RESUMO
Auxin is thought to be an important factor in the induction of galls by galling insects. We have previously shown that both galling and nongalling insects synthesize indole-3-acetic acid (IAA) from tryptophan (Trp) via two intermediates, indole-3-acetaldoxime (IAOx) and indole-3-acetaldehyde (IAAld). In this study, we isolated an enzyme that catalyzes the last step "IAAld â IAA" from a silk-gland extract of Bombyx mori. The enzyme, designated "BmIAO1", contains two 2Fe-2S iron-sulfur-cluster-binding domains, an FAD-binding domain, and a molybdopterin-binding domain, which are conserved in aldehyde oxidases. BmIAO1 causes the nonenzymatic conversion of Trp to IAAld and the enzymatic conversion of IAOx to IAA, suggesting that BmIAO1 alone is responsible for IAA production in B. mori. However, a detailed comparison of pure BmIAO1 and the crude silk-gland extract suggested the presence of other enzymes involved in IAA production from Trp. Abbreviations: BA: benzoic acid; CE: collision energy; CXP: collision cell exit potential; DP: declustering potential; IAA: indole-3-acetic acid; IBI1: IAA biosynthetic inhibitor-1; IAAld: indole-3-acetaldehyde; ICA: indole-3-carboxylic acid; IAOx: indole-3-acetaldoxime; IEtOH: indole-3-ethanol; LC-MS/MS: liquid chromatography-tandem mass spectrometry; Trp: tryptophan.
Assuntos
Aldeído Oxidase/metabolismo , Bombyx/fisiologia , Ácidos Indolacéticos/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Aldeído Oxidase/química , Aldeído Oxidase/isolamento & purificação , Sequência de Aminoácidos , Animais , Bombyx/anatomia & histologia , Bombyx/enzimologia , Catálise , Cromatografia/métodos , Eletroforese em Gel de Poliacrilamida , Homologia de Sequência de Aminoácidos , Espectrofotometria Ultravioleta/métodos , Especificidade por Substrato , Espectrometria de Massas em Tandem/métodosRESUMO
Using a mass-loss method, we investigated the solubility change of gallium nitride (GaN) in supercritical ammonia with mixed mineralizers [ammonium chloride (NH4Cl) + ammonium bromide (NH4Br) and NH4Cl + ammonium iodide (NH4I)]. The solubilities were measured over the temperature range 450-550 °C, at 100 MPa. The solubility increased with NH4Cl mole fraction at 450 °C and 100 MPa. The temperature dependence of the solubility curve was then measured at an equal mole ratio of the two mineralizers. The slope of the solubility-temperature relationship in the mixed mineralizer was between those of the individual mineralizers. These results show that the temperature dependence of the solubility of GaN can be controlled by the mineralizer mixture ratio. The results of the van't Hoff plot suggest that the solubility species were unchanged over the investigated temperature range. Our approach might pave the way to realizing large, high-quality GaN crystals for future gallium-nitride electronic devices, which are increasingly on demand in the information-based age.
RESUMO
In the course of our study of the biosynthetic pathway of auxin, a class of phytohormones, in insects, we proposed the biosynthetic pathway tryptophan (Trp)âindole-3-acetaldoxime (IAOx)âindole-3-acetadehyde (IAAld)âindole-3-acetic acid (IAA). In this study, we identified two branches in the metabolic pathways in the silkworm, possibly affecting the efficiency of IAA production: Trpâindole-3-pyruvic acidâindole-3-lactic acid and IAAldâindole-3-ethanol. We also determined the apparent conversion activities (2.05×10-7UmL-1 for TrpâIAA, 1.30×10-5UmL-1 for IAOxâIAA, and 3.91×10-1UmL-1 for IAAldâIAA), which explain why IAOx and IAAld are barely detectable as either endogenous compounds or metabolites of their precursors. The failure to detect IAAld, even in the presence of an inhibitor of the conversion IAAldâIAA, is explained by a switch in the conversion from IAAldâIAA to IAAldâIEtOH.
Assuntos
Bombyx/metabolismo , Ácidos Indolacéticos/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Triptofano/metabolismo , Animais , Vias BiossintéticasRESUMO
Insect galls are abnormal plant tissues induced by galling insects. The galls are used for food and habitation, and the phytohormone auxin, produced by the insects, may be involved in their formation. We found that the silkworm, a non-galling insect, also produces an active form of auxin, indole-3-acetic acid (IAA), by de novo synthesis from tryptophan (Trp). A detailed metabolic analysis of IAA using IAA synthetic enzymes from silkworms indicated an IAA biosynthetic pathway composed of a three-step conversion: Trp â indole-3-acetaldoxime â indole-3-acetaldehyde (IAAld) â IAA, of which the first step is limiting IAA production. This pathway was shown to also operate in gall-inducing sawfly. Screening of a chemical library identified two compounds that showed strong inhibitory activities on the conversion step IAAld â IAA. The inhibitors can be efficiently used to demonstrate the importance of insect-synthesized auxin in gall formation in the future.
Assuntos
Vias Biossintéticas , Bombyx/metabolismo , Inibidores Enzimáticos , Himenópteros/metabolismo , Ácidos Indolacéticos/metabolismo , Reguladores de Crescimento de Plantas/biossíntese , Animais , Ácidos Indolacéticos/antagonistas & inibidores , Ácidos Indolacéticos/isolamento & purificação , Reguladores de Crescimento de Plantas/antagonistas & inibidores , Reguladores de Crescimento de Plantas/isolamento & purificação , Triptofano/análogos & derivadosRESUMO
In this study, the aggregation and breakup behaviors of latex particles in shear flow confined between two parallel plates were investigated using an in situ observation apparatus with a laser scanning confocal microscope. To investigate the effects of shear rate and the gap width between two parallel plates on the size and structure of the aggregates in the steady state, the distributions of the projected cross-sectional area and perimeter-based fractal dimension of the aggregates were measured. As a result, the average size of the aggregates decreases as shear rate increases and the gap width decreases due to the hydrodynamic effect acting on the aggregates. The size distributions of the aggregates become narrow as the gap width decreases. In addition, the fractal dimension, that is, the structure of the aggregates, was almost independent of shear rate and the gap width and approximately 1.2, which suggests that the aggregates are relatively compact.