Effect of growth rate on histidine catabolism and histidase synthesis in Aerobacter aerogenes

A study was made of how the catabolism of a carbon and energy source is affected by the biosynthetic demands of growing bacterial cells. Cultures of Aerobacter aerogenes in l-histidine medium were grown in a chemostat at rates determined by the supply of either sulfate or a required amino acid, l-arginine. It was discovered that the rate at which these cells grow under a biosynthetic restriction determines both the rate and the pattern of histidine degradation. (i) Histidine catabolism is partially coupled to the growth rate. This coupling is achieved by catabolite repression of histidase (histidine ammonia lyase; EC 4.3.1.3.), and also by a slightly decreased in vivo function of this enzyme at low growth rates. (ii) The looseness of the coupling results in a direct relationship between growth rate and growth yield, and possibly is correlated with an altered pattern of carbon flow from histidine. (iii) Sudden decreases in growth rate cause total repression of histidase synthesis for substantial periods of time. (iv) Sudden release of biosynthetic restriction leads rapidly to an increase in the functioning of the cells' complement of histidase, an increase in the rate of synthesis of this enzyme, and an increase in the growth yield from histidine.     

Effect of Processing on the Microflora of Norwegian Seaweed Meal, with Observations on Sporendonema minutum (Høye) Frank and Hess

Meal from the brown seaweed Ascophyllum nodosum (L.) Le Jol. is mainly used as an animal feed supplement. Since moist weed often develops a marked mold growth and since little was known about the microflora of seaweed meal, a cultural procedure was developed to enumerate the populations of bacteria, yeasts, and molds of seaweed meals manufactured by different drying processes. The microflora could be supported by a variety of media varying in levels of nutrition and in the source and concentration of salts. Fresh weed contained less than 10(3) bacteria and less than 10(2) yeasts and molds per g (dry weight). The type and extent of microbial populations in seaweed meal appeared to be dependent upon the method of seaweed drying. Rotary drum-drying at temperatures decreasing from 800 to 80 C maintained or reduced the microbial populations to 10(3) organisms per g (dry weight). Although meals with high nutritional quality can be obtained with warm air- or rock-dried weed, these conditions can also permit bacterial and mold development. Extended rock-drying in variable weather conditions and prolonged storage of moist weed, both of which decrease the nutritional quality, also lead to high bacterial numbers and to a marked development of the halophilic brown mold Sporendonema minutum which attained populations of 10(8) viable spores per g of dried weed. A poultry diet containing 5% badly molded weed had no apparent toxic or growth-depressing effect when fed to chicks.     

Comparative control of a branch-point enzyme in microorganisms

Thirty-two genera of microorganisms were identified with one of six distinctive control patterns for the enzyme 3-deoxy-d-arabino-heptulosonate-7-phosphate synthetase. These patterns included sequential feedback inhibition, isoenzyme feedback inhibition, cumulative feedback inhibition, and three (apparent) simple one-effector patterns. Documentation is provided of an overwhelming tendency for control patterns to be strongly conserved among the member species of the various genera that were examined.     

Apparatus for monitoring the oxygen uptake and carbon dioxide production of fermentations

The requirements of the continuous analysis of effluent gas streams from aerated flash and tank fermentors are described, as are instrumental devices for measuring the oxygen and carbon dioxide content of fermentor gases. The use of a specially designed sequential gas sample for monitoring four fermentations simultaneously and a system for precise control of low air flow and pressure is explained. Equations for calculating carbon dioxide production or oxygen consumption rates and respiratory quotients are given. A discussion of the operating characteristics of a device for automatic translation of aeration data between fermentors is presented.     

Zur Heredität des Strabismus concomitans

697 children with concomitant strabism, all patients who were seen in the Ophthalmalogical university hospital and outpatient service of the Charité, Berlin, during a certain period, have been examined together with their parents and siblings. In a second series, 3398 12-years old school children (born 1953) from three urban districts of Berlin were examined at the occasion of a vaccination term, and 179 children with strabism were ascertained. Probands as well as their parents and siblings were examined thoroughly according to the criteria of strabism diagnostics. Incidence of squinting among siblings was increased markedly when compared with the population frequency, but did not reach the expectations under rthe assumption of a single, monomeric mode of inheritance. Admixture of phenocopies or new mutants could be excluded. Twin investigations (12 monocygotic and 27 dicygotic pairs) showed a manifestation rate of 94,1% in monocygotic pairs, corresponding to a 3 1/2 times higher concordance in monocygotic as compared with dicygotic twins. From the results discussed, a multifactorial genetic system in combination with a threshold effect seems to be the most likely genetic interpretation. An analysis of pedigrees shows that slight sensoric as well as motoric anomalies might combine in different and continuously varying quantities, contributing to the syndrom of concomitant strabism.     

A model for the stepwise radiation inactivation of the alpha 2-dimer of Na,K-ATPase

This study is a direct continuation of Jensen, J., and N?rby, J. G., (1988) J. Biol. Chem. 263, 18063-18070. A new model in which we propose that the in situ organization of the Na,K-ATPase alpha-subunit is an alpha 2-dimer and which describes the stepwise degradation by radiation inactivation of this assembly is presented on the basis of the following findings. Radiation inactivation size for alpha-peptide integrity, normal nucleotide, vanadate and ouabain binding, and K-pNPPase activity is close to m(alpha) = 112 kDa; for Na-ATPase activity it is 135 kDa and for Na,K-ATPase activity it increases from 140 to about 195 kDa with increasing assay ATP concentration (equal to increasing average turnover). Normal Tl+ occlusion had the same radiation inactivation size as Vmax for Na,K-ATPase, i.e. about 195 kDa. The binding experiments disclosed radiation-produced molecules with active binding sites but with a lower than normal affinity. Radiation inactivation size for the total binding capacity of ADP and ouabain was therefore smaller than the size of an alpha-peptide, namely about 70 kDa, and for total Tl+ occlusion it was down to 40 kDa. We can explain all these observations by using a new approach to target size analysis and by assuming a dimeric organization of the alpha-subunit. Each alpha-peptide is degraded stepwise by first destruction of either a 42- or a 70-kDa domain, and the partly damaged peptide may retain biochemical activity. We conclude that there is no role for the beta-subunit in catalysis and that the alpha-peptide is organized as an alpha 2-dimer in the membrane with each alpha-subunit being able to perform complete catalytic cycles (and probably also active transport), provided that it is stabilized by an adjacent alpha-peptide or a sufficiently large fragment thereof.     

Ca2+/calmodulin-dependent protein kinase enriched in cerebellar granule cells. Identification of a novel neuronal calmodulin-dependent protein kinase

Ca2+-sensitive protein kinases are thought to play a pivotal role in Ca2+-mediated neuronal communication. We describe here the cloning, purification, and characterization of a major Ca2+/calmodulin-dependent, brain-specific protein kinase which is particularly enriched in cerebellar granule cells. The enzyme is comprised of Mr 65,000 and 67,000 polypeptides which copurify to homogeneity and phosphorylate synapsin I. The protein kinase is coded for by two poly(A+) RNAs of 2.0 and 3.5 kilobases which probably derive from a single gene. Two cDNA inserts, one of 198 base pairs and one of 1225 base pairs, contain a total of 677 base pairs of the protein coding sequence which includes sequences homologous to other calmodulin-dependent protein kinases including part of the calmodulin-binding domain. The surprising presence of extended sequences which are enriched in glutamate residues may influence the subcellular distribution of this kinase. Immunohistochemical localization with an affinity-purified antibody reveals that whereas the enzyme is expressed in several neuronal subpopulations, it is exceptionally enriched in the granule cells of the cerebellum. The relevance of the biochemical, molecular, and histologic properties of this enzyme is discussed in the context of neuronal Ca2+ signaling.