Synthesis of mesoporous antimicrobial herbal nanomaterial-carrier for silver nanoparticles and antimicrobial sensing.

Herbal nanoparticles (HNPs) were introduced as a novel generation of antimicrobial nanoparticles. But in the battle against superbugs we need nanostructures with boosted antimicrobial potency. So in the current experiment, for the first time a green approach was developed for the silver functionalization of HNPs which were fabricated from an antimicrobial herb Thymus vulgaris. Silver functionalized HNPs (AgHNPs) were found to be mesoporous and were further fortified with antimicrobial compounds. The resulted structures were re-tested against MRSA and P. aeruginosa as superbugs. It was found that silver functionalization can provide eight-fold increase in the antimicrobial potency of HNPs. Moreover, MIC was reduced from 20 mg/mL to 2.5 mg/mL. Another eight-fold reduction in the MIC (0.3 mg/mL) was achieved by fortification with antimicrobial compounds. So, the antimicrobial potency of HNPs was successfully increase approximately up to 64-folds. Obtained results illustrated that silver functionalization and fortification with antimicrobial compounds can be considered as effective approaches to achieve HNPs with boosted antimicrobial potency. These nanostructures have the potency to be loaded with other antimicrobial compound such as antibiotics toward synergistic effects of AgNPs and antibiotics. Resulted nanostructures can be employed in the formulation of powerful topical and surface disinfectants against superbugs. Also, these particles can be considered as a next generation of boosted antimicrobial nanostructures.

Application of FeOOH Nano-Ellipsoids as a Novel Nano-Based Iron Supplement: an In Vivo Study.

The possibility of employing FeOOH nano-ellipsoids as a novel shape nano-based iron supplement was investigated. Ferrous sulfate and nano-ellipsoids were daily administered by gavage at low and high dosages. After 1 month of treatment, the hematologic parameters along with serum and organs' iron contents were measured. Liver enzymes, total serum bilirubin, and LDH level were assayed to evaluate any possible toxicity. More investigation was also performed by organ index calculation and also pathologic studies. It was found that nano-ellipsoids are an effective iron supplement to improve iron-related blood parameters. Interestingly, low-dose nano-ellipsoids were even more effective than high-dose ferrous sulfate. Nano-ellipsoids had no considerable impact on the liver enzymes and serum bilirubin. Meanwhile, high-dose ferrous sulfate significantly increases liver enzyme activity. The increased serum LDH was also the only concern in the groups that were treated with high-dose ferrous sulfate and nano-ellipsoids. Pathologic evaluations revealed some signs of liver inflammation after supplementation with high dose nano-ellipsoids and also ferrous sulfate. Overall, these data indicate FeOOH nano-ellipsoids as a novel shape iron supplement to be employed at low dosage but with greater beneficial effects than high-dose ferrous sulfate.

Probiotics/Prebiotics in Viral Respiratory Infections: Implication for Emerging Pathogens.

BACKGROUND: Viral respiratory infections could result in perturbation of the gut microbiota due to a probable cross-talk between lungs and gut microbiota. This can affect pulmonary health and the gastrointestinal system. OBJECTIVE: This review aimed to discuss the impact of probiotics/prebiotics and supplements on the prevention and treatment of respiratory infections, especially emerging pathogens. METHODS: The data were searched in PubMed, Scopus, Google Scholar, Google Patents, and The Lens-Patent using keywords of probiotics and viral respiratory infections in the title, abstract, and keywords. RESULTS: Probiotics consumption could decrease the susceptibility to viral respiratory infections, such as COVID-19 and simultaneously enhance vaccine efficiency in infectious disease prevention through the immune system enhancement. Probiotics improve the gut microbiota and the immune system via regulating the innate system response and production of anti-inflammatory cytokines. Moreover, treatment with probiotics contributes to intestinal homeostasis restitution under antibiotic pressure and decreasing the risk of secondary infections due to viral respiratory infections. Probiotics present varied performances in different conditions; thus, promoting their efficacy through combining with supplements (prebiotics, postbiotics, nutraceuticals, berberine, curcumin, lactoferrin, minerals, and vitamins) is important. Several supplements reported to enhance the probiotics' efficacy and their mechanisms as well as probiotics- related patents are summarized in this review. Using nanotechnology and microencapsulation techniques can also improve probiotics' efficiency. CONCLUSION: Given the global challenge of COVID-19, probiotic/prebiotic and following nutritional guidelines should be regarded seriously. Additionally, their role as an adjuvant in vaccination for immune response augmentation needs attention.

The effect of aeration and mixing in developing a dairy-based functional food rich in menaquinone-7.

Vitamin menaquinone-7 (MK-7) supplementation improves bone health and reduces the incidence of osteoporosis. Despite the recent developments in MK-7 fermentation using Bacillus subtilis natto, low fermentation yields, as well as complicated downstream processing steps, are still the main reasons for the expensive final product. To overcome these issues, developing a fermented dairy-based product rich in MK-7 by avoiding costly downstream steps and optimising the fermentation operating conditions to enhance the MK-7 concentration would be an alternative approach. The present study, therefore, aims to evaluate the role of agitation and aeration as the key operating conditions on MK-7 production by Bacillus subtilis natto using a milk media. The agitation and aeration rates of 525 RPM and 5 VVM were found to be the optimum levels leading to the production of 3.54 mg/L of MK-7. Further, the sensory evaluation was performed to compare the sensory properties of the freeze-dried fermented samples with non-fermented milk samples. The results illustrated that the fermented samples had a significant saltiness with intense aroma resulting in the less acceptability of them by the panellists.

Enterobacter sp. Mediated Synthesis of Biocompatible Nanostructured Iron-Polysaccharide Complexes: a Nutritional Supplement for Iron-Deficiency Anemia.

FDA has approved iron oxide nanoparticles (IONs) coated with organic compounds as a safe material with less toxic effects compared with the naked metal ions and nanoparticles. In this study, the biological and physicochemical characteristics of a nanostructured iron-polysaccharide complexes (Nano-IPC) biosynthesized by Enterobacter sp. were evaluated. Furthermore, the serum biochemical parameters, tissue iron level, red blood cell parameters, and organ ferritin of rats were measured for investigating the effect of the Nano-IPCs in comparison with FeSO4 as a supplement for iron deficiency. The biosafety data demonstrated 35% increment of viability in Hep-G2 hepatocarcinoma cell lines when treated with nanoparticles (500 µg/mL) for 24 h. Besides, iron concentration in serum and tissue as well as the expression of ferritin L subunit in animals treated with the Nano-IPCs supplement were meaningfully higher than the FeSO4-supplemented and negative control animals. Moreover, the expression level of ferritin H subunit and biochemical factors remained similar to the negative control animals in the Nano-IPC-supplemented group. These results indicated that Nano-IPCs can be considered as a nontoxic supplement for patients carrying iron-deficiency anemia (IDA).

Cis and trans isomers of the vitamin menaquinone-7: which one is biologically significant?

Recently, several studies have indicated that an adequate intake of menaquinone-7 (MK-7) offers numerous health benefits. However, the low availability of MK-7 in the diet necessitates the development of dietary supplements or functional food products to complement natural food sources and meet the daily intake requirements. Like most biological molecules, MK-7 can exist as geometric isomers that can occur in the cis, trans, and cis/trans forms; however, only the all-trans form is biologically significant. MK-7 is traditionally produced through bacterial fermentation, but various synthetic preparations have lately become available. The isomer composition in the final product is influenced by numerous factors, including the methods of production and purification, as well as particular environmental and storage conditions. The MK-7 profile obtained from the various production methods has not yet been elucidated, and the ideal method for the synthesis of the all-trans form of the vitamin is also debatable. Consequently, the quantification of the MK-7 profile of various products is necessary to develop an understanding of the factors that influence the proportion of isomers that are obtained in different preparations. Several possible methods exist for the quantification of MK-7 isomers, and of these, liquid chromatography in conjunction with mass spectrometry techniques appears to be the most promising. Evaluation of the isomer composition is an important consideration, as only the all-trans form sustains biological activity. Furthermore, knowledge of the prominent factors that influence the MK-7 composition may also enable their manipulation to obtain a more favorable MK-7 profile in the final product.

Controlled synthesis of iron oxyhydroxide (FeOOH) nanoparticles using secretory compounds from Chlorella vulgaris microalgae.

FeOOH nanoparticles are commonly synthesized at very high temperature and pressure that makes the process energy consuming and non-economic. Recently, novel approaches were developed for the fabrication of these particles at room temperature. But, the main problem with these methods is that the prepared structures are aggregates of ultra-small nanoparticles where no intact separate nanoparticles are formed. In this study, for the first time, secretory compounds from Chlorella vulgaris cells were employed for the controlled synthesis of FeOOH nanoparticles at room atmosphere. Obtained particles were found to be goethite (α-FeO(OH)) crystals. Controlled synthesis of FeOOH nanoparticles resulted in uniform spherical nanoparticles ranging from 8 to 17 nm in diameter with 12.8 nm mean particle size. Fourier-transform infrared and elemental analyses were indicated that controlled synthesized nanoparticles have not functionalized with secretory compounds of C. vulgaris, and these compounds just played a controlling role over the synthesis reaction.

Biofilm reactors as a promising method for vitamin K (menaquinone-7) production.

Menaquinone-7 (MK-7) is the most potent subtype of vitamin K with extraordinarily high half-life in the circulatory system. Therefore, MK-7 plays a critical role in promoting human wellbeing today. Studies on MK-7 every year show more and more magnificent benefits of it in preventing cardiovascular diseases and osteoporosis to battling cancer cells, Alzheimer's and Parkinson's diseases. Thus, it needs to be supplemented to daily diet for accumulative and long-term benefits. Chemical synthesis of MK-7 produces a significant cis-isomer form of it, which has no biological activity. Fortunately, due to its key role in electron transfer in bacteria, trans-MK-7 is biosynthesized by especially Gram-positive strains mainly Bacillus genus. Concordantly, MK-7 could be produced via solid or liquid state fermentation strategies. In either regime, when static fermentation is applied in the absence of agitation and aeration, operational issues arise such as heat and mass transfer inefficiencies. Thus, scaling up the process becomes a challenge. On the other hand, studies have indicated that biofilm and pellicle formation that occur in static fermentations are key characteristics for extracellular MK-7 secretion. Therefore, this review covers the most recent discoveries of the therapeutic properties of MK-7 and optimization attempts at increasing its biosynthesis in different media compositions and effective growth parameters as well as the cutting-edge use of biofilm reactors where B. subtilis cells have the infrastructures to form mature biofilm formations on plastic composite supports. Biofilm reactors therefore can provide robust extracellular MK-7 secretion while simultaneously enduring high agitation and aeration rates, which then address the scale-up and operational issues associated with static fermentation strategies.

Xanthan Gum Capped ZnO Microstars as a Promising Dietary Zinc Supplementation.

Zinc is one of the essential trace elements, and plays an important role in human health. Severe zinc deficiency can negatively affect organs such as the epidermal, immune, central nervous, gastrointestinal, skeletal, and reproductive systems. In this study, we offered a novel biocompatible xanthan gum capped zinc oxide (ZnO) microstar as a potential dietary zinc supplementation for food fortification. Xanthan gum (XG) is a commercially important extracellular polysaccharide that is widely used in diverse fields such as the food, cosmetic, and pharmaceutical industries, due to its nontoxic and biocompatible properties. In this work, for the first time, we reported a green procedure for the synthesis of ZnO microstars using XG, as the stabilizing agent, without using any synthetic or toxic reagent. X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM) were used to study the structure, morphology, and size of the synthesized ZnO structures. The results showed that the synthesized structures were both hexagonal phase and starlike, with an average particle size of 358 nm. The effect of different dosages of XG-capped ZnO nanoparticles (1⁻9 mM) against Gram-negative (Escherichia coli) and Gram-positive (Bacillus licheniformis, Bacillus subtilis, and Bacillus sphaericus) bacteria were also investigated. Based on the results, the fabricated XG-capped ZnO microstars showed a high level of biocompatibility with no antimicrobial effect against the tested microorganisms. The data suggested the potential of newly produced ZnO microstructures for a range of applications in dietary supplementation and food fortification.

Extracellular Production of a Potent and Chemically Resistant Nattokinase in Immobilized Escherichia coli Using Response Surface Methodology.

BACKGROUND: Nattokinase is a potent fibrinolytic protease, which is used as a drug for treatment or a supplement for preventing thrombosis besides various industrial applications. The present study aimed to produce a soluble nattokinase in low cost media, which has with high activity and resistance to metal ions, detergents, and organic solvents, and can be easily used in medicines or as detergents. METHODS: Generally, most of the native extracellular proteins, such as nattokinase from Bacillus subtilis, are lysed by secretory proteases. One way for solving this problem is to employ other hosts for nattokinase production. For producing secretory form of nattokinase from Bacillus subtilis, different factors such as a suitable host, optimized media for the maximum enzyme activity and easy purification are important. RESULTS: These factors are studied in this investigation. Escherichia coli BL21 (DE3), as a reliable host was selected for a high yield production of an extracellular recombinant nattokinase. A mature nattokinase gene from Bacillus subtilis 1023, was cloned in the expression vector pET22b by which the host was transformed. The recombinant nattokinase was expressed through induction with IPTG. The expressed protein was confirmed by SDS-PAGE, and its fibrinolytic activity was assayed on the fibrin plates. Afterwards, the enzyme was purified by Ni-NTA native affinity column. Different media components were evaluated for maximum nattokinase production and activity. The highest enzyme activity of 883.107 U/ml was obtained, when a medium approximately consisting of yeast extract (4.38 g/L), tryptone (4 g/L), K2HPO4 (1.61 g/L) and CaCl2 (0.01 g/L) was used. CONCLUSION: Entrapping the transformed host in calcium alginate could lead to more enzyme activity and decrease media cost.

Bio-reinforced self-healing concrete using magnetic iron oxide nanoparticles.

Immobilization has been reported as an efficient technique to address the bacterial vulnerability for application in bio self-healing concrete. In this study, for the first time, magnetic iron oxide nanoparticles (IONs) are being practically employed as the protective vehicle for bacteria to evaluate the self-healing performance in concrete environment. Magnetic IONs were successfully synthesized and characterized using different techniques. The scanning electron microscope (SEM) images show the efficient adsorption of nanoparticles to the Bacillus cells. Microscopic observation illustrates that the incorporation of the immobilized bacteria in the concrete matrix resulted in a significant crack healing behavior, while the control specimen had no healing characteristics. Analysis of bio-precipitates revealed that the induced minerals in the cracks were calcium carbonate. The effect of magnetic immobilized cells on the concrete water absorption showed that the concrete specimens supplemented with decorated bacteria with IONs had a higher resistance to water penetration. The initial and secondary water absorption rates in bio-concrete specimens were 26% and 22% lower than the control specimens. Due to the compatible behavior of IONs with the concrete compositions, the results of this study proved the potential application of IONs for developing a new generation of bio self-healing concrete.

Plant-Mediated Synthesis and Applications of Iron Nanoparticles.

Nanoscale iron particles have attracted substantial interest due to their unique physical and chemical properties. Over the years, various physical and chemical methods have been developed to synthesize these nanostructures which are usually expensive and potentially harmful to human health and the environment. Synthesis of iron nanoparticles (INPs) by using plant extract is now of great interest in order to develop a novel and sustainable approach toward green chemistry. In this method the chemical compounds and organic solvents are replaced with phytochemicals and aqueous matrixes, respectively. Similar to any chemical and biochemical reaction, factors such as reaction temperature, concentration of iron precursor, concentration of leaf extract, and reaction time have critical effects on the reaction yield. This review focuses on the novel approaches used for green synthesis of INPs by using plant resources. The currently available statistics including the factors affecting the synthesis process and potential applications of the fabricated nanoparticles are discussed. Recommendations are also given for areas of future research in order to improve the production process.

Optimization of Bacillus subtilis natto growth parameters in glycerol-based medium for vitamin K (Menaquinone-7) production in biofilm reactors.

Menaquinone-7 (MK-7) is the key form of vitamin K used as a dietary supplement and its production revolves around Bacillus subtilis natto. Current fermentation strategies, which suggest static fermentations without aeration and agitation, can be problematic for large scale MK-7 production due to biofilm formation. The use of biofilm reactors, therefore, is proposed in the present study, which could utilize both agitation and aeration without interrupting MK-7 secretion. In this study, biofilm reactors were constructed using the selected plastic composite support (PCS) and B. subtilis natto strain for MK-7 production. Using response surface methodology (RSM), optimum growth parameters including temperature, pH, and agitation were determined in a glycerol-based medium. Results were presented in a statistical model (R 2 = 0.90), leading to optimum growth conditions of temperature (35 °C), agitation (200 rpm) and pH (6.58). Model-predicted MK-7 concentration was validated and MK-7 concentration of 12.09 mg/L was produced in the biofilm reactor. The obtained concentration was 58% higher as compared to the suspended-cell culture (7.67 mg/L). The results of this study will provide a critical step towards improved industrial scale production of MK-7.

Iron oxide nanoparticles in modern microbiology and biotechnology.

Iron oxide nanoparticles (IONs) are one of the most developed and used nanomaterials in biotechnology and microbiology. These particles have unique physicochemical properties, which make them unique among nanomaterials. Therefore, many experiments have been conducted to develop facile synthesis methods for these particles and to make them biocompatible. Various effects of IONs on microorganisms have been reported. Depending on the microbial strain and nanoparticle (NP) concentration, IONs can stimulate or inhibit microbial growth. Due to the superparamagnetic properties of IONs, these NPs have used as nano sources of heat for hyperthermia in infected tissues. Antibiotic-loaded IONs are used for targeted delivery of chemical therapy direct to the infected organ and IONs have been used as a dirigible carrier for more potent antimicrobial nanomaterials such as silver nanoparticles. Magnetic NPs have been used for specific separation of pathogen and non-pathogen bacterial strains. Very recently, IONs were used as a novel tool for magnetic immobilization of microbial cells and process intensification in a biotechnological process. This review provides an overview of application of IONs in different microbial processes. Recommendations are also given for areas of future research.

Vitamin K series: current status and future prospects.

The Vitamin K series, particularly menaquinone, have been attracting research attention, due to the potential in reducing both osteoporosis and cardiovascular diseases. This review provides an overview of the types of vitamin K and their health benefits. This is followed by a critical review of the various biotechnological approaches used in the production of menaquinone, including solid and liquid state fermentations, extraction and recovery. The currently available market information is summarized and future growth prospects are discussed. Recommendations are also given for areas of future research in order to improve the production process for menaquinone and reduce costs.

Designing of an intensification process for biosynthesis and recovery of menaquinone-7.

A nutritional food rich in menaquinone-7 has a potential in preventing osteoporosis and cardiovascular diseases. The static fermentation of Bacillus subtilis natto is widely regarded as an optimum process for menaquinone-7 production. The major issues for the bulk production of menaquinone-7 are the low fermentation yield, biofilm formation and the use of organic solvents for the vitamin extraction. In this study, we demonstrate that the dynamic fermentation involving high stirring and aeration rates enhances the yield of fermentation process significantly compared to static system. The menaquinone-7 concentration of 226 mg/L was produced at 1,000 rpm, 5 vvm, 40 °C after 5 days of fermentation. This concentration is 70-fold higher than commercially available food products such as natto. Additionally, it was found that more than 80% of menaquinone-7 was recovered in situ in the vegetable oil that was gradually added to the system as an anti-foaming agent. The intensification process developed in this study has a capacity to produce an oil rich in menaquinone-7 in one step and eliminate the use of organic solvents for recovery of this compound. This oil can, therefore, be used for the preparation of broad range of supplementary and dietary food products rich in menaquinone-7 to reduce the risk of osteoporotic fractures and cardiovascular diseases.