Biosynthesis of isoprenoids. purification and properties of IspG protein from Escherichia coli

[Structure: see text]. The IspG protein is known to catalyze the transformation of 2-C-methyl-d-erythritol 2,4-cyclodiphosphate into 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate in the nonmevalonate pathway of isoprenoid biosynthesis. We have found that the apparent IspG activity in the cell extracts of recombinant Escherichia coli cells as observed by a radiochemical assay can be enhanced severalfold by coexpression of the isc operon which is involved in the assembly of iron-sulfur clusters. The recombinant protein was isolated by affinity chromatography under anaerobic conditions. With a mixture of flavodoxin, flavodoxin reductase, and NADPH as the reducing agent, stringent assay methods based on photometry or on 13C NMR detection of multiply 13C-labeled substrate/product ratios afforded catalytic activities greater than 60 nmol mg(-1) min(-1) for the protein "as isolated" (i.e., without reconstitution of any kind). Lower apparent activities were found using photoreduced deazaflavin as an artifactual electron donor, whereas dithionite was unable to serve as an artificial electron donor. The apparent Michaelis constant for 2-C-methyl-D-erythritol 2,4-cyclodiphosphate was 700 microM. The enzyme was inactivated by EDTA and could be reactivated by Mn2+. The pH optimum was at 9.0. The protein contained 2.4 iron ions and 4.4 sulfide ions per subunit. The replacement of any of the three conserved cysteine residues afforded mutant proteins which were devoid of catalytic activity and contained less than 6% of Fe2+ and less than 23% of S2- as compared to the wild-type protein. Sequence comparison indicates that putative IspG proteins of plants, the apicomplexan protozoan Plasmodium falciparum, and bacteria from the Bacteroidetes/Chlorobi group contain an insert of about 170-320 amino acid residues as compared with eubacterial enzymes.     

Corrigendum to “1/f Noise and intermittency due to diffusion of point defects in a semiconductor material” [Physica A 282 (2000) 108]

In the above-mentioned paper, the total rate of generation is defined by (unnumbered equation in Chapter 4.2 after Eq. (21)); in this equation the factor (1−c_d+) is missing. As a consequence, Eqs. (22)–(30) and Fig. 4 in the original paper have to be revised which is presented in this corrigendum. Numbering of references remains unchanged.     

Structure-Activity Relationship in Ambergris-Type Woody Odorants possessing a hydronaphthalene skeleton

A series of methyl hydronaphthyl alcohols, formates, acetates, and ketones were prepared and their odour properties evaluated. Minor structural changes, even opposite to the osmophoric group, were found to have major effects on the odour. ¹³C? NMR shift assignments of the hydronaphthyl derivatives are presented.     

Thin-Film Zinc/Manganese Dioxide Electrodes

Summary.  Thin-film electrodes allow the manufacturing of flat batteries of variable design. Their electric performance is better than that of customary cells because of a larger contact area between anode and cathode and better utilization of the electrochemically active materials. Bipolar thin-film electrodes for the rechargeable alkaline zinc/manganese dioxide system were assembled using graphite-filled plastics (high-density polyethylene and polyisobutylene). In a different approach, extremely thin electrodes were obtained using 25 μm thick foils made of non-conductive micro-porous polypropylene. The electroactive materials were electrolytically deposited into the pores of the previously metallized foil, providing the required conductive connection through the plastic matrix by themselves. Cycle behavior, cumulated capacities, and energies of batteries with up to two bipolar units were measured. At this early stage of development, batteries based on graphite-filled polymer foils showed better results with regard to storage capacity per unit area and to cycle life. Prototypes based on micro-porous polypropylene suffered from the relatively small fraction of pore volume available for the deposition of active material (about 38%) and from current collector corrosion. Received May 30, 2000. Accepted December 18, 2000     

Ideals and Filters in D-Posets

Ideals and filters in D-posets are defined. Therole of idempotent elements and atoms in the theory ofideals and filters is studied.     

A switching for all strongly regular collinearity graphs from polar spaces

We describe a general construction of strongly regular graphs from the collinearity graph of a finite classical polar spaces of rank at least 3 over a finite field of order q. We show that these graphs are non-isomorphic to the collinearity graphs and have the same parameters. For most of these parameters, the collinearity graphs were the only known examples, and so many of our examples are new.