Microbial metabolism of chlorosalicylates: effect of prolonged subcultivation on constructed strains

The hybrid strain Pseudomonas sp. WR4016 was subcultivated with increasing concentrations of 5-chlorosalicylate (510 mM) as sole carbon source over a period of 9 months. At intervals of approximately 3 months derivative strains WR4017, WR4018 and WR4019 were isolated which exhibited higher growth rates and increased substrate tolerance. Comparative analysis of the turnover rates of the key enzymes in chlorosalicylate degradation showed that the adaptation process did not result from structural modifications of these proteins. Instead, balanced over-production of the salicylate hydroxylase and catechol 1,2-dioxygenase prevented the accumulation of toxic chlorocatechols and accounted for the reduction of the doubling times with 4- or 5-chlorosalicylate. A comparative analysis of a genetically engineered chlorosalicylate degrader PL300-1 showed similar regulatory patterns as the most advanced isolate WR4019 from the adaptation series.     

Molecular basis for the special properties of proteins and enzymes from Halobacterium marismortui

Abstract Three proteins from Halobacterium marismortui , malate dehydrogenase (hMDH), glutamate dehydrogenase (hGDH) and ferredoxin (hFD) were purified and characterized with respect to their molecular masses, amino acid composition and, for hFD only, primary structure. Striking features of halophilic proteins are: the high excess of acidic over basic residues; acidic clusters in the sequence. Low-salt concentration causes inactivation and changes in structural parameters of hMDH and hGDH. Reactivation of hMDH involves long-lived stable intermediates. The salt concentration optimum of enzymic activity is independent of salt nature. The high capacity of halophilic proteins to retain water and salt is due to unique molecular properties, studied by physico-chemical techniques.     

Reproductive structures and phylogenetic significance of extant primitive Angiosperms

A brief survey is presented on fossil reproductive structures of early Angiosperms from the Lower and mid-Cretaceous and at the same time on the reproductive structures of those extant Angiosperms which resemble most closely these fossils and which seem to be especially primitive also on other grounds: a first group (Degeneriaceae, Himantandraceae, Eupomatiaceae, Austrobaileyaceae) possessing relatively complicated and conspicuous flowers with elaborated inner staminodes, a second group (Chloranthaceae, Trimeniaceae, Amborellaceae) possessing small and relatively simple, inconspicuous flowers with peculiar features in the carpels, and a third group (Winteraceae) possessing flowers with unusual variability in organ number and size. The three groups exhibit a certain diversity in pollination biology, although cantharophily seems to prevail, however different the cantharophily character syndromes may be between the groups. In the extant primitiveMagnoliidae variability occurs on other morphological levels than in the higher advanced Angiosperms. This has to be taken into consideration in evaluations of the systematic relationships of the various groups of theMagnoliidae. Presumably often their relationships are closer than it may appear at first sight. This is also true for the three groups here discussed.     

Dental microwear features as an indicator for plant food in early hominids: A preliminary study of enamel

Studies of dental microwear have shown qualitative and quantitative differences related with specific diets of mammals. Here we report a classification of the features of microwear and the results of a comparative study of the characteristics of wear produced by plant food. Species which are mainly folivorous or herbivorous show common features useful for providing information about the diet of early hominids at Garusi and Laetoli (Tanzania), Hadar (Ethiopia) and Olduvai (Tanzania).     

Mutant forms of staphylococcal nuclease with altered patterns of guanidine hydrochloride and urea denaturation

Eleven mutant forms of staphylococcal nuclease with one or more defined amino acid substitutions have been analyzed by solvent denaturation by using intrinsic fluorescence to follow the denaturation reaction. On the basis of patterns observed in the value of m--the rate of change of log Kapp (the apparent equilibrium constant between the native and denatured states) with denaturant concentration--these proteins can be grouped into two classes. For class I mutants, the value of m with guanidine hydrochloride is less than the wild-type value and is either constant or increases slightly with increasing denaturant; the value of m with urea is also less than wild type but shows a marked increase with increasing denaturant concentration, often approaching but never exceeding the wild-type value. For class II mutants, m is constant and is greater than wild type in both denaturants, with the increase being consistently larger in guanidine hydrochloride than in urea. When double or triple mutants are constructed from members of the same mutant class, the change in m is usually the sum of the changes produced by each mutation in isolation. One plausible explanation for these altered patterns of denaturation is that chain-chain or chain-solvent interactions in the denatured state have been modified--interactions which appear to involve hydrophobic groups.     

Partial purification,subunit structure and thermal stability of the photochemical reaction center of the thermophilic green bacterium Chloroflexus aurantiacus

Spectrally pure reaction center preparations from Chloroflexus aurantiacus have been obtained in a stable form; however, the product contained several contaminating polypeptides. The reaction center pigment molecules (probably three bacteriochlorophyll a and three bacteriopheophytin a molecules) are associated with two polypeptides (M_r = 30000 and 28000) in a reaction center complex of M_r = 52000. No carotenoid is present in the complex. These data together with previous spectral data suggest that the Chloroflexus reaction center represents a more primitive evolutionary form of the purple bacterial reaction center, and that it has little if any relationship to the green bacterial component. A reaction center preparation from Rhodopseudomonas sphaeroides R26 was fully denatured at 50°C while the Chloroflexus reaction center required higher temperatures (70–75°C) for complete denaturation. Thus, an intrinsic membrane protein of a photosynthetic thermophile has been demonstrated to have greater thermal stability than the equivalent component of a mesophile.     

Light-driven proton transport of bacteriorhodopsin incorporated into long-term stable liposomes of a polymerizable sulfolipid

The chromoprotein bacteriorhodopsin from Halobacterium halobium has been incorporated into liposomes made of a fully synthetic, polymerizable lipid. Bacteriorhodopsin is found to be active in these polymer liposomes. The advantage in the use of such polymer systems concerning long-term stability in comparison with liposomes made of natural lipid is demonstrated.     

Intracellular recordings from nonspiking interneurons in a semiintact,tethered walking insect

Nonspiking interneurons were investigated in a tethered, walking insect, Carausius morosus, that was able to freely perform walking movements. Experiments were carried out with animals walking on a lightweight, double-wheel treadmill. Although the animal was opened dorsally, the walking system was left intact. Intracellular recordings were obtained from the dorsal posterior neuropil of the mesothoracic ganglion. Nonspiking inter-neurons, in which modulations of the membrane potential were correlated with the walking rhythm, were described physiologically and stained with Lucifer Yellow. Interneurons are demonstrated in which membrane potential oscillations mirror the leg position or show correlation with the motoneuronal activity of the protractor and retractor coxae muscles during walking. Other interneurons showed distinct hyperpolarizations at certain important trigger points in the step cycle, for example, at the extreme posterior position. Through electrical stimulation of single, nonspiking interneurons during walking, the motoneuronal activity in two antagonistic muscles—protractor and retractor coxae—could be reversed and even the movement of the ipsilateral leg could be influenced. The nonspiking interneurons described appear to be important premotor elements involved in walking. They receive, integrate, and process information from different leg proprioceptors and drive groups of leg motoneurons during walking.     

微管的冷稳定性与植物抗寒性关系的研究

利用间接免疫荧光的细胞化学技术对番茄、黄瓜、菠菜、甜菜及小麦等不同抗寒性植物微管的冷稳性进行了比较研究。结果指出,不抗寒的喜温性植物番茄和黄瓜的气孔保卫细胞的微管在0℃—1℃冷处理3小时即解聚;属于中等抗寒性植物的菠菜和甜菜幼苗经秋季低温锻炼后,其气孔保卫细胞的微管在0℃和—5℃低温处理3小时,均不发生解聚;具有较强抗寒性的冬小麦品种农大139幼苗在2—3℃低温锻炼期间,微管结构保持完整,经过15天低温锻炼的幼苗在-8℃冰冻处理3小时,微管也不受破坏。这些结果表明,微管的冷稳性与植物的抗寒性成正相关。