Effect of mode of operation of bioreactors on the biosynthesis of penicillin amidase in Escherichia coli

Different operational mode of bioreactors influence the biosynthesis of the enzyme and related products as well as the growth of industrial microorganisms. This communication deals with the effect of mode of operation of various bioreactors with different geometric configurations, viz., batch (includes commercially available batch stirred tank, and custom-designed cylindrical and tapered reactors), batch-fed, continuous flow stirred tank reactors on the biosynthesis of penicillin amidase in Escherichia coli. Experimental findings show that the biosynthesis of penicillin amidase in E. coli show a little variation among batch reactor modes and significant variation on the continuous mode of operation. Further analysis show that the different reactor modes also influence periplasmic localization of the enzyme in the cell.     

Cloning and characterization of theNeisseria gonorrhoeae MS11folC gene

The gene coding for folylpoly-(γ)-glutamate synthetase (FPGS)-dihydrofolate synthetase (DHFS) ofNeisseria gonorrhoeae (Ngo) has been cloned by functional complementation of anEscherichia coli folC mutant (SF4). The sequence encodes a 224-residue protein of 46.4 kDa. It shows 46% identity to theE. coli FPGS-DHFS and 29% identity to the PFGS ofLactobacillus casei. Sequence comparisons between the three genes reveal regions of high homology, including ATP binding sites required for bifunctionality, all of which may be important for FPGS activity. In contrast toL. casei FPGS, theE. coli andNgo enzymes share some additional regions which may be essential for DHFS activity. The products ofNgo folC and flanking genes were monitored by T7 promoter expression. Interestingly, deletion of the upstreamfolI gene, which encodes a 16.5 kDa protein, abolishes the capacity offolC to complementE. coli SF4 to the wild-type phenotype. The ability to complement can be restored byfolI providedin trans. UnlikefolC mutants, gonococcalfolI mutants are viable and display no apparent phenotype. Thus, in contrast toE. coli, Ngo folC is expressed at a sufficiently high level from its own promoter, in the absence of FolI. This study provides the first insights into the genetic complexity of one-carbon metabolism inNgo.     

Recoverability of heat-injured Bacillus spores by lysozyme and EDTA or alkaline thioglycollate

The D_95°C value of Bacillus thuringiensis spores plated in the presence of lysozyme increased from 3.0 min to 3.6 min by post-treatment of heat-injured spores with 50mm EDTA. In the case of Bacillus alvei and Bacillus polymyxa spores D-values decreased from 4.9 to 4.3 min and from 4.7 to 4.1 min respectively. Post-treatment of heat-injured spores treated with alkaline thioglycollate increased D_95°C values of Bacillus alvei from 4.2 to 5.3 min, B. thuringiensis 3.6 to 4.7 min, and Bacillus polymyxa from 4.2 to 5.0 min when spores were plated in the presence of lysozyme. Electron micrographs of heat-injured B. alvei spores treated with sodium thioglycollate indicated that the coat layers of the treated spores were granulated and less intact than the control spores.     

Population differentiation and evolution in the common guillemot Uria aalge

Common (Uria aalge) and Brünnich's guillemots (U. Iomvia) are colonial seabirds that nest in temperate to arctic oceans throughout the Northern hemisphere. They are very similar in the characteristics of ecology, demography and life history that are thought to determine the extent of differentiation among populations, yet geographic variation in morphology is notably greater in common guillemots. Despite evidence of strong natal philopatry, previous analyses of allozymes and the mitochondrial cytochrome b gene revealed little genetic differentiation among North Atlantic colonies of Brünnich's guillemots. To determine if the more extensive morphological variability in common guillemots reflects greater genetic variability, we sequenced part of the cytochrome b gene for 160 common guillemots from 10 colonies distributed throughout the Northern hemisphere. Genotype frequencies and phylogenetic relationships among genotypes both indicated that Atlantic and Pacific populations are genetically distinct. Genetic divergence among genotypes suggested that differentiation of these populations has resulted from separation by Pleistocene glaciers and the Bering Landbridge, as well as by currently unsuitable breeding habitat in the Arctic Ocean. Cytochrome b genotype frequencies also differed among Atlantic colonies, and appeared to define a cline similar to that described for morphological characters. Analyses of sequence variation suggested that this variation probably results from secondary contact between two refugial populations from the Pleistocene glaciations, rather than from isolation by distance or selection. In contrast, the Atlantic population of Brünnich's guillemots appears to have arisen through recent expansion of a single homogeneous refugial population.     

Actin' like actin?

The most biologically significant property of actin is its ability to self-associate and form two-stranded polymeric microfilaments. In living cells, these micro filaments form the actin cytoskeleton, essential for maintenance of the shape, passive mechanical properties and active motility of eukaryotic cells. Recently discovered actin-related proteins (ARPs) appear to share a common ancestor with conventional actin. At present, six classes of ARPs have been discovered, three of which have representatives in diverse species across eukaryotic phyla and may share functional characteristics with conventional actin. The three most ubiquitous ARPs are predicted to share a common core structure with actin and contain all the residues required for ATP binding. Surface residues involved in protein protein interactions, however, have diverged. Models of these proteins based on the atomic structure of actin provide some clues about how ARPs interact with each other, with conventional actin and with conventional actin-binding proteins.     

Changes in TNF and IL-6 production after diphtheria toxoid vaccination: drug modulation of the cytokine levels

The effect of vaccination with diphtheria toxoid (AD-M) on TNF and IL-6 production has been studied in humans. In the present study it was demonstrated that immunization with AD-M resulted in changes of in vitro TNF and IL-6 production by peripheral blood mononuclear cells. TNF release was suppressed but IL-6 production was stimulated. On the other hand, serum levels of TNF were markedly increased over a period of 3 weeks. It was also demonstrated that the postvaccinal cytokine production disturbances may be corrected by pretreatment with a new synthetic hexapeptid (Imunofan((R))). It is possible that the imunofan treatment could prevent some postvaccinal complications.     

Seasonality ofOrmia depletaand Limits to Its Spread

Ormia depleta(Wiedemann) (Diptera: Tachinidae), a South American fly that homes on the calling songs of its hosts, was brought from Piracicaba, Brazil; colonized; and released in Florida for the biological control ofScapteriscusspp. mole crickets. It became established at some release sites and rapidly spread through most of peninsular Florida. The seasonal distributions of phonotactic females in Florida differed from the single summer peak recorded at Rio Claro and Piracicaba, Brazil (23° S). Near Bradenton (27° N), spring and fall peaks with a summer hiatus were quickly established and remain evident. At two sites near Gainesville (30° N), a strong fall peak and a modest-to-none spring peak developed. Peaks of phonotactic females seem to follow times of peak host availability and sometimes coincide with minima of host availability. Limits to geographical expansion ofO. depletaare poorly understood. At Gainesville, the site of earliest introduction, establishment was rapid and soundtrap catches increased for the first 3 years, exceeding 1000 per year at one site. Catches then declined for 3 years, almost reaching zero in 1994. On the other hand, annual sound-trap catches ofO. depletaat Bradenton exceeded 1000 in <2 years and have remained above that level for 5 years. Winters in Gainesville are more severe than those in Bradenton, but minimum winter temperatures in Gainesville were substantially lower duringO. depleta's increase than during its decline. Perhaps mild winters cause the flies to become active early—only to starve for lack of adequate winter nectar sources.O. depletamay yet spread to the northern limits of itsScapteriscushosts as present populations adapt to new environments. Or farther spread may require that new strains be introduced from more southerly sites in South America.     

Biology and life cycle ofAtelocauda koae,an unusual demicyclic rust

Atelocauda koae, a rust of the native HawaiianAcacia koa, is considered as a demicyclic species, having spermogonial, aecial, and telial states, but is unusual in production of aeciospores simultaneously with teliospores rather than consecutively. Host inoculation with spores of each state separately confirmed that the life cycle was perpetuated by the telial state, but the aeciospores, while capable of germination and stomal penetration, did not produce detectable infection. This rust therefore behaves as a microcyclic species, and appears to be in evolutionary transition toward this reduced state. Teliospores produced vestigial, permanently attached basidiosopores which germinated to produce infective hyphae. The hyphae entered the host either through stomata or penetrated the epidermis directly, with the latter method being more common. Unusual nuclear associated with teliospore germination, in which meiosis occurs in more than one diploid nucleus was observed, in confirmation of an earlier study.