Vertical distribution of three species of eriophyid mites on tea in south India

Two eriophyid mites,Acaphylla theae andCalacarus carinatus, have been known for some time as important pests of tea. In recent years a third eriophyid,Acaphyllisa parindiae, has also become abundant. The vertical distribution of these three species on tea is examined.     

Preparation of S-(N6,N6-dimethyladenosyl)-L-methionine

4′,5′-Unsaturated nucleosides are obtained by the action of 1,5-diazabicyclo-[5.4.0]undec-5-ene on N¹- and N³-(methyl 2,3,4-tri-O-acetyl-β-d-glucopyranosyluronate)-5-fluorouracil. The ²H₁ conformation of N¹- and N³-(methyl 4-deoxy-α-l-threo-hex-4-enopyranosyluronate)-5-fluorouracil has been established by ¹H-n.m.r. and c.d. methods. Interaction of the heterocyclic base and the double bond of the sugar moiety is demonstrated.     

Studies on the size and shape of rabbit intestinal glucoamylase-maltase complex.

Rabbit intestinal glucoamylase-maltase was examined in detail with respect to its molecular weight, sedimentation, diffusion and viscosity. It is a large asymmetrical molecule, with a molecular weight of 750 000-760 000. Its appearance under the electron microscope supports the idea that it is a long string (62.0 nm) consisting of eight beads of diameter 6.0 nm each and a surface-to-surface interbead distance of approx. 2.0 nm. The shape of the enzyme derived from its hydrodynamic behaviour by using the string-of-spherical-beads model originally proposed by Kuhn [(1932) Z. Phys. Chem. Abt. A 161, 1-32] and later modified by Shulman [(1953) J. Am. Chem. Soc. 75, 5846-5852] fits moderately well with the electron-microscopic picture. The beads might represent about six subunits, and the absence of sulphur from the enzyme and the inability to dissociate the enzyme by conventional methods indicate the possibility of unusual covalent cross-linking between the subunits and between the beads.     

An Evaluation of the Metachromasia of Bromophenol Blue

The heterogeneity of bromophenol blue from different commercial sources was revealed by paper chromatography. Isopropanol:ammonia:water (20:1:2) as the solvent system gave the best separation. A variety of impurities: violet, pink, light blue and yellow coloured ones were observed. Two of the yellow fractions showed a spectral shift to red in the presence of ammonia vapour. The respone of the main dye component with the anionic chromotropes such as heparin and hyaluronate was found to be metachromatic similar to that exhibited by the dye solution and not due to a polychromatic effect. The metachromatic effect was blocked by FeCl₃ as in the case of cationic dye metachromasy. The observed metachromatic colour is not one of the colours which characterize those resulting from changes caused by pH.     

Side reactions in the synthesis of phosphotyrosine-containing peptides

Summary A series of phosphopeptides Tyr(PO₃H₂)-Val-Pro-Xxx-Leu (Xxx=Met, Met(O), Nle, Dab or Cys), derived from the native platelet-derived growth factor- receptor (PDGF-) sequence, has been prepared to study their interaction with the src-homology 2 (SH2) domains of the p85 subunit of PI3 kinase. The phosphopeptides were synthesized using Fmoc methodology incorporating N-Boc dibenzyl-protected phosphotyrosine (Boc-Tyr[PO₃(Bzl)₂]) as the N-terminal amino acid, since the benzyl groups can be removed during resin cleavage with TFA. Only peptides containing methionine were found to exist partially as S-benzyl sulfonium salts after TFA cleavage from the resin. The desired peptide could be obtained from the S-benzyl sulfonium salt by hydrogenolysis.     

Engineered nickel bioaccumulation in Escherichia coli by NikABCDE transporter and metallothionein overexpression

Mine wastewater often contains dissolved metals at concentrations too low to be economically extracted by existing technologies, yet too high for environmental discharge. The most common treatment is chemical precipitation of the dissolved metals using limestone and subsequent disposal of the sludge in tailing impoundments. While it is a cost-effective solution to meet regulatory standards, it represents a lost opportunity. In this study, we engineered Escherichia coli to overexpress its native NikABCDE transporter and a heterologous metallothionein to capture nickel at concentrations in local effluent streams. We found the engineered strain had a 7-fold improvement in the bioaccumulation performance for nickel compared to controls, but also observed a drastic decrease in cell viability due to metabolic burden or inducer (IPTG) toxicity. Growth kinetic analysis revealed the IPTG concentrations used based on past studies lead to growth inhibition, thus delineating future avenues for optimization of the engineered strain and its growth conditions to perform in more complex environments.     

Crystal structure of the hydroxylase component of methane monooxygenase from Methylosinus trichosporium OB3b.

Methane monooxygenase (MMO), found in aerobic methanotrophic bacteria, catalyzes the O2-dependent conversion of methane to methanol. The soluble form of the enzyme (sMMO) consists of three components: a reductase, a regulatory "B" component (MMOB), and a hydroxylase component (MMOH), which contains a hydroxo-bridged dinuclear iron cluster. Two genera of methanotrophs, termed Type X and Type II, which differ markedly in cellular and metabolic characteristics, are known to produce the sMMO. The structure of MMOH from the Type X methanotroph Methylococcus capsulatus Bath (MMO Bath) has been reported recently. Two different structures were found for the essential diiron cluster, depending upon the temperature at which the diffraction data were collected. In order to extend the structural studies to the Type II methanotrophs and to determine whether one of the two known MMOH structures is generally applicable to the MMOH family, we have determined the crystal structure of the MMOH from Type II Methylosinus trichosporium OB3b (MMO OB3b) in two crystal forms to 2.0 A resolution, respectively, both determined at 18 degrees C. The crystal forms differ in that MMOB was present during crystallization of the second form. Both crystal forms, however, yielded very similar results for the structure of the MMOH. Most of the major structural features of the MMOH Bath were also maintained with high fidelity. The two irons of the active site cluster of MMOH OB3b are bridged by two OH (or one OH and one H2O), as well as both carboxylate oxygens of Glu alpha 144. This bis-mu-hydroxo-bridged "diamond core" structure, with a short Fe-Fe distance of 2.99 A, is unique for the resting state of proteins containing analogous diiron clusters, and is very similar to the structure reported for the cluster from flash frozen (-160 degrees C) crystals of MMOH Bath, suggesting a common active site structure for the soluble MMOHs. The high-resolution structure of MMOH OB3b indicates 26 consecutive amino acid sequence differences in the beta chain when compared to the previously reported sequence inferred from the cloned gene. Fifteen additional sequence differences distributed randomly over the three chains were also observed, including D alpha 209E, a ligand of one of the irons.