Expression and characterization of heparinase II with MBP tag from a novel strain, Raoultella NX-TZ-3-15.

The enzymes are biological macromolecules that biocatalyze certain biochemical reactions without undergoing any modification or degradation at the end of the reaction. In this work, we constructed a recombinant novel Raoultella sp. NX-TZ-3-15 strain that produces heparinase with a maltose binding tag to enhance its production and activity. Additionally, MBP-heparinase was purified and its enzymatic capabilities are investigated to determine its industrial application. Moreover, the recombinant plasmid encoding the MBP-heparinase fusion protein was effectively generated and purified to a high purity. According to SDS-PAGE analysis, the MBP-heparinase has a molecular weight of around 70 kDa and the majority of it being soluble with a maximum activity of 5386 U/L. It has also been noted that the three ions of Ca2 + , Co2 + , and Mg2 + can have an effect on heparinase activities, with Mg2 + being the most noticeable, increasing by about 85%, while Cu2 + , Fe2 + , Zn2 + having an inhibitory effect on heparinase activities. Further investigations on the mechanistic action, structural features, and genomes of Raoultella sp. NX-TZ-3-15 heparinase synthesis are required for industrial-scale manufacturing.

Cloning and Expression of Heparinase Gene from a Novel Strain Raoultella NX-TZ-3-15.

Heparin is a class of highly sulfated, acidic, linear, and complex polysaccharide that belongs to the heparin/heparan sulfate (HS) glycosaminoglycans family. Enzymatic depolymerization of heparin by heparinases is a promising strategy for the production of ultra-low molecular weight heparins (ULMWHs) as anticoagulants. In the present study, a novel heparinase-producing strain Raoultella NX-TZ-3-15 was isolated and identified from soil samples. Herein, the heparinase gene MBP-H1 was cloned to the pBENT vector to enable expression in Escherichia coli. The optimized conditions made the activity of recombinant heparinase reach the highest level (2140 U/L). The overexpressed MBP-H1 was purified by affinity chromatography and a purity of more than 90% was obtained. The condition for biocatalysis was also optimized and three metal ions Ca2+, Co2+, and Mg2+ were utilized to activate the reaction. In addition, the kinetics regarding the new fusion heparinase was also determined with a Vm value of 11.29 µmol/min and a Km value of 31.2 µmol/L. In short, due to excellent Km and Vmax, the recombinant enzyme has great potential to be used in the clinic in medicine and industrial production of low or ultra-low molecule weight heparin.

Effect of Squid Cartilage Chitosan Molecular Structure on the Properties of Its Monofilament as an Absorbable Surgical Suture.

Suture is an important part of surgery, and wounds closing after surgery remains a challenge for postoperative care. Currently, silk, linen fiber, and cotton are available in the market as non-absorbable suture biomaterials. So, there is an urgent need to develop a novel suture with advantageous characteristics compared to the ones available on the market. In present study, a series of ultra-high molecular weight chitosan with different DD and MV were prepared from squid cartilage by alkaline treatment and ultrasonic degradation. The corresponding chitosan monofilaments were prepared by a wet spinning process and were characterized as sutures. The effects of the DD and MV of chitosan on the properties of its monofilament were studied, including surface morphology, mechanical property, swelling ratio, ash content, in vitro enzymatic degradation, and in vitro cytotoxicity. According to the results, AS-85 was chosen to be the best suitable as an absorbable surgical suture, which was spun from squid cartilage chitosan with DD~85% and MV~1.2 × 106. The outcome of the present study might derive tremendous possibilities for the utilization of squid cartilage ß-chitin for biomedical applications.

Immunomodulation Potential of Probiotics: A Novel Strategy for Improving Livestock Health, Immunity, and Productivity.

Over the past decade, the use of probiotics as feed supplements in animal production has increased considerably due to the ban on antibiotic growth promoters in livestock. This review provides an overview of the current situation, limitation, and prospects for probiotic formulations applied to livestock. Recently, the use of probiotics in livestock has been suggested to significantly improve their health, immunity, growth performance, nutritional digestibility, and intestinal microbial balance. Furthermore, it was reported that the use of probiotics in animals was helpful in equilibrating their beneficial microbial population and microbial turnover via stimulating the host immune response through specific secretions and competitive exclusion of potentially pathogenic bacteria in the digestive tract. Recently, there has been great interest in the understanding of probiotics targeted diet and its ability to compete with harmful microbes and acquire their niches. Therefore, the present review explores the most commonly used probiotic formulations in livestock feed and their effect on animal health. In summary, this article provides an in-depth knowledge about the formulation of probiotics as a step toward a better alternative to antibiotic healthy growth strategies.

Techno-functional properties and immunomodulatory potential of exopolysaccharide from Lactiplantibacillus plantarum MM89 isolated from human breast milk.

An exopolysaccharide, designated as MM89-EPS, was isolated from Lactiplantibacillus plantarum MM89. It was comprised of glucose and mannose molecules with an average molecular weight of 138 kDa. FTIR and NMR spectra showed that MM89-EPS had characteristic polysaccharide functional groups. MM89-EPS displayed excellent water solubility and capacities to retain water and oil due to its porous structure. MM89-EPS exhibited no significant cytotoxicity on RAW264.7 cells and showed strong immunomodulatory activity by increasing phagocytosis, acid phosphatase activity, and cytokine production in RAW264.7 cells. Furthermore, an in vivo study revealed that splenic indices, intestinal IgA levels, serum cytokine levels, and lymphocyte proliferation were increased in an MM89-EPS-treated cyclophosphamide-induced immunosuppressed mouse model. To summarize, our results indicate that MM89-EPS can efficiently enhance the immunostimulatory activity of immune cells and an immunosuppressed mouse model. Hence, MM89-EPS may be use as a potential source of immunomodulatory agent in various food products.

Cyanobacteria derived compounds: Emerging drugs for cancer management.

The wide diversity of cyanobacterial species and their role in a variety of biological activities have been reported in the previous few years. Cyanobacteria, especially from marine sources, constitutes a major source of biologically active metabolites that have gained great attention especially due to their anticancer potential. Numerous chemically diverse metabolites from various cyanobacterial species have been recognized to inhibit the growth and progression of tumor cells through the induction of apoptosis in many different types of cancers. These metabolites activate the apoptosis in the cancer cells by different molecular mechanisms, however, the dysregulation of the mitochondrial pathway, death receptors signaling pathways, and the activation of several caspases are the crucial mechanisms that got considerable interest. The array of metabolites and the range of mechanisms involved may also help to overcome the resistance acquired by the different tumor types against the ongoing therapeutic agents. Therefore, the primary or secondary metabolites from the cyanobacteria as well as their synthetic derivates could be used to develop novel anticancer drugs alone or in combination with other chemotherapeutic agents. In this study, we have discussed the role of cyanobacterial metabolites in the induction of cytotoxicity and the potential to inhibit the growth of cancer cells through the induction of apoptosis, cell signaling alteration, oxidative damage, and mitochondrial dysfunctions. Moreover, the various metabolites produced by cyanobacteria have been summarized with their anticancer mechanisms. Furthermore, the ongoing trials and future developments for the therapeutic implications of these compounds in cancer therapy have been discussed.

Role of Food Antioxidants in Modulating Gut Microbial Communities: Novel Understandings in Intestinal Oxidative Stress Damage and Their Impact on Host Health.

Dietary components have an important role on the structure and function of host gut microbial communities. Even though, various dietary components, such as carbohydrates, fats, proteins, fibers, and vitamins, have been studied in depth for their effect on gut microbiomes, little attention has been paid regarding the impact of several food antioxidants on the gut microbiome. The long-term exposure to reactive oxygen species (ROS) can cause microbial dysbiosis which leads to numerous intestinal diseases such as microbiota dysbiosis, intestinal injury, colorectal cancers, enteric infections, and inflammatory bowel diseases. Recently, it has been shown that the food derived antioxidant compounds might protect the host from intestinal oxidative stress via modulating the composition of beneficial microbial species in the gut. The present review summarizes the impact of food antioxidants including antioxidant vitamins, dietary polyphenols, carotenoids, and bioactive peptides on the structure as well as function of host gut microbial communities. Several in vitro, animal model, and clinical studies indicates that food antioxidants might modify the host gut microbial communities and their health status. However, still further clarification is needed as to whether changes in certain microbial species caused by food additives may lead to changes in metabolism and immune function.

An overview of chia seed (Salvia hispanica L.) bioactive peptides' derivation and utilization as an emerging nutraceutical food.

Chia (S. hispanica L.) is an annual herbaceous plant that has gained popularity for its seeds of high-quality vegetative proteins, richest contents of omega-3 polyunsaturated fatty acids (Ω-3 PUFA), soluble dietary fiber, and great gelling ability, as well as its high contents of bioactive peptides of antioxidative and nutraceutical potential for many other clinical biomarkers. Such health protective bioactive peptides should be utilized for supplementation in the food and nutrition industries. This review was therefore designed to align the researches done on chia bioactive peptide's derivation, processing, consumption and to identify their antioxidative and nutraceutical potential for various disease biomarkers. The evidence gathered is fairly compelling for the health-promising nutraceutical and clinical potential of chia seed bioactive peptides as antioxidants, dipeptidyl peptidase-IV inhibitors (DPP4), angiotensin-converting enzyme (ACE) inhibitors, and anti-inflammatory drugs. Their assimilation into everyday diets has the potential to open new doors in health departments and food sectors.

Jawbones Scaffold Constructed by TGF-ß1 and BMP-2 Loaded Chitosan Microsphere Combining with Alg/HA/ICol for Osteogenic-Induced Differentiation.

Bone scaffolds based on multi-components are the leading trend to address the multifaceted prerequisites to repair various bone defects. Chitosan is the most useable biopolymer, having excellent biological applications. Therefore, in the present study, the chitosan microsphere was prepared by the ion-gel method; transforming growth factor ß (TGF-ß1) and bone morphogenetic protein 2 (BMP-2) were loaded onto it and then combined with alginate/hyaluronic acid/collagen (Alg/HA/ICol) to construct a jawbones scaffold. The Alg/HA/ICol scaffolds were characterized by FTIR and SEM, and the water content, porosity, tensile properties, biocompatibility, and osteogenic-induced differentiation ability of the Alg/HA/ICol jawbones scaffolds were studied. The results indicate that a three-dimensional porous jawbone scaffold was successfully constructed having 100-250 µm of pore size and >90% of porosity without cytotoxicity against adipose-derived stem cells (ADSCs). Its ALP quantification, osteocalcin expression, and Von Kossamineralized nodule staining was higher than the control group. The jawbones scaffold constructed by TGF-ß1 and BMP-2 loaded chitosan microsphere combining with Alg/HA/ICol has potential biomedical application in the future.

Therapeutic potential of Moringa oleifera seed polysaccharide embedded silver nanoparticles in wound healing.

Wound healing is a complicated process that influences patient's life quality. Plant-based polysaccharide has recently gained interest in its use in wound dressing materials because of its biological compatibility, natural abundance, and ideal physiochemical properties. The present study reveals the potential of polysaccharide isolated from Moringa oleifera seed (MOS-PS) and its nanocomposite with silver (MOS-PS-AgNPs) as alternative materials for wound dressing. First, MOS-PS was isolated and structurally characterized by TLC, HPLC, FTIR, NMR, and GPC analyses. A green and simple method was used to synthesize AgNPs using MOS-PS as a stabilizing and reducing agent. The size, morphology, and structure of the MOS-PS-AgNPs were characterized by UV-Vis spectroscopy, X-ray diffraction, scanning electron microscopy, transmission electron microscopy, and zeta potential analysis. The results showed that the MOS-PS-AgNPs were spherically shaped, having no cytotoxicity toward mouse fibroblasts cells and promoting their in-vitro migration. Moreover, the MOS-PS-AgNPs displayed strong anti-microbial activity against wound infectious pathogenic bacteria. Finally, the MOS-PS-AgNPs were used for dressing animal wounds and its preliminary mechanism was studied by RT-PCR and histological analysis. The results showed that the MOS-PS-AgNPs can promote wound contraction and internal tissue growth well. Overall, our results indicated that the MOS-PS-AgNPs might be an excellent candidate for use as an optimal wound dressing material.

Isolation and functional characterization of exopolysaccharide produced by Lactobacillus plantarum S123 isolated from traditional Chinese cheese.

During the past few years, there are growing interests in the potential use of exopolysaccharide (EPS) in the food industry as an efficient biopolymer because of its exceptional biological features. Therefore, the aim of the present study is EPS production by Lactobacillus Plantarum S123 (S123 EPS), its partial structural and biopotential characterization. The results from this study suggested that the major portion of S123 EPS has an amorphous sponge-like structure with partial crystalline nature. The FTIR and NMR results suggested that the S123 EPS consists of carbonyl and hydroxyl groups, respectively. Furthermore, the results of technological as well as biotechnological characterization suggested that the S123 EPS was exhibited excellent antibacterial activity against Gram-positive (7.2 mm) and Gram-negative bacteria (11.5 mm), DPPH radical scavenging activity (> 65%), water holding capacity (326.6 ± 0.5%), oil holding capacity (995.3 ± 0.2%), flocculation (89.5 ± 0.6%), and emulsifying (80.1 ± 1.1%) activities. Overall, the present results suggested that due to the highly porous structure and efficient biotechnological potential, S123 EPS from Lactobacillus plantarum S123 (L. plantarum S123) can be used in the functional food product.

Virucidal activity of Moringa A from Moringa oleifera seeds against Influenza A Viruses by regulating TFEB.

Influenza A viruses (IAVs) are highly contagious pathogens infecting human and numerous animals. The viruses cause millions of infection cases and thousands of deaths every year, making IAVs a continual threat to global health. Our study demonstrated the virucidal activity of Moringa A as a new compound from Moringa oleifera seeds against IAVs. It inhibits virus replication in host cells and protects infected cells from the cytopathic effect induced by IAVs. The EC50andEC90 values of Moringa A for IAVs were 1.27 and 5.30 µM, respectively, when RAW264.7 cells were infected at MOI of 1. The different treatment experiments revealed that Moringa A has a significant inhibitory effect on the IAVs both before and afterdrug addition. Moringa A was observed to decrease the inflammatory cytokines TNF-α, IL-6, IL-1ß, and IFN-ß in H1N1 infected RAW264.7 cells. Finally, Moringa A was found to inhibit the expression and nuclear transfer of the cellular protein transcription factor EB (TFEB) and weaken the autophagy in infected cells, which could be an important antiviral mechanism. Our study demonstrates Moringa A has potent antiviral activity against IVAs, which could be due to the autophagy inhibition property.

Chitin/chitosan derivatives and their interactions with microorganisms: a comprehensive review and future perspectives.

Chitosan, obtained as a result of the deacetylation of chitin, one of the most important naturally occurring polymers, has antimicrobial properties against fungi, and bacteria. It is also useful in other fields, including: food, biomedicine, biotechnology, agriculture, and the pharmaceutical industries. A literature survey shows that its antimicrobial activity depends upon several factors such as: the pH, temperature, molecular weight, ability to chelate metals, degree of deacetylation, source of chitosan, and the type of microorganism involved. This review will focus on the in vitro and in vivo antimicrobial properties of chitosan and its derivatives, along with a discussion on its mechanism of action during the treatment of infectious animal diseases, as well as its importance in food safety. We conclude with a summary of the challenges associated with the uses of chitosan and its derivatives.

Antibacterial and antioxidant activity of exopolysaccharide mediated silver nanoparticle synthesized by Lactobacillus brevis isolated from Chinese koumiss.

Recently, silver nanoparticles gain significant attention due to their applications in various fields. The aim of present study was to develop the eco-friendly, cost effective, and simple method to biosynthesized the silver nanoparticle using sliver nitrate as precursor. In this study, we investigated the physical characterization and biotechnological applications of biosynthesized silver nanoparticle using exopolysaccharide of probiotic Lactobacillus brevis MSR104 isolated from Chinese koumiss. Biosynthesized silver nanoparticles were characterized using the fourier-transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy, X-ray diffraction analysis, and elemental analyzer. The achieved results indicate that silver nanoparticles varied in sized with an average size of 45 nm. The X-ray diffraction analysis results showed that the silver nanoparticles have a crystalline nature. The results of antimicrobial assay indicated that the silver nanoparticles exhibited outstanding antimicrobial activity in dose dependent manner against both Gram's negative as well as Gram's positive. The antioxidant results indicate that the silver nanoparticles showed excellent scavenging rate against DPPH free radicals (81.4 ±â€¯1.2%) and nitric oxide free radicals (75.06 ±â€¯0.4%). Furthermore, the results of MTT assay revealed that the AgNPs significantly reduced the percentage of live HT-29 cells at higher concentration. This study concluded that the newly synthesized silver nanoparticles have antibacterial, antioxidant, and anticancer applications in agricultural and food industries.

Anti-tumor potential of cell free culture supernatant of Lactobacillus rhamnosus strains isolated from human breast milk.

Lactobacilli rhamnosus has been characterized as a probiotic and plays an important role in human health by stimulating the supplement of nutrients, preventing the colonization of pathogens, and influencing the immune system. This study investigated the anticancer activity of the three Lactobacillus rhamnosus strains SHA111, SHA112, and SHA113 isolated from human breast milk. The cell-free supernatant of a liquid culture of the three strains showed excellent antioxidant activities against DPPH free radicals, superoxide anion radicals, and hydroxyl radicals; furthermore, significant anticancer activity was found on cervix cancer cells (HeLa) via cytotoxicity and induction of apoptosis. RT-qPCR and western blot analysis showed the induction of apoptosis was achieved via the up-regulation of BAD, BAX, Caspase3, Caspase8, Caspase9, and down-regulation of BCL-2 genes in HeLa cells. The results suggest that these strains have potential anticancer capability.

Correction to: Characterization, the Antioxidant and Antimicrobial Activity of Exopolysaccharide Isolated from Poultry Origin Lactobacilli.

The original version of this article unfortunately contained mistakes. Replacements are needed on the following figures and captions.

Chitosan and its derivatives: synthesis, biotechnological applications, and future challenges.

Chitosan is a naturally occurring biodegradable as well as a non-toxic polymer generated from chitin through alkaline deacetylation reaction, and it is insoluble in organic/inorganic solvents and water. Furthermore, chitosan is one of the most plentiful cationic polymers in natural surroundings. Due to its non-toxicity and biocompatibility, chitosan is extensively employed in industrial, biomedical, food, pharmaceutical, environmental, and agricultural industry. Chitosan-based biomaterials exhibit great potential in various biotechnological applications, such as anti-hypertensive therapy, anti-oxidant, anti-microbial, anti-allergic, immunostimulant, cancer therapy, delivery of genetic materials, delivery of bone morphogenetic type-2, wound healing, treatment of wastewater, hypocholesterolemic, and bio-imaging. Therefore, this review mainly focuses on the biotechnological potential of chitosan and its derivatives as well as presents the potential of chitosan-based biomaterial/pharmaceutical for the prevention of various life-threating chronic disorders.

Characterization, the Antioxidant and Antimicrobial Activity of Exopolysaccharide Isolated from Poultry Origin Lactobacilli.

The natural antioxidant agent is urgently needed to prevent the negative effects of newly generated free radicals and chronic disorders. Recently, the microbial exopolysaccharide (EPS) is currently used as a potential biopolymer due to its unique biological characteristics. In this study, the biological potential was carried out on the EPSs produced by Lactobacillus reuteri SHA101 (EPS-lr) and Lactobacillus vaginalis SHA110 (EPS-lvg) isolated from gut cecum samples of healthy poultry birds (hen). As results, the EPS-lr and EPS-lvg showed the emulsifying activity of 37.8 ± 1.6% and 27.8 ± 0.5% after the 360 h, respectively. The scanning electron microscopy analysis of EPS-lr and EPS-lvg demonstrated a smooth surface with a compact structure. The both EPSs exhibited strong antibacterial activity against E. coli and Salmonella typhimurium in vitro. In additions, at 4 mg/mL concentration, the EPS-lr and EPS-lvg samples showed potent antioxidant activity regarding hydroxyl radical DPPH (2,2-diphenyl-1-picrylhydrazyl) radical, superoxide anion radical and reducing power at OD700 nm. Furthermore, the EPS-lr and EPS-lvg (600 µg/mL) possessed antitumor activity against colon cancer (Caco-2) cell after 72 h. The results suggested that these EPSs would have great potential in the application of antitumor and antioxidant foods, biomedicine, and pharmaceutics.