ALBERT

Notes: Abstract A comparative study was made of the regulation of the synthesis of methanol dissimilating enzymes inkloeckera sp. 2201 andHansenula polymorpha using chemostat and batch growth conditions and methanol or glucose as carbon sources. During growth in methanol-limited chemostat cultures similar enzyme patterns for alcohol oxidase, catalase, formaldehyde dehydrogenase and formate dehydrogenase in the two yeasts were found. When growing in batch culture with glucoseH. polymorpha, but notKloeckera sp. 2201, was found to produce ethanol which might affect the synthesis of these enzymes.

Notes: Ab initio MO calculations, using both minimal (STO-3G) and extended (Roos-Siegbahn) basis sets are reported for the systems methanethiol-imidazole, methanethiol-imidazole-formaldehyde, and methanethiol-imidazole-formamide, which, together with a point-change representation of a long α-helix, form models for the active site of papain. It is shown that the large electric field exerted by the helix in the active-site region is responsible for the presence of the essential residues Cys 25 and His 159 in the form of an ion pair RS- ··· ImH+, which is crucial for a recently proposed mechanism for the catalytic action of the enzyme. Also, an explanation is given for the anomalies in measured pK values for these residues. Detailed studies on the (sub)systems show that minimal basis sets lack the flexibility necessary for describing the type of proton transfer involved. We conclude that α-helices are essential parts of enzymes and that they play a significant role in the catalytic process.

Notes: Me3SiCl bildet mit N-Methylimidazol (NMI) eine bei Raumtemperatur beständige, kristalline 1:1 Verbindung. Nach der Röntgenstrukturanalyse entsteht unter Spaltung der Si—Cl-Bindung und Anlagerung eines NMI-Rings eine Verbindung ionischer Struktur [Me3Si(NMI)]+Cl- 1. Die Reaktion von Me2HSiCl mit NMI (Molverhältnis 1:2) führt unter Spaltung der Si—Cl-Bindung und Koordination von zwei NMI-Ringen zur Verbindung [Me2HSi(NMI)2]+Cl- 2. Bei der analogen Umsetzung von Me2SiCl2 mit NMI (Molverhältnis 2:1) entsteht eine Verbindung, die Me2SiCl2 und NMI im Molverhältnis 1:2 enthält. Bei der Sublimation werden Einkristalle der Verbindung [Me2Si(NMI)3]2+ 2 Cl-· NMI 3 erhalten.

Notes: One of the basic prerequisites for rational and high quality production of plastic parts is a tool layout tailored to the production process. To date, both design and construction have generally been based on values acquired by experience. This first, necessitates highly qualified personnel and second, involves what is frequently time consuming and costly finishing work. Experience acquired so far with computer-aided layout of injection molds shows that even a designer with little experience reaches the target more quickly and more reliably. At the same time he is able to draw on the results of intricate calculation and simulation methods which he was unable to apply in the past for reasons of time alone. This paper thus sets out the possibilities currently open and the experience available for computer-aided mold layout. The chief point of focus here is a system for the layout of injection molds. Working on from this system, however, the potentials for computer application are presented for blow mold- and foaming mold-design. After finding the mold principle, mold layout essentially divides up into two major areas, namely dimensioning calculations (CAE) and compilation of production documents (CAD). In dimensioning, the different functional elements of the mold are calculated. The aids that have been developed and the potentials of computer-aided dimensioning are presented with examples from the fields of rheological, thermal, and mechanical mold layout. Computer-aided rheological layout divides up into two steps. The first gives information on qualitative filling behavior (filling picture, flow paths) and the second provides quantitative results (pressures, shear stresses, temperatures). Computer-aided thermal layout similarly divides up into part steps. These are a rough overall energy balance, a rough layout of the tempering system, a segmented layout, and a homogeneity check, which involves simulating the temperature conditions in the mold by means of difference methods. When it comes to mechanical layout of the mold, programs are available for deformation calculations on basic cases and these will frequently be sufficient. For more complex cases of loading and deformation, a finite element program is used. Graphic data processing units can be used to supply extra facilities - first, to provide an aid for the simulation programs in dimensioning and second, to rationalize the compilation of the production documents. An illustration of a CAD workplace is given, incorporating the necessary computer configuration and peripherals. Compiling production documents is essentially a problem of variant design. The variants in this case are the individual components of the mold and a number of standardized accessories that can be called up as “standardized components.” The mold cavity, however, always has to be a free design. All programs are dialogue driven and are in a standardized manner so that even designers with no data processing experience can use the computer as an aid. The CAD/CAE system presented duly fulfils all these requirements. It allows the designer, at a single computer work station, to carry out both simulation and dimensioning calculations, to obtain information on material data, and to compile production documents on the basis of variant and free design. This provides the designer with a readily manageable aid and makes a considerable contribution towards improving the design result. Finally, the capacity of different computer concepts and the CAE/CAD/CAM systems on the market are discussed. For the future it will be possible to establish a computer-aided link between the different areas of design, from development of the molded part, via mold design and production, through setting the processing parameters of the injection molding machine.

Notes: Abstract The growth of Hansenula polymorpha and Kloeckera sp. 2201 with a mixture of glucose and methanol (38.8%/61.2%, w/w) and the regulation of the methanol dissimilating enzymes alcohol oxidase, catalase, formaldehyde dehydrogenase and formate dehydrogenase were studied in chemostat culture, as a function of the dilution rate. Both organisms utilized and assimilated glucose and methanol simultaneously up to dilution rates of 0.30 h-1 (H. polymorpha) and 0.26h-1, respectively (Kloeckera sp. 2201) which significantly exceeded μmax found for the two yeasts with methanol as the only source of carbon. At higher dilution rates methanol utilisation ceased and only glucose was assimilated. Over the whole range of mixed-substrate growth both carbon sources were assimilated with the same efficiency as during growth with glucose or methanol alone. In cultures of H. polymorpha, however, the growth yield for glucose was lowered by the unmetabolized methanol at high dilution rates. During growth on both carbon sources the repression of the synthesis of all catabolic methanol enzymes which is normally caused by glucose was overcome by the inductive effect of the simultaneously fed methanol. In both organisms the synthesis of alcohol oxidase was found to be regulated differently as compared to catalase, formaldehyde and formate dehydrogenase. Whereas increasing repression of the synthesis of alcohol oxidase was found with increasing dilution rates as indicated by gradually decreasing specific activities of this enzyme in cell-free extracts, the specific activities of this enzyme in cell-free extracts, the specific activities of catalase and the dehydrogenases increased with increasing growth rates until repression started. The results indicate similar patterns of the regulation of the synthesis of methanol dissimilating enzymes in different methylotrophic yeasts.

Notes: Abstract The influence of the composition of methanol/glucose-mixtures as only sources of carbon and energy on growth and regulation of the synthesis of enzymes involved in methanol-dissimilation was studied under chemostat conditions at a fixed dilution rate with the methylotrophic yeasts Hansenula polymorpha and Kloeckera sp. 2201. Both carbon sources were found to be utilized completely independently of the composition of the C1/C6 mixture. Using mixtures of 14C-labelled methanol and glucose the growth yield for glucose was found to be constant for all C1/C6-mixtures tested and both yeasts. The growth yield for methanol, however, was reduced by up to 25% when the proportion of methanol in the inflowing medium was lower than 20% (w/w with respect to glucose) for H. polymorpha and 50% (w/w with respect to glucose) for Kloeckera sp. 2201 respectively. During growth with C1/C6-mixtures containing higher C1-proportions of methanol regular growth yields for methanol were recorded which corresponded to the growth yields found with methanol as the only carbon source. The regulation of the synthesis of the enzymes of the dissimilatory pathway for methanol was found to be under multiple control. Although glucose was present in the medium methanol had a positive effect on the synthesis of these enzymes. Thus, in addition to derepression induction by methanol was also observed. This inductive effect was found to increase with increasing proportions of methanol in the mixture. Depending on the enzyme, 10–40% methanol in the mixture resulted in a maximal induction with enzyme specific activities equal to those found in cells grown with methanol as the only carbon source. No further enhancements in enzyme specific activities were observed during growth on mixtures containing more than 40% methanol.

Notes: Thermo-oxidative ageing processes were investigated in films (100 μm thick) made from isotactic polypropylene with different antioxidant concentrations between 0% and 0.1% using isothermal long-term differential thermal analysis (ILDTA). The end of the oven life of polypropylene coincided with the loss of mechanical properties which was confirmed by tensile tests at temperatures far below the crystallite melting range. Already prior to the beginning of autocatalytic decomposition, ILDTA experiments permits conclusions to be drawn on the thermal history of the specimens and statements made regarding the end of thermo-oxidative stability of the polymer, which is accompanied by a deterioration in the mechanical properties.In consequence of the correlation between oxidation time and antioxidant concentration the local antioxidant concentration in sheets or films can be measured by ILDTA. Thus, investigating the diffusion of an antioxidant in polypropylene using ILDTA is possible.

Notes: Infrared spectroscopy was used for the determination of the base-pairing content of four specific tRNAs in deuterium oxide solution. Infrared spectra were obtained in the 1750-1550 cm-1 region at various temperatures ranging from about 15 to 90°C. Melting curves were constructed by plotting the molar extinction coefficient at ν = 1657 cm-1 versus temperature. These transition curves enabled us to determine the ranges of temperature which correspond to the ordered (partially double-stranded) or randomly coiled structure of the tRNA. For a set of wavenumbers the extinction coefficients at these temperatures were used for the calculation of the base-pairing content. The procedure employed here is based on a method described earlier by Thomas [(1969) Biopolymers 7, 325-334]. For the conditions selected for this investigation (Mg2+-free D2O-buffer; 0.01M tris-DCl, 0.015M NaCl, pD 7.5) the results of this determination agree within the limits of errors with the number of base pairs predicted by the cloverleaf model.

Notes: Recently, it was suggested that parallel β-sheets have a significant dipole moment, in contrast to antiparallel sheets. Ab initio molecular-orbital (MO) calculations on parallel and antiparallel β-strands of tetra(Gly) show that they have very similar charge distributions. Interaction energies between two and three strands of tetra(Gly), obtained using the direct reaction field Hamiltonian, show that a particular choice of point charges is probably not crucial for calculating interactions within β-sheets, but that it might be for calculating interactions between these sheets and other parts of a protein, in particular, α-helices. The point-charge representation of our MO-SCF results will probably reduce the hazard of introducing artefacts in electrostatic calculations of protein conformational energies, provided the short-range interactions are treated in a more realistic way, i.e., such that intra- and interchain induction effects are included.