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LsSAT2 (serine acetyltransferase in Lathyrus sativus) is the rate-limiting enzyme in biosynthesis of ß-N-oxalyl-l-α,ß-diaminopropionic acid (ß-ODAP), a neuroactive metabolite distributed widely in several plant species including Panax notoginseng, Panax ginseng, and L. sativus. The enzymatic activity of LsSAT2 is post-translationally regulated by its involvement in the cysteine regulatory complex in mitochondria via interaction with ß-CAS (ß-cyanoalanine synthase). In this study, the binding sites of LsSAT2 with the substrate Ser were first determined as Glu290, Arg316, and His317 and the catalytic sites were determined as Asp267, Asp281, and His282 via site-directed/truncated mutagenesis, in vitro enzymatic activity assay, and functional complementation of the SAT-deficient Escherichia coli strain JM39. Furthermore, the C-terminal 10-residue peptide of LsSAT2 is confirmed to be critical to interact with LsCAS, and Ile336 in C10 peptide is the critical amino acid. These results will enhance our understanding of the regulation of LsSAT2 activities and the biosynthesis of ß-ODAP in L. sativus.

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To reduce packaging-induced stress of long cavity length high-power single emitter semiconductor laser, the relationship between the stress and the wavelength shift was deduced on the basis of the theory that the stress can change the band gap. A method was developed for quantitatively calculating the stress by measuring the emission spectrum of the laser under pulse conditions. The results show that the soldering quality is a critical factor affecting thermal stress. The difference in stress can exceed 300 MPa due to the difference in soldering quality. By optimizing the reflowing soldering curve of the laser, the stress of the laser drops from 129.7 to 53.4 MPa. This method can also effectively solve the problem that the stress varies with storage time. This work demonstrates that the measurement and analysis of the emission spectrum of the laser can provide a useful method to study packaging stress of the high-power single emitter semiconductor laser. It is also an available means to evaluate and analyze soldering quality.

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Recent evidence has demonstrated that both copper amine oxidase (CuAO; EC 1.4.3.6) and phospholipase D (PLD; EC 3.1.4.4) are involved in abscisic acid (ABA)-induced stomatal closure. In this study, we investigated the interaction between CuAO and PLD in the ABA response. Pretreatment with either CuAO or PLD inhibitors alone or that with both additively led to impairment of ABA-induced H2O2 production and stomatal closure in Vicia faba. ABA-stimulated PLD activation could not be inhibited by the CuAO inhibitor, and CuAO activity was not affected by the PLD inhibitor. These data suggest that CuAO and PLD act independently in the ABA response. To further examine PLD and CuAO activities in ABA responses, we used the Arabidopsis mutants cuaoζ and pldα1. Ablation of guard cell-expressed CuAOζ or PLDα1 gene retarded ABA-induced H2O2 generation and stomatal closure. As a product of PLD, phosphatidic acid (PA) substantially enhanced H2O2 production and stomatal closure in wide type, pldα1, and cuaoζ. Moreover, putrescine (Put), a substrate of CuAO as well as an activator of PLD, induced H2O2 production and stomatal closure in WT but not in both mutants. These results suggest that CuAO and PLD act independently in ABA-induced stomatal closure.

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H(2)O(2) is an essential signal in absicic acid (ABA)-induced stomatal closure. It can be synthesized by several enzymes in plants. In this study, the roles of copper amine oxidase (CuAO) in H(2)O(2) production and stomatal closure were investigated. Exogenous ABA stimulated apoplast CuAO activity, increased H(2)O(2) production and [Ca(2+)](cyt) levels in Vicia faba guard cells, and induced stomatal closure. These processes were impaired by CuAO inhibitor(s). In the metabolized products of CuAO, only H(2)O(2) could induce stomatal closure. By the analysis of enzyme kinetics and polyamine contents in leaves, putrescine was regarded as a substrate of CuAO. Putrescine showed similar effects with ABA on the regulation of H(2)O(2) production, [Ca(2+)](cyt) levels, as well as stomatal closure. The results suggest that CuAO in V. faba guard cells is an essential enzymatic source for H(2)O(2) production in ABA-induced stomatal closure via the degradation of putrescine. Calcium messenger is an important intermediate in this process.

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In order to analyze the relationship between polyamine oxidative degradation induced by light and the Lignin synthesis in cell walls, the activities of diamine oxidases and peroxidase, the contents of H2O2 and lignin, and the growth of hypocotyls in soybean [Glycine max (Linn.) Merr.] grown under tight or in darkness were investigated. In comparison with the dark treatment, light irradiation significantly inhibited the growth of soybean hypocotyls and promoted the activities of diamine oxidases and peroxidase as well as the accumulation of H2O2 and lignin. Treatments with the different concentrations of diamine oxidase inhibitors (2-hydroxyethylhydrazine and aminoguanidine) under the light condition inhibited diamine oxidase activity, and decreased the contents of H2O2 and lignin. The results provide evidence for the hypothesis that light irradiation could promote the accumulation of H2O2 and lignin in cell walls by activating polyamine oxidative degradation mediated by diamine oxidases.

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