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The present study assesses the Paenibacillus sp. D9 lipopeptide biosurfactant synthesis in cheap substrates including functional properties and applicability for varying biotechnological processes. Different experimental setups were made for oil dispersion, heavy metals removals from contaminated environments, and washing performance. The study revealed surface tension activities of 31.7–32.7 mN/m, and maximum biosurfactant yield of more than 8 g/L. Removals of 85.90%, 98.68%, 99.97%, 63.28%, 99.93%, and 94.22% were obtained for Ca, Cu, Fe, Mg, Ni, and Zn, respectively from acid mine effluents. In comparison with chemical surfactants, there was pronounced removal of heavy metals from wastewater, contaminated sands, and vegetable matter, as well as improved oil dispersing activity. A comparative study revealed that biosurfactant was more efficient (> 60%) for removal of tomato sauce and coffee stains than chemical surfactants (< 50%). Thus, lipopeptide biosurfactants are green biomolecules reducing hazards and contaminations within the environment. The future use of this lipopeptide biosurfactant is greatly promising in biotechnology.

Interests in biosurfactant in industrial and environmental applications have increased considerably in recent years, owing to their potential benefits over synthetic counterparts. The present study aimed at analyzing the stability and oil removal efficiency of a new lipopeptide biosurfactant produced by Paenibacillus sp. D9 and its feasibility of its use in biotechnological applications. Paenibacillus sp. D9 was evaluated for optimal growth conditions and improved production yield of lipopeptide biosurfactant with variations in different substrate parameters such as carbon (C), nitrogen (N), C:N: ratio, metal supplements, pH, and temperature. Enhanced biosurfactant production was observed when using diesel fuel and ammonium sulfate as carbon and nitrogen source respectively. The maximum biosurfactant yield of 4.11 g/L by Paenibacillus sp. D9 occurred at a C/N ratio of 3:1, at pH 7.0, 30 °C, 4.0 mM MgSO 4 , and 1.5% inoculum size. The D9 biosurfactant was found to retain surface-active properties under the extreme conditions such as high thermal, acidic, alkaline, and salt concentration. The ability to emulsify further emphasizes its potential usage in biotechnological application. Additionally, the lipopeptide biosurfactant exhibited good performance in the degradation of highly toxic substances when compared with chemical surfactant, which proposes its probable application in biodegradation, microbial-enhanced oil recovery or bioremediation. Furthermore, the biosurfactants were effective in a test to stimulate the solubilization of hydrophobic pollutants in both liquid environments removing 49.1 to 65.1% diesel fuel including hydrophobic pollutants. The study highlights the usefulness of optimization of culture parameters and their effects on biosurfactant production, high stability, improved desorption, and solubilization of hydrophobic pollutants.

Bacillus tropicus is a recently identified subspecies of the Bacillus cereus group of bacteria that have been shown to possess genes associated with antimicrobial resistance (AMR) and identified as the causative agent for anthrax-like disease in Chinese soft-shelled turtles. In addition, B. tropicus has demonstrated great potential in the fields of bioremediation and bioconversion. This article describes the comparative genomics of a Bacillus phage vB_Btc-RBClinn15 (referred to as RBClin15) infecting the recently identified B. tropicus AOA-CPS1. RBClin15 is a temperate phage with a putative parABS partitioning system as well as an arbitrium system, which are presumed to enable extrachromosomal genome maintenance and regulate the lysis/lysogeny switch, respectively. The temperate phage RBClin15 has been sequenced however, was erroneously deposited as a plasmid in the NCBI GenBank database. A BLASTn search against the GenBank database using the whole genome sequence of RBClin15 revealed seven other putative temperate phages that were also deposited as plasmids in the database. Comparative genomic analyses shows that RBClin15 shares between 87 and 92% average nucleotide identity (ANI) with the seven temperate phages from the GenBank database. All together RBClin15 and the seven putative temperate phages share common genome arrangements and < 29% protein homologs with the closest phages, including 0105phi7-2. A phylogenomic tree and proteome-based phylogenetic tree analysis showed that RBClin15 and the seven temperate phages formed a separate branch from the closest phage, 0105phi7-2. In addition, the intergenomic similarity between RBClin15 and its closely related phages ranged between 0.3 and 47.7%. Collectively, based on the phylogenetic, and comparative genomic analyses, we propose three new species which will include RBClin15 and the seven temperate phages in the newly proposed genus Theosmithvirus under Caudoviricetes .

Phosphopantetheinyl transferases are of tremendous enthusiasm inferable from their fundamental parts in activating polyketide, fatty acid, and non-ribosomal peptide synthetase enzymes and additionally an increasing number of biotechnological applications. The present study reports the identification of sfp gene from the Paenibacillus sp. D9, which encompasses 693 bp encoding a 230-amino acid protein with a molecular weight of 25.3 kDa. The amino acid sequence Paenibacillus sp. D9 Sfp revealed more than 90% sequence identity to other Sfp proteins from other Paenibacillus. The sfp gene was cloned and recovered efficiently using affinity chromatography with maximal specific phosphopantetheinyl transferase activity at an optimal pH of 8.0 and temperature of 30 °C. The enzyme also exhibited stability under a wide-ranging pH and temperature. The presence of Zn2+, Cu2+, and Fe2+ ions improved the enzymatic activity, while other metals such as Ni2+, Co2+, and Mg2+ had inhibitory effects. The introduction of EDTA also displayed no inhibition. Kinetic parameters were obtained having values of 4.52 mg/mL, 35.33 U/mg, 3.64 s−1, and 0.104 mM−1 s−1 for Km, Vmax, kcat, and kcat/Km, respectively. The biosurfactant synthesized by the recombinant BioSp was found to be surface active, reducing the surface tension to 33.7 mN/m on the glucose substrate after 5 days of incubation at 37 °C. The recombinant Escherichia coli strain also exhibited an improvement in biosurfactant yield (1.11 g/L) when contrasted with 0.52 g/L from Paenibacillus sp. D9. High esterase activity of 2.55 IU/mL using p-nitrophenyl acetate was observed on the recombinant strain, as the protein connected with the release of the biosurfactant was observed to be an esterase. The characteristics of improved biosurfactant and esterase synthesis by hyper-producing recombinant strain possess numerous values from biotechnology standpoint.

Current research energies are fixated on the synthesis of environmentally friendly and non-hazardous products, which include finding and recognizing biosurfactants that can substitute synthetic surfactants. Microbial biosurfactants are surface-active compounds synthesized intracellularly or extracellularly. To use biosurfactants in various industries, it is essential to understand scientific engagements that demonstrate its potentials as real advancement in the 21st century. Other than applying a substantial effect on the world economic market, engineered hyper-producing microbial strains in combination with optimized cultivation parameters have made it probable for many industrial companies to receive the profits of 'green' biosurfactant innovation. There needs to be an emphasis on the worldwide state of biosurfactant synthesis, expression of biosurfactant genes in expressive host systems, the recent developments, and prospects in this line of research. Thus, molecular dynamics with respect to genetic engineering of biosynthetic genes are proposed as new biotechnological tools for development, improved synthesis, and applications of biosurfactants. For example, mutant and hyper-producing recombinants have been designed efficaciously to advance the nature, quantity, and quality of biosurfactants. The fastidious and deliberate investigation will prompt a comprehension of the molecular dynamics and phenomena in new microorganisms. Throughout the decade, valuable data on the molecular genetics of biosurfactant have been produced, and this solid foundation would encourage application-oriented yields of the biosurfactant production industry and expand its utilization in diverse fields. Therefore, the conversations among different interdisciplinary experts from various scientific interests such as microbiology, biochemistry, molecular biology, and genetics are indispensable and significant to accomplish these objectives.

Due to the continued persistence of waterborne viral-associated infections, the presence of enteric viruses is a concern. Notwithstanding the health implications, viral diversity and abundance is an indicator of water quality declination in the environment. The aim of this study was to evaluate the presence of viruses (bacteriophage and enteric viruses) in a highly polluted, anthropogenic-influenced river system over a 6-month period at five sampling points. Cytopathic-based tissue culture assays revealed that the isolated viruses were infectious when tested on Hep-G2, HEK293 and Vero cells. While transmission electron microscopy (TEM) revealed that the majority of the viruses were bacteriophages, a number of presumptive enteric virus families were visualized, some of which include Picornaviridae, Adenoviridae, Polyomaviridae and Reoviridae. Finally, primer specific nested polymerase chain reaction (nested-PCR)/reverse transcription-polymerase chain reaction (RT-PCR) coupled with BLAST analysis identified human adenovirus, polyomavirus and hepatitis A and C virus genomes in river water samples. Taken together, the complexity of both bacteriophage and enteric virus populations in the river has potential health implications. Finally, a systematic integrated risk assessment and management plan to identify and minimize sources of faecal contamination is the most effective way of ensuring water safety and should be established in all future guidelines.

Background Anti-idiotypic antibodies have garnered significant attention in biotechnology and immunology due to their unique ability to mimic specific epitopes on target antigens, thereby serving as functional analogues. This property makes them valuable tools for various applications. In this study, we aimed to isolate an anti-idiotypic single domain antibody against Cry2Aa from a naive phage-display library and investigate its structural and functional mimicry of the Cry2Aa toxin. Results An anti-idiotypic single domain antibody (sdAb) specific for the Cry2Aa toxin was successfully isolated from a naive phage-display library. Sequence analysis revealed a 57.1% identity between the epitopes on Cry2Aa mimicked by the sdAb and Cry2Aa. The sdAb could compete with Cry2Aa toxin for binding to anti-Cry2Aa F(ab’) 2 fragments and potential Cry2Aa receptors, including aminopeptidase N5 (APN5), vacuolar-type proton ATPase subunit A (V-ATPase A), and toxin-binding region (CR9-CR11) of cadherin-like protein (Cad-TBR) from Plutella xylostella . Molecular docking simulations indicated that the complementarity determining regions 2 (CDR2) and CDR3 of the antibody played critical roles in binding to these receptors and alanine mutant binding studies also proved that CDR2 and CDR3 played critical roles in receptor binding. These results indicated that the Cry2Aa anti-idiotypic sdAb has the potential to characterize a similar pattern of molecular interactions as Cry2Aa toxin. Conclusions The findings from this study indicate that the isolated Cry2Aa anti-idiotypic sdAb mimics the molecular interaction pattern of the Cry2Aa toxin with its midgut receptors. The anti-idiotypic sdAb offers new potential for developing novel insect control strategies.

The poor intracellular uptake and non-specific binding of anticancer drugs into cancer cells are the bottlenecks in cancer therapy. Nanocarrier platforms provide the opportunities to improve the drug efficacy. Here we show a carbon-based nanomaterial nanodiamond (ND) that carried paclitaxel (PTX), a microtubule inhibitor, and cetuximab (Cet), a specific monoclonal antibody against epidermal growth factor receptor (EGFR), inducing mitotic catastrophe and tumor inhibition in human colorectal cancer (CRC). ND-PTX blocked the mitotic progression, chromosomal separation, and induced apoptosis in the CRC cells; however, NDs did not induce these effects. Conjugation of ND-PTX with Cet (ND-PTX-Cet) was specifically binding to the EGFR-positive CRC cells and enhanced the mitotic catastrophe and apoptosis induction. Besides, ND-PTX-Cet markedly decreased tumor size in the xenograft EGFR-expressed human CRC tumors of nude mice. Moreover, ND-PTX-Cet induced the mitotic marker protein phospho-histone 3 (Ser10) and apoptotic protein active-caspase 3 for mitotic catastrophe and apoptosis. Taken together, this study demonstrated that the co-delivery of PTX and Cet by ND enhanced the effects of mitotic catastrophe and apoptosis in vitro and in vivo , which may be applied in the human CRC therapy.

BackgroundThe objective of this study was to evaluate the efficacy and safety of induction chemotherapy (ICT), GOFL (gemcitabine, oxaliplatin plus fluorouracil (5-FU)/leucovorin) versus modified FOLFIRINOX (irinotecan, oxaliplatin plus 5-FU/leucovorin), followed by concurrent chemoradiotherapy (CCRT) in locally advanced pancreatic adenocarcinoma (LAPC).MethodsChemo-naive patients with measurable LAPC were eligible and randomly assigned to receive biweekly ICT with either mFOLFIRINOX or GOFL for 3 months. Patients without systemic progression would have 5-FU- or gemcitabine-based CCRT (5040 cGy/28 fractions) and were then subjected to surgery or continuation of chemotherapy until treatment failure. The primary endpoint was 9-month progression-free survival (PFS) rate.ResultsBetween July 2013 and January 2019, 55 patients were enrolled. After ICT, 21 (77.8%) of 27 patients who received mFOLFIRINOX and 17 (60.7%) of 28 patients who received GOFL completed CCRT. Of them, one and five had per-protocol R0/R1 resection. On intent-to-treat analysis, the 9-month PFS rate, median PFS and overall survival in mFOLFIRINOX and GOFL arms were 30.5% versus 35.9%, 6.6 (95% confidence interval: 5.9–12.5) versus 7.6 months (3.9–12.3) and 19.6 (13.4–22.9) versus 17.9 months (13.4–23.9), respectively. Grade 3–4 neutropenia and diarrhoea during induction mFOLFIRINOX and GOFL were 37.0% versus 21.4% and 14.8% versus 3.6%, respectively.ConclusionInduction GOFL and mFOLFIRINOX followed by CCRT provided similar clinical outcomes in LAPC patients.Clinicaltrial.gov identifierNCT01867892.

Background Mycoplasma hyopneumoniae ( M. hyopneumoniae ) is the etiological agent of enzootic pneumonia, a highly infectious swine respiratory disease that distributed worldwide. The pathogenesis and virulence factors of M. hyopneumoniae are not fully clarified. As an important virulence factor of bacteria, nicotinamide adenine dinucleotide (NADH) oxidase (NOX) participates in host-pathogen interaction, however, the function of NOX involved in the pathogenesis of M. hyopneumoniae is not clear. Results In this study, significant differences in NOX transcription expression levels among different strains of M. hyopneumoniae differed in virulence were identified , suggesting that NOX may be correlated with M. hyopneumoniae virulence. The nox gene of M. hyopneumoniae was cloned and expressed in Escherichia coli , and polyclonal antibodies against recombinant NOX (rNOX) were prepared. We confirmed the enzymatic activity of rNOX based on its capacity to oxidize NADH to NAD + . Flow cytometry analysis demonstrated the surface localization of NOX, and subcellular localization analysis further demonstrated that NOX exists in both the cytoplasm and cell membrane. rNOX was depicted to mediate adhesion to immortalized porcine bronchial epithelial cells (hTERT-PBECs). Pre-neutralizing M. hyopneumoniae with anti-rNOX antibody resulted in a more than 55% reduction in the adhesion rate of high- and low-virulence M. hyopneumoniae strains to hTERT-PBECs. Moreover, a significant difference appeared in the decline in CCU 50 titer between virulent (168) and virulence-attenuated (168L) strains. NOX not only recognized and interacted with host fibronectin but also induced cellular oxidative stress and apoptosis in hTERT-PBECs. The release of lactate dehydrogenase by NOX in hTERT-PBECs was positively correlated with the virulence of M. hyopneumoniae strains. Conclusions NOX is considered to be a potential virulence factor of M. hyopneumoniae and may play a significant role in mediating its pathogenesis.