A facile strategy for preparation of Fe3O4 magnetic nanoparticles using Cordia myxa leaf extract and investigating its adsorption activity in dye removal.

This study demonstrates the successful, facile, and cost-effective preparation of magnetic Fe3O4 nanoparticles (MNPs) via green procedure using Cordia myxa leaf extracts for efficient adsorption of methylene blue (MB) as a model of organic pollutant. The formation of Fe3O4 NPs was confirmed by a range of spectroscopy and microscopy techniques including FT-IR, XRD, FE-SEM, TEM, EDS, VSM, TGA, and BET-BJH. The synthesized spherical nanoparticles had a high specific surface area of 115.07 m2/g with a mesoporous structure. The formed Fe3O4 MNPs exhibited superparamagnetic behavior with saturation magnetization of 49.48 emu/g. After characterization, the adsorptive performance of the synthesized MNPs toward MB was evaluated. To achieve the maximum removal efficiency, the effect of key parameters such as adsorbent dosage (MNPs), initial adsorbate concentration, pH, and contact time on the adsorption process was evaluated. A maximum adsorption capacity of 17.79 mg/g was obtained, after one-hour incubation at pH 7.5. From the pHPZC of 7.1 of the synthesized adsorbent, the electrostatic attraction between MB and Fe3O4 NPs plays an important role in the adsorption process. The adsorption experimental data showed the closest match with the pseudo-second-order kinetic and Langmuir isotherm. The prepared Fe3O4 NPs were easily recovered by an external magnet and could be reused several times. Therefore, the synthesized MNPs seem to be excellent adsorbents for the removal of MB from aqueous solution.

Immobilization of l-asparaginase on chitosan nanoparticles for the purpose of long-term application.

Asparaginase holds significant commercial value as an enzyme in the food and pharmaceutical industries. This study examined the optimum and practical use of the l-asparaginase derived from Pseudomonas aeruginosa HR03. Specifically, the study focused on the effectiveness of the stabilized enzyme when applied to chitosan nanoparticles. The structure, size, and morphology of chitosan nanoparticles were evaluated in relation to the immobilization procedure. This assessment involved the use of several analytical techniques, including FT-IR, DLS, SEM, TEM, and EDS analysis. Subsequently, the durability of the enzyme that has been stabilized was assessed by evaluating its effectiveness under extreme temperatures of 60 and 70 °C, as well as at pH values of 3 and 12. The findings indicate that incorporating chitosan nanoparticles led to enhanced immobilization of the l-asparaginase enzyme. This improvement was observed in terms of long-term stability, stability under crucial temperature and pH conditions, as well as thermal stability. In addition, the optimum temperature increased from 40 to 50 °C, and the optimum pH increased from 8 to 9. Enzyme immobilization led to an increase in Km and a decrease in kcat compared to its free counterpart. Because of its enhanced long-term stability, l-asparaginase immobilization on chitosan nanoparticles may be a potential choice for use in industries that rely on l-asparaginase enzymes, particularly the pharmaceutical and food industries.

Polyamidoamine Dendrimers Functionalized with ZnO-Chitosan Nanoparticles as an Efficient Surface for L-asparaginase Immobilization.

In this study, the third-generation polyamidoamine dendrimer was functionalized with a 5-amino-1H-tetrazole heterocycle to load the synthesis enzyme and its surface groups. Then, chitosan was attached to the dendrimer by a suitable linker, and finally, zinc oxide nanoparticles were inserted into dendrimer cavities to increase loading. FTIR, FESEM, TEM, and DLS analysis showed that this new dendrimer has specific branches, and ZnO nanoparticles were spread between the branches and connected with the branches and chitosan biopolymer. Also proved the presence of stabilized L-asparaginase enzyme and ZnO nanoparticles in the designed system. Furthermore, the extent of L-asparaginase enzyme loading and release was investigated in the laboratory with a dialysis bag. Examining the toxicity of the new third-generation polyamidoamine (PAMAM) dendrimeric nanocarrier based on chitosan-zinc oxide biopolymer (PAMAM-G3@ZnO-Cs nanocarrier) on the Jurkat cell line (human acute lymphoblastic leukemia) at pH 7.4 showed that this nanocarrier effectively encapsulates the drug L-asparaginase and slowly releases it and also preventing the growth of cancer cells. The activity of the loaded enzyme in the nanocarrier and the free enzyme was calculated. During the investigations, it was found that the enzyme attached to the nanocarrier is more stable than the free enzyme at optimal pH and temperature and at high temperatures, acidic and basic pHs. Vmax and Km values were lower for loaded enzymes. The synthesized PAMAM-G3@ZnO-Cs nanocarrier can be a promising candidate in the pharmaceutical industry and medical science for cancer treatment due to its biocompatibility, non-toxicity, stability, and slow release of L-asparaginase.

An emphatic study on the luciferin-luciferase bioluminescence system of Benthosema pterotum.

Approximately 80% of luminous organisms live in the oceans, and considerable diversity of life dependence on bioluminescence has been observed in marine organisms. Among vertebrates, luminous fish species are the only group of vertebrates that have the ability to emit bioluminescent light. Meanwhile, the lantern fish family (Myctophidae), with 33 genera all of which have the ability to emit light, is considered the most prominent family among the luminous fish of the deep oceans and seas. Lantern fish Benthosema pterotum has bioluminescence properties due to the presence of photophores scattered in its ventral-lateral region. However, no research has been performed on its bioluminescence system and light emission mechanism. The present research aimed to assess the type of bioluminescence, pigment, photoprotein, or luciferin-luciferase system in B. pterotum. In order to determine the type of light-emitting system in B. pterotum species, several specific experiments were designed and performed. It was shown that the light emission system in B. pterotum species is categorized into the luciferin-luciferase type. Conducting this research was not only innovative, but it also could be the beginning of further research in the field of marine biochemistry and production of the recombinant active forms of enzymes for industrial, commercial, medical, and pharmaceutical purposes.

Application of enzymes for targeted removal of biofilm and fouling from fouling-release surfaces in marine environments: A review.

Biofouling causes adverse issues in underwater structures including ship hulls, aquaculture cages, fishnets, petroleum pipelines, sensors, and other equipment. Marine constructions and vessels frequently are using coatings with antifouling properties. During the previous ten years, several alternative strategies have been used to combat the biofilm and biofouling that have developed on different abiotic or biotic surfaces. Enzymes have frequently been suggested as a cost-effective, substitute, eco-friendly, for conventional antifouling and antibiofilm substances. The destruction of sticky biopolymers, biofilm matrix disorder, bacterial signal interference, and the creation of biocide or inhibitors are among the catalytic reactions of enzymes that really can successfully prevent the formation of biofilms. In this review we presented enzymes that have antifouling and antibiofilm properties in the marine environment like α-amylase, protease, lysozymes, glycoside hydrolase, aminopeptidases, oxidase, haloperoxidase and lipases. We also overviewed the function, benefits and challenges of enzymes in removing biofouling. The reports suggest enzymes are good candidates for marine environment. According to the findings of a review of studies in this field, none of the enzymes were able to inhibit the development of biofilm by a site marine microbial community when used alone and we suggest using other enzymes or a mixture of enzymes for antifouling and antibiofilm purposes in the sea environment.

A Novel, Highly Potent NADPH-Dependent Cytochrome P450 Reductase from Waste Liza klunzingeri Liver.

The use of marine enzymes as catalysts for biotechnological applications is a topical subject. Marine enzymes usually display better operational properties than their animal, plant or bacterial counterparts, enlarging the range of possible biotechnological applications. Due to the fact that cytochrome P450 enzymes can degrade many different toxic environmental compounds, these enzymes have emerged as valuable tools in bioremediation processes. The present work describes the isolation, purification and biochemical characterization of a liver NADPH-dependent cytochrome P450 reductase (CPR) from the marine fish Liza klunzingeri (LkCPR). Experimental results revealed that LkCPR is a monomer of approximately 75 kDa that is active in a wide range of pH values (6-9) and temperatures (40-60 °C), showing the highest catalytic activity at pH 8 and 50 °C. The activation energy of the enzyme reaction was 16.3 kcal mol-1 K-1. The KM values for cytochrome C and NADPH were 8.83 µM and 7.26 µM, and the kcat values were 206.79 s-1 and 202.93 s-1, respectively. LkCPR displayed a specific activity versus cytochrome C of 402.07 µmol min-1 mg1, the highest activity value described for a CPR up to date (3.2-4.7 times higher than the most active reported CPRs) and showed the highest thermostability described for a CPR. Taking into account all these remarkable catalytic features, LkCPR offers great potential to be used as a suitable biocatalyst.

Marine enzymes: Classification and application in various industries.

Oceans are regarded as a plentiful and sustainable source of biological compounds. Enzymes are a group of marine biomaterials that have recently drawn more attention because they are produced in harsh environmental conditions such as high salinity, extensive pH, a wide temperature range, and high pressure. Hence, marine-derived enzymes are capable of exhibiting remarkable properties due to their unique composition. In this review, we overviewed and discussed characteristics of marine enzymes as well as the sources of marine enzymes, ranging from primitive organisms to vertebrates, and presented the importance, advantages, and challenges of using marine enzymes with a summary of their applications in a variety of industries. Current biotechnological advancements need the study of novel marine enzymes that could be applied in a variety of ways. Resources of marine enzyme can benefit greatly for biotechnological applications duo to their biocompatible, ecofriendly and high effectiveness. It is beneficial to use the unique characteristics offered by marine enzymes to either develop new processes and products or improve existing ones. As a result, marine-derived enzymes have promising potential and are an excellent candidate for a variety of biotechnology applications and a future rise in the use of marine enzymes is to be anticipated.

Recombinant protein expression: Challenges in production and folding related matters.

Protein folding is a biophysical process by which proteins reach a specific three-dimensional structure. The amino acid sequence of a polypeptide chain contains all the information needed to determine the final three-dimensional structure of a protein. When producing a recombinant protein, several problems can occur, including proteolysis, incorrect folding, formation of inclusion bodies, or protein aggregation, whereby the protein loses its natural structure. To overcome such limitations, several strategies have been developed to address each specific issue. Identification of proper protein refolding conditions can be challenging, and to tackle this high throughput screening for different recombinant protein folding conditions can prove a sound solution. Different approaches have emerged to tackle refolding issues. One particular approach to address folding issues involves molecular chaperones, highly conserved proteins that contribute to proper folding by shielding folding proteins from other proteins that could hinder the process. Proper protein folding is one of the main prerequisites for post-translational modifications. Incorrect folding, if not dealt with, can lead to a buildup of protein misfoldings that damage cells and cause widespread abnormalities. Said post-translational modifications, widespread in eukaryotes, are critical for protein structure, function and biological activity. Incorrect post-translational protein modifications may lead to individual consequences or aggregation of therapeutic proteins. In this review article, we have tried to examine some key aspects of recombinant protein expression. Accordingly, the relevance of these proteins is highlighted, major problems related to the production of recombinant protein and to refolding issues are pinpointed and suggested solutions are presented. An overview of post-translational modification, their biological significance and methods of identification are also provided. Overall, the work is expected to illustrate challenges in recombinant protein expression.

Exploring in vitro effect of silver nanoparticles and Holothuria parva extracts on kinetics and stability of α-amylase.

Diabetes is a chronic metabolic disorder characterized by elevated blood glucose levels. Major limitations of synthetic drugs, including high cost, efficacy, and adverse side effects, have prompted researchers to seek more effective and low-cost alternative therapies with fewer adverse effects. Marine life forms are considered the most important sources of biologically active natural products due to their secondary metabolites. In this research, sea cucumber Holothuria parva was collected from coastal areas of Bandar Lengeh, Hormozgan, Iran, and was then subjected to extraction. The results showed that compounds extracted from Holothuria parva had a stimulatory effect on enzyme activity, and in the presence of these compounds, the Vmax value of the enzyme was increased about two times, while the Km value was reduced. The phosphate buffer form of extracts had the greatest impact on enzyme activity. Upon an increase in the concentration of silver nanoparticles (AgNPs), the α-amylase activity was inhibited in parallel. Silver nanoparticles exhibited the highest enzyme inhibition with an IC50 of 0.86 mg/ml. Silver nanoparticles showed anti-α-amylase activity and had the ability to decrease intestinal glucose uptake in diabetic individuals when prescribed as a novel supplementary medicine.

Zinc sulfide-chitosan hybrid nanoparticles as a robust surface for immobilization of Sillago sihama α-amylase.

In the present study, zinc sulfide-chitosan hybrid nanoparticles synthesized by chemical deposition were used as a matrix for the immobilization of purified α-amylase extracted from Sillago sihama (Forsskal, 1775). In this regard, the size and morphological structure of zinc sulfide-chitosan hybrid nanoparticles before and after the stabilization process were evaluated using FT-IR, DLS methods, as well as SEM and TEM electron microscopy, and EDS analyses. Then, the efficiency of the immobilized enzyme was measured in terms of temperature, optimal pH, stability at the critical temperature, and pH values. Immobilization of α-amylase on zinc sulfide -chitosan hybrid nanoparticles increased the long-term stability, as well as its endurance to critical temperatures and pH values; however, the optimal temperature and pH values of the enzyme were not altered following the immobilization process. The kinetic parameters of the enzyme were also changed during immobilization. Enzyme immobilization increased the Km, whereas decreased the catalytic efficiency (Kcat / Km) of the immobilized enzyme compared with the free enzyme. These results are very important as, in most cases, enzyme immobilization reduces the activity and catalytic efficiency of enzymes. The nano-enzyme produced in this study, due to its high temperature, and pH stability, could be a good candidate for industrial applications, especially in the food industry.

Purification and characterization of a robust thermostable protease isolated from Bacillus subtilis strain HR02 as an extremozyme.

AIM: Since the hot water of Genow, a village in Isin rural district in the central district of Bandar Abbas, Hormozgan Province, Iran, has a rich source of thermophilic bacteria, the current study aimed to find a new thermophilic protease enzyme with suitable properties to be used in different industries. METHODS AND RESULTS: Water and sediment samples were collected from the hot water of Genow, and finally, 20 colonies were isolated. Among these isolated colonies, two bacterial strains grew on the skim milk agar medium, and a clear halo was formed around the colony, which was accurately identified by 16S rRNA gene sequence analysis. The comparison of 16S rRNA gene sequence analyses of isolated strains HR01 and HR02 with registered sequences of 16S rRNA genes in NCBI showed that the two isolates had the most similarity to Bacillus sonorensis and Bacillus subtilis, respectively. Among the two bacterial strains, the highest enzymatic activity was observed in B. subtilis strain HR02, from which the protease purification process was performed. A putative native B. subtilis strain HR02 protease (BSHR02PR) was purified by the UNO Q-6 ionic exchange chromatography method. Biochemical analyses revealed a monomeric enzyme, BsHR02Pro, with a molecular weight of 25 kDa, showing the maximum activity at 70°C and pH 8.0. Moreover, the purified enzyme was stable up to 80 °C and in a pH range of 6.0-12.0. The steady-state kinetic analysis for colloidal casein showed that the Km , Vmax and kcat values of the purified enzyme were 25.7 µM, 93.2 µM min-1 and 2.18 s-1 , respectively. CONCLUSION: The hot water of Genow is a rich source of protease-producing bacteria. Sediments are a better source for the isolation of these types of bacteria than spring water. Overall, our results demonstrated a potential bacterial enzyme BsHR02Pro as a suitable catalyst to be used in the various industries.

Production of a Novel Marine Pseudomonas aeruginosa Recombinant L-Asparaginase: Insight on the Structure and Biochemical Characterization.

The present study focused on the cloning, expression, and characterization of L-asparaginase of marine Pseudomonas aeruginosa HR03 isolated from fish intestine. Thus, a gene fragment containing the L-asparaginase sequence of Pseudomonas aeruginosa HR03 isolated from the fish intestine was cloned in the pET21a vector and then expressed in Escherichia coli BL21 (DE3) cells. Thereafter, the recombinant L-asparaginase (HR03Asnase) was purified by nickel affinity chromatography, and the enzymatic properties of HR03Asnase, including the effects of pH and temperature on HR03Asnase activity and its kinetic parameters, were determined. The recombinant enzyme HR03Asnase showed the highest similarity to type I L-asparaginase from Pseudomonas aeruginosa. The three-dimensional (3D) modeling results indicate that HR03Asnase exists as a homotetramer. Its molecular weight was 35 kDa, and the maximum activity of the purified enzyme was observed at pH8 and at 40 °C. The km and Vmax of the enzyme obtained with L-asparagine as substrate were 10.904 mM and 3.44 × 10-2 mM/min, respectively. The maximum activity of HR03Asnase was reduced by 50% at 90 °C after 10-min incubation; however, the enzyme maintained more than 20% of its activity after 30-min incubation. This enzyme also maintained almost 50% of its activity at pH 12 after 40-min incubation. The evaluation of pH and temperature stability of HR03Asnase showed that the enzyme has a wide range of activity, which is a suitable characteristic for its application in different industries. Overall, the results of the present study indicate that marine sources are promising biological reservoirs for enzymes to be used for biotechnological purposes, and marine thermostable HR03Asnase is likely a potential candidate for its future usage in the pharmaceutical and food industries.

Marine Cellulases and their Biotechnological Significance from Industrial Perspectives.

Marine microorganisms represent virtually unlimited sources of novel biological compounds and can survive extreme conditions. Cellulases, a group of enzymes that are able to degrade cellulosic materials, are in high demand in various industrial and biotechnological applications, such as in the medical and pharmaceutical industries, food, fuel, agriculture, and single-cell protein, and as probiotics in aquaculture. The cellulosic biopolymer is a renewable resource and is a linearly arranged polysaccharide of glucose, with repeating units of disaccharide connected via ß-1,4-glycosidic bonds, which are broken down by cellulase. A great deal of biodiversity resides in the ocean, and marine systems produce a wide range of distinct, new bioactive compounds that remain available but dormant for many years. The marine environment is filled with biomass from known and unknown vertebrates and invertebrate microorganisms, with much potential for use in medicine and biotechnology. Hence, complex polysaccharides derived from marine sources are a rich resource of microorganisms equipped with enzymes for polysaccharides degradation. Marine cellulases' extracts from the isolates are tested for their functional role in degrading seaweed and modifying wastes to low molecular fragments. They purify and renew environments by eliminating possible feedstocks of pollution. This review aims to examine the various types of marine cellulase producers and assess the ability of these microorganisms to produce these enzymes and their subsequent biotechnological applications.

Industrial applications of immobilized nano-biocatalysts.

Immobilized enzyme-based catalytic constructs could greatly improve various industrial processes due to their extraordinary catalytic activity and reaction specificity. In recent decades, nano-enzymes, defined as enzyme immobilized on nanomaterials, gained popularity for the enzymes' improved stability, reusability, and ease of separation from the biocatalytic process. Thus, enzymes can be strategically incorporated into nanostructured materials to engineer nano-enzymes, such as nanoporous particles, nanofibers, nanoflowers, nanogels, nanomembranes, metal-organic frameworks, multi-walled or single-walled carbon nanotubes, and nanoparticles with tuned shape and size. Surface-area-to-volume ratio, pore-volume, chemical compositions, electrical charge or conductivity of nanomaterials, protein charge, hydrophobicity, and amino acid composition on protein surface play fundamental roles in the nano-enzyme preparation and catalytic properties. With proper understanding, the optimization of the above-mentioned factors will lead to favorable micro-environments for biocatalysts of industrial relevance. Thus, the application of nano-enzymes promise to further strengthen the advances in catalysis, biotransformation, biosensing, and biomarker discovery. Herein, this review article spotlights recent progress in nano-enzyme development and their possible implementation in different areas, including biomedicine, biosensors, bioremediation of industrial pollutants, biofuel production, textile, leather, detergent, food industries and antifouling.

Emerging trends in environmental and industrial applications of marine carbonic anhydrase: a review.

Biocatalytic conversion of greenhouse gases such as carbon dioxide into commercial products is one of the promising key approaches to solve the problem of climate change. Microbial enzymes, including carbonic anhydrase, NAD-dependent formate dehydrogenase, ribulose bisphosphate carboxylase, and methane monooxygenase, have been exploited to convert atmospheric gases into industrial products. Carbonic anhydrases are Zn2+-dependent metalloenzymes that catalyze the reversible conversion of CO2 into bicarbonate. They are widespread in bacteria, algae, plants, and higher organisms. In higher organisms, they regulate the physiological pH and contribute to CO2 transport in the blood. In plants, algae, and photosynthetic bacteria carbonic anhydrases are involved in photosynthesis. Converting CO2 into bicarbonate by carbonic anhydrases can solidify gaseous CO2, thereby reducing global warming due to the burning of fossil fuels. This review discusses the three-dimensional structures of carbonic anhydrases, their physiological role in marine life, their catalytic mechanism, the types of inhibitors, and their medicine and industry applications.

Estimation of carbon pools in the biomass and soil of mangrove forests in Sirik Azini creek, Hormozgan province (Iran).

Despite the increasing interest in mangroves as one of the most carbon-rich ecosystems, arid mangroves are still poorly investigated. We aimed to improve the knowledge of biomass and soil carbon sequestration for an arid mangrove forest located at the Azini creek, Sirik, Hormozgan Province (Iran). We investigated the biomass and organic carbon stored in the above and belowground biomass for three different regions selected based on the composition of the principal species: (1) Avicennia marina, (2) mixed forest of A. marina and Rhizophora mucronata, and (3) R. mucronata. Topsoil organic carbon storage to 30 cm depth was also estimated for each analyzed area. Biomass carbon storage, considering both aboveground (AGB) and belowground biomass (BGB), was significantly different between the cover areas. Overall, the mean forest biomass (MFB) was 283.1 ± 89 Mg C ha-1 with a mean C stored in the biomass of 128.9 ± 59 Mg C ha-1. Although pure Rhizophora stand showed the lowest value of above and below tree carbon (AGC + BGC); 17.6 ± 1.9 Mg C ha-1), soil organic carbon stock in sites under Rhizophora spp. was significantly higher than in the site with pure stand of Avicennia spp. Overall, forest soil stored the highest proportion of Sirik mangrove ecosystem organic carbon (59%), with a mean value of 188.3 ± 27 Mg C ha-1. These results will contribute to broaden the knowledge and the dataset available, reducing the uncertainties related to estimates and modeling of carbon pools in arid mangrove ecosystem, which also represent an important climatic threshold of mangrove worldwide distribution.

Improved thermal stability of phytase from Yersinia intermedia by physical adsorption immobilization on amino-multiwalled carbon nanotubes.

Phytase is used in poultry diets to hydrolyze and release of phytate-bound phosphorus. Immobilization on nanomaterials optimizes enzyme's thermal stability and reusability. This study aimed to immobilize the recombinant phytase from Yersinia intermedia on the surface of amino-multi-walled carbon nanotubes (amino-MWCNTs) by physical adsorption. For this, zeta potential measurement, FTIR spectroscopic analysis, scanning electron microscope (SEM), kinetic as well as thermodynamic parameters were used to characterize immobilized phytase on amino-MWCNTs. According to results, the optimum temperature of the immobilized phytase increased from 50 to 70 °C and also thermal and pH stability improved considerably. Moreover, immobilization led to an increase in the value of Km and kcat from 0.13 to 0.33 mM and 2220 to 2776 s-1, respectively. In addition, the changes in activation energy of thermal inactivation (ΔE#a (D)), the free energy of thermal inactivation (ΔG#D) and the enthalpy of thermal inactivation (ΔH#D) for immobilized phytase increased by +11.05, +24.7 and +11.4 kj/mole, respectively, while the value of the change in the entropy of thermal inactivation (ΔS#D) decreased by - 0.04 kj/mole.K. Overall, our results showed that adsorption immobilization of phytase on amino-MWCNTs increases thermal, pH and storage stability as well as some of kinetic parameters.

Nano-organic supports for enzyme immobilization: Scopes and perspectives.

A variety of organic nanomaterials and organic polymers are used for enzyme immobilization to increase enzymes stability and reusability. In this study, the effects of the immobilization of enzymes on organic and organic-inorganic hybrid nano-supports are compared. Immobilization of enzymes on organic support nanomaterials was reported to significantly improve thermal, pH and storage stability, acting also as a protection against metal ions inhibitory effects. In particular, the effects of enzyme immobilization on reusability, physical, kinetic and thermodynamic parameters were considered. Due to their biocompatibility with low health risks, organic support nanomaterials represent a good choice for the immobilization of enzymes. Organic nanomaterials, and especially organic-inorganic hybrids, can significantly improve the kinetic and thermodynamic parameters of immobilized enzymes compared to macroscopic supports. Moreover, organic nanomaterials are more environment friendly for medical applications, such as prodrug carriers and biosensors. Overall, organic hybrid nanomaterials are receiving increasing attention as novel nano-supports for enzyme immobilization and will be used extensively.

Identification of a novel tailor-made chitinase from white shrimp Fenneropenaeus merguiensis.

Fenneropenaeus merguiensis (commonly named banana shrimp) is one of the most important farmed crustacean worldwide species for the fisheries and aquaculture industry. Besides its nutritional value, it is a good source of chitinase, an enzyme with excellent biological and catalytic properties for many industrial applications. In the present study, a putative chitinase-encoding cDNA was synthesized from mRNA from F. merguiensis hepatopancreas tissue. Subsequently, the corresponding cDNA was cloned, sequenced and functionally expressed in Escherichia coli, and the recombinant F. merguiensis chitinase (rFmCHI) was purified by His-tag affinity chromatography. The bioinformatics analysis of aminoacid sequence of rFmCHI displayed a cannonical multidomain architecture in chitinases which belongs to glycoside hydrolase family 18 (GH18 chitinase). Biochemical characterization revealed rFmCHI as a monomeric enzyme of molecular weight 52 kDa with maximum activity at 40 °C and pH 6.0 Moreover, the recombinant enzyme is also stable up to 60 °C, and in the pH range 5.0-8.0. Steady-state kinetic studies for colloidal chitin revealed KM, Vmax and kcat values of 78.18 µM, 0.07261 µM. min-1 and 43.37 s-1, respectively. Overall, our results aim to demonstrate the potential of rFmCHI as suitable catalyst for bioconversion of chitin waste.

Induced dysregulation of ACE2 by SARS-CoV-2 plays a key role in COVID-19 severity.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the cause of COVID-19, is reported to increase the rate of mortality worldwide. COVID-19 is associated with acute respiratory symptoms as well as blood coagulation in the vessels (thrombosis), heart attack and stroke. Given the requirement of angiotensin converting enzyme 2 (ACE2) receptor for SARS-CoV-2 entry into host cells, here we discuss how the downregulation of ACE2 in the COVID-19 patients and virus-induced shift in ACE2 catalytic equilibrium, change the concentrations of substrates such as angiotensin II, apelin-13, dynorphin-13, and products such as angiotensin (1-7), angiotensin (1-9), apelin-12, dynorphin-12 in the human body. Substrates accumulation ultimately induces inflammation, angiogenesis, thrombosis, neuronal and tissue damage while diminished products lead to the loss of the anti-inflammatory, anti-thrombotic and anti-angiogenic responses. In this review, we focus on the viral-induced imbalance between ACE2 substrates and products which exacerbates the severity of COVID-19. Considering the roadmap, we propose multiple therapeutic strategies aiming to rebalance the products of ACE2 and to ameliorate the symptoms of the disease.