The influence of the ion content of whey on the pH-activity profile of the β-galactosidase from Aspergillus oryzae

The pH-dependence of the reaction kinetics of lactase (β-galactosidase) from Aspergillus oryzae in different reaction media is presented in terms of a two-parameter model. The lactase from A. oryzae seems to have replaced the A. niger lactase on the market owing to a better activity/price ratio and may be utilised for lactose hydrolysis in acid as well as in neutral milk products. Its pH optimum is around pH 4.5. However, in the neutral pH-range its activity depended strongly on the salt content of the substrate solution. For example, its activity in whey (pH 6.5) fell to only 30% of its expected activity in a pure lactose solution at the same pH. The whey effect was the same for both soluble and immobilised lactase. The two parameter kinetic model, which included a term for competitive product inhibition gave excellent agreement with experimental data, and may thus be useful for the prediction of reactor performance with this enzyme.     

Yeast exo-β-glucanases can be used as efficient and readily detectable reporter genes in Saccharomyces cerevisiae

Yeast exo-1,3-β-glucanases are secretable proteins whose function is basically trophic and may also be involved in cell wall glucan hydrolytic processes. Since fluorescein di(β-D -glucopyranoside) is a fluorogenic substrate detectable and quantifiable by flow cytometry, it was used for testing the ability of the EXG1 gene product of Saccharomyces cerevisiae and its homologous gene in Candida albicans to function as reporter genes. These open reading frames were coupled to different promoters in multicopy plasmids, and exoglucanase activity quantified at flow cytometry. Exoglucanases were found to be useful tools for the study of promoter regions in S. cerevisiae. This technique has the advantage over other reporter gene systems—such as β-galactosidase fusions—that it does not require permeabilization of yeast cells and therefore it allows the recovery of viable cells—by sorting—after flow cytometry analysis.     

钒冶炼焙烧添加剂选择研究

对小型钒冶炼厂焙烧工艺所用添加剂进行改进的可能性进行了探讨，研究了几种常用添加剂的焙烧条件，分析比较了其性能，提出用ＮａＣｌ－Ｎａ２ＣＯ３作焙烧添加剂替代ＮａＣｌ可大幅度减少大气污染，提高冶钒转化率；且不改变工艺流程，无需设备投资，具有较好的经济效益和环境效益。     

Yeast sequencing reports. Isolation and sequence analysis of a gene from the linear DNA plasmid pPAC1–2 of Pichia acaciae that shows similarity to a killer toxin gene of Kluyveromyces lactis

The toxin-encoding linear plasmid systems found in Pichia acaciae and Kluyveromyces lactis yeasts appear to be quite similar, both in function and structural organization. By Southern hybridization, a linear plasmid of P. acaciae, pPac1–2, was found to hybridize to the second open reading frame (ORF2) of K. lactis plasmid pGKL1, known to encode the α and β subunits of the K. lactis toxin. A 1·7 kbp segment of pPac1–2 DNA was cloned, sequenced and shown to contain four regions of strong homology to four similarly oriented regions of K. lactis ORF2. This 1·7 kbp fragment also contained an ORF of 1473 bp that could encode a protein of ～ 55·8 kDa. Like the α subunit gene of K. lactis ORF2, a very hydrophobic region occurs at the N-terminus, perhaps representing a signal sequence for transport out of the cell. Unlike K. lactis ORF2, however, the encoded polypeptide is much smaller and lacks a recognizable domain common to chitinases. The structure of a toxin that includes the translation product of this P. acaciae ORF would likely be quite different from that of the K. lactis toxin. Analysis of the upstream region of the P. acaciae ORF revealed an upstream conserved sequence identical to that found before ORFs 8 and 9 of pGKL2. A possible hairpin loop structure, as has been described for each of the four K. lactis pGKL1 ORFs, was found just upstream of the presumed start codon. The similarity of the promoter-like elements found in the linear plasmid genes of these diverse yeasts reinforces the idea of the existence of a unique, but highly conserved, expression system for these novel plasmids. The sequence has been deposited in the GenBank data library under Accession Number U02596.     

非线性编辑中特效图文素材的应用

Cool 3D是一款特效图文制作软件，适用于动画片段、3D文字、图形等制作场合。Cool 3D中制作的特效素材应用于非线性编辑中，可以丰富素材的表现形式，提高节目编辑制作的整体质量。     

Thermally stimulated discharge current studies in pure and fe-doped polystyrene films

Studies were made on the thermally stimulated discharge currents (TSDCs) in pure (undoped) and Fe-doped polystyrene films as a function of polarizing field, polarizing temperature and dopant concentration. While undoped films exhibited a single peak, doped films showed two peaks one at low temperatures and another at high temperatures. The low temperature peak, which exhibits a shift towards lower temperatures with increasing dopant concentration, is attributed to the relaxation of the main chain, while the high temperature peak, which shows a tendency to shift towards higher temperatures with dopant concentration, is due to space charge polarization. The TSDCs were higher for low dopant concentrations than their undoped counterparts, while for high concentrations of the dopant, the TSDCs decreased. Formation of charge transfer complexes at low dopant concentrations and molecular aggregates at higher dopant concentrations are suggested as the possible reasons for this behaviour.     

Purification and renaturation of recombinant human lymphotoxin (tumour necrosis factor beta) expressed in Escherichia coli as inclusion bodies

High level expression of recombinant human tumour necrosis factor β (rh TNF-β) in Escherichia coli results in the formation of two portions of protein, namely soluble active protein and insoluble protein which is inactive and aggregates in the form of inclusion bodies (IBs). In this study, a procedure for purification and renaturation of rh TNF-β from inclusion bodies has been designed and verified experimentally with a product purity of more than 90% and a recovery of about 30%. The procedure includes washing of IBs with specific wash buffer (Triton X-100/EDTA/lysozyme/PMSF), their solubilization with 8 mol dm^?3 alkaline urea, purification with ion-exchange columns, refolding with renaturation buffer and finally concentration and desalination with an ultrafiltration membrane. The characteristics of the renatured protein were identical with those of purified protein from the soluble fraction as demonstrated by (1) SDS-PAGE, (2) cytotoxic activity on mouse L929 cells, (3) N-terminal amino acid sequence, and (4) gel filtration chromatography.     

Synthesis and characterisation of N-toluyl methacrylatoethyl carbamates,homopolymers and copolymers with methyl methacrylate

The paper describes the synthesis of N-2/4-toluyl methacrylatoethyl carbamates using 2/4-toluyl isocyanate and 2-hydroxyethyl methacrylate. Homopolymerisation and copolymerisation of these novel monomers with methyl methacrylate was carried out using benzoyl peroxide as an initiator and tetrahydrofuran as solvent. Photopolymerisation of N-4-toluyl methacrylatoethyl carbamate could be carried out without the use of photosensitiser. Structural characterisation of copolymers was done using ¹H-NMR. Thermal stability of copolymers was evaluated in a nitrogen atmosphere by dynamic thermogravimetry.     

The fracture stress of chemical vapour deposited diamond

The factors which control the fracture stress of chemical vapour deposited diamond have been studied using the 3-point bend geometry. Fracture stress values of 300–800 MPa for the growth side and 600–1200 MPa for the nucleation side were recorded for samples of thickness 0.4–2.4 mm. A Weibull modulus of 23 was calculated for the growth surface data, showing that the fracture stress variability was low for a brittle material. A theory based on these results demonstrates that the material behaviour is remarkably simple, depending only on the grain size and the sample thickness, regardless of wide variations in other parameters such as optical transmission and stress state. The paper also contains a possible explanation for this well-defined behaviour based on microstress variations resulting from differences in defect density in different growth sectors within a grain.