Changes in molecular weight of transacylated pectin catalyzed by tomato and citrus pectinesterases as determined by gel permeation chromatography

The changes in molecular masses of pectin in 0.5% pectin-pectinesterase (PE) mixtures (2 units/mL) incubated at various temperatures, pH values, and NaCl levels for 30 min were observed by a Toyopearl TSK HW-65 (F) gel permeation chromatography. The molecular mass of pectin was remarkably increased from 103 to 266 kDa when the incubation temperature of pectin-tomato PE was increased from 25 to 45 degrees C. A further increase in molecular mass was observed when a pectin-citrus PE mixture was incubated at 65 degrees C. The values of pH and NaCl levels were also crucial to the transacylation activity of PEs. Reaction at pH 7.5 with tomato PE and citrus PE remarkably expanded the molecular mass of pectin to 410 and 670 kDa, respectively. The NaCl level of 0.3-0.5 and 0.3 M was favorable for the transacylation reaction of tomato PE and citrus PE, respectively. Only high methoxylpectin was the suitable substrate for PE to conduct the transacylation reaction.     

Mode of de-esterification of alkaline and acidic pectin methyl esterases at different pH conditions

Highly esterified citrus pectin was de-esterified at pH 4.5 and 8.0 by a fungal pectin methyl esterase (PME) that was shown to have an acidic isoelectric pH (pI) and an acidic pH optimum and by a plant PME that was characterized by an alkaline pI and an alkaline pH optimum. Interchain and intrachain de-esterification patterns were studied by digestion of the pectin products with endo-polygalacturonase and subsequent analysis using size exclusion and anion-exchange chromatography. No effect of pH was observed on the de-esterification mode of either of the two enzymes. Acidic, fungal PME converted pectin according to a multiple-chain mechanism, with a limited degree of multiple attack at the intrachain level, both at pH 4.5 and at pH 8.0. A multiple-attack mechanism, with a high degree of multiple attack, was more appropriate to describe the action mode of alkaline, plant PME, both at pH 4.5 and at pH 8.0.     

Effects of cryostabilizers, low temperature, and freezing on the kinetics of the pectin methylesterase-catalyzed de-esterification of pectin

The kinetics of the pectin methylesterase (PME)-catalyzed de-esterification of pectin was studied at 25 degrees C in the presence of sucrose, fructose, maltodextrin (DE = 16.5-19.5), and carboxymethylcellulose at different concentrations and in the presence of maltodextrin and sucrose at different concentrations in a temperature range between +25 and -4 degrees C in subcooled and frozen states. The objective was to determine whether the reaction is diffusion-controlled, to gain insight about the factors determining the diffusion of the reactants, and to determine the effect of the carbohydrates, low temperature, and freezing on the structural conformation of the enzyme. The results indicate that the PME-catalyzed de-esterification of pectin is diffusion-controlled. Nevertheless, the diffusion is not controlled by the macroviscosity of the reaction medium, but rather by the microviscosity experienced by the diffusants. Low temperature in the temperature range studied does not affect the structural conformation of the enzyme, while freezing seems to have some effect.     

Thermal inactivation of pectin methylesterase,polygalacturonase, and peroxidase in tomato juice

Thermal inactivation kinetics have been determined for pectin methylesterase (PME), polygalacturonase (PG), and peroxidase (POD) in tomato juice. Two parameters, the inactivation rate constant (k) at a reference temperature and the activation energy for inactivation (E(a)), were determined for each enzyme. For PME and PG, the k and E(a) values reported here do not agree with those in several previously published reports. These differences can be explained either by the differences in pH values used for inactivation determinations or by inadequacies in the heating methods used in some previous studies. POD showed simple first-order inactivation kinetics and was less thermally stable than either PME or PG. When different cultivars of tomatoes were evaluated, there was no difference in the thermal inactivation kinetics of these enzymes.     

柑橘果实发育中果胶酸钙、草酸钙和果胶动态的研究

以单性结实的龟井蜜柑和自花结实的鄂柑1号橘为试材,对整个果实发育期的子房(幼果)、果皮和果肉的果胶酸钙、草酸钙和果胶含量变化进行了测定。结果表明,1)两品种子房(幼果)果胶酸钙含量呈类似的下降趋势;草酸钙则相反,龟井花后趋下降,而鄂柑1号却明显上升;而且鄂柑1号子房(幼果)果胶酸钙、草酸钙和果胶含量均相对较高。2)在果实增大期内,两品种果皮和果肉的果胶酸钙含量均出现显著上升,对应果皮草酸钙含量虽有波动但居相对较高水平,而果肉草酸钙则趋明显下降。3)两品种果皮和果肉水溶性果胶含量均在增大期内呈显著上升,对应原果胶含量均相对较高,进入增大后期均明显下降。     

Antioxidant activity of tomato products as studied by model reactions using xanthine oxidase, myeloperoxidase, and copper-induced lipid peroxidation

The antioxidant content and activity of commercial tomato products differing in variety and processing were studied. Two procedures for extracting hydrophilic and lipophilic antioxidants, namely, two-step 0.1 M phosphate buffer (pH 3.0 and 7.4) extraction and tetrahydrofuran extraction followed by petroleum ether fractionation, were developed. Carotenoids (lycopene, beta-carotene, and lutein) and ascorbic acid were analyzed by HPLC with spectrophotometric and electrochemical detectors, respectively. Total phenolics were determined by using the Folin-Ciocalteu reagent. The antioxidant activity was studied by the following three model systems: (a) the xanthine oxidase (XOD)/xanthine system, which generates superoxide radical and hydrogen peroxide; (b) the myeloperoxidase (MPO)/NaCl/H(2)O(2) system, which produces hypochloric acid; and (c) the linoleic acid/CuSO(4) system, which promotes lipid peroxidation. Results showed that the hydrophilic and lipophilic fractions of all tomato products were able to affect model reactions, whatever reactive oxygen species and catalysts were used to drive oxidation. In the XOD/xanthine system both the hydrophilic and lipophilic fractions displayed an inhibitory activity. The hydrophilic fractions were more effective (I(50) ranging from 680 to 3200 microg, dry weight) than the lipophilic fractions (I(50) ranging from 4000 to 7750 microg, dry weight). In the MPO/NaCl/H(2)O(2) system the hydrophilic fractions inhibited oxidation (I(50) ranging from 2300 to 2900 microg, dry weight), whereas the lipophilic fractions had a lower inhibitory effect at the same concentration. Conversely, in the copper-catalyzed lipid peroxidation only the lipophilic fractions were effective (I(50) ranging from 1030 to 2100 microg, dry weight), whereas the hydrophilic fractions had a pro-oxidant effect in the same concentration range. The extent of inhibition varied according to the tomato sample in the superoxide and hydrogen peroxide generating system and in lipid peroxidation, but was substantially the same in the HClO generating system. Fresh tomato varieties differed considerably in the antioxidant activities of their hydrophilic and lipophilic fractions. Processed tomatoes showed a significantly lower antioxidant activity than fresh tomatoes in their hydrophilic fractions but had a high antioxidant activity in their lipophilic fractions. Because the oxidative reactions produced by the above-mentioned model systems are also involved in the pathogenesis of several chronic diseases, the antioxidant activity of tomato fractions might be related to their in vivo activity. Hence, these measurements may be used for optimizing tomato technologies.     

微波加热法提取柚果皮果胶的工艺（简报）

对新鲜柚果皮中果胶进行微波提取,采用咔唑比色法测定提取液中的果胶含量.探讨了料液比、微波功率、微波处理时间、盐析条件等对果胶得率和半乳糖醛酸含量的影响,运用L9(34)正变试验对微波加热提取果胶的工艺条件进行了优化.结果表明:微波功率对果胶得率有极显著影响,微波处理时间影响较小.较佳工艺是:液料比(V/m)8:1,调pH值为2.0,微波功率640W,处理时间8 min,盐析饱和硫酸铝用量与酸萃取液比例为3:5,此条件下提取柚皮果胶得率为4.457％,果胶的半乳糖醛酸含量为42.58％．     

Waste water from citrus processing as a source of hesperidin by concentration on styrene-divinylbenzene resin

This paper describes a procedure for recovering hesperidin from the waste water of orange juice processing, namely, yellow water, by concentration of diluted extracts on styrene-divinylbenzene resin. Turbid raw material flowing out from centrifuges of essential oil separation contains considerable amount of hesperidin ( approximately 1 g/L) mainly associated with solid particles. Yellow water was treated with calcium hydroxide until pH 12 to solubilize hesperidin, filtered, neutralized at pH 6, and loaded on resin up to saturation. Desorption with 10% ethanol aqueous solutions at different NaOH concentrations (0.23-0.92 M) assured high concentration of hesperidin in selected fractions (10-78 g/L), from which it precipitated in high yield and purity immediately after acidification at pH 5. Best results were obtained using 0.46 M NaOH as eluent: 71.5% of the adsorbed hesperidin was desorbed in 300 mL, with an overall 64% yield of isolated product at 95.4% purity (HPLC). These experiments can constitute a useful starting point for an industrial application.     

Nitrogen availability to citrus seedlings from urea and from mineralization of citrus leaf or compost

A pot experiment was conducted using a Candler fine sand (hyperthermic, uncoated, Typic Quartzipsamments) amended with either citrus leaves or compost, to measure the nitrogen (N) mineralization and its availability to two citrus rootstock seedlings. A rapid increase in NH₄‐N concentration was evident in the soil amended with citrus leaves as compared to compost during the initial 14 to 20 d. Subsequently, the concentration of NH₄‐N decreased in the citrus leaves amended soil. The extractable NO₃‐N concentration was greater in the soil amended with citrus leaves as compared to compost, throughout the 270 d duration of the study. The N concentrations and N uptake by Cleopatra mandarin (CM) and Swingle citrumelo (SC) seedlings grown in citrus leaf amended soil were very similar to those in urea amended soil. Therefore, mineralization of N from dry ground citrus leaves was quite rapid. The N concentrations in both rootstock seedlings were much lower in the compost amended and unamended soils as compared to those in either citrus leaves or urea amended soils. Rapid mineralization of N from cirrus leaves added to sandy soil, resulting in an increased availability of N, suggested that the contribution of N from shed leaf mineralization must be considered while developing N rate recommendations for improving N use efficiency.     

Accumulation and availability of copper in citrus grove soils as affected by fungicide application

Purpose  

Copper is a trace element of environmental concern. Repeated applications of Cu-containing fungicides have resulted in a large scale of Cu contamination in agricultural soils. However, limited information is available regarding Cu accumulation and availability in soils under citrus production in the Indian River Area, South Florida, which has received increasing amounts of Cu fungicides to control canker and other diseases. The purpose of this study was to investigate Cu transformation, availability, and mobility in soils as affected by external Cu loading and soil properties.     

Growth and elemental composition of tomato as affected by fungicides and nitrogen sources

Tomato (Lycopersicon esculentum Mill.) plant growth and elemental composition were evaluated using three NH₄‐N:NO₃‐N form ratios with or without the fungicide benomyl, captan, lime‐sulfur, nitrapyrin, or terrazole in a greenhouse soil culture study. Nitrogen was applied weekly for 5 weeks providing a total of 115 mg N/kg. Each fungicide was applied at 0.25 mg/kg 3 days before transplanting, followed by 3 weekly applications with each N treatment. The largest shoot and root dry weights were obtained with 1:1 N form ratio. With 1:1 N ratio treatment, all chemicals significantly increased plant growth resulting in lower element concentrations relative to the untreated control. However, growth of plants receiving either 1:0 or 0:1 N ratio treatment was not affected by nitrapyrin and terrazole, but was restricted by benomyl, captan, or lime‐sulfur. Overall, elemental concentrations in the tissues of plants receiving either N form was related to the fungicide treatment.     

Nonenzymatic degradation of citrus pectin and pectate during prolonged heating: effects of pH, temperature, and degree of methyl esterification

The underlying mechanisms governing nonenzymatic pectin and pectate degradation during thermal treatment have not yet been fully elucidated. This study determined the extent of nonenzymatic degradation due to beta-elimination, acid hydrolysis, and demethylation during prolonged heating of citrus pectins and its influence on physicochemical properties. Solutions of citrus pectins, buffered from pH 4.0 to 8.5, were heated at 75, 85, 95, and 110 degrees C for 0-300 min. Evolution of methanol and formation of reducing groups and unsaturated uronides were monitored during heating. Molecular weight and viscosity changes were determined through size exclusion chromatography and capillary viscometry, respectively. Results showed that at pH 4.5, the activation energies of acid hydrolysis, beta-elimination, and demethylation are 95, 136, and 98 kJ/mol, respectively. This means that at this pH, acid hydrolysis occurs more rapidly than beta-elimination. Furthermore, the rate of acid hydrolysis is diminished by higher levels of methyl esterification. Also, citrus pectin (93% esterified) degrades primarily via beta-elimination even under acidic conditions. Acid hydrolysis and beta-elimination caused significant reduction in relative viscosity and molecular weight.     

旱地长期培肥土壤脲酶和碱性磷酸酶动力学及热力学特征研究

研究了连续25年长期培肥试验条件下土壤脲酶和碱性磷酸酶酶促反应的动力学和热力学特征,从酶学角度揭示长期培肥的效应。结果表明,长期培肥增加了脲酶和碱性磷酸酶酶促反应的Vmax、Vmax/Km和k值;降低了Ea、△G、△H和△S值,说明培肥能提高酶促反应速度、减小活化自由能、加快土壤中物质的生物循环过程。酶促反应动力学参数和热力学参数与土壤性质相关分析表明,酶促反应动力学参数大多依赖于土壤化学性状,基于动力学参数的土壤肥力指标体系可评价土壤肥力水平,且U-Vmax、P-Km、P-Vmax可作为土壤肥力的重要指标。     

Mixture approach for optimizing lycopene extraction from tomato and tomato products

A simple mixture process design based on the comparison of both quadratic and special cubic models and involving three mixture components (hexane/acetone/ethanol) as a solution for extracting lycopene from raw tomato, tomato sauce, and tomato paste was used to confirm the hypothesis that lycopene extraction rates are a function of the solvent used during the extraction process. Conventional criteria (p 相似文献   

Analysis of limonoid glucosides from citrus by electrospray ionization liquid chromatography-mass spectrometry

An electrospray ionization liquid chromatography-mass spectrometry (ESI-LC-MS) method for the detection and quantitation of limonoid glucosides has been developed. Negative ions [M - H(+)](-) characteristic of six limonoid glucosides can be detected and quantified from selected ion monitoring chromatograms using carminic acid as an internal standard. The described method has been applied to the analysis of limonoid glucoside content in various liquid and solid Citrus spp. samples as well as complex mixtures of partially purified limonoid glucosides. Rapid and sensitive qualitative screening of samples for limonoid glucosides can also be accomplished with slight modifications of the method.     

Effects of pH on chemical stability and de-esterification of fenoxaprop-ethyl by purified enzymes, bacterial extracts, and soils

De-esterification is an initial step in the metabolism of certain herbicides, for example, fenoxaprop-ethyl [(+/-)-ethyl 2-[4-[(6-chloro-2-benzoxaolyl)oxy]phenoxy]propanoate] (FE). The ethyl-ester bond cleavage of FE to fenoxaprop acid (FA) by purified enzymes, crude bacterial enzyme preparations, and soils was investigated. In similar experiments fluorescein diacetate (FDA) was used as an alternative substrate. FE stability was pH sensitive in acidic buffered solutions; that is, below pH 4.6, rapid nonenzymatic hydrolysis of the benzoxazolyl-oxy-phenoxy ether linkage occurred, forming 6-chloro-2,3-dihydro-benzoxazol-2-one (CDHB) and ethyl 4-hydroxyphenoxypropanoate or 4-hydroxyphenoxypropanoate. With porcine esterase and cell-free Pseudomonas fluorescens extracts, activity on FE and FDA was most rapid at pH 7.6-8.6 but decreased 80-90% at pH 5.6. Yeast (Candida cylindrica) lipase-mediated de-esterification of FE and FDA was not as sensitive to pH; that is, activity at pH 4.6 was 70% of that at pH 7.6. Short-term incubations (20 h) were conducted in eight soils (pH 4.5-6.9) treated with (14)C-chlorophenyl ring-labeled FE (2 mg kg(-)(1)). In the most acidic soils (pH 4.4-4.5) 25% of the (14)C was recovered as FA, versus 30-40% in moderately acid soils (pH 5.0-5.6) and 55% in neutral soils (pH 6.8-6.9). There was a similar correlation between soil pH and FDA de-esterification. CDHB was formed in all acidic soils with levels 4-fold greater in pH 4.4-4.5 soils than in pH 5. 0-5.6 soils. CDHB was not formed in neutral soils. Results demonstrate some chemical hydrolysis (benzoxazolyl-oxy-phenoxy ether linkage) of FE in acid soils, the sensitivity of enzymatic de-esterification of FE to pH, and the potential of FDA as a colorimetric indicator for esterase hydrolysis of FE.     

Cleavage of 14C-labeled glycine and its polycondensation with pyrogallol as catalyzed by birnessite

Deamination as well as decarboxylation of amino acids and their polycondensation with polyphenols catalyzed by Mn(IV) oxide have been reported. However, the mechanisms of cleavage of amino acids and their polycondensation with polyphenols by soil inorganic components are not well understood. The objective of this study was, thus, to investigate the enhanced abiotic decarboxylation of carboxyl group and dealkylation of alkyl group of glycine by birnessite (δ-MnO₂, 0.2–2 μm) both in the presence and absence of pyrogallol and the resultant formation of humic polycondensates in systems free of microbial activity. The degradation of ¹⁴C-labeled glycine through either carboxyl or alkyl group on the birnessite surface was demonstrated. Birnessite enhanced the dealkylation of alkyl C and especially decarboxylation of carboxyl C of glycine. Furthermore, it promoted the interaction of glycine and pyrogallol and the resulting simultaneous decarboxylation and dealkylation of glycine, ring cleavage of pyrogallol, and polycondensation. Carboxyl C and especially alkyl C of glycine were incorporated into the humic polycondensates. This study indicates that structural configuration and functionality of amino acids and polyphenols merits closer attention in understanding mechanisms of humification processes catalyzed by soil mineral colloids.     

Uptake and distribution of selenium in tomato plants as affected by genotype and sulphate supply

Aim of this work was to investigate if the variation among tomato genotypes in selenium (Se) uptake and accumulation observed in short term experiments are maintained over longer growth periods and if there is a positive correlation in shoot between sulphur (S) accumulation and Se accumulation across different genotypes or if higher tissue S results in greater feedback inhibition of Se uptake. Two experiments were carried out under greenhouse conditions and different genotypes of Lycopersicon lycopersicum (UC82B and LA2711), Lycopersicon pennellii (LA716) and Lycopersicon peruvianum (LA2157) were grown until fruit ripening. The results obtained in the two experiments confirmed that sulphate in the growth solution reduced selenate uptake by plants and increased the S content of the leaves. Under low sulphate treatment there was a clear correlation (R²=0.82) between leaf S content and shoot Se content across genotypes in both experiments, indicating that the overall activity of the S transport systems also determines Se transport. Selenium was translocated from shoot to fruit, but the edible portion of the plant contained much less total Se than the inedible plant parts. The difference in Se content between the low and the high sulphate treatments was significantly higher in shoot than in root, confirming that the Se translocation from root to shoot is probably more affected by high sulphate supply than Se uptake by root. In the first experiment the genotype LA716 showed ah higher Se, accumulation together with higher S content in leaves, indicating a marked ability of this genotype to absorb ions from substrate. In the second experiment UC82B appeared to be more capable to accumulate Se and S rather than LA2711 and LA2157. In both experiments Lycopersicon peruvianum appeared to be less affected by the high concentration of ions in the growth solution and to be able to reduce ion uptake than Lycopersicon lycopersicum and Lycopersicon pennellii.     

木质纤维素酸催化制备糠醛的工艺及机理研究进展

糠醛是一种重要的生物质基平台化合物,国内外学者针对生物质产糠醛展开诸多研究,尤其是酸催化水解领域。该文综述了糠醛制备工艺在不同时期的情况,阐述并评价了目前新颖的同步产糠醛与纤维素基化学品工艺。对酸(稀布朗斯特酸和路易斯金属盐)催化木糖和半纤维素的反应动力学进行系统归纳,并阐述了相关机理的研究进展。最后,对现在研究热点——酸/有机溶剂作用体系中有机溶剂的作用机制进行归纳,并对计算化学在其中的最新研究情况进行总结。该文旨在为学者开展生物质产糠醛的研究提供信息,有利于学者进行选择性研究。