Impact of interspecific competition and drought on the allocation of new assimilates in trees.

In trees, the interplay between reduced carbon assimilation and the inability to transport carbohydrates to the sites of demand under drought might be one of the mechanisms leading to carbon starvation. However, we largely lack knowledge on how drought effects on new assimilate allocation differ between species with different drought sensitivities and how these effects are modified by interspecific competition. We assessed the fate of (13) C labelled assimilates in above- and belowground plant organs and in root/rhizosphere respired CO2 in saplings of drought-tolerant Norway maple (Acer platanoides) and drought-sensitive European beech (Fagus sylvatica) exposed to moderate drought, either in mono- or mixed culture. While drought reduced stomatal conductance and photosynthesis rates in both species, both maintained assimilate transport belowground. Beech even allocated more new assimilate to the roots under moderate drought compared to non-limited water supply conditions, and this pattern was even more pronounced under interspecific competition. Even though maple was a superior competitor compared to beech under non-limited soil water conditions, as indicated by the changes in above- and belowground biomass of both species in the interspecific competition treatments, we can state that beech was still able to efficiently allocate new assimilate belowground under combined drought and interspecific competition. This might be seen as a strategy to maintain root osmotic potential and to prioritise root functioning. Our results thus show that beech tolerates moderate drought stress plus competition without losing its ability to supply belowground tissues. It remains to be explored in future work if this strategy is also valid during long-term drought exposure.

Evidence for two fatty alcohol oxidases in the biosurfactant-producing yeast Candida (Torulopsis) bombicola.

Fatty alcohol oxidase activities in Candida (Torulopsis) bombicola ATCC 22214, which produces large amounts of glycolipids consisting of omega- and (omega-1)-hydroxyfatty acids and a sugar moiety, occurred only in the microsomal fraction whether the cells had grown on n-alkanes, carbohydrates or a mixture of the two. High activities occurred in glucose-grown cells. Aliphatic alcohols from octanol to hexadecanol were oxidized with two maxima in activities for decanol and for tetradecanol. Differences in their pH optima and in temperature stability suggest two separate enzymes are present. Long chain diols, but not omega-hydroxyfatty acids, were also oxidized.

Formation and physiological role of biosurfactants produced by hydrocarbon-utilizing microorganisms. Biosurfactants in hydrocarbon utilization.

Microbial growth on water-insoluble carbon sources such as hydrocarbons is accompanied by metabolic and structural alterations of the cell. The appearance of surface-active compounds (biosurfactants) in the culture medium or attached to the cell boundaries is often regarded as a prerequisite for initial interactions of hydrocarbons with the microbial cell. Under this point of view, biosurfactants produced by hydrocarbon-utilizing microorganisms, their structures and physico-chemical properties are reviewed. The production of such compounds is mostly connected with growth limitation in the late logarithmic and the stationary growth phase, in which specific enzymes are induced or derepressed. Addition of purified biosurfactants to microbial cultures resulted in inhibitory as well as in stimulatory effects on growth. Therefore, a more differentiated view of microbial production of surface-active compounds is proposed. Biosurfactants should not only be regarded as prerequisites of hydrocarbon uptake, but also as secondary metabolic products.

Selective and rapid solubilization of the microbial membrane enzyme aldehyde dehydrogenase.

An improved solubilization procedure for the membrane-bound quinoprotein aldehyde dehydrogenase from Acetobacter rancens CCM 1774 was established. After the first solubilization of membrane enzymes by Brij 35 which provided important extraction of membrane proteins other than aldehyde dehydrogenase, the application of Trition X-100 resulted in an almost 20-fold purification of quinoprotein aldehyde dehydrogenase. The optimal solubilization was closely connected with definite detergent/protein ratios.

Effects of detergents and phospholipids on the pyridine nucleotide-independent aldehyde dehydrogenase from membranes of Acetobacter rancens.

The pyridine nucleotide-independent aldehyde dehydrogenase solubilized and purified from membranes of Acetobacter rancens CCM 1774 requires the presence of detergents for activity. While several detergents could stimulate the enzyme activity the stability of the enzyme-detergent complexes was rather low. Phospholipid substitution experiments revealed the reversibility of the loss of activity caused by phospholipid removal. Enzyme-phospholipid complexes generated from a complex phospholipid fraction exhibited an increased stability. The predominant effect of phosphatidylglycerol on both enzyme activity and stability is discussed in terms of essential boundary layer phospholipids.

Petite mutants of sophorolipid-producing Candida yeasts.

Treatment of both Candida apicola IMET 43747 and Candida bombicola ATCC 22214 with N-methyl-N'-nitro-N-nitroso guanidine resulted in formation of auxotrophic mutants and cytochrome oxidase negative mutants. The deficiency of intact cytochrome oxidase did not affect the ability to produce sophorose lipid. This indicates that intact mitochondrial energy supplying system is not necessary for the production of the extracellular sophorose lipids by both Candida yeasts.

Phenol degradation by Acinetobacter calcoaceticus NCIB 8250.

Acinetobacter calcoaceticus NCIB 8250 utilizes phenol as sole source of carbon and energy via an ortho-cleavage pathway. The presence of ethanol in mixed substrate cultivations repressed the utilization of phenol. In fed batch cultivation the phenol tolerance was increased at least 2-fold. Maximum degradation rates of 150 mg phenol/(1 h) and 280 mg phenol/(g h), respectively were observed. Phenol hydroxylase is induced by its substrate and in parallel the catechol-1,2-dioxygenase is detectable. The presence of active phenol hydroxylase is strongly connected with the phenol degradation. Using a spectrophotometric enzyme assay the partially purified phenol hydroxylase was characterized with respect to kinetic parameters. The apparent Km values for phenol, FAD and NADPH were estimated to be 147 microM, 35 microM and 416 microM, respectively. Both FAD and NADPH were essential for maximum activity of the cytoplasmically localized enzyme. No substrate inhibition of phenol hydroxylase by phenol was observed up to 0.8 mM. The pH and temperature optima were pH 7.8 and 33 degrees C, respectively. The partially purified enzyme showed a broad substrate specificity. It hydroxylated the three isomeric cresols, chlorophenols and methylated chlorophenols. Pyrogallol, 3,4-dihydroxy-L-phenylalanine and resorcinol were oxygenated with higher rates than phenol. With the exception of phenol all other enzyme substrates tested did not serve as growth substrates.

Crotonobetaine reductase from Escherichia coli--a new inducible enzyme of anaerobic metabolization of L(-)-carnitine.

Crotonobetaine reductase from Escherichia coli 044 K74 is an inducible enzyme detectable only in cells grown anaerobically in the presence of L(-)-carnitine or crotonobetaine as inducers. Enzyme activity was not detected in cells cultivated in the presence of inducer plus glucose, nitrate, gamma-butyrobetaine or oxygen, respectively. Fumarate caused an additional stimulation of growth and an increased expression of crotonobetaine reductase. The reaction product, gamma-butyrobetaine, was identified by autoradiography. Crotonobetaine reductase is localized in the cytoplasm, and has been characterized with respect to pH (pH 7.8) and temperature optimum (40-45 degrees C). The Km value for crotonobetaine was determined to be 1.1 x 10(-2M). gamma-Butyrobetaine, D(+)-carnitine and choline are inhibitors of crotonobetaine reduction. For gamma-butyrobetaine (Ki = 3 x 10(-5M)) a competitive inhibition type was determined. Various properties suggest that crotonobetaine reductase is different from other reductases of anaerobic respiration.

Kinetics of enzymatic lysis, formation and regeneration of protoplasts ofCandida (Torulopsis) apicola.

The optimal conditions for protoplast formation ofCandida apicola were by using an enzyme fromArthrobacter sp. in combination with 2-mercaptoethanol. The kinetic data support the two-layered structure model of cell wall for this yeast but the structure of the cell wall depended on the age of cells and culture conditions. To regenerate the protoplasts, the type of osmotic stabilizer was important: sorbitol gave 16 to 30% regeneration. Electron microscopy revealed the presence of vesicles in the sections of protoplasts and whole cells ofCandida apicola grown in production medium and producing glycolipids. In sections of whole cells, vesicle-like structures are located in the periplasmic space and in protoplasts they can either be attached to, or released from, the cell surface. These vesicles are thought to be involved in the transport of the surface-active glycolipids and in the protection of the cell against denaturing effects.

Human skin fibroblasts in vitro differentiate along a terminal cell lineage.

Secondary mitotic human skin fibroblast populations in vitro underwent 53 +/- 6 cumulative population doublings (CPD) in 302 +/- 27 days. When the growth capacity of the mitotic fibroblasts is exhausted, and if appropriate methods are applied, the fibroblasts differentiate spontaneously into postmitotic fibroblast populations, which were kept in stationary culture for up to 305 +/- 41 additional days. Mitotic and postmitotic fibroblast populations are heterogeneous populations with reproducible changes in the proportions of mitotic fibroblasts F I, F II, and F III, and postmitotic fibroblasts F IV, F V, F VI, and F VII. This process makes it evident that the fibroblasts differentiate spontaneously along a seven-stage terminal cell lineage F I-F II-F III-F IV-F V-F VI-F VII. Shifts in the frequencies of the mitotic and postmitotic fibroblasts in mass populations are accompanied by alterations in the [35S]methionine polypeptide pattern of the developing mass populations. The [35S]methionine polypeptide patterns of homogeneous subpopulations of F I, F II, F III, F IV, F V, and F VI isolated from heterogeneous mass populations reveal that the six fibroblast morphotypes studied express their cell-type-specific [35S]methionine polypeptide pattern in the heterogeneous mass populations.