L vitamin B1, and 20 g/L agar; the optimum pH was 6.5, and the optimum culture temperature was 25 °C. The optimal liquid culture medium was 26.2 g/L glycerol, 11.1 g/L yeast extract, 1 g/L MgSO4·7H2O, 1 g/L KH2PO4, and 10 mg/L vitamin B1; the mycelia grew well at pH 6.6 and 25 °C. The average biological efficiencies of fruiting bodies on wheat and rice as culture media were 1.880% and 2.115%, respectively; the polysaccharide contents of fruiting bodies on the two media were 6.635% and 9.264%, respectively, while the adenosine contents were 0.145% and 0.150%, respectively. This study provides a valuable reference for further artificial cultivation and utilization of B. caledonica by investigating its biological characteristics, cultivation conditions for artificial domestication, and polysaccharide and adenosine contents in cultivated fruiting bodies.

Epoxidation of perillyl alcohol by engineered bacterial cytochrome P450 BM3

Perillyl alcohol (POH) is a secondary metabolite of plants. POH and its derivatives are known to be effective as an anticancer treatment. In this study, oxidative derivatives of POH, which are difficult to synthesize chemically, were synthesized using the engineered bacterial cytochrome P450 BM3 (CYP102A1) as a biocatalyst. The activity of wild-type (WT) CYP102A1 and 29 engineered enzymes toward POH was screened using a high-performance liquid chromatography. They produced one major product. Among them, the engineered CYP102A1 M601 mutant with seven mutations (R47L/F81I/F87V/E143G/L150F/L188Q/E267V) showed the highest conversion, 6.4-fold higher than the WT. Structure modeling using AlphFold2 and PyMoL suggests that mutations near the water channel may be responsible for the increased catalytic activity of the M601 mutant. The major product was identified as a POH-8,9-epoxide by gas chromatography-mass spectrometry and nuclear magnetic resonance analysis. The optimal temperature and pH for the product formation were 35 °C and pH 7.4, respectively. The kcat and Km of M601 were 540min−1 and 2.77mM, respectively. To improve POH-8,9-epoxide production, substrate concentration and reaction time were optimized. The optimal condition for POH-8,9-epoxide production by M601 was 5.0mM POH, pH 7.4, 35 ℃, and 6h reaction, which produced the highest concentration of 1.72mM. Therefore, the biosynthesis of POH-8,9-epoxide using M601 as a biocatalyst is suggested to be an efficient and sustainable synthetic process that can be applied to chemical and pharmaceutical industries.

Enzymatic glycosylation of aloesone performed by plant UDP-dependent glycosyltransferases.

Aloesone is a bioactive natural product and biosynthetic precursor of rare glucosides found in rhubarb and some aloe plants including Aloe vera. This study aimed to investigate biocatalytic aloesone glycosylation and more than 400 uridine diphosphate-dependent glycosyltransferase (UGT) candidates, including multifunctional and promiscuous enzymes from a variety of plant species were assayed. As a result, 137 selective aloesone UGTs were discovered, including four from the natural producer rhubarb. Rhubarb UGT72B49 was further studied and its catalytic constants (kcat=0.00092±0.00003s-1, KM=30±2.5μM) as well as temperature and pH optima (50°C and pH7, respectively) were determined. We further aimed to find an efficient aloesone glycosylating enzyme with potential application for biocatalytic production of the glucoside. We discovered UGT71C1 from Arabidopsis thaliana as an efficient aloesone UGT showing a 167-fold higher catalytic efficiency compared to that of UGT72B49. Interestingly, sequence analysis of all the 137 newly identified aloesone UGTs showed that they belong to different phylogenetic groups, with the highest representation in groups B, D, E, F and L. Finally, our study indicates that aloesone C-glycosylation is highly specific and rare, since it was not possible to achieve in an efficient manner with any of the 422 UGTs assayed, including multifunctional GTs and 28 known C-UGTs.