Obtainment of chelating agents through the enzymatic oxidation of lignins by phenol oxidase

Oxidation of lignin obtained from acetosolv and ethanol/water pulping of sugarcane bagasse was performed by phenol oxidases: tyrosinase (TYR) and laccase (LAC), to increase the number of carbonyl and hydroxyl groups in lignin, and to improve its chelating capacity. The chelating properties of the original and oxidized lignins were compared by monitoring the amount of Cu²⁺ bound to lignin by gel permeation chromatography. The Acetosolv lignin oxidized with TYR was 16.8% and with LAC 21% higher than that of the original lignin. For ethanol/water lignin oxidized with TYR was 17.2% and with LAC 18% higher than that of the original lignin.     

Comparison of physical and chemical properties of high pressure- and heat-treated Lychee (Litchi chinensis Sonn.) in syrup

Lychee usually has white flesh, but its flesh is very sensitive to thermal and enzymatic impairment and ultimately changes in color. This investigation was intended to study the magnitude of color change in lychee by high pressure and thermal processing. The lychee was packed in syrup prior to being processed. Pressurized lychee was performed at 600 MPa at 30°C or 50°C for 20 min, while the pasteurized sample was heated at 90°C for 3 min. It was found that pressurization induced lower color L*, a* and b* values, including low anthocyanin content. For enzymatic activities, high pressure could reduce the activity of polyphenoloxidase by 33–51%, whereas pasteurization markedly reduced that activity by 90%.     

超高压处理对多酚氧化酶活性的影响

 研究了超高压中温协同处理对砀山梨汁中多酚氧化酶活性的影响,实验压力为0.1~500 MPa,温度为20~60 ℃。此外,考察了不同pH值（3~7）和保压时间（2~34 min）超高压处理对酶活性的影响。实验结果分析表明：在处理温度为50 ℃、保压时间为10 min和梨汁pH值为5的条件下,200~300 MPa处理梨汁时多酚氧化酶被激活,活性表现最高;500 MPa时酶的活性下降到75.3%。协同温度为30 ℃处理梨汁时,酶的活性反而增大;30 ℃以后,酶活性随温度升高而迅速降低;有效协同高压处理的温度为40 ℃。随着保压时间的延长,梨汁中过氧化物酶的活性减小;18 min以前下降速度较快些,之后下降速度变缓。pH在5~6之间,酶的残留活性最大;pH值为6时,梨汁中多酚氧化酶最为耐压。     

A validated solid-liquid extraction method for the HPLC determination of polyphenols in apple tissues Comparison with pressurised liquid extraction

A solid-liquid extraction procedure followed by reversed-phase high-performance liquid chromatography (RP-HPLC) coupled with a photodiode array detector (DAD) for the determination of polyphenols in freeze-dried apple peel and pulp is reported. The extraction step consists in sonicating 0.5 g of freeze-dried apple tissue with 30 mL of methanol-water-acetic acid (30:69:1, v/v/v) containing 2 g of ascorbic acid/L, for 10 min in an ultrasonic bath. The whole method was validated, concluding that it is a robust method that presents high extraction efficiencies (peel: >91%, pulp: >95%) and appropriate precisions (within day: R.S.D. (n = 5) <5%, and between days: R.S.D. (n = 5) <7%) at the different concentration levels of polyphenols that can be found in apple samples. The method was compared with one previously published, consisting in a pressurized liquid extraction (PLE) followed by RP-HPLC-DAD determination. The advantages and disadvantages of both methods are discussed.     

蘑菇多酚氧化酶的性质及其催化反应动力学的研究

本文用二次丙酮法从磨菇中提取多酚氧化酶，用分光光度法测定多酚氧化酶催化邻苯二酚的氧化反应。实验结果表明：酶作用的最佳条件是pH等于7、温度30℃，反应活化能等于21．20kJ．mol-1。     

Resonance Scattering Effect of Dopamine Product Particle and Its Application to Polyphenoloxidase Activity Assay

In pH 7.2 Na₂HPO₄‐NaH₂PO₄ buffer solution, polyphenoloxidase (PPO) could catalyze the oxidization of dopamine (DA) to form polymer particles in dark‐red color, which exhibited a strong resonance scattering (RS) peak at 780 nm. In the chosen conditions, as the PPO activity increased, the RS intensity at 780 nm increased linearly. The increased RS intensity (ΔI_{780 nm}) was linear to the PPO activity in the range of 0.10–6.0 U·mL^?1, with a regression equation of ΔI_{780 nm}?96.6C+15.1, a relative coefficient of 0.9987 and a detection limit of 0.06 U·mL^?1 PPO. The proposed method was applied to detecting PPO activity in potato sample with satisfactory results.     

水溶性壳聚糖浸种对小麦幼苗抗性相关酶活性的影响

利用0.5 g/L的5 种水溶性壳聚糖分别对小麦进行浸种处理,在幼苗发育的5 个时期对叶内苯丙氨酸解氨酶、多酚氧化酶、过氧化物酶和超氧化物歧化酶的活性进行测定.壳聚糖处理降低叶内苯丙氨酸解氨酶活性.多酚氧化酶活性变化不显著,初期酶活性较对照高,随幼苗发育先降后升.壳聚糖浸种处理显著提高叶内过氧化物酶和超氧化物歧化酶活性.5种分子量中以10×103的壳聚糖C浸种处理小麦幼苗叶内抗性相关酶活性强.     

多酚氧化酶催化反应的抑制动力学及机理

用丙酮自香蕉皮中提取多酚氧化酶，用氧电极法测得酶促儿茶酚氧化反应的表现活化能为２３．６ｋＪ·ｍｏｌ－１，米氏常数为１．５８×１０－２ｍｏｌ／Ｌ，测试了９种化合物对酶活性的抑制效果。以苯甲酸为抑制剂研究了抑制动力学及机理，测出蕉皮酚酶中含有铜离子，它起着活化反应物的作用，抑制剂与铜离子的结合抑制了酶的催化活性。     

Tyrosinase versus Catechol Oxidase: One Asparagine Makes the Difference

Tyrosinases mediate the ortho‐hydroxylation and two‐electron oxidation of monophenols to ortho‐quinones. Catechol oxidases only catalyze the oxidation of diphenols. Although it is of significant interest, the origin of the functional discrimination between tyrosinases and catechol oxidases has been unclear. Recently, it has been postulated that a glutamate and an asparagine bind and activate a conserved water molecule towards deprotonation of monophenols. Here we demonstrate for the first time that a polyphenoloxidase, which exhibits only diphenolase activity, can be transformed to a tyrosinase by mutation to introduce an asparagine. The asparagine and a conserved glutamate are necessary to properly orient the conserved water in order to abstract a proton from the monophenol. These results provide direct evidence for the crucial importance of a proton shuttle for tyrosinase activity of type 3 copper proteins, allowing a consistent understanding of their different chemical reactivities.     

Biochemical alternation and silicon accumulation in groundnut (Arachis hypogaea L.) in response to root and foliar application of two sources of silicon

Abstract

Silica deposits in leaves of groundnut ranged from 3.23 to 6.16% and 2.61 to 6.52% when sodium meta silicate was applied as foliar spray at the rate of 0.5, 1.0, 2.0, 3.5 and 5.0%, and soil application at the rate of 5.0, 7.5, 10.0, 15 and 20% and 4.44 to 11.94% and 3.57 to 9.33% when calcium silicate was applied through leaves and soil at the same doses, respectively in contrast to 2.0% in plants without silica fertilizer applications. Similarly, polyphenol oxidase (PPO) activity was highest (43.26 units) in plant system, in case of foliar spray of calcium silicate at the rate of 5.0% and 33.80 units with reference to foliar spray of meta silicate at the rate of 5.0% whereas it was only 19.10 units in untreated check. Among the two sources of silica fertilizers tested, foliar spray of calcium silicate at 5.0% was effective in facilitating the maximum deposition of silica content apart from increasing the activities of PPO in groundnut leaves.