Enhancing the expression of recombinant small laccase in Pichia pastoris by a double promoter system and application in antibiotics degradation.

Low-expression levels remain a challenge in the quest to use the small laccase (rSLAC) as a viable catalyst. In this study, a recombinant Pichia pastoris strain (rSLAC-GAP-AOX) producing rSLAC under both AOX and GAP promoters (located in two different plasmids) was generated and cultivated in the presence of methanol and mixed feed (methanol:glycerol). Induction with methanol resulted in a maximum laccase activity of 1200 U/L for rSLAC-GAP-AOX which was approximately 2.

Enzymatic treatment of phenolic pollutants by a small laccase immobilized on APTES-functionalised magnetic nanoparticles.

Unlabelled: In this study, we have successfully synthesized magnetic nanoparticles (MNPs), functionalised them by silanization and used them for the covalent immobilization of a recombinant small laccase (rSLAC) from . The immobilized recombinant laccase (MNP-rSLAC) was subsequently used for the treatment of phenol, 4-chlorophenol (4-CP) and 4-fluorophenol (4-FP). The enzyme completely degraded 80 µg/mL of the selected phenolic compounds within 2 h in the presence of a natural mediator, acetosyringone.

Crystal structure of a thermophilic fungal cyanase and its implications on the catalytic mechanism for bioremediation.

Cyanase catalyzes the bicarbonate-dependent degradation of cyanate to produce ammonia and carbon dioxide, and ammonia is a considerable alternative nitrogen source. Strikingly, the cyanase from the thermophilic fungus Thermomyces lanuginosus (Tl-Cyn) has the highest catalytic efficiency reported among these enzymes. However, its molecular mechanism of action is not clearly understood, because currently there is no structural information available on fungal cyanases.

Simultaneous removal of heavy metals and cyanate in a wastewater sample using immobilized cyanate hydratase on magnetic-multiwall carbon nanotubes.

Global environmental problems allied with waste management require novel approaches for the simultaneous removal of heavy metals and other associated compounds including cyanate. In this study, iron-oxide filled multi-walled carbon nanotubes (m-MWCNTs) were successfully synthesized and characterized by field emission gun scanning electron microscopy (FEGSEM), high-resolution transmission electron microscopy (HRTEM) and X-ray diffraction (XRD). The m-MWCNTs were amino-functionalized for the covalent immobilization of a recombinant cyanate hydratase (rTl-Cyn), and were characterized by fourier transform infrared (FTIR) spectroscopy.

A novel strategy for the efficient removal of toxic cyanate by the combinatorial use of recombinant enzymes immobilized on aminosilane modified magnetic nanoparticles.

Cyanase detoxifies cyanate by transforming it to ammonia and carbon dioxide in a bicarbonate-dependent reaction, however, dependence on bicarbonate limits its utilization in large-scale applications. A novel strategy was therefore developed for overcoming this bottleneck by the combined application of cyanase (rTl-Cyn) and carbonic anhydrase (rTl-CA). The synergistic effect of rTl-Cyn and rTl-CA could reduce the dependence of bicarbonate by 80%, compared to using rTl-Cyn alone.

Secretory expression of recombinant small laccase from Streptomyces coelicolor A3(2) in Pichia pastoris.

This work reports for the first time the secretory expression of the small laccase (SLAC) from Streptomyces coelicolor A3(2) in Pichia pastoris. Using an AOX1 promoter and α factor as a secretion signal, the recombinant P. pastoris harbouring the laccase gene (rSLAC) produced high titres of extracellular laccase (500 ± 10 U/l), which were further increased seven fold by pre-incubation at 80 °C for 30 min.

Synthesis, molecular docking, antimicrobial, antioxidant and toxicity assessment of quinoline peptides.

A series of quinoline based peptides were synthesized by a one-pot reaction through Ugi-four component condensation of lipoic acid, cyclohexyl isocyanide, aniline derivatives and 2-methoxy quinoline-3-carbaldehyde derivatives under microwave irradiation. The products were obtained in excellent yields and high purity. Solvent optimization and the effect of microwave irradiation with various powers were also observed.

Expression of a novel recombinant cyanate hydratase (rTl-Cyn) in Pichia pastoris, characteristics and applicability in the detoxification of cyanate.

A recombinant Pichia pastoris harbouring the cyanate hydratase gene (rTl-Cyn) from the thermophilic fungus Thermomyces lanuginosus SSBP yielded a high titre of extracellular cyanate hydratase (100±13UmL) which was ∼10-fold higher than the native fungal strain. The purified rTl-Cyn had a molecular mass of ∼20kDa on SDS-PAGE, with K, V, k and k/K values of 0.34mM, 2857.

Recombinant HAP Phytase of the Thermophilic Mold Sporotrichum thermophile: Expression of the Codon-Optimized Phytase Gene in Pichia pastoris and Applications.

The codon-optimized phytase gene of the thermophilic mold Sporotrichum thermophile (St-Phy) was expressed in Pichia pastoris. The recombinant P. pastoris harboring the phytase gene (rSt-Phy) yielded a high titer of extracellular phytase (480 ± 23 U/mL) on induction with methanol.

Characteristics of Recombinant Phytase (rSt-Phy) of the Thermophilic mold Sporotrichum thermophile and its applicability in dephytinizing foods.

Sporotrichum thermophile produces very low titres of phytase (St-Phy) extracellularly, which is acidstable, thermostable, and protease insensitive with broad substrate specificity, and therefore, the gene encoding phytase (St-Phy) has been cloned and expressed in E. coli. The purified recombinant phytase (rSt-Phy) has the molecular mass of 55 kDa with Km and Vmax (calcium phytate), kcat and kcat/Km of 0.

Green rapid biogenic synthesis of bioactive silver nanoparticles (AgNPs) using Pseudomonas aeruginosa.

The present work was focused on isolating a bacterial strain of Pseudomonas sp. with the ability to synthesise AgNPs rapidly. A strain of Pseudomonas aeruginosa designated JO was found to be a potential candidate for rapid synthesis of AgNPs with a synthesis time of 4h in light, at room temperature which is a shorter time period noticed for the synthesis when compared to the previous reports Biosynthesis of AgNPs was achieved by addition of culture supernatant with aqueous silver nitrate solution (1 mM).

Effects of novel brominated flame retardants on steroidogenesis in primary porcine testicular cells.

Brominated flame retardants are chemicals with fire quenching properties which are extensively used in manufacturing. Historically, less regulated use of legacy brominated flame retardants (BFRs) for a number of years has resulted in ubiquitous contamination of the environment. As a result, some of the more persistent BFRs have been phased out and are being replaced by a next generation of brominated compounds for which there is little toxicological data.

Pharmacokinetics, urinary excretion and plasma protein binding of ofloxacin in water buffalo calves (Bubalus bubalis).

Pharmacokinetics and urinary excretion of an intravenous dose of 5 mg.kg-1 ofloxacin were investigated in water buffalo calves. Plasma concentrations of ofloxacin were determined by high-performance liquid chromatography.

Pharmacokinetics following intravenous administration and pharmacodynamics of cefquinome in buffalo calves.

Disposition following single intravenous injection (2 mg/kg) and pharmacodynamics of cefquinome were investigated in buffalo calves 6-8 months of age. Drug levels in plasma were estimated by high-performance liquid chromatography. The plasma concentration-time profile following intravenous administration was best described by a two-compartment open model.