Catalytic and Thermodynamic Characterization of Endoglucanase (CMCase) from <Emphasis Type="Italic">Aspergillus oryzae</Emphasis> cmc-1

Monomeric extracellular endoglucanase (25 kDa) of transgenic koji (Aspergillus oryzae cmc-1) produced under submerged growth condition (7.5 U mg⁻¹ protein) was purified to homogeneity level by ammonium sulfate precipitation and various column chromatography on fast protein liquid chromatography system. Activation energy for carboxymethylcellulose (CMC) hydrolysis was 3.32 kJ mol⁻¹ at optimum temperature (55 °C), and its temperature quotient (Q ₁₀) was 1.0. The enzyme was stable over a pH range of 4.1–5.3 and gave maximum activity at pH 4.4. V _max for CMC hydrolysis was 854 U mg⁻¹ protein and K _m was 20 mg CMC ml⁻¹. The turnover (k _cat) was 356 s⁻¹. The pK _a1 and pK _a2 of ionisable groups of active site controlling V _max were 3.9 and 6.25, respectively. Thermodynamic parameters for CMC hydrolysis were as follows: ΔH* = 0.59 kJ mol⁻¹, ΔG* = 64.57 kJ mol⁻¹ and ΔS* = −195.05 J mol⁻¹ K⁻¹, respectively. Activation energy for irreversible inactivation ‘E _a(d)’ of the endoglucanase was 378 kJ mol⁻¹, whereas enthalpy (ΔH*), Gibbs free energy (ΔG*) and entropy (ΔS*) of activation at 44 °C were 375.36 kJ mol⁻¹, 111.36 kJ mol⁻¹ and 833.06 J mol⁻¹ K⁻¹, respectively.     

Isolation and structure determination of a new indene derivative from endophytic fungus Aspergillus flavipes Y-62

As part of the search for naturally derived secondary metabolites, a novel indene derivative, compound 1, together with nine known metabolites (2–10) have been purified from an ethyl acetate extract of the plant-associated fungus Aspergillus flavipes Y-62, isolated from Suaeda glauca (Bunge) Bunge which was collected along Zhoushan coast, Zhejiang province, East China. The structure of the new compound 1 was elucidated by extensive use of spectroscopic techniques like 1D, 2D NMR, and HR-TOF-MS, while the known metabolites were established based on both spectral methods as well as by comparison with the previous literature. Compound 1 exhibited antimicrobial activities against the gram-negative pathogen Pseudomonas aeruginosa and Klebsiella pneumoniae with equal MIC values of 32 µg/ml.     

Location of delamination in laminated composite plates by pulsed laser holography

The location of delamination in composite laminates based on their vibration characteristics is presented in this paper. Composite materials are widely used as structural materials for aerospace engineering, because of its excellent mechanical properties such as light weight, high stiffness and anti-corrosion characteristics. This paper uses pulsed electronic speckle pattern interferometry (Pulsed-ESPI) to perform non-destructive evaluation of carbon fiber reinforced plastic (CFRP) laminate plates containing various sizes of artificial defects located at the centre of the specimen, based on vibration characteristic using double-pulsed Ruby laser with a pulse duration of 30 ns. The pulsed separation can be adjusted in between 2 and 800 μs. The main advantage of this technique over conventional ESPI is the ability to carry out measurements even under harsh environmental conditions. From the results, it seems to be effective for the detection of defects in various kinds of composite materials. In this paper, the basic principles of the technique are described briefly.     

Modeling primary substrate controlled biotransformation and transport of halogenated aliphatics in porous media

In situ biorestoration is a groundwater remediation technique in which the indigenous aquifer bacteria are stimulated by injecting compounds to provide carbon source and energy. Stimulated bacteria may transform the target contaminants such as tetrachloroethylene (PCE) and trichloroethylene (TCE) into intermediate products. In this study, we developed a model to simulate the substrate-limited biotransformation of the halogenated solvents present in anoxic groundwater by sequential reductive dehalogenation under methanogenic conditions. The model consists of conservation of mass equations for the primary substrate, immobile indigenous biomass, organic solvents such as PCE and TCE, and their intermediate products trichloroethylene, dichloroethylene, and vinyl chloride. The utilization of primary substrate and the biotransformation of organic solvents are assumed to follow Monod kinetics. The limiting factor on bacterial growth is assumed to be the primary substrate. The microbial yield coefficient is determined from the stoichiometric equation describing the anaerobic process. The model is solved by using a finite difference technique. Results are presented for three different case studies: continuous injection of primary substrate (acetate), single-pulse injection, and double-pulse injection. The single-pulse or double-pulse injection techniques were found to be more effective than continuous injection of primary substrate. Double-pulse technique reduces the clogging of injection wells caused by excessive microbial growth around boreholes and achieves a more uniform distribution of microbial growth in the subsurface. In all cases target compounds were effectively removed. The results, however, indicate substantial levels of intermediate product accumulation. Numerical results of a simplified model which assumes an abundance of primary substrate and a constant population of biomass, compare favorably with experimental data reported in the literature.     

A New Dimeric Secoiridoid Glycoside from the Leaves of Olea ferruginea Royle

A new dimeric secoiridoid glycoside, trivially named oleferrugine B ( 1 ), has been isolated from the AcOEt‐soluble part of the MeOH extract of leaves of Olea ferruginea Royle . The structure of the isolated compound was established on the basis of ESI‐MS fragmentation patterns, and 1D‐ and 2D‐NMR techniques, including ¹H‐ and ¹³C‐NMR, HSQC, ¹H,¹H‐COSY, HMBC, and NOESY experiments, and by comparison with literature data.     

Structural diversities and preliminary antimicrobial studies of 1-((<Emphasis Type="Italic">E</Emphasis>)-(pentylimino)methyl)naphthalen-2-ol and its metal complexes

Schiff bases and their metal complexes have a number of biological activities such as antidepressant, analgesic, antimicrobial, antiviral, and antitumor. In the present studies, the 1-((E)-(pentylimino)methyl)naphthalen-2-ol ligand is prepared by the reaction of 2-hydroxynaphthaldehyde with n-amyl amine. The ligand results in CoL₃, NiL₂ and CuL₂ complexes when reacted with the cobalt acetate, nickel acetate, and copper acetate salts respectively. The complexes along with their ligand are fully characterized by X-ray diffraction studies and biologically screened. The ligand structure reveals that it is a zwitterion species where the iminic nitrogen atom is protonated and the C–O bond shows a high double bond character. The cobalt complex has an octahedral geometry around the metal center and each nitrogen atom is in the trans position to the oxygen donor site. On the other hand, the copper complex shows a centrosymmetric square planar coordination, and in this case, the nitrogen sites are trans to each other. The complexes and the ligand are found to be more potent than the standard drugs used against some microbes in their preliminary biological studies.     

1H NMR, mass spectroscopic studies and color fastness properties of methyl-4,6-dihydroxy-2-oxo-1-phenyl-5-arylazopyridine-3-carboxylate, part III: new disperse dyes

1H NMR and mass spectroscopic studies of new disperse dyes synthesized from methyl-4,6-dihydroxy-2-oxo-1-phenyl pyridine-3-carboxylate. Application of the dyes on polyester fabric and color fastness properties are also studied. The dyes are also found to possess DPPH free radical scavenging activity.     

Design and characterization of an engineered gp41 protein from human immunodeficiency virus-1 as a tool for drug discovery

Two new proteins of approximately 70 amino acids in length, corresponding to an unnaturally-linked N- and C-helix of the ectodomain of the gp41 protein from the human immunodeficiency virus (HIV) type 1, were designed and characterized. A designed tripeptide links the C-terminus of the C-helix with the N-terminus of the N-helix in a circular permutation so that the C-helix precedes the N-helix in sequence. In addition to the artificial peptide linkage, the C-helix is truncated at its N-terminus to expose a region of the N-helix known as the “Trp-Trp-Ile” binding pocket. Sedimentation, crystallographic, and nuclear magnetic resonance studies confirmed that the protein had the desired trimeric structure with an unoccupied binding site. Spectroscopic and centrifugation studies demonstrated that the engineered protein had ligand binding characteristics similar to previously reported constructs. Unlike previous constructs which expose additional, shallow, non-conserved, and undesired binding pockets, only the single deep and conserved Trp-Trp-Ile pocket is exposed in the proteins of this study. This engineered version of gp41 protein will be potentially useful in research programs aimed at discovery of new drugs for therapy of HIV-infection in humans.     

Trace elements in human fingernails: Measurement by proton-induced X-ray emission

The method of proton particle-induced X-ray emission (proton PIXE) has been employed to study the trace element composition of human fingernails. The samples were colleted from 51 subjects randomly selected from a working community of about 500 adults and they were analyzed by the thick-target external beam technique of the PIXE method. The samples were exposed to the proton beam as 1-mm thick pellets and irradiated with 2 MeV protons having 20 nA beam intensity. For 40 C irradiations, the concentration of fourteen elements, K, Ca, Ti, Mn, Fe, Ni, Cu, Zn, As, Se, Br, Rb, Sr and Pb, were measured by comparison with a calibration obtained from the NBS orchard leaf standard (SRM 1571). Some anomalous cases have been revealed from this study and they are attributed to environmental factors. The frequency distributions of the elements are presented and the results compared with available data.This research received financial support from the International Atomic Energy Agency, Vienna, under the research contract RC/2536/RBl.