An algal photoenzyme converts fatty acids to hydrocarbons.

Although many organisms capture or respond to sunlight, few enzymes are known to be driven by light. Among these are DNA photolyases and the photosynthetic reaction centers. Here, we show that the microalga Chlorella variabilis NC64A harbors a photoenzyme that acts in lipid metabolism. This enzyme belongs to an algae-specific clade of the glucose-methanol-choline oxidoreductase family and catalyzes the decarboxylation of free fatty acids to n-alkanes or -alkenes in response to blue light. Crystal structure of the protein reveals a fatty acid-binding site in a hydrophobic tunnel leading to the light-capturing flavin adenine dinucleotide (FAD) cofactor. The decarboxylation is initiated through electron abstraction from the fatty acid by the photoexcited FAD with a quantum yield >80%. This photoenzyme, which we name fatty acid photodecarboxylase, may be useful in light-driven, bio-based production of hydrocarbons.

Photoregulation process of sorghum leaf phosphoenolpyruvate carboxylase: study with monoclonal antibodies.

Monoclonal antibodies were produced against the G isozyme subunit of PEP carboxylase (PEPC) from Sorghum leaves by the hybridoma technique. More than 400 antibodies-producing hybridomas to PEPC were produced from the fusion of spleen cells from immunized mice with NS1 myeloma cells. By using an ELISA, three hybridomas (91-G, 83-G, 49-EG) were selected. Monoclonal antibodies were subsequently characterized in a Western experiment; Mabs 83-G and 91-G were found to be highly specific to the G isozyme whereas Mab 49-EG recognized both forms (E and G isozymes) of the enzyme. Addition of Mabs to the enzyme preparation did not modify its catalytic activity nor its activation by glycine. Use of these probes provided direct and definite evidence of the specific enhancing effect of light on the G form and on its corresponding mRNA.

mRNA-dependent regulation of UDP-apiose synthase activity in irradiated plant cells.

Large changes in the rate of synthesis of UDP-apiose synthase, an enzyme of the flavonoid glycoside pathway, were observed both in vivo and in vitro following irradiation of previously dark-grown cell suspension cultures of parsley (Petroselinum hortense). Irradiation of the cells with ultraviolet light for 2 1/2 h caused a large increase in UDP-adipose synthase mRNA activity which continued for several hours during the subsequent dark period. A sharp peak in this activity was followed by a rapid exponential decline. The timing of changes in mRNA activity was very similar for UDP-apiose synthase and chalcone synthase, a previously investigated enzyme of the flavonoid glycoside pathway, but differed from that for phenylalanine ammonia-lyase, an enzyme of general phenylpropanoid metabolism. Thus, the present data provide further evidence for a differential regulation of the two groups of enzymes and a high degree of co-ordination within each group. The expected light-induced changes in UDP-apiose synthase activity were calculated from the observed changes in the rate of synthesis, allowing for an apparently constant rate of degradation (t 1/2 = 17 h). The resulting time course was in agreement with experimental data, suggesting that the changes in enzyme activity were caused by corresponding changes in the rate of synthesis. A protein of unknown function (Protein X), which is tightly associated with UDP-apiose synthase and co-purifies with the enzyme at a molar ratio of 1:1, is shown to be regulated independently of the enzymes of the flavonoid glycoside pathway.

Dietary modification of UV-induced epidermal ornithine decarboxylase.

The effect of several dietary antioxidant supplements upon ultraviolet light-induced ornithine decarboxylase activity was determined. Hairless mice received diets supplemented with either butylated hydroxytoluene, disulfiram, phenobarbital, glutathione (reduced), or a special antioxidant mixture for 2 weeks before irradiation with FS-20 fluorescent sun lamps. Epidermal ornithine decarboxylase activity, the induction of which is thought to be a necessary component of skin tumor promotion, was determined at designated post-irradiation periods. Significant inhibition of ornithine decarboxylase induction was found in epidermis from animals receiving diets containing butylated hydroxytoluene, the antioxidant mixture, or disulfiram whereas no significant effects were noted in animals receiving reduced glutathione or phenobarbital. Butylated hydroxytoluene, at physiological concentrations, had no effect upon ornithine decarboxylase activity when added directly to the reaction mixture. Nor did this compound, when provided in the diet of animals, evoke a notable effect upon 12-0-tetra-decanoylphorbol-13-acetate induced ornithine decarboxylase. The latter finding suggests that dietary butylated hydroxytoluene inhibition of ultraviolet light-induced ornithine decarboxylase is a response related directly to the degree of irradation insult rather than a general effect upon the processes associated with carcinogenic promotion.

[Light activation of the enzyme ribulose diphosphate carboxylase]. / Aktivirovanie svetom fermenta ribulozodifosfatkarboksilazy.

Some activation mechanisms of RuDP-carboxylase under illumination are considered. It is assumed on the grounds of experimental data that under illumination thylacoid membranes pass from one phase (I) to another (II). Kinetics of this transition is taken as similar to that of surface phase transition of the first kind. It is assumed that the number of enzyme molecules passing from the inactive to active state is proportional to the area of thylacoid membranes in the II phase (as the time function). A change in the stroma determined by the kinetics of transition of thylocoid membranes in the transition process leads to a change of the mean time between two successive catalytic acts of the enzyme activated molecules. The obtained kinetic curve of RuDP-carboxylating reaction in the transition process under illumination coincides with the curve obtained in [1] by expanding the experimental curves of the values of labels 14C, 32P of 3PGA to constituents.

Degradation of biochemical activity in soil sterilized by dry heat and gamma radiation.

The activities of three enzymes present in soil, phosphatases, urease, and decarboxylase, were monitered as indicators of the loss of biochemical information occurring when soil was sterilized by dry heat (0.08% relative humidity), gamma radiation, or a combination of both. More enzymatic activity was retained in soil sterilized by a long exposure to dry heat at relatively low temperature (8 weeks at 100.5 degrees C) than by a shorter exposure to a higher temperature (2 weeks at 124.5 degrees C). No enzymatic activity was detectable in soil sterilized by an even higher temperature (4 days at 148.5 degrees C). Soil sterilized with 7.5 Mrads of radiation retained much higher enzymatic activity than with heat sterilization. Combining sublethal doses of heat radiation effectively sterilized the soil and yielded enzymatic activities higher than those of soil sterilized by dry heat alone but lower than those of soil sterilized by radiation.