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.     

Transacylation and de-esterification reactions of pectin as catalyzed by pectinesterases from tomato and citrus

The optimal conditions for the de-esterification reaction of tomato pectinesterase (PE) and citrus PE was 0.1-0.2 M NaCl and at pH 7.5-8.5, 65 degrees C, almost identical to those for the transacylation reaction as observed by turbidity (absorbance at 400 nm) change. Among the PEs tested, pea pod PE presented the most remarkable catalysis on the transacylation reaction, and 1.5% pectin solution was determined to be suitable for this reaction. Low methoxy pectin with a DE (degree of esterification) of 31% displayed a slow turbidity increase, revealing that the extent of DE was influential on the transacylation. Besides citrus pectin, apple pectin was also proved to progress transacylation reaction by PEs from tomato and citrus sources as apparently observed by turbidity method.     

Encapsulation of quercetin and myricetin in cyclodextrins at acidic pH

The in vitro formation of quercetin- and myricetin-cyclodextrin inclusion complexes in acidic medium has been characterized using the enzymatic system horseradish peroxidase, which oxidizes those flavonols in the presence of H2O2. The presence of cyclodextrins (CDs) in the reaction medium inhibited flavonol oxidation due to the complexation of the flavonol in the hydrophobic cavity of CDs. This inhibitory effect depends on the complexation constant Kc between flavonol and the CD type used. The Kc for quercetin and myricetin with the different types of CD used was calculated by nonlinear regression of the inhibition curves obtained in the presence of CDs. In both cases (quercetin and myricetin), the Kc values obtained followed the order hydroxypropyl-beta-CDs > maltosyl-beta-CDs > beta-CDs, reflecting the greater affinity of modified cyclodextrins for the studied flavonols compared with their parental beta-CDs. Moreover, the complexation efficiency (CE) values for HP-beta-CDs and quercetin or myricetin were calculated (267.4 and 5.3, respectively), indicating that HP-beta-CDs are more efficient for the complexation of quercetin than myricetin in the studied conditions, despite of the K c values being very similar in both cases.     

Solvent-mediated disruption of bovine casein micelles at alkaline pH

The disruption of casein micelles at alkaline pH was investigated using turbidity measurements. The rate and extent of disruption of casein micelles at alkaline pH (8.0-11.0) increased with pH. Furthermore, the extent of alkaline disruption increased with increasing temperature (5-40 degrees C). Preheating milk for 10 min at 90 degrees C did not influence the extent of alkaline disruption of casein micelles, suggesting that whey proteins do not influence the alkaline disruption process. Levels of ionic calcium and serum calcium and phosphate decreased in a logarithmic fashion with increasing pH, indicating precipitation of calcium phosphate onto the casein micelles. A mechanism for alkaline disruption of casein micelles is proposed, in which increasing the milk pH improves the solvent quality for the caseins, thereby leading to the disruption of casein micelles into their constituent nanoclusters; increases in the net-negative charge on the caseins on increasing pH may contribute to micellar dissociation.     

不同酸性物质对石灰性土壤的酸化效果研究

采用室内模拟试验研究了酸性物质对石灰性土壤的酸化效果。结果表明,磷酸能有效地降低石灰性土壤的pH,要使土壤pH由8.89降至6.0左右,最佳酸用量为3.27g/100g土。对于pH较高的石灰性土壤,只用低pH(pH=5.5)的酸水溶液在短时间内降低土壤pH是不切实际的。磷酸二氢铵不仅具有很好的酸化效果,且具有很好的缓冲性能,每100g土壤加入2.3g磷酸二氢铵可使石灰性土壤pH从8.89降低到6.39,并能维持较长的酸性环境。施用5%的硫磺可在30d内使土壤pH降低到7.0～7.5之间。     

Interactions of high methoxyl pectin with whey proteins at oil/water interfaces at acid pH

The interactions between whey protein isolate (WPI) and high methoxyl pectin (HMP) at pH 3.5 were investigated in situ using ultrasound (US) and diffusing wave spectroscopy (DWS). HMP was added to 10% oil-in-water emulsions containing 1% WPI. At neutral pH, no protein-pectin interactions were observed as both molecules are negatively charged, while at pH 3.5 bridging flocculation occurred via electrostatic interactions. Four different stages were distinguished during the addition of HMP in WPI-stabilized emulsions at pH 3.5. At a concentration below a critical value, no interactions were observed. At concentrations >0.02% HMP, a change in the l factor indicated a change in the ordering of the emulsion droplets, influenced by long-range interactions. At higher concentrations (in the range between 0.04 and 0.06% HMP), attenuation showed significant changes in the surface of the oil droplets, changes which affected the droplet-droplet interactions. At pectin concentrations >0.05%, attenuation of sound and 1/l* decreased, while velocity of sound and particle size increased, as a result of bridging flocculation. These results demonstrated for the first time that methods such as US and DWS combined permit the observation of the early stages of the interactions between two biopolymers at the interface. This is significant in light of increasing efforts in engineering complex interfacial layers.     

Reactions of zinc with iron-oxide coated calcite surfaces at alkaline pH

Both iron oxides and carbonate minerals, such as calcite, can sorb zinc (Zn), and therefore are important in controlling the solution concentration and availability of Zn to plants growing in calcareous soil. When present together, interactions between these components affect their sorption behaviour. We investigated changes in the reactions of Zn with calcite at alkaline pH, as the calcite surface was progressively coated by iron oxide. Coated calcite surfaces were prepared that had from 0.05 to 1.45% iron oxide. The initial concentration of Zn and the amount of iron oxide on the calcite were the most critical factors affecting adsorption, precipitation of solid phases, and the desorbability of sorbed Zn. For pure calcite at small initial Zn concentrations (< 2.5 × 10^?5 m ) adsorption was dominant; with increasing concentration, precipitation of hydrozincite (ZHC) became more important. With increasing amounts of iron oxide the amount of Zn adsorbed increased, the desorbability of the Zn decreased, and precipitation became progressively less evident, and at 1.45% iron oxide content there was no evidence of any precipitation of ZHC. The calculated maximum adsorption attributable to the iron oxide coating was inversely proportional to the thickness of the oxides on the calcite, and greatly exceeded that of iron oxide as a separate phase. The common occurrence of iron‐coated carbonates in calcareous soils and their capacity to adsorb Zn contributes to the problems of Zn deficiency, for which these soils are noted.     

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.     

Proton release by roots of Medicago murex and Medicago sativa growing in acidic conditions, and implications for rhizosphere pH changes and nodulation at low pH

Medicago murex nodulates faster and produces more nodules than Medicago sativa in acidic sandy soils. Experiments using a ‘root mat’ approach and videodensitometry examined pH changes in the rhizospheres of nitrate-fed plants of M. murex and M. sativa. Using the ‘root mat’ approach with soil disks of pH 4.49, M. sativa cv. Aquarius acidified its rhizosphere by approximately 0.2-0.4 pH-units within 4 d, while M. murex cv. Zodiac did not acidify its rhizosphere. Rates of H⁺ release were higher from M. sativa than from M. murex. Videodensitometry of roots embedded in agarose of pH 4.5 showed that the mature parts of the tap-root of both species exuded OH⁻ ions, but was approximately twofold more in M. murex than in M. sativa. Consequently, young parts of the M. sativa rhizosphere were less alkaline than that of M. murex. It is suggested that the difference in nodulation response between the two species at low pH may be related to the different patterns of rhizosphere acidification: the stronger rhizosphere acidification of M. sativa being less favourable for survival and growth of Sinorhizobium medicae. The higher rate of rhizosphere acidification by M. sativa roots may be related to its genetic characteristics including greater relative root growth rate and greater sensitivity to acidity in comparison to M. murex.     

Effects of acidic meltwater on chemical conditions at nearshore spawning sites

Chemical conditions at lake charr (Salvelinus namaycush) spawning sites were monitored during snowmelt in low alkalinity lakes in Ontario, Canada. Embryos within the interstitial water of the spawning substrate were exposed to abrupt and potentially toxic levels of H⁺ and inorganic Al as acidic runoff water inundated the shallow nearshore sites. Early runoff events, those that occurred while the lake ice was still snow-covered and under-ice water temperatures were <2°C, appeared to be most threatening, because of deep penetration of the runoff water. These highly site-specific events exhibit wide temporal and spatial variability.     

Recolonization of methyl bromide sterilized soils under four different field conditions

Summary The course of recovery in biological activity was assessed in the top 5 cm of undisturbed soil cores (29.7 cm diameter, 30 cm deep) that had been fumigated in the laboratory with methyl bromide. The cores were returned to their original pasture and forest sites, two with a moderate and two with a high rainfall, and untreated soils at all sites served as baselines. Sampling took place over 166 days (midsummer to midwinter). Microbial biomass (as measured by fumigation-extraction and substrate-induced respiration procedures) and dehydrogenase activity both recovered rapidly, but remained consistently lower in the fumigated than in untreated samples at both forest sites and at the moister of the two pasture sites. Bacterial numbers also recovered rapidly. Fungal hyphal lengths were, on average over 166 days, 25% lower in the fumigated soils. Levels of mineral N were initially highest in the fumigated soils, but declined with time. Fumigation generally had no detectable effects on the subsequent rates of net N mineralization and little effect on nitrification rates. Fumigation almost totally eliminated protozoa, with one to three species being recovered on day 0; the numbers recovered most rapidly in the moist forest soil and slowly in the dry pasture soil. The recoionization rate of protozoan species was similar in all soils, with species numbers on day 110 being 33 and 34 in the fumigated and untreated soils, respectively. Nematodes were eliminated by fumigation; recolonization was first detected on day 26 but by day 166, nematode numbers were still lower in fumigated than in untreated soils, the abundance being 10 and 62 g^-1 soil and diversity 10 and 31 species, respectively. Overall, the results suggest that protozoan and nematode populations and diversities could provide a useful medium-term ecological index of the recovery in comprehensive soil biological activity following major soil pollution or disturbance.     

Hydrolytic products and kinetics of triazophos in buffered and alkaline solutions with different values of pH

The hydrolysis of triazophos was studied in buffered solutions in the range of pH 4-10 and in sodium hydroxide solutions with pH values up to 12. The results showed that the degradation of triazophos in the above solutions followed simple pseudo-first-order kinetics. At 35 degrees C, the rate constants in buffered solutions ranged from 0.0222 d(-1) at pH 4 to 0.5357 d(-1) at pH 10, and increased to 0.6251 h(-1) in 0.01 mol/L sodium hydroxide solution. The results also indicated that the base-catalysis was more important than acid-catalysis in the hydrolysis of triazophos. On the basis of the Arrhenius plot, the calculated activation energy (E(a)) and the frequency factor (A) for the hydrolysis of triazophos in buffered solution of pH 10 were 78.6 kJ/mol and 1.13 x 10(13) d(-1), respectively. Hydrolytic products of triazophos in buffered solutions of pH 4 and 10, as well as in sodium hydroxide solution of pH 11, were identified as their corresponding trimethylsilyl derivatives with a gas chromatography-mass spectrometer (GC-MS). The possible hydrolytic pathways of triazophos were also proposed.     

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.     

微波辅助提取中pH值与脱色对苹果果胶的影响

为了得到高得率、高品质的苹果果胶,该文利用不同pH值的盐酸溶液对苹果渣中果胶进行微波辅助提取（Microwave-assisted extraction, MAE）,之后对果胶提取液进行大孔树脂XAD-16HP脱色,研究了pH值与脱色对果胶得率和品质的影响。结果表明：微波辅助提取工艺中随着pH值的升高,果胶得率、半乳糖醛酸质量分数和总离子含量减少,酯化度、黏均分子量和总多酚增加,褐变度无显著变化;大孔树脂吸附脱色后果胶褐变度、总多酚、彩度C*值显著下降,色调角H°值显著增加,半乳糖醛酸质量分数和酯化度没有     

Critical pH and exchangeable Al of four acidic soils derived from different parent materials for maize crops

Purpose

The purpose of this study is to determine the critical soil pH, exchangeable aluminum (Al), and Al saturation of the soils derived from different parent materials for maize.

Materials and methods

An Alfisol derived from loess deposit and three Ultisols derived from Quaternary red earth, granite, and Tertiary red sandstone were used for pot experiment in greenhouse. Ca(OH)₂ and Al₂(SO₄)₃ were used to adjust soil pH to target values. The critical soil pH was obtained by two intersected linear lines of maize height, chlorophyll content, and yield of shoot and root dry matter changing with soil pH.

Results and discussion

In low soil pH, Al toxicity significantly decreased plant height, chlorophyll content, and shoot and root dry matter yields of maize crops. The critical values of soil pH, exchangeable Al, and Al saturation varied with soil types. Critical soil pH was 4.46, 4.73, 4.77, and 5.07 for the Alfisol derived from loess deposit and the Ultisol derived from Quaternary red earth, granite, and Tertiary red sandstone, respectively. Critical soil exchangeable Al was 2.74, 1.99, 1.93, and 1.04 cmol_ckg^?1 for the corresponding soils, respectively. Critical Al saturation was 5.63, 12.51, 14.84, and 15.16% for the corresponding soils.

Conclusions

Greater soil cation exchange capacity and exchangeable base cations led to lower critical soil pH and higher critical soil exchangeable Al and Al saturation for maize.

     

Impacts of elemental S applied under various temperature and moisture regimes on pH and available P in acidic, neutral and alkaline soils

We evaluated the effect of elemental S (S⁰) under three moisture (40, 60, 120% water-filled pore space; WFPS) and three temperature regimes (12, 24, 36°C) on changes in pH and available P (0.5 N NaHCO₃-extractable P) concentrations in acidic (pH 4.9), neutral (pH 7.1) and alkaline (pH 10.2) soils. Repacked soil cores were incubated for 0, 14, 28 and 42 days. Application of S⁰ did not alter the trends of pH in acidic and neutral soils at all moisture regimes but promoted a decrease in the pH of alkaline soil under aerobic conditions (40%, 60% WFPS). Moisture and temperature had profound effects on the available P concentrations in all three soils, accumulation of available P being greatest under flooded conditions (120% WFPS) at 36°C. Application of S⁰ in acidic, neutral and alkaline soils resulted in the net accumulation of 16.5, 14.5 and 13 g P g^–1 soil after 42 days at 60% WFPS, but had no effect under flooded conditions. The greatest available P accumulations in the respective soils were 19, 19.5 and 20 g P g^–1 soil (equivalent to 38, 41, 45 kg P ha^–1) with the combined effects of 36°C, 60% WFPS and applied S⁰. The results of our study revealed that oxidation of S⁰ lowered the pH of alkaline soil (r=–0.88, P<0.01), which in turn enhanced available P concentrations. Also, considering the significant relationship between the release of sulphate and accumulation of P, even in acidic soil (r=0.92, P<0.01) and neutral soil (r=0.85, P<0.01) where the decrease in pH was smaller, it is possible that the stimulatory effect of sulphate on the availability of P was due to its concurrent desorption from the colloidal surface, release from fixation sites and/or mineralization of organic P. Thus, in the humid tropics and irrigated subtropics where high moisture and temperature regimes are prevalent, the application of S⁰ could be beneficial not only in alleviating S deficiency in soils but also for enhancing the availability of P in arable soils, irrespective of their initial pH.     

Do stream invertebrates accumulate aluminium at low pH conditions?

Acidified surface waters often show elevated aluminium (Al) levels, detrimental to fish and some invertebrates. Whether Al can accumulate in benthic invertebrates, with time and/or along the food chain, is not clear. To test this, benthic invertebrates, representing different functional feeding groups, were collected in spring from streams, with different acidity and Al concentrations. Weight-specific Al content was determined with an AAS. At localities with pH ≈ 4, high Al contents (≈ 1 mg inorg-Al g^?1 af dw) were found in shredders and/or deposit feeders (Asellus aquaticus, Nemoura sp., and limnephilids), while the predator Isoperla grammatica contained only ≈ 0.3 mg Al g^?1, and the “filtering predator” Plectrocnemia conspersa almost no Al. Also at pH ≈ 6 Nemoura sp. and limnephilids showed significantly higher Al contents than did the predators Isoperla grammatica and Rhyacophila nubila, Al concentrations of the animals were often higher at pH 4 than at pH 6. Thus, no evidence of any food chain accumulation (or biomagnification) of Al could be validated. Accordingly, this study gives no support that the high concentrations of Al in fish and birds are due to their feeding on benthic invertebrates at low pH conditions. It was also found that animals that inhabit and/or consume benthic detritus as food contain highest Al levels.     

不同pH条件下腐植酸对土壤中砷形态转化的影响

为了探索在不同pH条件下腐植酸对土壤中砷形态转化及生物毒性的影响,应用油菜盆栽试验,检测砷加入土壤后15 d、45 d、90d土壤中AE-As(水溶态、可交换态和碳酸盐结合态)Fe,Mn-As(铁/锰氧化物结合态),O,S-As(有机物及硫化物结合态),Res-As(残渣态)的含量及油菜生物量.结果表明,外源水溶态砷加入土壤后均迅速向相对稳定的形态转化,15 d时A3H1(pH9.5,腐殖酸用量为0g/kg)和A3H4(pH9.5,腐殖酸用量20 g/kg)处理的AE-As含量分别为13.40 mg/kg、9.23 mg/kg,转化率分别为78.7％、82.51％;在90 d时,A3H4处理的AE-As、Fe,Mn-As含量分别为6.28 mg/kg、1.23 mg/kg,仅为处理A3H1处理的53.9％和10.7％,O,S-As、Res-As含量分别为20.24 mg/kg、41.21 mg/kg,是处理A3H1的165％和127％.说明腐植酸更有利于土壤中AE-As向其他相对稳定的形态转化,主要体现在O,S-As的增加,腐植酸对碱性土壤中砷向较稳定形态转化的促进作用更加显著,而且与腐植酸的施用量呈正相关.外源砷在碱性条件下对植物的毒害更大,腐植酸用量在10 g/kg土时就可以有效降低砷的毒害.     

Stabilizing behavior of soy soluble polysaccharide or high methoxyl pectin in soy protein isolate emulsions at low pH

The stability of emulsions prepared with soy protein isolates was investigated as a function of pH in the presence of two negatively charged polysaccharides: high methoxyl pectin (HMP) and soy soluble polysaccharide (SSPS). Both polysaccharides are composed of a backbone which contains galacturonic acid but, when added to soy protein isolate-stabilized emulsions, SSPS showed a different behavior than that of HMP. At neutral pH and above a critical concentration of stabilizer (0.05%), HMP caused flocculation of the emulsion droplets via a depletion mechanism. On the other hand, the emulsions containing a similar amount of SSPS did not show creaming or flocculation. At acidic pH (<4.0) the addition of pectin caused extensive droplet aggregation, while no aggregation was observed with the addition of SSPS. The differences in the stabilization behavior between the two polysaccharides can be attributed to their differences in charge, neutral sugars side chains, and molecular weight.