富士苹果多酚氧化酶特性研究

本文从富士苹果中提取和部分纯化多酚氧化酶,并对其特性进行研究。以邻苯二酚为作用底物,该酶最适pH为50,在pH50～80范围内有较高的稳定性。最适温度为30℃,在60℃以上迅速失活。该酶对不同的酚类物质表现出不同的底物专一性,由高至低的趋势依次为邻苯二酚、焦性没食子酸、DL-多巴、酪氨酸,其中对酪氨酸的活力为零。浓度为04mmol/L的VC、L-半胱氨酸及浓度为03mmol/L的亚硫酸氢钠,可完全抑制该酶活性。     

In vivo and in vitro protective role of proline

Accumulation of proline in response to environmental stresses seems tobe widespread among plants. To elucidate the role of proline in plantresponses,in vivo and in vitro, we studied theeffect of proline on catalase (CAT; EC 1.11.1.6), peroxidase (POD; EC 1.11.1.7)and polyphenol oxidase (PPO; EC 1.14.18.1). In vivo, thesethree enzymes were activated by proline, while CAT and POD were activated andPPO was inactivated by NaCl. In vitro, CAT and POD wereactivated and PPO was inactivated by proline. Proline appeared to protect thesethree enzyme activities. The significance of these findings with regard toenvironmental stress-induced proline accumulation in vivois discussed. The ability of proline to activate the enzymes may suggest alimited conformational change. These results are important for characterisationof metabolic responses to environmental stresses and can be used as a stressindicator.     

Effect of ultrasound combined with malic acid on the activity and conformation of mushroom (Agaricus bisporus) polyphenoloxidase

Polyphenoloxidase (PPO) plays an important role in the browning of vegetables, fruits and edible fungi. The effects of ultrasound, malic acid, and their combination on the activity and conformation of mushroom (Agaricus bisporus) PPO were studied. The activity of PPO decreased gradually with the increasing of malic acid concentrations (5–60 mM). Neither medium concentrations (10, 20, 30 mM) malic acid nor individual ultrasound (25 kHz, 55.48 W/cm²) treatment could remarkably inactivate PPO. However, the inactivation during their combination was more significant than the sum of ultrasound inactivation and malic acid inactivation. The inactivation kinetics of PPO followed a first-order kinetics under the combination of ultrasound and malic acid. The conformation of combination treated PPO was changed, which was reflected in the decrease of α-helix, increase of β-sheet contents and disruption of the tertiary structure. Results of molecular microstructure showed that ultrasound broke large molecular groups of PPO into small ones. Moreover, combined treatment disrupted the microstructure of PPO and molecules were connected together.     

Biochemistry of ginseng root tissues affected by rusty root symptoms

Ginseng rusty root, a disorder of unknown cause (s), in which reddish-brown to orange-brown areas develop on the surface of field-grown roots, was studied at the cellular and biochemical levels. Using light microscopy, the affected areas were shown to comprise of the epidermis and underlying 6-8 cell layers of the cortical tissues. Rusty root areas ranged from small clusters of 3-4 cells to larger expanding areas of >80 cells. These cells appeared golden-brown and stained a bluish-green with Toluidine Blue indicating the presence of phenolic compounds. Energy-dispersive X-ray spectroscopy and atomic emission spectrometry of affected epidermal cells revealed a significant accumulation of Fe, Al, Si, Mg and other cations when compared to adjoining healthy cells. The concentrations of the six most common ginsenosides found in ginseng roots (Rg(1), Re, Rb(1), Rc, Rb(2), and Rd) were reduced by 40-50% in rusty root-affected epidermal and cortical tissues when compared to adjacent healthy tissues. Total phenolic compounds were increased by up to threefold in affected tissues and HPLC analysis revealed significantly higher levels of quercetin, cinnamic acid, vanillic acid, p-coumaric acid, benzoic acid, chlorogenic acid and catechin. In vitro phenolic-metal binding assays confirmed that phenolic compounds were able to sequester positively-charged metal ions, in particular Fe, to form a phenolic-metal ion complex. In ginseng callus cultures, accumulation of phenolic compounds was increased threefold within 12 h of treatment with chitosan (1%), and to a lesser extent by wounding. Specific defense enzymes, namely phenylalanine ammonia-lyase (PAL, E.C. 4. 3. 1. 5.), polyphenoloxidase (PPO, E.C. 1. 10. 3. 1.) and peroxidase (POD, E.C. 1. 11. 1. 7.), were also significantly enhanced in treated callus tissues and in rusty root tissues. On field-grown ginseng roots, application of chitosan induced symptoms similar to rusty root, whereas wounding and ethylene treatments did not. Based on these results, rusty root symptoms on ginseng are proposed to result from an induction of host defense responses, especially phenolic production, in epidermal and underlying cortical cells. This induction is likely due to attempted invasion by as-yet uncharacterized chitin-containing soil fungi, which were observed in many of the affected cells. Subsequent oxidation of phenolic compounds and sequestration of metal ions, in particular Fe, appear to be largely responsible for the symptoms observed.     

Biochemical genetic basis of downy mildew resistance in pearl millet

Summary The study of phenolic content and activities of peroxidase and polyphenoloxidase in relation to the degree of downy mildew infection of 12 pearl millet cultivars revealed that these were linearly related to the degree of resistance at both the 30 and 50 day growth stages. Useful electrophoretic differences in peroxidase and polyphenoloxidase were also observed with respect to the expression of resistance.