The antibiofilm potential of a heteropolysaccharide produced and characterized from the isolated marine bacterium Glutamicibacter nicotianae BPM30.

Biofilms are the significant causes of 80% of chronic infections in the oral cavity, urinary tract, biliary tube, lungs, gastrointestinal tract, and so on to the general public. Treatment of pathogenic biofilm using bacterial exopolysaccharides (EPS) is an effective and promising strategy. In the present work, a marine bacterium was isolated, studied for exopolysaccharide production, and tested for its antibiofilm activity. Approximately 1.31 ± 0.07 g/L of a purified extracellular polysaccharide was produced and characterized from the isolated marine bacterium Glutamicibacter nicotianae BPM30. The hydrolyzed EPS contains multiple monosaccharides such as rhamnose, fructose, glucose, and galactose. The EPS demonstrated potential antibiofilm activity on four tested pathogens in a concentration-dependent mode. The antibiofilm activity of the purified EPS was studied by crystal violet assay and fluorescence staining method. Comparative inhibition results obtained for the tested strains are 93.25% ± 5.25 and 88.56% ± 2.25 for K. pneumoniae; 92.65% ± 7.6 and 98.33% ± 0.85 for P. aeruginosa; 90.36% ± 6.3 and 52.08% ± 7.74 for S. typhi; 84.62% ± 5.6 and 77.90% ± 5.90 for S. dysenteriae. The results of the present work demonstrated the antibiofilm potential of EPS, which could be helpful in the invention of novel curative approaches in battling bacterial biofilm-related medical complications.

Kinetics of dye decolorization using heterogeneous catalytic system with immobilized Achromobacter xylosoxidans DDB6.

Textile effluents containing toxic dyes must be treated effectively before discharge to prevent adverse environmental impacts. Traditional physical and chemical treatment methods are costly and generate secondary pollutants. In contrast, biological treatment is a more suitable, clean, versatile, eco-friendly, and cost-effective technique for treating textile effluent. It is well established that indigenous microbial populations present in effluents can effectively degrade toxic dyes. In this regard, Achromobacter xylosoxidans DDB6 was isolated from the effluent sample to decolorize crystal violet (CV), Coomassie brilliant blue (CBB), and alizarin red (AR) by 67.20%, 28.58%, and 20.41%, respectively. The growth parameters of A. xylosoxidans DDB6 in media supplemented with 100 ppm of various dyes were determined using the modified Gompertz growth model. The immobilized cells in calcium alginate beads showed apparent decolorization rate constant of 0.27, 0.18, and 0.13 h-1 for CV, CBB, and AR, respectively. The immobilized cells in a packed bed reactor with an optimum flow rate of 0.5 mL/min were used to treat 100 ppm of CV with a percentage decolorization of 79.47% after three cycles. Based on the findings, A. xylosoxidans DDB6 could be effectively used for decolorization of various dyes.

Heterogeneous biocatalytic system for effective decolorization of textile dye effluent.

The current physicochemical methods for decolorizing toxic synthetic dyes are not sustainable to halt the environmental damage as they are expensive and often produce concentrated sludge, which may lead to secondary disposal problems. Biocatalysis (microbes and/or their enzymes) is a cost-effective, versatile, energy-saving and clean alternative. The most common enzymes involved in dye degradation are laccases, azoreductases and peroxidases. Toxic dyes could be converted into less harmful byproducts through the combined action of many enzymes or the utilization of whole cells. The action of whole cells to treat dye effluents is either by biosorption or degradation (aerobic or anaerobic). Using immobilized cells or enzymes will offer advantages such as superior stability, persistence against harsh environmental conditions, reusability and longer half-lives. This review envisages the recent strategies of immobilization and bioreactor considerations with the immobilized system as the effective treatment of textile dye effluents. Packed bed reactors are the most popular heterogeneous biocatalytic reactors for dye decolorization due to their efficiency and cost-effectiveness.

Oceanimonas sp. BPMS22-derived protein protease inhibitor induces anti-leishmanial immune responses through macrophage M2 to M1 repolarization.

The present study aimed to validate the potential of a novel serine protein protease inhibitor (PPI), purified from marine Oceanimonas sp. BPMS22, induced M2 to M1 repolarization of the macrophages to treat visceral leishmaniasis (VL). Peptide mass fingerprint of the purified trypsin digested PPI peptide was obtained using matrix-assisted laser desorption ionization-time of flight combined with tandem mass spectrometry (MALDI-TOF MS/MS) and the sequence was used to construct a 3D protein model by homology modelling. The IC50 of PPI were 25.28 ± 1.675 µg/mL and 0.415 ± 0.015 µg/mL against promastigotes and intracellular amastigotes, respectively, indicating the host-directed therapy using PPI. The PPI enhanced the effector molecule i.e., nitric oxide (NO), and dampened the arginase activity in a dose-dependent manner. In vitro studies revealed that the BPMS22-derived PPI significantly (p < 0.05) decreased the mRNA expressions of M2 markers (FIZZ-1, YM-1, CD206, Arg-1) and increased the mRNA expressions of M1 markers (iNOS, IL-1ß, IL-12) in rIL-4 + rIL-10 induced M2 macrophages. Interestingly, the BPMS22-derived PPI also significantly (p < 0.05) decreased the FIZZ-1, YM-1, CD206, and Arg-1; significantly (p < 0.05) increased iNOS, IL-12, and IFN-γ mRNA expression in L. donovani -infected murine macrophages, alongside the decreased parasite load in it. Hence, PPI has the potential to repolarize the cytokines (rIL-4 + rIL-10) pre-stimulated and L. donovani-infected M2 macrophages to M1 phenotype in vitro. A decrease in parasite burden after treatment with PPI indicated the acceleration of the parasite killing by enhancing the macrophage effector functions. Further, in vivo PPI treatment reduced hepatic and splenic Leishman donovan units (LDU) up to 93.34 % and 87.63 %, respectively. This was followed by a surge in pro-inflammatory cytokines and dampening anti-inflammatory cytokines (p < 0.01), which exhibited anti-VL immunity. These observations might open new perspectives on PPI in macrophage repolarization to treat VL.

Maximizing the direct recovery and stabilization of cellulolytic enzymes from Trichoderma harzanium BPGF1 fermented broth using carboxymethyl inulin nanoparticles.

The carboxymethylated inulin (CMI) nanoparticles prepared by the salt out method was demonstrated to harvest cellulolytic enzymes (Ez) directly from the clarified fermented broth of Trichoderma harzanium BPGF1. The formation of CMI nanoparticles and entrapment of Ez in CMI was confirmed by scanning electron microscopy and Fourier transform infrared spectroscopy, respectively. A factorial design was developed to maximize enzymes recovery directly from the fermented broth. A maximum of 71.68 ± 8.61% cellulolytic enzymes was recovered using 20 mg/L inulin, 2 M sodium chloroacetate at 80 °C for 2 h. The resultant CMIEz nanohybrid displayed excellent activity in broad pH and temperature. Moreover, CMIEz was reusable for >30 cycles without losing efficiency. The real-time application of CMIEz was demonstrated by hydrolyzing acid pretreated corncob. High-pressure liquid chromatography revealed that the hydrolyzed corncob contained cellobiose, glucose, galactose, xylose, mannose, and arabinose. The results highlight that carbohydrate nanoparticles was useful in engulfing enzymes directly from the fermentation broth.

Potential Anticoagulant Activity of Trypsin Inhibitor Purified from an Isolated Marine Bacterium Oceanimonas Sp. BPMS22 and its Kinetics.

Protease inhibitors control major biological protease activities to maintain physiological homeostasis. Marine bacteria isolated from oligotrophic conditions could be taxonomically distinct, metabolically unique, and offers a wide variety of biochemicals. In the present investigation, marine sediments were screened for the potential bacteria that can produce trypsin inhibitors. A moderate halotolerant novel marine bacterial strain of Oceanimonas sp. BPMS22 was isolated, identified, and characterized. The effect of various process parameters like salt concentration, temperature, and pH was studied on the growth of the bacteria and production of trypsin inhibitor. Further, the trypsin inhibitor was purified to near homogeneity using anion exchange, size exclusion, and affinity chromatography. The purified trypsin inhibitor was found to competitively inhibit trypsin activity with an inhibition coefficient, Ki, of 3.44 ± 0.13 µM and second-order association rate constant, kass, of 1.08 × 103 M-1 S-1. The proteinaceous trypsin inhibitor had a molecular weight of approximately 30 kDa. The purified trypsin inhibitor showed anticoagulant activity on the human blood samples.

Modeling of growth kinetics for an isolated marine bacterium, Oceanimonas sp. BPMS22 during the production of a trypsin inhibitor.

Protease inhibitors significantly control physiologically relevant protease activities. Protease inhibitors from marine microbial sources are unique due to their rough living environmental conditions. In the present study, a protein protease inhibitor (PI) was produced from marine Oceanimonas sp. BPMS22. Seven different media were screened for the growth of the bacterium and production of PI. Different carbon and nitrogen sources were screened and optimized for the specific protease inhibitor activity. Three different growth models were checked for the best fit of the bacterial growth. A modified Gompertz model was selected as the best model for the growth of Oceanimonas sp. BPMS22 with the maximum specific growth rate of 0.165 hr-1 and doubling time of 4.2 hr. The production of PI takes place during the non-growing phase of the bacterial growth. A kinetic model for the production of PI during non-growing phase was used for studying various process parameters. From the model, the maximum trypsin inhibitor formation rate of 0.3802 IU per mg of biomass per hour was observed at 49.91 hr.

Microbial serine protease inhibitors and their therapeutic applications.

Serine protease inhibitors, inhibit serine proteases either partially or completely after forming complexes with their respective proteases. Protease actions are significant for many physiological pathways found in living forms and any anomalies may lead to numerous physiological complications. Each cell or organism has its own mechanism for controlling these protease actions. It is often regulated by the action of inhibitors or activators. Among the proteases, serine proteases are the most common that are involved in many life and death processes. Selective inhibitors of physiologically relevant proteases can be used as a lead compound for the drug development. Therefore, it is imperative to identify small peptides and proteins that selectively inhibit serine proteases from various sources. Microbes can be considered as a major source of diverse serine protease inhibitors since they have the prominent and diverse domain in nature. Most of the microbial serine protease inhibitors are intracellular and few are extracellular. Microbes produce protease inhibitors for protection against its own proteases or against other environmental factors. The status and future prospects of microbial serine protease inhibitors and their therapeutic benefits in treating cancer, blood coagulation disorders and viral infections, are reviewed here.

Immobilization of levan-xylanase nanohybrid on an alginate bead improves xylanase stability at wide pH and temperature.

Despite the sustainable availability, levan, a fructose based natural polysaccharide has not received significant attention in the development of enzyme immobilization technology. Herein, we prepared levan-xylanase (LXy) nanohybrid and characterized by scanning electron microscopy, particle size analyzer and zeta potential. To prevent the enzyme leakage from the nanohybrid, LXy was immobilized onto an alginate beads (NaAlg). Immobilization yield was optimized using a statistical method, central composite design. A maximum immobilization yield of 95.3% was achieved at 2.13% (w/v) of sodium alginate, 2.14% (w/v) of calcium chloride, 64min of curation time and 1.4mm bead size. Immobilized LXy retains nearly 80% of the enzyme activity at a wide range of temperature (20-90°C) and pH (3-10). Immobilization of LXy onto NaAlg increases the activation energy from 28.50Jmol-1K-1 to 39.38Jmol-1K-1. Collectively, this result implies that LXy immobilized onto NaAlg increases the enzyme stability and retains its activity.

Synthesis of fibrinolytic active silver nanoparticle using wheat bran xylan as a reducing and stabilizing agent.

A facile synthesis of highly stable silver nanoparticles (AgNPs) was reported using a biopolymer, xylan as both a reducing and stabilizing agent. Xylan was isolated from waste biomass, wheat bran (WB) by alkaline treatment and was characterized by Fehling's test, dinitrosalicylic acid assay, FTIR, (1)H NMR and (13)C NMR. The synthesized nanoparticles were characterized by UV-Vis spectroscopy and transmission electron microscopy. The nanoparticles were polydispersed with the size ranging from 20 to 45 nm. The synthesized WB-xylan AgNPs showed excellent free radical scavenging activity. In addition, WB-xylan AgNPs showed fibrinolytic activity as evidenced by the zone of clearance in fibrin plate assay. The biomedical potential of the WB-xylan AgNPs was demonstrated by dissolution of preformed blood clots. These results suggest that the development of xylan-metal nanoparticle composite would be feasible to treat thrombus related diseases.