Rapid Biosynthesis Research Articles

Rapid Biosynthesis of Gold Nanoparticles Using Aqueous-ethanoic Leaf Extract of Heartleaf Moonseed: Characterization and Effect of pH on its Synthesis

Background: Gold nanoparticles (GNPs) are of tremendous interest due to their wide application range in the field of therapeutics and diagnostics. However, its potential is limited by its mode of synthesis utilizing different chemicals which are costly, hazardous and often labor-intensive. Nevertheless, the GNPs synthesized through green methods by using plant extracts can overcome these limitations and have wider acceptance. Keywords: Gold nanoparticles, medicinal plant, zeta potential, X-ray diffraction, fourier transforms infrared spectroscopy, transmission electron microscopy.

Biological synthesis of gold and silver chloride nanoparticles by Glycyrrhiza uralensis and in vitro applications

The current study highlights the rapid biosynthesis of gold nanoparticles (Gu–AuNps) and silver chloride nanoparticles (Gu–AgClNps) by aqueous root extract of Glycyrrhiza uralensis, a medicinal plant. G. uralensis has been reported for anticancer and hepatoprotective effects. The reduction of chloroauric acid and silver nitrate by the Glycyrrhiza root extract prompted the formation of Gu–AuNps and Gu–AgClNps within 4 and 40 min at 80 °C, respectively. The complete reaction did not require supplemental reducing and stabilizing agents, which demonstrated green synthesis. Field emission transmission electron microscopy (FE-TEM) revealed the spherical shape of Gu–AuNps and Gu–AgClNps. X-ray diffraction (XRD) showed face-centred cubic structure of Gu–AuNps and Gu–AgClNps with average crystallite size 12.25 nm and 8.01 nm, respectively. The biosynthesized Gu–AgClNps served as competent antimicrobial agent against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Salmonella enterica. Additionally, Gu–AuNps and Gu–AgClNps were analyzed for their catalytic ability to reduce methylene blue as model test pollutant. Likewise, both nanoparticles possessed free radical scavenging activity against 2,2-diphenyl-1-picrylhydrzyl (DPPH). Moreover, in vitro cytotoxicity in murine macrophage (RAW264.7) and human breast cancer (MCF7) cells were evaluated. Thus, the study proposes a green synthesis of Gu–AuNps and Gu–AgClNps by G. uralensis extract and in vitro biological applications.

English

Biological extracts have recently shown a great potential for rapid biosynthesis of silver nanoparticles (AgNPs) with antibacterial activity. AgNPs were synthetized by reduction effect of secondary metabolites produced by the Nigrospora sp. fungus, from Moringa oleifera stem as a reducing agent and silver nitrate (AgNO3) (1 mM) as salt precursor. The synthesis of nanoparticles (NPs) was monitored through analysis of the UV-Vis spectroscopy absorption in the 436 to 440 nm range indicating the presence of AgNPs in the colloidal aqueous solutions. Fourier transform infrared spectroscopy (FTIR) spectra were performed to identify the compounds responsible for the bio reduction of the Ag+. The morphology and sizes of AgNPs were characterized by scanning electron microscopy (SEM), energy dispersive of X-ray spectroscopy (EDS), dynamic light scattering (DLS), and colloid stability by zeta potential measurements. The NPs obtained were spherical in shape with size in the 3 to 70 nm range. Antibacterial activity was confirmed by evaluation of their effect against Escherichia coli, Klebsiella cloacae and Staphylococcus epidermidis. The proposed green synthesis of AgNPs from secondary metabolites produced by the Nigrospora sp. fungus from M. oleifera stem can be strongly recommended as a potential method for biomedical application. Key words: Silver nanoparticles, secondary metabolites, green chemistry, antibacterial activity.

Rapid Biosynthesis of Silver Nanoparticles Based on Flocculation and Reduction of an Exopolysaccharide from Arthrobacter sp. B4: Its Antimicrobial Activity and Phytotoxicity

Silver nanoparticles (AgNPs) were rapidly synthesized using an exopolysaccharide from Arthrobacter sp. B4 (B4-EPS). The optimum condition for AgNPs synthesis was under the concentration of 5 g/L B4-EPS and 1 mM AgNO3 at 80°C between pH 7.0 and 8.0. The resulting AgNPs displayed a face-centred-cubic structure with the size range from 9 nm to 72 nm. Further analysis showed that flocculation and reduction of B4-EPS played a pivotal role in the formation of AgNPs. Furthermore, these nanoparticles exhibited great stability, excellent antimicrobial activity, and low phytotoxicity. The aforementioned data provide a feasible and efficient approach for green synthesis of AgNPs using microbial polysaccharides with flocculation and reduction activity, which will be promising in medical filed.

Mechanistic insight into the rapid one-step facile biofabrication of antibacterial silver nanoparticles from bacterial release and their biogenicity and concentration-dependent in vitro cytotoxicity to colon cells

Rapid biosynthesis of silver nanoparticles from Gram +ve and Gram −ve bacterial strains and their hiogenicity dependent antibacterial and cytntoxicity.

Rapid Biosynthesis of AgNPs Using Soil Bacterium Azotobacter vinelandii With Promising Antioxidant and Antibacterial Activities for Biomedical Applications

Silver nanoparticles (AgNPs) are applied in various fields from electronics to biomedical applications as a result of their high surface-to-volume ratio. Even though different approaches are available for synthesis of AgNPs, a nontoxic method for the synthesis has not yet been developed. Thus, this study focused on developing an easy and ecofriendly approach to synthesize AgNPs using Azotobacter vinelandii culture extracts. The biosynthesized nanoparticles were further characterized by ultraviolet–visible (UV–Vis) spectroscopy, x-ray diffraction (XRD), Fourier transform infrared (FTIR), energy-dispersive spectrum, particle size distribution (PSD), and transmission electron microscopy (TEM). UV absorption noticed at 435 nm showed formation of AgNPs. The XRD pattern showed a face-centered cubic structure with broad peaks of ~28.2°, ~32.6°, ~46.6°, ~55.2°, ~57.9°, and ~67.8°. The FTIR confirmed the involvement of various functional groups in the biosynthesis of AgNPs. The PSD and TEM analyses showed spherical, well-distributed nanoparticles with an average size of 20–70 nm. The elemental studies confirmed the existence of pure AgNPs. The bacterial extract containing extracellular enzyme nitrate reductase converted silver nitrate into AgNPs. AgNPs significantly inhibited the growth of pathogenic bacteria such as Streptomyces fradiae (National Collection of Industrial Microorganisms (NCIM) 2419), Staphylococcus aureus (NCIM 2127), Escherichia coli (NCIM 2065), and Serratia marcescens (NCIM 2919). In addition, biosynthesized AgNPs were found to possess strong antioxidant activity. Thus, the results of this study revealed that biosynthesized AgNPs could serve as a lead in the development of nanomedicine.

Biologically synthesized silver nanoparticles by aqueous extract of Satureja intermedia C.A. Mey and the evaluation of total phenolic and flavonoid contents and antioxidant activity

Developing low cost and environmentally friendly methods for metallic nanoparticles is an increasing need. Using plants towards synthesis of nanoparticles are beneficial with the presence of bio-molecules in plants, which can act as capping/stabilizing and reducing agents. In the present attempt, we describe rapid biosynthesis of silver nanoparticles by Satureja intermedia C. A. Mey (Lamiaceae) aqueous extract. Synthesized nanoparticles were characterized by UV–Visible spectroscopy, X-ray diffraction (XRD), transmission electron microscopy (TEM) and the chemical groups in plant extract were detected by Fourier Transform Infra-Red (FT-IR) spectroscopy. The XRD study showed crystalline nature and face cubic center shape for nanoparticles. TEM study showed that the mean diameter and standard deviation for the silver nanoparticles were 29.29 ± 28.18 nm. Total phenolic and flavonoid contents and radical scavenging activity of the aqueous extract and SNPs/extract mixture, were also evaluated in this study. It can be concluded that the aerial parts of S. intermedia is a good source of phenolic compounds, a potent antioxidant and a valuable choice for bio-reduction and biosynthesis of silver nanoparticles.

Biosynthesis of gold nanoparticles using Capsicum annuum var. grossum pulp extract and its catalytic activity

Biological synthesis approach has been regarded as a green, eco-friendly and cost effective method for nanoparticles preparation without any toxic solvents and hazardous bi-products during the process. This present study reported a facile and rapid biosynthesis method for gold nanoparticles (GNPs) from Capsicum annuum var. grossum pulp extract in a single-pot process. The aqueous pulp extract was used as biotic reducing agent for gold nanoparticle growing. Various shapes (triangle, hexagonal, and quasi-spherical shapes) were observed within range of 6–37nm. The UV–Vis spectra showed surface plasmon resonance (SPR) peak for the formed GNPs at 560nm after 10min incubation at room temperature. The possible influences of extract amount, gold ion concentration, incubation time, reaction temperature and solution pH were evaluated to obtain the optimized synthesis conditions. The effects of the experimental factors on NPs synthesis process were also discussed. The produced gold nanoparticles were characterized by transform electron microscopy (TEM), X-ray diffraction (XRD), energy dispersive X-ray (EDS) and Fourier Transform infrared spectroscopy (FTIR). The results demonstrated that the as-obtained GNPs were well dispersed and stable with good catalytic activity. Biomolecules in the aqueous extract were responsible for the capping and stabilization of GNPs.

Biomimetic synthesis of antimicrobial silver nanoparticles using in vitro-propagated plantlets of a medicinally important endangered species: Phlomis bracteosa.

In vitro-derived cultures of plants offer a great potential for rapid biosynthesis of chemical-free antimicrobial silver nanoparticles (AgNPs) by enhancing their phytochemical reducing potential. Here, we developed an efficient protocol for in vitro micropropagation of a high-value endangered medicinal plant species, Phlomis bracteosa, in order to explore its biogenic potential in biomimetic synthesis of antimicrobial AgNPs. Murashige and Skoog medium supplemented with 2.0 mg/L thidiazuron was found to be more efficient in inducing optimum in vitro shoot regeneration (78%±4.09%), and 2.0 mg/L indole-3-butyric acid was used for maximum root induction (86%±4.457%). Antimicrobial AgNPs were successfully synthesized by using aqueous extract (rich in total phenolics and flavonoids content) of in vitro derived plantlets of P. bracteosa. Ultraviolet–visible spectroscopy of synthesized AgNPs showed characteristic surface plasmon band in the range of 420–429 nm. The crystallinity, size, and shape of the AgNPs were characterized by X-ray diffraction and scanning electron microscopy. Face-centered cubic AgNPs of almost uniform spherical size (22.41 nm) were synthesized within a short time (1 hour) at room temperature. Fourier-transform infrared spectroscopy revealed that the polyphenols were mainly responsible for reduction and capping of synthesized AgNPs. Energy dispersive X-ray analysis further endorsed the presence of elemental silver in synthesized AgNPs. These biosynthesized AgNPs displayed significantly higher bactericidal activity against multiple drug-resistant human pathogens. The present work highlighted the potent role of in vitro-derived plantlets of P. bracteosa for feasible biosynthesis of antimicrobial AgNPs, which can be used as nanomedicines in many biomedical applications.

Photo-induced rapid biosynthesis of silver nanoparticle using aqueous extract of Xanthium strumarium and its antibacterial and antileishmanial activity

The current work describes the biosynthesis of stable AgNPs using aqueous extract of Xanthium strumarium (AEX) which act as both reducing as well as a stabilizing agent. The biosynthesis was confirmed by UV–visible spectroscopy where the presence of SPR band at λmax 436nm corresponded to the existence of AgNPs in reaction mixture. The optimum conditions for biosynthesis of AgNPs were 30min of sunlight exposure time, 3.0% (v/v) of AEX inoculum dose and 3.5mM AgNO3 concentration. The synthesized AgNPs was characterized by HRTEM, SAED, FESEM, EDX, XRD, AFM, and FTIR which showed potent antibacterial and antileishmanial activity.

Optimization of process parameters for the rapid biosynthesis of hematite nanoparticles

Hematite (α-Fe2O3) nanoparticles are widely used in various applications including gas sensors, pigments owing to its low cost, environmental friendliness, non-toxicity and high resistance to corrosion. These nanoparticles were generally synthesized by different chemical methods. In the present study, nanoparticles were synthesized rapidly without heat treatment by biosynthesis approach using culture supernatant of Bacillus cereus SVK1. The physiochemical parameters for rapid synthesis were optimized by using UV–visible spectroscopy. The time taken for hematite nanoparticle synthesis was found to increase with the increasing concentration of the precursor. This might be due to the inadequate proportion of quantity of biomolecules present in the culture supernatant to the precursor which led to delayed bioreduction. Greater quantities of culture supernatant with respect to precursor lead to rapid synthesis of hematite nanoparticles. The nucleation of the hematite nucleus happens more easily when the solution pH was less than 10. The optimum parameters identified for the rapid biosynthesis of hematite nanoparticles were pH9, 37°C (temperature) and 1mM ferric chloride as precursor. The particles were well crystallized hexagonal structured hematite nanoparticles and are predominantly (110)-oriented. The synthesized nanoparticles were found to contain predominantly iron (73.47%) and oxygen (22.58%) as evidenced by Energy Dispersive X-ray analysis. Hematite nanoparticles of 15–40nm diameters were biosynthesized in 48h under optimized conditions, compared to 21days before optimization.

Sunlight mediated synthesis of silver nanoparticles by a novel actinobacterium (Sinomonas mesophila MPKL 26) and its antimicrobial activity against multi drug resistant Staphylococcus aureus

Synthesis of silver nanoparticles using microorganism are many, but there are only scanty reports using actinobacteria. In the present study, the actinobacterium of the genus Sinomonas was reported to synthesis silver nanoparticles for the first time. A photo-irradiation based method was developed for the synthesis of silver nanoparticles, which includes two day old cultural supernatant of novel species Sinomonas mesophila MPKL 26 and silver nitrate solution, exposed to sunlight. The preliminary synthesis of silver nanoparticles was noted by the color change of the solution from colorless to brown; the synthesis was further confirmed using UV–visible spectroscopy which shows a peak between 400 and 450nm. Spherical shape silver nanoparticles of size range 4–50nm were synthesized, which were characterized using transmission electron microscopy. The Fourier transform infrared spectroscopy result indicates that, the metabolite produced by the novel species S. mesophila MPKL 26 was the probable reducing/capping agent involved in the synthesis of silver nanoparticles. The synthesized silver nanoparticles maintained consistent shape with respect to different time periods. The synthesized silver nanoparticles were evaluated for the antimicrobial activity against multi drug resistant Staphylococcus aureus which show good antimicrobial activity. The method developed for synthesis is easy, requires less time (20min) and produces spherical shape nanoparticles of size as small as 4nm, having good antimicrobial activity. Hence, our study enlarges the scope of actinobacteria for the rapid biosynthesis of silver nanoparticles and can be used in formulating remedies for multi drug resistant S. aureus.

Facile synthesis of silver nanoparticles using Euphorbia antiquorum L. latex extract and evaluation of their biomedical perspectives as anticancer agents

This study reveals the rapid biosynthesis of silver nanoparticles (EAAgNPs) using aqueous latex extract of Euphorbia antiquorum L as a potential bioreductant. Synthesized EAAgNPs generate the surface plasmonic resonance peak at 438nm in UV–Vis spectrophotometer. Size and shape of EAAgNPs were further characterized through transmission electron microscope (TEM) which shows well-dispersed spherical nanoparticles with size ranging from 10 to 50nm. Energy dispersive X-ray spectroscopic analysis (EDAX) confirms the presence of silver (Ag) as the major constituent element. X-ray diffraction (XRD) pattern of EAAgNPs corresponding to (111), (200), (220) and (311) planes, reveals that the generated nanoparticles were face centered cubic crystalline in nature. Interestingly, fourier-transform infrared spectroscopy (FTIR) analysis shows the major role of active phenolic constituents in reduction and stabilization of EAAgNPs. Phyto-fabricated EAAgNPs exhibits significant antimicrobial and larvicidal activity against bacterial human pathogens as well as disease transmitting blood sucking parasites such as Culex quinquefasciatus and Aedes aegypti (IIIrd instar larvae). On the other hand, in vitro cytotoxicity assessment of bioformulated EAAgNPs has shown potential anticancer activity against human cervical carcinoma cells (HeLa). The preliminary biochemical (MTT assay) and microscopic studies depict that the synthesized EAAgNPs at minimal dosage (IC50=28μg) triggers cellular toxicity response. Hence, the EAAgNPs can be considered as an environmentally benign and non-toxic nanobiomaterial for biomedical applications.

Extracellular Biofabrication, Characterization, and Antimicrobial Efficacy of Silver Nanoparticles Loaded on Cotton Fabrics Using Newly IsolatedStreptomycessp. SSHH-1E

Biological method for silver nanoparticles synthesis has been developed to obtain cost effective, clean, nontoxic, and ecofriendly size-controlled nanoparticles. The objective of this study is extracellular biosynthesis of antimicrobial AgNPs using cell-free supernatant of a localStreptomycessp. strain SSHH-1E. Different medium composition and fermentation conditions were screened for maximal AgNPs biosynthesis using Plackett-Burman experimental design and the variables with statistically significant effects were selected to study their combined effects and to find out the optimum values using a Box-Behnken design. The synthesized AgNPs were characterized using UV-visible spectroscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, and energy dispersive X-ray spectroscopy. Rapid biosynthesis of AgNPs was achieved by addition of 1 mM AgNO3solution to the cell-free supernatant. The produced particles showed a single surface plasmon resonance peak at 400 nm by UV-Vis spectroscopy which confirmed the presence of AgNPs.Streptomycessp. SSHH-1E was identified asStreptomyces narbonensisSSHH-1E. Transmission electron microscopy study indicated that the shape of AgNPs is spherical and the size is ranging from 20 to 40 nm. Fourier transform infrared spectroscopy analysis provides evidence for proteins as possible reducing and capping agents. Furthermore, the biosynthesized AgNPs significantly inhibited the growth of medically important pathogenic Gram-positive and Gram-negative bacteria and yeast. The maximum biosynthesis of AgNPs was achieved at initial pH of 8, peptone of 0.5 g, and inoculum age of 48 h. The statistical optimization resulted in a 4.5-fold increase in the production of AgNPs byStreptomyces narbonensisSSHH-1E.

Rapid Biosynthesis of Gold Nanoparticles by the Extracellular Secretion ofBacillus niabensis45: Characterization and Antibiofilm Activity

The present study demonstrated that the extracellular biosynthesis of gold nanoparticles (GNPs) usingB. niabensis45 may be mediated by a cyclic peptide (P2). The molecular weight of P2 was determined to be about 1122 Da by MALDI-TOF-MS and ESI-MS. A novel protocol for rapid biosynthesis of GNPs using P2 was developed. The results showed that GNP synthesis could be completed in a wide range of temperatures (40–100°C) and pH (6.0–10.0) within few minutes when 9 mL of P2 (2 mg/mL) and 1 mL of HAuCl4solution (2 mM) were mixed together. The synthesized GNPs were further characterized. Energy dispersive spectroscopy (EDS) and X-ray diffraction (XRD) analysis confirmed the presence of elemental gold and crystalline structure of the GNPs, respectively. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) revealed the formation of spherical metallic GNPs. The size distribution of GNPs calculated using ImageJ software was found to be 10–20 nm. And these GNPs showed excellent antibiofilm activity againstPseudomonas aeruginosaPAO1 andStaphylococcus aureusATCC25923. The results revealed microbial cyclic peptides could be used as synthesis of GNPs which had potent antibiofilm potential.

Green Synthesis and Antibacterial Activities of Silver Nanoparticles Using Extracellular Laccase of <i>Lentinus edodes</i>

This study reports the multi-step mutagenesis of Lentinus edodes towards optimization of the production of laccase and novel application of laccase in the biosynthesis of silver nanoparticles (AgNPs) which could be used to develop an eco-friendly method for the rapid biosynthesis of AgNPs. The wild strain of L. edodes was subjected to UV irradiation at 254 nm and the resultant viable mutant was further treated with acridine orange, a chemical mutagen. The strains were evaluated for the production of laccase and the crude laccase of the UV mutant (UV10) was used for the green synthesis of AgNPs. The particles were characterized by UV-Visible spectroscopy, Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM). Laccase activities of wild, UV10 and UV10ACR8 strains of L. edodes were obtained as 2.6, 10.6 and 2.8 U/ml/min respectively after 7 days of fermentation, showing laccase yield improvement of 4.08-fold for UV10 mutant. UV-Visible spectroscopy indicated the formation of AgNPs at absorption band of 430 nm. FTIR result indicated that proteins were responsible for AgNP synthesis, while SEM analysis confirmed the formation of walnut-shaped nanoparticles with size range of 50-100 nm. The biosynthesized nanoparticles revealed effective inhibition against clinical isolates of Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae. To the best of the authors’ knowledge, this result represents the first report on the biosynthesis of AgNPs using L. edodes metabolite. The report adds to the growing relevance of L. edodes as potential industrially viable organism, used for diverse biotechnological applications.

Green Synthesis and Antibacterial Activities of Silver Nanoparticles Using Extracellular Laccase of <i>Lentinus edodes</i>

This study reports the multi-step mutagenesis of Lentinus edodes towards optimization of the production of laccase and novel application of laccase in the biosynthesis of silver nanoparticles (AgNPs) which could be used to develop an eco-friendly method for the rapid biosynthesis of AgNPs. The wild strain of L. edodes was subjected to UV irradiation at 254 nm and the resultant viable mutant was further treated with acridine orange, a chemical mutagen. The strains were evaluated for the production of laccase and the crude laccase of the UV mutant (UV10) was used for the green synthesis of AgNPs. The particles were characterized by UV-Visible spectroscopy, Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM). Laccase activities of wild, UV10 and UV10ACR8 strains of L. edodes were obtained as 2.6, 10.6 and 2.8 U/ml/min respectively after 7 days of fermentation, showing laccase yield improvement of 4.08-fold for UV10 mutant. UV-Visible spectroscopy indicated the formation of AgNPs at absorption band of 430 nm. FTIR result indicated that proteins were responsible for AgNP synthesis, while SEM analysis confirmed the formation of walnut-shaped nanoparticles with size range of 50-100 nm. The biosynthesized nanoparticles revealed effective inhibition against clinical isolates of Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae. To the best of the authors’ knowledge, this result represents the first report on the biosynthesis of AgNPs using L. edodes metabolite. The report adds to the growing relevance of L. edodes as potential industrially viable organism, used for diverse biotechnological applications.

Precursor-Directed Combinatorial Biosynthesis of Cinnamoyl, Dihydrocinnamoyl, and Benzoyl Anthranilates in Saccharomyces cerevisiae.

Biological synthesis of pharmaceuticals and biochemicals offers an environmentally friendly alternative to conventional chemical synthesis. These alternative methods require the design of metabolic pathways and the identification of enzymes exhibiting adequate activities. Cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates are natural metabolites which possess beneficial activities for human health, and the search is expanding for novel derivatives that might have enhanced biological activity. For example, biosynthesis in Dianthus caryophyllus is catalyzed by hydroxycinnamoyl/benzoyl-CoA:anthranilate N-hydroxycinnamoyl/ benzoyltransferase (HCBT), which couples hydroxycinnamoyl-CoAs and benzoyl-CoAs to anthranilate. We recently demonstrated the potential of using yeast (Saccharomyces cerevisiae) for the biological production of a few cinnamoyl anthranilates by heterologous co-expression of 4-coumaroyl:CoA ligase from Arabidopsis thaliana (4CL5) and HCBT. Here we report that, by exploiting the substrate flexibility of both 4CL5 and HCBT, we achieved rapid biosynthesis of more than 160 cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates in yeast upon feeding with both natural and non-natural cinnamates, dihydrocinnamates, benzoates, and anthranilates. Our results demonstrate the use of enzyme promiscuity in biological synthesis to achieve high chemical diversity within a defined class of molecules. This work also points to the potential for the combinatorial biosynthesis of diverse and valuable cinnamoylated, dihydrocinnamoylated, and benzoylated products by using the versatile biological enzyme 4CL5 along with characterized cinnamoyl-CoA- and benzoyl-CoA-utilizing transferases.

EKylation: Addition of an Alternating-Charge Peptide Stabilizes Proteins.

For nearly 40 years, therapeutic proteins have been stabilized by chemical conjugation of polyethylene glycol (PEG), but recently zwitterionic materials have proved to be a more effective substitute. In this work, we demonstrate that genetic fusion of alternating-charge extensions consisting of anionic glutamic acid (E) and cationic lysine (K) is an effective strategy for protein stabilization. This bioinspired "EKylation" method not only confers the stabilizing benefits of poly(zwitterions) but also allows for rapid biosynthesis of target constructs. Poly(EK) peptides of different predetermined lengths were appended to the C-terminus of a native β-lactamase and its destabilized TEM-19 mutant. The EK-modified enzymes retained biological activity and exhibited increased stability to environmental stressors such as high temperature and high-salt solutions. This one-step strategy provides a broadly applicable alternative to synthetic polymer conjugation that is biocompatible and degradable.

Rapid biosynthesis of silver nanoparticles using Crotalaria verrucosa leaves against the dengue vector Aedes aegypti: what happens around? An analysis of dragonfly predatory behaviour after exposure at ultra-low doses

Aedes aegypti is a primary vector of dengue, a mosquito-borne viral disease infecting 50–100 million people every year. Here, we biosynthesised mosquitocidal silver nanoparticles (AgNP) using the aqueous leaf extract of Crotalaria verrucosa. The green synthesis of AgNP was studied by UV–vis spectroscopy, SEM, EDX and FTIR. C. verrucosa-synthesised AgNPs were toxic against A. aegypti larvae and pupae. LC50 of AgNP ranged from 3.496 ppm (I instar larvae) to 17.700 ppm (pupae). Furthermore, we evaluated the predatory efficiency of dragonfly nymphs, Brachydiplax sobrina, against II and III instar larvae of A. aegypti in an aquatic environment contaminated with ultra-low doses of AgNP. Under standard laboratory conditions, predation after 24 h was 87.5% (II) and 54.7% (III). In an AgNP-contaminated environment, predation was 91 and 75.5%, respectively. Overall, C. verrucosa-synthesised AgNP could be employed at ultra-low doses to reduce larval population of dengue vectors enhancing predation rates of dragonfly nymphs.