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Thermophilic actinomycetes are commonly found in extreme environments and can thrive and adapt to extreme conditions. These organisms exhibit substantial variation and garnered significant interest due to their remarkable enzymatic activities. This study evaluated the potential of Streptomyces griseorubens NBR14 and Nocardiopsis synnemataformans NBRM9 strains to produce thermo-stable amylase via submerged fermentation using wheat and bean straw. The Box-Behnken design was utilized to determine the optimum parameters for amylase biosynthesis. Subsequently, amylase underwent partial purification and characterization. Furthermore, the obtained hydrolysate was applied for ethanol fermentation using Saccharomyces cerevisiae. The optimal parameters for obtaining the highest amylase activity by NBR14 (7.72 U/mL) and NBRM9 (26.54 U/mL) strains were found to be 40 and 30 °C, pH values of 7, incubation time of 7 days, and substrate concentration (3 and 2 g/100 mL), respectively. The NBR14 and NBRM9 amylase were partially purified, resulting in specific activities of 251.15 and 144.84 U/mg, as well as purification factors of 3.91 and 2.69-fold, respectively. After partial purification, the amylase extracted from NBR14 and NBRM9 showed the highest activity level at pH values of 9 and 7 and temperatures of 50 and 60 °C, respectively. The findings also indicated that the maximum velocity (Vmax) for NBR14 and NBRM9 amylase were 57.80 and 59.88 U/mL, respectively, with Km constants of 1.39 and 1.479 mM. After 48 h, bioethanol was produced at concentrations of 5.95 mg/mL and 9.29 mg/mL from hydrolyzed wheat and bean straw, respectively, through fermentation with S. cerevisiae. Thermophilic actinomycetes and their α-amylase yield demonstrated promising potential for sustainable bio-ethanol production from agro-byproducts.
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Actinobacteria , Amilases , Etanol , Fermentação , Saccharomyces cerevisiae , Temperatura , Triticum , Etanol/metabolismo , Amilases/metabolismo , Concentração de Íons de Hidrogênio , Cinética , Actinobacteria/metabolismo , Actinobacteria/enzimologia , Saccharomyces cerevisiae/metabolismo , Hidrólise , Streptomyces/enzimologia , Streptomyces/metabolismo , Estabilidade EnzimáticaRESUMO
Currently, heavy metal-resistant (HMR) marine actinomycetes have attracted much attention worldwide due to their unique capabilities. In this study, 27 marine-derived actinomycetes were isolated from coastal beaches in the Arabian Gulf of Al-Jubail in Saudi Arabia and screened for resistance to 100 mg/L of the heavy metals Cd2+, Cr6+, Cu2+, Fe2+, Pb2+, and Ni2+ using different assay techniques. Six isolates were selected as HMRs, of which two isolates, JJB5 and JJB11, exhibited the highest maximum tolerance concentrations (200- > 300 mg/L). Both isolates were the highest among six-HMR screened for their biodegradation potential of plastics low-density polyethylene, polystyrene, and polyvinyl chloride, recording the highest weight loss (15 ± 1.22 - 65 ± 1.2%) in their thin films. They also showed the highest biodegradability of the pesticides acetamiprid, chlordane, hexachlorocyclohexane, indoxacarb and lindane, indicating promising removal capacities (95.70-100%) for acetamiprid and indoxacarb using HPLC analysis. Additionally, the cell-free filtrate (CFF) of both isolates displayed the highest antimicrobial activity among the six-HMR screened against a variety of microbial test strains, recording the highest inhibition zone diameters (13.76 ± 0.66 - 26.0 ± 1.13 mm). GCâMS analyses of the ethyl acetate extract of their CFFs revealed the presence of diverse chemical compounds with a multitude of remarkable biological activities. Based on their spore morphology and wall-chemotype, they were assigned to the nocardioform-actinomycetes. Furthermore, their phenotypic characteristics, together with 16S rRNA gene sequencing (OR121525-OR121526), revealed them as Nocardia harenae JJB5 and Amycolatopsis marina JJB11. Our results suggest that marine HMR actinomycetes are promising candidates for various biotechnological applications.
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Biodegradação Ambiental , Metais Pesados , Testes de Sensibilidade Microbiana , Nocardia , RNA Ribossômico 16S , Metais Pesados/metabolismo , RNA Ribossômico 16S/genética , Nocardia/isolamento & purificação , Nocardia/genética , Nocardia/metabolismo , Arábia Saudita , Antibacterianos/farmacologia , Filogenia , Actinobacteria/metabolismo , Actinobacteria/isolamento & purificação , Actinobacteria/genética , Actinobacteria/classificação , Poluentes Químicos da Água/metabolismo , Água do Mar/microbiologia , Praguicidas/metabolismo , Farmacorresistência BacterianaRESUMO
Actinomycetes are an attractive source of lignocellulose-degrading enzymes. The search for actinomycetes producing extremozyme cellulase using cheap lignocellulosic waste remains a priority goal of enzyme research. In this context, the extremophilic actinomycete NBRM9 showed promising cellulolytic activity in solid and liquid assays. This actinomycete was identified as Nocardiopsis synnemataformans based on its phenotypic characteristics alongside phylogenetic analyses of 16S rRNA gene sequencing (OQ380604.1). Using bean straw as the best agro-waste, the production of cellulase from this strain was statistically optimized using a response surface methodology, with the maximum activity (13.20 U/mL) achieved at an incubation temperature of 40 °C, a pH of 9, an incubation time of 7 days, and a 2% substrate concentration. The partially purified cellulase (PPC) showed promising activity and stability over a wide range of temperatures (20-90 °C), pH values (3-11), and NaCl concentrations (1-19%), with optimal activity at 50 °C, pH 9.0, and 10% salinity. Under these conditions, the enzyme retained >95% of its activity, thus indicating its extremozyme nature. The kinetics of cellulase showed that it has a Vmax of 20.19 ± 1.88 U/mL and a Km of 0.25 ± 0.07 mM. The immobilized PPC had a relative activity of 69.58 ± 0.13%. In the in vitro microtiter assay, the PPC was found to have a concentration-dependent anti-biofilm activity (up to 85.15 ± 1.60%). Additionally, the fermentative conversion of the hydrolyzed bean straw by Saccharomyces cerevisiae (KM504287.1) amounted to 65.80 ± 0.52% of the theoretical ethanol yield. Overall, for the first time, the present work reports the production of extremozymatic (thermo, alkali-, and halo-stable) cellulase from N. synnemataformans NBRM9. Therefore, this strain is recommended for use as a biotool in many lignocellulosic-based applications operating under harsh conditions.
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The demand for the functionalization of additive materials based on bacterial cellulose (BC) is currently high due to their potential applications across various sectors. The preparation of BC-based additive materials typically involves two approaches: in situ and ex situ. In situ modifications entail the incorporation of additive materials, such as soluble and dispersed substances, which are non-toxic and not essential for bacterial cell growth during the production process. However, these materials can impact the yield and self-assembly of BC. In contrast, ex situ modification occurs subsequent to the formation of BC, where the additive materials are not only adsorbed on the surface but also impregnated into the BC pellicle, while the BC slurry was homogenized with other additive materials and gelling agents to create composite films using the casting method. This review will primarily focus on the in situ and ex situ functionalization of BC then sheds light on the pivotal role of functionalized BC in advancing biomedical technologies, wound healing, tissue engineering, drug delivery, bone regeneration, and biosensors.
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Celulose , Engenharia Tecidual , Celulose/metabolismo , Bactérias/metabolismo , Materiais Biocompatíveis/metabolismoRESUMO
Climate change is one of the main challenges, and it poses a tough challenge to the agriculture industry globally. Additionally, greenhouse gas (GHG) emissions are the main contributor to climate change; however, croplands are a prominent source of GHG emissions. Yet this complex challenge can be mitigated through climate-smart agricultural practices. Conservation tillage is commonly known to preserve soil and mitigate environmental change by reducing GHG emissions. Nonetheless, there is still a paucity of information on the influences of conservation tillage on wheat yield, soil properties, and GHG flux, particularly in the semi-arid Dingxi belt. Hence, in order to fill this gap, different tillage systems, namely conventional tillage (CT) control, straw incorporation with conventional tillage (CTS), no-tillage (NT), and stubble return with no-tillage (NTS), were laid at Dingxi, Gansu province of China, under a randomized complete block design with three replications to examine their impacts on yield, soil properties, and GHG fluxes. Results depicted that different conservative tillage systems (CTS, NTS, and NT) significantly (p < 0.05) increased the plant height, number of spikes per plant, seed number per meter square, root yield, aboveground biomass yield, thousand-grain weight, grain yield, and dry matter yield compared with CT. Moreover, these conservation tillage systems notably improved the soil properties (soil gravimetric water content, water-filled pore space, water storage, porosity, aggregates, saturated hydraulic conductivity, organic carbon, light fraction organic carbon, carbon storage, microbial biomass carbon, total nitrogen, available nitrogen storage, microbial biomass nitrogen, total phosphorous, available phosphorous, total potassium, available potassium, microbial counts, urease, alkaline phosphatase, invertase, cellulase, and catalase) while decreasing the soil temperature and bulk density over CT. However, CTS, NTS, and NT had non-significant effects on ECe, pH, and stoichiometric properties (C:N ratio, C:P ratio, and N:P ratio). Additionally, conservation-based tillage regimes NTS, NT, and CTS significantly (p < 0.05) reduced the emission and net global warming potential of greenhouse gases (carbon dioxide, methane, and nitrous oxide) by 23.44, 19.57, and 16.54%, respectively, and decreased the greenhouse gas intensity by 23.20, 29.96, and 18.72%, respectively, over CT. We conclude that NTS is the best approach to increasing yield, soil and water conservation, resilience, and mitigation of agroecosystem capacity.
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Microbial enzymes are crucial catalysts in various industries due to their versatility and efficiency. The microbial enzymes market has recently expanded due to increased demand for many reasons. Among them are eco-friendly solutions, developing novel microbial strains with enhanced enzymes that perform under harsh conditions, providing sustainability, and raising awareness about the benefits of enzyme-based products. By 2030, the global enzyme market is expected to account for $525 billion, with a growth rate of 6.7â¯%. L-asparaginase and L-glutaminase are among the leading applied microbial enzymes in antitumor therapy, with a growing market share of 16.5â¯% and 9.5â¯%, respectively. The use of microbial enzymes has opened new opportunities to fight various tumors, including leukemia, lymphosarcoma, and breast cancer, which has increased their demand in the pharmaceutical and medicine sectors. Despite their promising applications, commercial use of microbial enzymes faces challenges such as short half-life, immunogenicity, toxicity, and other side effects. Therefore, this review explores the industrial production, purification, formulation, and commercial utilization of microbial enzymes, along with an overview of the global enzyme market. With ongoing discoveries of novel enzymes and their applications, enzyme technology offers promising avenues for cancer treatment and other therapeutic interventions.
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Therapeutic proteins have been employed for centuries and reached approximately 50â¯% of all drugs investigated. By 2023, they represented one of the top 10 largest-selling pharma products ($387.03 billion) and are anticipated to reach around $653.35 billion by 2030. Growth hormones, insulin, and interferon (IFN α, γ, and ß) are among the leading applied therapeutic proteins with a higher market share. Protein-based therapies have opened new opportunities to control various diseases, including metabolic disorders, tumors, and viral outbreaks. Advanced recombinant DNA biotechnology has offered the production of therapeutic proteins and peptides for vaccination, drugs, and diagnostic tools. Prokaryotic and eukaryotic expression host systems, including bacterial, fungal, animal, mammalian, and plant cells usually applied for recombinant therapeutic proteins large-scale production. However, several limitations face therapeutic protein production and applications at the commercial level, including immunogenicity, integrity concerns, protein stability, and protein degradation under different circumstances. In this regard, protein-engineering strategies such as PEGylation, glycol-engineering, Fc-fusion, albumin conjugation, and fusion, assist in increasing targeting, product purity, production yield, functionality, and the half-life of therapeutic protein circulation. Therefore, a comprehensive insight into therapeutic protein research and findings pave the way for their successful implementation, which will be discussed in the current review.
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Peptídeos , Humanos , Peptídeos/química , Peptídeos/uso terapêutico , Animais , Viroses/tratamento farmacológico , Viroses/prevenção & controle , Proteínas Recombinantes/uso terapêutico , Engenharia de Proteínas/métodos , Antivirais/uso terapêutico , VírusRESUMO
The global increase in multidrug-resistant (MDR) bacteria has inspired researchers to develop new strategies to overcome this problem. In this study, 23 morphologically different, soil-isolated actinomycete cultures were screened for their antibacterial ability against MDR isolates of ESKAPE pathogens. Among them, isolate BOGE18 exhibited a broad antibacterial spectrum, so it was selected and identified based on cultural, morphological, physiological, and biochemical characteristics. Chemotaxonomic analysis was also performed together with nucleotide sequencing of the 16S rRNA gene, which showed this strain to have identity with Streptomyces lienomycini. The ethyl acetate extract of the cell-free filtrate (CFF) of strain BOGE18 was evaluated for its antibacterial spectrum, and the minimum inhibitory concentration (MIC) ranged from 62.5 to 250 µg/ml. The recorded results from the in vitro anti-biofilm microtiter assay and confocal laser scanning microscopy (CLSM) of sub-MIC concentrations revealed a significant reduction in biofilm formation in a concentration-dependent manner. The extract also displayed significant scavenging activity, reaching 91.61 ± 4.1% and 85.06 ± 3.14% of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis( 3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), respectively. A promising cytotoxic ability against breast (MCF-7) and hepatocellular (HePG2) cancer cell lines was obtained from the extract with IC50 values of 47.15 ± 13.10 and 122.69 ± 9.12 µg/ml, respectively. Moreover, based on gas chromatography-mass spectrometry (GC-MS) analysis, nine known compounds were detected in the BOGE18 extract, suggesting their contribution to the multitude of biological activities recorded in this study. Overall, Streptomyces lienomycini BOGE18-derived extract is a good candidate for use in a natural combating strategy to prevent bacterial infection, especially by MDR pathogens.
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Actinobacteria , Actinobacteria/metabolismo , Antioxidantes/farmacologia , Antioxidantes/química , RNA Ribossômico 16S/genética , Antibacterianos/química , Testes de Sensibilidade Microbiana , Extratos Vegetais/farmacologia , Extratos Vegetais/químicaRESUMO
The biocontrol approach using beneficial microorganisms to control crop diseases is becoming an essential alternative to chemical fungicides. Therefore, new and efficient biocontrol agents (BCA) are needed. In this study, a rhizospheric actinomycete isolate showed unique and promising antagonistic activity against three of the most common phytopathogenic fungi, Fusarium oxysporum MH105, Rhizoctonia solani To18, and Alternaria brassicicola CBS107. Identification of the antagonistic strain, which was performed according to spore morphology and cell wall chemotype, suggested that it belongs to the Nocardiopsaceae. Furthermore, cultural, physiological, and biochemical characteristics, together with phylogenetic analysis of the 16S rRNA gene (OP869859.1), indicated the identity of this strain to Nocardiopsis alba. The cell-free filtrate (CFF) of the strain was evaluated for its antifungal potency, and the resultant inhibition zone diameters ranged from 17.0 ± 0.92 to 19.5 ± 0.28 mm for the tested fungal species. Additionally, the CFF was evaluated in vitro to control Fusarium wilt disease in Vicia faba using the spraying method under greenhouse conditions, and the results showed marked differences in virulence between the control and treatment plants, indicating the biocontrol efficacy of this actinomycete. A promising plant-growth promoting (PGP) ability in seed germination and seedling growth of V. faba was also recorded in vitro for the CFF, which displayed PGP traits of phosphate solubilization (48 mg/100 ml) as well as production of indole acetic acid (34 µg/ml) and ammonia (20 µg/ml). This study provided scientific validation that the new rhizobacterium Nocardiopsis alba strain BH35 could be further utilized in bioformulation and possesses biocontrol and plant growth-promoting capabilities.
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Actinobacteria , Antifúngicos , Antifúngicos/farmacologia , Filogenia , RNA Ribossômico 16S/genética , Actinobacteria/genética , Doenças das Plantas/prevenção & controle , Doenças das Plantas/microbiologiaRESUMO
During potato chips manufacturing, large amounts of wastewater and potato powder wastes are produced. The wastewater obtained at washing after cutting the peeled potatoes into slices was analyzed, and a large quantity of organic compounds and minerals such as starch (1.69%), protein (1.5%), total carbohydrate (4.94%), reducing sugar (0.01%), ash (0.14%), crude fat (0.11%), Ca (28 mg/L), Mg (245 mg/L), Fe (45.5 mg/L), and Zn (6.5 mg/L) were recorded; these wastes could be considered as valuable by-products if used as a fermentation medium to increase the value of the subsequent products and to exceed the cost of reprocessing. In this study, we used wastewater and potato powder wastes as a growth medium for pigment and biomass production by Monascus purpureus (Went NRRL 1992). The response surface methodology was used to optimize total pigment and fungal biomass production. The influence of potato powder waste concentration, fermentation period, and peptone concentration on total pigment and biomass production was investigated using the Box-Behnken design method with 3-factors and 3-levels. The optimal production parameters were potato powder waste concentration of 7.81%, fermentation period of 12.82 days, and peptone concentration of 2.87%, which produced a maximum total pigment of 29.86 AU/ml that include, respectively, a maximum biomass weight of 0.126 g/ml and the yield of pigment of 236.98 AU/g biomass. The pigments produced were used as coloring agents for ice lolly. This study has revealed that the ice lolly preparations supplemented with these pigments received high acceptability. Finally, we recommend using wastewater and potato powder wastes for pigment and biomass production, which could reduce the cost of the pigment production process on an industrial scale in the future.
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PURPOSE: The purpose of this study is to evaluate the short-term results of lateral closing wedge osteotomy with medial hemiplateau elevation for the management of severe infantile Blount's disease. MATERIALS AND METHODS: In this prospective study, 11 cases of severe Blount's disease (Langenskiold stages five and six) were managed in the period between January 2017 and January 2020. Double osteotomy technique was applied, namely a metaphyseal closing wedge and a medial hemiplateau elevation, through a single midline incision. Fixation was achieved by a medial anatomical locked plate. Patients were evaluated clinically according to a modified version of paediatric outcomes data collection Instrument (PODCI) and radiologically by measuring the angle between the tibial and the femoral shaft, the mechanical axis deviation (MAD) and the angle of the medial tibial plateau (MTP) depression. RESULTS: The average follow-up period was 2 years. Healing of the osteotomies was achieved in all cases after the index operation within an average of 3 months. Based on our modification of the PODCI score, five cases had an excellent outcome, five were good, and one case ended with a fair outcome. No major complications were encountered in this study. CONCLUSION: The management of severe Blount's disease by acute correction using the aforementioned technique has been proven to achieve acceptable clinical and radiological outcomes without significant complications. LEVEL OF EVIDENCE: Level IV case series study. HOW TO CITE THIS ARTICLE: Nada AA, Hammad ME, Eltanahy AF, et al. Acute Correction and Plate Fixation for the Management of Severe Infantile Blount's Disease: Short-term Results. Strategies Trauma Limb Reconstr 2021;16(2):78-85.
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OBJECTIVE: To investigate intestinal motility changes due to uremia, and the effect of pretreatment with erythropoietin. METHODS: This randomized control study was conducted in the Department of Physiology, Faculty of Medicine, Ain Shams University, Cairo, Egypt from September 2010 to July 2011. Forty adult female Wistar albino rats were allocated into 3 groups: control group, gentamicin-treated group, receiving intraperitoneal gentamicin sulphate (100 mg/kg for 5 days), and erythropoietin-gentamicin-treated group, receiving subcutaneous erythropoietin (1000 IU/kg for 3 days) prior to gentamicin injection. Isolated segments of duodenum and descending colon was subjected to in vitro motility study. Plasma creatinine and urea were assayed. RESULTS: Induction of acute renal failure by gentamicin treatment resulted in a significant decrease in frequency of contraction of the duodenum and descending colon, an increase in the average duration of contraction of the duodenum, and a significant decrease in the average force of contraction in the descending colon. Moreover, the average force of contraction in response to acetylcholine was significantly decreased in the duodenum. The erythropoietin-gentamicin-treated group revealed a significant decrease in plasma creatinine and urea, and a significant increase in the duodenal average force of contraction and motility index, and colonic frequency. The duodenal absolute and average forces of contraction after acetylcholine increased significantly. CONCLUSION: Acute uremia impairs small and large intestinal motility, probably due to uremic toxins and autonomic dysfunction. Erythropoietin pretreatment protected against intestinal dysmotility through the improvement of renal function and its neurotropic action.