Secretory Expression and Characterization of Chinese <Emphasis Type="Italic">Narcissus</Emphasis> GNA-Like Lectin in <Emphasis Type="Italic">Pichia pastoris</Emphasis>

Narcissus tazetta lectin (NTL) is a GNA-like lectin, which has a wide potential application in medicine, agriculture, and glycobiology. In the present paper, the codon-optimized ntl gene was transformed into the yeast Pichia pastoris; SDS–PAGE gel and western blotting analysis revealed that the recombinant lectin was expressed successfully in Pichia yeast. The similarity between the recombinant NTL and the native NTL was confirmed by circular dichroism (CD) and hemagglutination assay further. In the 5-L scale fermentator, the protein yield was as high as 1.2 g/L after fermentation for 96 h. In addition, the effect of metal ions (K⁺, Mg²⁺, Ca²⁺, and Cu²⁺), acid, and alkaline on hemagglutinating activity of NTL was tested, which provided biochemical characterizations of the mannose-binding lectin from Chinese Narcissus.     

Production and Biotechnological Application of Extracellular Alkalophilic Lipase from Marine Macroalga-Associated <Emphasis Type="Italic">Shewanella algae</Emphasis> to Produce Enriched C<Subscript>20-22</Subscript><Emphasis Type="Italic">n</Emphasis>-3 Polyunsaturated Fatty Acid Concentrate

An extracellular alkalophilic lipase was partially purified from heterotrophic Shewanella algae (KX 272637) associated with marine macroalgae Padina gymnospora. The enzyme possessed a molecular mass of 20 kD, and was purified 60-fold with a specific activity of 36.33 U/mg. The enzyme exhibited V_max and K_m of 1000 mM/mg/min and 157 mM, respectively, with an optimum activity at 55 °C and pH 10.0. The catalytic activity of the enzyme was improved by Ca²⁺ and Mg²⁺ ions, and the enzyme showed a good tolerance towards organic solvents, such as methanol, isopropanol, and ethanol. The purified lipase hydrolyzed the refined liver oil from leafscale gulper shark Centrophorus squamosus, yielding a total C_20-22 n-3 PUFA concentration of 34.99% with EPA + DHA accounting the major share (34% TFA), after 3 h of hydrolysis. This study recognized the industrial applicability of the thermostable and alkalophilic lipase from marine macroalga-associated bacterium Shewanella algae to produce enriched C_20-22 n-3 polyunsaturated fatty acid concentrate.     

Purification and characterization of an extracellular lipolytic enzyme from the fermented fish-originated halotolerant bacterium, <Emphasis Type="Italic">Virgibacillus alimentarius</Emphasis> LBU20907

A halotolerant Virgibacillus alimentarius LBU20907 isolated from fermented fish (Budu) was found to be an efficient producer of extracellular halophilic lipase enzyme. The enzyme was purified 5.99-fold with a 0.15% final yield to homogeneity by ammonium sulfate precipitation, followed by dialysis, Toyopearl DEAE-650 M ion exchange chromatography, Toyopearl butyl-650 M hydrophobic interaction chromatography, and Toyopearl-HW 55 F gel filtration chromatography. SDS-PAGE of purified lipase exhibited a homogenous single band with a very high molecular weight of 100 kDa. The properties of purified lipase revealed maximum activity at pH 7.0 and 40 °C. It was also highly stable in a pH range of 6.0–7.0, retaining more than 90% activity for 24 h. It was stable at the temperature of 30–50 °C and maintained more than 80% activity for 16 h. The purified lipase performing the maximal activity in the presence of 20.0% NaCl indicated halophilic enzyme properties. Its lipolytic activity was highest against p-nitrophenyl palmitate. The lipase activity was found to be enhanced in hexane. The enzyme activity was stimulated in the presence of Zn²⁺, Ca²⁺, Mg²⁺, and Sr²⁺; while, it was completely inhibited by Ba²⁺ and Co²⁺. The enzyme had a K _m and V _max of 108.0 mg and 79.1 U mL^?1, respectively.     

Bioconversion of cellulose and simultaneous production of thermoactive exo- and endoglucanases by <Emphasis Type="Italic">Fusarium oxysporum</Emphasis>

Saccharification of cellulose is a promising method for production of biofuels. However, low bioconversion efficiency of cellulose to soluble sugars is a major challenge. In this study, a cellulolytic strain of Fusarium oxysporum was cultivated on pure cellulosic substrates (avicel, α-cellulose, carboxymethylcellulose and methylcellulose) and conversion efficiency into glucose was investigated. Production of exo- and endoglucanases during the bioconversion process was evaluated. Influence of pH on saccharification of cellulose and enzyme production by F. oxysporum were determined. Highest yield of glucose (1.76 μmol/ml) was obtained from F. oxysporum on methyl cellulose at 192 h under basal conditions. Liberated glucose under optimized condition of pH 6.0 at 96 h of fermentation was 2.12 μmol/ml with maximum production of exo- and endoglucanases (23.70 and 34.72 U/mg protein, respectively). The crude exo- and endoglucanases had optimum activities at pH 8.0, 70 °C and pH 7.0, 50 °C, respectively. The enzymes were stable over pH of 4.0–7.0 with relative residual activity above 60% after 1 h incubation. Exoglucanase activity was enhanced by Ca²⁺ and Cu²⁺ at 5 mM and Mg²⁺ at 10 mM. Endoglucanase activity was greatly enhanced in the presence of Mn²⁺, Ca²⁺, Mg²⁺, Cu²⁺ and Fe³⁺ at 5 and 10 mM. Activities of both enzymes were inhibited in the presence of Hg²⁺ at 5 and 10 mM. Results show that F. oxysporum possessed good cellulolytic enzyme system for efficient conversion of cellulose. Exhibited thermotolerance of exoglucanase with the striking tolerance of endoglucanase to metal ions demonstrate potentials of enzymes for biofuel industry.     

Dissolved air flotation for treating wastewater of the nuclear industry: preliminary results

Preliminary testing of dissolved air flotation (DAF) for wastewater treatment is presented. A combined coagulation-flocculation/DAF column system is used to remove oil and ⁶⁰Co from nuclear industry wastewater. In this work, operational conditions and coagulant/flocculant concentrations are optimized by varying pH. Determinations of air-solids ratio (G/S), retention time (θ), pressure (P), volume of depressurized air–water mixture (V), turbidity and ⁶⁰Co concentrations are reported. The effect of the treatment on the efficiency of separation of oily residues is also discussed. The results establish that the coagulant/flocculant system, formed by a modified polyamine (25 mgL^?1) and a slightly cationic polyacrylamide (1.5 mgL^?1), under specific operational conditions (pH = 7, mixing intensity Im₁ = 300 s^?1 and Im₂ = 30 s^?1), allowed the destabilization of colloidal matter, resulting in resistant flocs. It was concluded that by using G/S = 0.3, θ = 15 min, P = 620 kPa and V = 0.0012 m³, the greatest percentage removals of oil, turbidity, total cobalt and ⁶⁰Co were obtained. These preliminary results then show that dissolved air flotation represents a good alternative for treatment of nuclear industry wastewater contaminated with radionuclides.     

Synthesis of water soluble metalloporphyrin-cored amphiphilic star block copolymer photocatalysts for an environmental application

Two series of water-soluble metalloporphyrin-cored amphiphilic star block copolymers were synthesized by controlled radical polymerizations such as atom transfer radical polymerization (ATRP) and reversible addition fragmentation chain transfer (RAFT), which gave eight amphiphilic block copolymer arm chains consisting of poly(n-butyl acrylate-b-poly(ethylene glycol) methyl ether methacylate) (PnBA-b-PEGMEMA, M_n,GPC = 78,000, M_w/M_n = 1.2, 70 wt% of PPEGMEMA) and poly(styrene-b-2-dimethylamino ethyl acrylate) (PS-b-PDMAEA, M_n,GPC = 83,000, M_w/M_n = 1.2, 67 wt% of PDMAEA), yielding porphyrin(Pd)-(PnBA-b-PPEGMEMA)₈ and porphyrin(Pd)-(PS-b-PDMAEA)₈, respectively. Obtained metalloporphyrin polymer photocatalysts were homogeneously solubilized in water to apply to the removal of chlorophenols in water, and was distinguished from conventional water-insoluble small molecular metalloporphyrin photocatalysts. Notably, we found that the water-soluble star block copolymers with hydrophobic–hydrophilic core–shell structures more effectively decomposed the chlorophenol, 2,4,6-trichlorophenol (2,4,6-TCP), in water under visible light irradiation (k = 1.39 h^?1, t_1/2 = 0.5 h) in comparison to the corresponding water-soluble star homopolymer, because the hydrophobic core near the metalloporphyrin effectively captured and decomposed the hydrophobic chlorophenols in water.     

Microbial Transformation of Curcumin to Its Derivatives with a Novel <Emphasis Type="Italic">Pichia kudriavzevii</Emphasis> ZJPH0802 Strain

Curcumin, a polyphenolic compound, has shown a wide range of pharmacological activities and has been widely used as a food additive. However, the clinical use of curcumin is limited to some extent because of its poor water solubility and low bioavailability. To overcome these problems, many approaches have been attempted and structural modification of curcumin by microbial transformation has been proven to be an alternative. In this study, we isolated a novel yeast strain Pichia kudriavzevii ZJPH0802 from a soil sample, which is capable of converting curcumin to its derivatives. The transformed products by this strain were evaluated by HPLC, (+) electrospray ionization (ESI)-MSⁿ, and ¹H nuclear magnetic resonance methods. Compared with controls, two new peaks of the transformed broth appeared at retention times of 26 min (I) and 62 min (II) by HPLC analysis. The two transformed products were then further identified by (+) ESI-MSⁿ. The spectrum showed that compound I had an accurate [M+H+NH₃]⁺ ion at m/z 392, [M+H]⁺ ion at m/z 375, [M+H–H₂O]⁺ ion at m/z 357, and (+) ESI-MS³ spectrum showed that ion at m/z 357 could further form fragment ions at m/z 339, 177, and 163; compound II had an accurate [M+H]⁺ ion at m/z 373, [M+H–H₂O]⁺ ion at m/z 355, and (+) ESI-MS³ spectrum showed that ion at m/z 355 could further form fragment ions at m/z 219, 179, 177, 163, and 137. These two transformed products thereby were confirmed as hexahydrocurcumin (I) and tetrahydrocurcumin (II).     

Overproduction,Purification and Characterization of Adenylate Deaminase from <Emphasis Type="Italic">Aspergillus oryzae</Emphasis>

Adenylate deaminase (AMPD, EC 3.5.4.6) is an aminohydrolase that widely used in the food and medicine industries. In this study, the gene encoding Aspergillus oryzae AMPD was cloned and expressed in Escherichia coli. Induction with 0.75 mM isopropyl β-d-l-thiogalactopyranoside resulted in an enzyme activity of 1773.9 U/mL. Recombinant AMPD was purified to electrophoretic homogeneity using nickel affinity chromatography, and its molecular weight was calculated as 78.6 kDa. Purified AMPD exhibited maximal activity at 35 °C, pH 6.0 and 30 mM K⁺, with apparent K _m and V _max values of 2.7 × 10^?4 M and 77.5 μmol/mg/min under these conditions. HPLC revealed that recombinant AMPD could effectively catalyse the synthesis of inosine-5′-monophosphate (IMP) with minimal by-products, indicating high specificity and suggesting that it could prove useful for IMP production.     

Purification and Characterization of Agarase from Marine Bacteria <Emphasis Type="Italic">Acinetobacter</Emphasis> sp. PS12B and Its Use for Preparing Bioactive Hydrolysate from Agarophyte Red Seaweed <Emphasis Type="Italic">Gracilaria verrucosa</Emphasis>

Acinetobacter strain PS12B was isolated from marine sediment and was found to be a good candidate to degrade agar and produce agarase enzyme. The extracellular agarase enzyme from strain PS12B was purified by ammonium sulfate precipitation followed by DEAE-cellulose ion-exchange chromatography. The specific activity of the crude enzyme which was 1.52 U increased to 45.76 U, after two-stage purification, with an enzyme yield of 9.76%. Purified enzyme had a molecular mass of 24 kDa. The optimum pH and temperature for activity of purified agarase were found to be 8.0 and 40 °C, respectively. The K_m and V_max values for agarase were 4.69 mg/ml and 0.5 μmol/min, respectively. Treatment with EDTA reduced the agarase activity by 58% at 5 mM concentration. The enzyme activity was stimulated by the presence of Fe²⁺, Mn²⁺, and Ca2⁺ ions while reducing reagents (β-mercaptoethanol and dithiothreitol, DTT) enhanced its activity by 30–40%. The purified agarase exhibited tolerance to both detergents and organic solvents. Major hydrolysis products of agar were DP4 and also a mixture of longer oligosaccharides DP6 and DP7. The enzyme hydrolysed seaweed (Gracilaria verrucosa) exhibited strong antioxidant activity in vitro. Successful hydrolysis of seaweed indicates the potential use of the enzyme to produce seaweed hydrolysate having health benefits as well as the industrial application like the production of biofuels.     

<Emphasis Type="Italic">cis</Emphasis>-Dioxomolybdenum(VI) complexes of sterically encumbered phenol-based tetradentate N<Subscript>2</Subscript>O<Subscript>2</Subscript> ligands: Structural,spectroscopic, and electrochemical studies

Two cis-dioxomolybdenum(VI) complexes [MoO₂L] (L: L ¹, 2 and L: L ², 3) in a phenol-based sterically encumbered N₂O₂ ligand environment have been synthesized, and their crystallographic characterizations are reported. The orange crystals of 2 are monoclinic, space group P2₁/a with unit cell dimensions as a=16.2407(17) Å, b=7.2857(8) Å, c=18.400(2) Å, β=98.002(9)°, Z=4, and d _cal=1.486 g cm^?3. The light orange crystals of 3, however, are orthorhombic, space group, Pbcn, with unit cell dimensions a=8.3110(12) Å, b=12.637(3) Å, c=34.673(5) Å, Z=4, and d _cal=1.187 g cm^?3. The structures were refined by a full-matrix least-squares procedure on F ² to a final R=0.046 (0.055 for 3) using 4944 (3677) all independent data. In both the cases, the Mo atom exists in a distorted octahedral geometry defined by a N₂O₄ donor set, which features a cis-Mo(–O)₂ and a trans-Mo(OPh)₂ arrangement. Compound 2 undergoes a quasireversible one-electron reduction at ?1.3 V vs Ag/AgCl reference due to Mo^VIO₂/Mo^VO₂ electron transfer and thus providing a rare example of steric solution to the comproportionation–dimerization problem encountered frequently in the development of valid biomimetic models for the active sites of oxomolybdenum enzymes.     

Production of <Emphasis Type="Italic">R</Emphasis>-Mandelic Acid Using Nitrilase from Recombinant <Emphasis Type="Italic">E. coli</Emphasis> Cells Immobilized with Tris(Hydroxymethyl)Phosphine

Recombinant Escherichia coli cells harboring nitrilase from Alcaligenes faecalis were immobilized using tris(hydroxymethyl)phosphine (THP) as the coupling agent. The optimal pH and temperature of the THP-immobilized cells were determined at pH 8.0 and 55 °C. The half-lives of THP-immobilized cells measured at 35, 40, and 50 °C were 1800, 965, and 163 h, respectively. The concentration of R-mandelic acid (R-MA) reached 358 mM after merely 1-h conversion by the immobilized cells with 500 mM R,S-mandelonitrile (R,S-MN), affording the highest productivity of 1307 g L^?1 day^?1 and the space-time productivity of 143.2 mmol L^?1 h^?1 g^?1. The immobilized cells with granular shape were successfully recycled for 60 batches using 100 mM R,S-MN as substrate at 40 °C with 64% of relative activity, suggesting that the immobilized E. coli cells obtained in this study are promising for the production of R-MA.     

Synthesis,antifungal and insecticidal activity of novel [1,2,4]triazolo[4,3-<Emphasis Type="Italic">a</Emphasis>]pyridine derivatives containing a sulfide substructure

A series of [1,2,4]triazolo[4,3-a]pyridine derivatives bearing a sulfide substructure was designed, synthesized and characterized via ¹H·NMR, ¹³C·NMR, IR and elemental analyses. Bioassay Results indicated some of the derivatives displayed good fungicidal activity on Rhizoctonia cerealis, moderated insecticidal activity against Plutella xylostella and good insecticidal activity on Helicoverpa armigera. The inhibitory effects of compounds 4g and 4u against Rhizotonia cerealis were 70.9% at 50 μg mL^?1; the IC₅₀ values of compounds 4d and 4s against Plutella xylostella were 43.87 and 50.75 μg mL^?1, respectively. And the IC₅₀ values of compounds 4d, 4q, and 4s on Helicoverpa armigera were 58.3, 77.14 and 65.31 μg mL^?1, respectively, which were better than that of commercial chlorpyrifos (103.77 μg mL^?1).     

Solution Thermodynamics of <Emphasis Type="Italic">N</Emphasis>,<Emphasis Type="Italic">N</Emphasis><Superscript>/</Superscript>-Ethylenebis-(salicylideneiminato)-diaquochromium(III) Chloride in Aqueous Dimethylsulphoxide

The densities, viscosities and refractive indices of N,N ^/-ethylene-bis(salicylideneiminato)-diaquochromium(III) chloride, [Cr(salen)(H₂O)₂]Cl, in aqueous dimethylsulfoxide (DMSO) with different mass fractions (w ₂ = 0.20, 0.40, 0.60, 0.80 and 1.00) of DMSO were determined at 298.15, 308.15 and 318.15 K under atmospheric pressure. From measured densities, viscosities and refractive indices the apparent molar volumes (V _φ ), standard partial molar volume (V _φ ⁰ ), the slope (S _V ^* ), standard isobaric partial molar expansibility (φ _E ⁰ ) and its temperature dependence (?φ _E ⁰ /?T) _p , the viscosity B-coefficient, its temperature dependence (?B/?T), solvation number (S _n ) and apparent molar refractivity (R _D ^φ ), etc., were calculated and discussed on the basis of ion–ion and ion–solvent interactions. These results revealed that the solutions are characterized by ion–solvent interactions rather than by ion–ion interactions and the complex behaves as a long range structure maker. Thermodynamics of viscous flow was discussed in terms of transition state theory.     

Electrical conductivity and ionic association of lithium and sodium perchlorates in tetrahydrofuran

The total limiting molar electrical conductivities of ions and triads of ions and the association constants of ions with the formation of ion pairs and triads of ions were calculated from the concentration dependences of the electrical conductivity of solutions of lithium and sodium perchlorates in tetrahydrofuran at 278.15–318.15 K with the use of the method specially developed earlier. The experimental total limiting electrical conductivities were used to calculate the limiting molar electrical conductivities and attraction friction factors of separate ions (Li⁺, Na⁺, ClO ₄ ^? , Li₂ClO ₄ ⁺ , Na₂ClO ₄ ⁺ , Li(ClO₄) ₂ ^? , and Na(ClO₄) ₂ ^? ). The constants of ion association into ion pairs were used to calculate the Gibbs energy of non-Coulomb interionic interaction (ΔG*_+?), and the constants of association into triads of ions, to determine the a ₃ distance parameter between the centers of the ion and the dipole of the ion pair. Positive ΔG*_+?), values and deviations of the experimental a ₃ value from the distance parameter calculated theoretically (a ₃ ⁰ ) for the triad of ions (Δa ₃ = a ₃ ? a ₃ ⁰ ) were related to non-Coulomb repulsion in the region of overlap of the solvation shells of ions and the influence of temperature and ion charge density on this repulsion.     

Recovery of Phenolic Acid and Enzyme Production from Corn Silage Biologically Treated by <Emphasis Type="Italic">Trametes versicolor</Emphasis>

Corn silage is used as high-energy forage for dairy cows and more recently for biogas production in a process of anaerobic co-digestion with cow manure. In this work, fresh corn silage after the harvest was used as a substrate in solid-state fermentations with T. versicolor with the aim of phenolic acid recovery and enzyme (laccase and manganese peroxidase) production. During 20 days of fermentation, 10.4-, 3.4-, 3.0-, and 1.8-fold increments in extraction yield of syringic acid, vanillic acid, p-hydroxybenzoic acid, and caffeic acid, respectively, were reached when compared to biologically untreated corn silage. Maximal laccase activity was gained on the 4th day of fermentation (V.A. = 180.2 U/dm³), and manganese peroxidase activity was obtained after the 3rd day of fermentation (V.A. = 30.1 U/dm³). The addition of copper(II) sulfate as inducer during solid state fermentation resulted in 8.5- and 7-fold enhancement of laccase and manganese peroxidase activities, respectively. Furthermore, the influence of pH and temperature on enzyme activities was investigated. Maximal activity of laccase was obtained at T = 50 °C and pH = 3.0, while manganese peroxidase is active at temperature range T = 45–70 °C with the maximal activity at pH = 4.5.     

Synthesis,characterization, antimicrobial activity and ultrastructure of the affected bacteria of new quinoline compounds

The reaction on 8-hydroxy quinoline-7-aldehyde azo compounds (HL _n ) (where n = 1–5) with 4-amino-1,2-dihydro-2,3-dimethyl-1-phenylpyrazol-5-one to obtain HL _n (where n = 6–10) have been characterized by means of TLC, melting point and spectral data, such as IR, ¹H NMR, mass spectra and thermal studies. The X-ray diffraction patterns of two starting materials 8-hydroxy quinoline-7-aldehyde (start 1), 4-amino-1,2-dihydro-2,3-dimethyl-1-phenylpyrazol-5-one (start 2) and the ligands (HL_5,10) are investigated in powder form. All the ligands have been screened for their antimicrobial activity against four local bacterial species, two Gram-positive bacteria (Bacillus cereus and Staphylococcus aureus) and two Gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae) as well as against four local fungi; Aspergillus niger, Alternaria alternata, Penicillium italicum and Fusarium oxysporium. The results show that the azo ligands (HL _n ) (where n = 1–5) have no antimicrobial activity against bacteria and fungi while most azomethine ligands (HL _n ) (where n = 6–10) are good antibacterial agents against E. coli and K. pneumoniae as well as antifungal agents against P. italicum and A. alternata. The results were compared to standard substances (start 1) and (start 2). Among the azomethine ligands, HL₁₀ was the most effective against the most microorganisms tested. The size of clear zone was ordered as p-(OCH₃ < CH₃ < H < Cl < NO₂) as expected from Hammett’s constant (σ ^R ). Also, the ultrastructure study of the affected bacteria confirmed that HL₈ is good antibacterial agent against E. coli and S. aureus.     

Bioaccumulation of <Superscript>137</Superscript>Cs and <Superscript>60</Superscript>Co by bacteria isolated from spent nuclear fuel pools

This paper is focused on a characterization of bacterial contamination in pool water of the interim spent fuel storage (JAVYS Inc.) in Slovak Republic and on bioaccumulation of ¹³⁷Cs and ⁶⁰Co by isolated bacteria. Bacterial community in pool water is kept on very low level by extremely low concentration of solutes in deionized water and by the efficient water filtration system. Based on standard methods and sequencing of 16S rDNA four pure bacterial cultures were identified as Kocuria palustris, Micrococcus luteus, Ochrobactrum spp. and Pseudomonas aeruginosa. Isolated aerobic bacteria were able to bioaccumulate ¹³⁷Cs and ⁶⁰Co in laboratory experiments. The mechanism of Co and Cs binding involve rapid interactions with anionic groups of the components of cell surface and in the case of Cs⁺ ions is followed by transport processes across cytoplasm membranes and by intracellular distribution. The maximum specific uptake of Cs⁺ after 48 h cultivation in mineral medium (MM) reached 7.54 ± 0.48 μmol g^?1 dw (Ochrobactrum spp.), 19.6 ± 0.1 μmol g^?1 dw (M. luteus) and 20.1 ± 2.2 μmol g^?1 dw (K. palustris). The maximum specific uptake of Co²⁺ after 24 h cultivation in MM reached 31.1 ± 3.5 μmol g^?1 dw (Ochrobactrum spp.), 86.6 ± 12.2 μmol g^?1 dw (M. luteus) and 16.9 ± 1.2 μmol g^?1 dw (K. palustris). These results suggest that due to the long lasting uptake of ¹³⁷Cs, ⁶⁰Co and other radionuclides by biofilm in pool water high specific radioactivities (Bq m^?2) can be expected on stainless steel walls of pools.     

Ambident chemosensors based on benzo[<Emphasis Type="Italic">h</Emphasis>]chromen-2-one

Schiff bases derived from 7-hydroxy-4-methyl-2-oxobenzo[h]chromene-8-carbaldehyde in solution exist as equilibrium mixtures of benzoid and quinoid tautomers. The fraction of the quinoid tautomer increases with rise in solvent polarity. The Schiff base containing a benzo-15-crown-5 fragment on the nitrogen atom was shown to be a new ambident chemosensor capable of selectively binding transition metal cations via reaction at the o-hydroxyaldehyde imine fragment and alkaline-earth metals via host-guest interaction with the crown ether moiety. This compound exhibits a pronounced sensor activity toward Mg²⁺ and Ba²⁺ ions and is a selective naked-eye fluorescent chemosensor for Cu²⁺ and Co²⁺ ions.     

Production,Purification and Characterisation of a Potential Fibrinolytic Protease from Endophytic <Emphasis Type="Italic">Xylaria curta</Emphasis> by Solid Substrate Fermentation

The present investigation highlights the optimal conditions for production of a non-toxic, bi-functional fibrinolytic enzyme xylarinase produced by endophytic fungus Xylaria curta by solid substrate fermentation using rice chaff medium. The purified enzyme is a monomeric protein with a molecular mass of ～33 kDa. The enzyme exhibits cleavage of Aα and Bβ chains of fibrin(ogen) and has no effect on γ chain. The optimal fibrinolytic activity of the enzyme was observed at 35 °C and pH 8. The fibrinolytic activity was enhanced in the presence of Ca²⁺, whereas it was completely inhibited in the presence of Fe²⁺ and Zn²⁺ ions and inhibitors like EDTA and EGTA suggesting it to be a metalloprotease. The K _m and V _max of the enzyme for azocasein were 326 μM and 0.13 μM min^?1. The N-terminal sequence of the enzyme (SNGPLPGGVVWAG) was same when compared to xylarinase isolated from culture broth of X. curta. Thus, xylarinase could be exploited as a potent clot busting enzyme which could be produced on large scale using solid substrate fermentation.