Evaluation of Food Conservation Technologies for Potato Cubes

Potato cubes of cultivar Innovator were subjected to different food conservation treatments: osmotic dehydration and antioxidants (OD + A), edible coating (E), edible coating with osmotic dehydration and antioxidants (EC + OD + A), immersion in antioxidants (A) and microwave blanching (B). The quality of the products was evaluated by measuring dry matter content, colour, browning index, total phenols and antioxidant capacity and polyphenol oxidase (PPO) activity. Treatments with A resulted in a lower a* colour parameter (red to green) but increased total phenols and antioxidant activity. Those treatments with OD and A together resulted in even larger increases in total phenols and antioxidant activity and also reduced PPO to negligible activity. OD increased dry matter content in OD + A treatments. B increased the browning index (BI) and colour parameters a* and ΔE (total change). As a consequence, potato cubes treated with osmotic dehydration and antioxidants had improved quality characteristics and these treatments are appropriate for minimally processed vegetables.     

Oxidation of ortho‐diphenols in red clover with and without polyphenol oxidase (PPO) activity and their role in PPO activation and inactivation

Oxidation of phenol to quinone with its subsequent binding and complexing with protein in red clover (Trifolium pratense L.) when fed to ruminant livestock has been shown to improve nitrogen‐use efficiency and the deposition of polyunsaturated fats into animal products. This oxidation has, almost exclusively, been attributed to the activity of the enzyme group polyphenol oxidases (PPOs) and, specifically, catecholase during conservation. However, during conservation, temporal inactivation of PPO occurs and oxidation of phenolic substrate may occur through other mechanisms not associated with PPO. T₁ progeny of a cross between a red clover clone having normal levels of foliar PPO activity and a transgenic PPO‐silenced red clover plant with undetectable levels of PPO activity were divided into two populations based on the presence of the silencing transgene and lack of measurable PPO activity (PPO?) or the absence of the silencing transgene and normal levels of PPO activity (PPO+). This material was subsequently used to determine the relevant extent of phenolic oxidation in the presence and absence of PPO activity under two damage regimes (heavy and light). PPO+ and PPO? material was passed through a garden shredder as the light‐damaged regime, or frozen at ?20°C as the heavy damaged regime. Material was left at room temperature and sampled at regular intervals for the determination of PPO enzyme activity (active and total), PPO activation (conversion from latent to active forms) and formation of protein‐bound phenol (PBP) as a measure of oxidation. Experiments with red clover leaf extracts were carried out to determine the mechanism for the temporal activation and inactivation (loss of enzymatic activity) of PPO during wilting. PBP formation was evident in both PPO? and PPO+ treatments with the response further catalysed during the first 2 h by extent of damage that increased the activation of PPO, but which also resulted in a more rapid temporal inactivation of PPO. PBP formation, PPO activation and PPO inactivation were all shown to be mediated predominantly by quinone binding. A role of non‐PPO oxidation in PBP formation in wilted red clover is reported, showing the importance of o‐diphenolic substrate concentration and not just PPO activity to increase nitrogen use efficiency and the deposition of polyunsaturated fats into animal products.     

小麦面粉白度与色泽因子配合力及相关分析

为探讨面粉白度与色泽相关因子的遗传特性及相互关系,选择11个在面粉白度等性状方面具有不同特点的小麦品种作亲本,按5×6不完全双列杂交设计组配杂交组合,对30个F₁杂种和11个亲本的面粉白度与色泽相关因子进行了分析。结果表明,面粉白度、黄色素含量、多酚氧化酶活性在亲本表型值与其一般配合力效应值间存在显著的正相关。在亲本表型值间和一般配合力效应值间,面粉白度与黄色素含量均呈显著负相关,与多酚氧化酶活性间无显著相关性。面粉白度、黄色素含量和多酚氧化酶活性的广义遗传力和狭义遗传力都较高,主要受基因加性效应影响,适合在早代进行选择。德麦3号、济麦22号和济麦20号都能较好地降低后代的黄色素含量和多酚氧化酶活性,是小麦白度育种中较适宜选择的亲本。在优质中强筋小麦育种中,不应过分追求面粉的白度和亮度,但可通过对低多酚氧化酶活性和低黄色素含量的选择,培育面粉色泽优良的小麦新品种。     

茶树多酚氧化酶基因的克隆及其序列比较

根据已经发表的多酚氧化酶基因中的保守序列 ,设计兼并引物 ,利用nest-PCR技术 ,首次克隆了茶树多酚氧化酶基因。该基因已经在GenBank上登录 ,登录号为AF2 69192。序列分析表明 ,茶树多酚氧化酶基因与其它植物中的多酚氧化酶基因有较高的相似性 ,尤其在铜离子结合部位 ,所有的植物基本上一致。进化树分析表明 ,茶树的多酚氧化酶可以与大多数的木本植物聚合成一个组     

多酚氧化酶活性基因等位变异在陕西小麦品种中的分布

为了解陕西小麦品种控制多酚氧化酶(Polyphenol oxidase,PPO)活性的主效基因组成.利用PPO 18和PPO 29标记时138份推广品种的PPO-2A和PP(9-2D位点的等位变异进行了分子检测.结果表明,在PPO2A住点,含Ppo-A1a(高PPO活性)和Ppo-A1b(低PPO活性)等位变异类型的品种比例分别为47.1%和52.9%;在PPO2D位点含PpoD1a(中等低PPO活性)和PpcrD1b(中等高PPO活性)等位变异类型的品种频率分剐为30.4%和69.6%;含Ppo-A1a/Ppo-D1a(较高PPO活性)、PpoAla/PpoD1b(最高PPO活性)、Ppo-A1b/Ppo-D1a(最低PPO活性)、Ppo-A1b/Ppo-D1b(较低PPO活性)等住变异组合类型品种的频率依次为10.1%、37.0%,20.3%和32.6%;不同地区不同等位变异类型及其组合的分布比例也不同.总体来看,陕西低PPO活性的小麦品种比例偏低,选育低PPO活性品种应成为当前陕西小麦品质改良的主要目标之一.     

多酚氧化酶活性与小麦抗穗发芽的关系及抗穗发芽新品种秦麦3号的选育

为更好地选育抗穗发芽小麦新品种,解决小麦生产中的穗发芽损失问题,以18个穗发芽抗性不同的小麦品种(系)为试材,以微喷人工降雨法诱导穗发芽,测定了发芽和未发芽籽粒的多酚氧化酶(PPO)活性、酚含量和吸水率,并分析了他们与穗发芽抗性之间的关系。结果表明,PPO活性、吸水率均与穗发芽率呈显著正相关,即PPO活性、吸水率越低,穗发芽抗性越强;而酚含量与穗发芽率无相关性。同时,以PPO活性为抗穗发芽能力的筛选指标,从“昌农921×东方红3号”的杂交后代中培育出了高抗穗发芽的小麦新品种秦麦3号。     

短波紫外线和低温处理对采后芒果蒂腐病病原菌抑菌效果和生理指标的影响

以‘台农一号’芒果为材料,研究不同剂量短波紫外线和贮藏温度对采后芒果蒂腐病病原菌（Pestalotiopsis mangiferae）病斑直径、总酚和类黄酮含量、丙二醛、可溶性蛋白质及过氧化物酶、多酚氧化酶活性的影响。结果表明：4.9 kJ/m ²辐照能显著抑制病斑直径的扩展,诱导总酚、类黄酮含量的升高,降低可溶性蛋白质的消耗,提高过氧化物酶、多酚氧化酶活性的活力,抑制丙二醛的生成;9.8 kJ/m ²辐照会破坏植物细胞损伤,削弱类黄酮、可溶性蛋白质、过氧化物酶、多酚氧化酶上的诱导作用。与25 ℃相比,13 ℃下贮藏可抑制病斑直径的扩展,维持较高的总酚、类黄酮含量,抑制丙二醛的生成,延迟过氧化物酶、多酚氧化酶活力的达到峰值的时间。4.9 kJ/m ²,13 ℃的处理对芒果蒂腐病抑制和品质维持最好。     

做青过程中重要品质关联酶活性变化研究

以鸠坑种的夏茶对夹二、三叶新梢为原料,研究了两种做青方法过程中多酚氧化酶(PPO)、过氧化物酶(POD)的活性变化。结果表明两种做青方法PPO的活性在摇青阶段均呈现高→低→高的变化,且在第三次摇青后活性达到最高;而POD以鲜叶中的活性为最高,整个做青过程中略有下降但不明显。丙酮粉法比匀浆法测定酶活性更为稳定。     

Molecular and physico-chemical evaluation of enzymatic browning of whole meal and dough in a collection of tetraploid wheats

Many studies have demonstrated the role of polyphenol oxidase (PPO; E.C. 1.14.18.1) in darkening of wheat products. Even slight browning of pasta is a major hindrance to consumer acceptance. The aim of this research was to evaluate the variability of PPO activity, browning level, and protein content in a collection of more than 100 genotypes of tetraploid wheat, including cultivars and landraces. A molecular approach was followed to evaluate the efficiency of marker PPO18, discovered and used in common wheat, in detecting tetraploid wheat genotypes with low browning level. The data showed a significant genotype influence on the activity of the PPO enzyme, which was correlated with brown index of whole meal and dough, and with protein content. On the whole, the cultivars showed lower PPO activity, brown index, and protein content than landraces, but wide variability was present. Marker PPO18 detected four different alleles for Ppo-A1. The allele Ppo-A1f identified genotypes with high PPO activity and brown index, while the Ppo-A1b and null alleles were associated with the opposite characteristics. Hence, this marker can be used to select new tetraploid wheat cultivars with low browning level.     

Polyphenol oxidase and peroxidase activities of apricots (Prunus armeniaca L.) as affected by cultivar,location, and year of harvest

A total of 22 samples of apricots belonging to ten cultivars from two locations and picked in the years 1975–78 were assayed for polyphenol oxidase (PPO) and partly for peroxidase (POD) activities, heat resistance of these enzymes,o-dihydroxy phenol (ODP) content, and initial rate of enzymatic browning (BA). Of these characteristics, ODP content showed the greatest and BA the least variation. PPO activity relative to fresh weight was found to depend on the year of harvest rather than on the cultivar, while POD activity relative to fresh weight was cultivar dependent. The values of BA and ODP content were markedly influenced by both cultivar and year. For a given cultivar, PPO and POD activities relative to fresh weight as well as BA were higher in the samples picked in 1976 at one location than in those picked in the other years at the other location.The initial rate of browning of apricots was primarily dependent on the ODP content as shown by linear regression analysis with two independent variables. The relative influence of ODP content as compared with that of PPO activity related to fresh weight was the stronger the higher the value of the ratio of PPO-ODP, i.e. the smaller the values of the endogenous substrate available to the enzyme in the tissues. This suggests substrate depletion to be the limiting factor in the enzymatic browning of apricots.Heat inactivation of PPO was found to be a simple exponential process, while that of POD a biphasic one. PPO was less heat resistant than POD: total inactivation occurred after 10-min heat treatment at 75°C and 85°C, respectively.     

打顶对烤烟主要化学成分及多酚氧化酶活性的影响

为探讨烤烟品质的顶端调控措施,研究了不同打顶措施对烟叶多酚氧化酶活性和主要化学成分的影响。结果表明,与不打顶处理相比,打顶抹权明显促进了烟碱的合成以及在烟叶中的积累,有增加还原糖含量的趋势,能减少后期烟叶中钾素的流失,从而提高了上、中部叶的钾含量,有效提高了成熟期多酚氧化酶的活性。     

毛栓菌多酚氧化酶酶学性质及茶黄素的酶促合成研究

研究从毛栓菌中分离得到的多酚氧化酶(PPO)用于茶黄素的体外氧化制备。结果表明毛栓菌胞外PPO较适的反应温度范围在28~36℃之间;最适pH值为5.2;恒温反应30min内均表现出较高的活性和稳定性;50%的硫酸铵可以沉淀粗酶液中96.0%的PPO。将毛栓菌PPO加入到儿茶素反应液中进行双液相反应,可得到10.19%的茶黄素,与茶鲜叶来源的PPO相比,毛栓菌PPO制备茶黄素的总含量偏低,但其中TF-3-G的比例较高,达到总茶黄素的68.10%,占脂型茶黄素总量的92.08%。     

外源IBA对裸花紫珠插穗营养物质含量及抗氧化酶活性的影响

以药用植物裸花紫珠(Callicarpa nudiflora)一年生半木质化枝条为材料,使用吲哚丁酸(IBA,1 500 mg/L)处理插穗,以清水为对照,每隔6 d取样1次,测定生根过程中插穗基部可溶性糖、淀粉、可溶性蛋白含量及吲哚乙酸氧化酶(IAAO)、多酚氧化酶(PPO)、过氧化物酶(POD)活性的动态变化。结果表明,扦插后0~12 d,1 500 mg/L IBA处理增加了插穗内可溶性蛋白含量,降低了淀粉含量,可溶性糖含量先降后升,IAAO、POD、PPO活性增加;扦插后12~30 d,淀粉及可溶性糖含量逐渐降低,可溶性蛋白含量先升后降,但仍都高于对照,IAAO和PPO活性逐渐下降,POD活性先降后增;扦插30 d后,可溶性糖、蛋白、淀粉含量逐步趋于稳定,IAAO活性趋于稳定,而PPO和POD活性逐渐降低。总之,较高浓度的可溶性糖、可溶性蛋白,低浓度的淀粉,较高的IAAO、POD和PPO活性有利于裸花紫珠插穗不定根形成和发育。     

硝酸镍诱导水稻白叶枯病抗性与防卫反应酶活性的变化

用1．0和2．0mmol／L的硝酸镍处理水稻幼苗2d后，再用稻白叶枯细菌(Xanthomonns oryzae pw.oryzae)挑战接种，14d后调查病情，发现病斑长分别比对照下降了42．1％和48．0％，说明硝酸镍能诱导水稻幼苗对白叶枯病的抗性。生理指标测定及同工酶电泳结果表明，两种浓度的硝酸镍处理均能明显提高叶片中POD、PPO和PAL活性以及木质素含量，大部分POD同工酶带和4条PPO同工酶带增强，提示硝酸镍对水稻抗白叶枯病的诱导效应可能与POD、PPO和PAL活性以及木质素含量升高有关。     

温度、pH对马铃薯多酚氧化酶活性的影响

对加工型马铃薯试管苗、茎段愈伤组织、块茎及块茎芽在不同温度、不同 pH条件下PPO活性的检测结果表明 :pH对马铃薯PPO活性具有明显的影响 ,块茎芽以及愈伤组织PPO活性均在 pH 5～ 5 5之间最高 ,分别为 94 2 0及 2 0 4 0 (0 0 1ΔOD/min) ,而试管苗在培养基 pH为 8时PPO活性最高 (19 80 (0 0 1ΔOD/min) )。不同温度下不同品种PPO活性表现不同 ,甘农薯 1号、Atlantic试管苗PPO活性在 18～ 2 5℃之间达到高峰 ,分别为 16 80和 37 2 0 (0 0 1ΔOD/min) ;Shep ody、Snowden试管苗在 5℃下PPO活性较高 ;除此之外 ,不同品种贮藏块茎PPO活性均在 2 5℃表现最高。     

茶树PPO融合蛋白真核表达载体的构建与表达

以迎霜品种茶树基因组为模板,利用高保真聚合酶pfu,通过PCR方法扩增得到茶树PPO基因的编码区序列。通过设计酶切引物将其成功融合至真核表达载体pPICZa中,构建出茶多酚氧化酶的融合蛋白真核表达载体pPICZa-PPO,并将其转化进入巴斯德毕赤酵母（Pichia pastoris）GS115中,成功筛选出多个阳性转化子。对阳性转化子进行甲醇诱导,采用Western-Blotting方法在培养液中成功检测到诱导表达的目的蛋白。酶活检测结果也显示诱导产物具有较高的相对酶活性,能够正确发挥酶蛋白的生物功能。     

DEHP胁迫对高/低累积邻苯二甲酸酯品种水稻抗氧化酶系统的影响

以前期筛选获得的邻苯二甲酸酯(PAEs)高/低累积基因型水稻(Oryza sativa L.)品种(培杂泰丰/丰优丝苗)进行土培试验,对比研究邻苯二甲酸二(2-乙基己基)酯(DEHP)污染胁迫下2个品种水稻体内过氧化物酶(POD)和多酚氧化酶(PPO)活性及丙二醛(MDA)含量变化,以揭示DEHP胁迫下2个品种的生理生化和DEHP累积差异。结果显示,2个品种水稻体内DEHP含量随土壤污染浓度增加而增加,但生物量下降,而且培杂泰丰地上部的生物量敏感性响应指数和DEHP含量比丰优丝苗的高,说明前者对DEHP的耐受性更强。2个品种水稻体内丙二醛含量基本保持不变,但过氧化物酶和多酚氧化酶活性则随土壤DEHP浓度的增加而升高,说明水稻可能通过提高过氧化物酶和多酚氧化酶活性以降低DEHP胁迫。丰优丝苗的多酚氧化酶活性较培杂泰丰的高,可能影响到DEHP在水稻体内降解进而导致前者体内DEHP含量较低。     

丰水梨多酚氧化酶基因的克隆与原核表达

通过PCR方法克隆丰水梨PPO基因全长,并将其登录Genebank,登录号JQ861265。基因全长1782 bp,无内含子,编码的PPO属于亲水性蛋白质,无跨膜结构,含有593个氨基酸,分子量约为65.8 k Da,理论等电点为8.4。N端含有一段由47个氨基酸组成的转运肽。去除转运肽的成熟PPO分子量为60.8 k Da,理论等电点为6.69。PPO中含有两个铜离子结合区,主要位于PPO二级结构中的α-螺旋区域中。原核诱导目的蛋白在诱导3~6 h后积累量较大。诱导蛋白(PPO前体和成熟PPO)均能氧化儿茶素形成茶黄素。     

福建主要乌龙茶品种PPO酶活特性的研究

对四种乌龙茶茶树鲜叶的多酚氧化酶(PPO)进行粗提,测定并比较不同品种酶的特性。结果表明:在邻苯二酚与被测酶液的反应体系中,当底物邻苯二酚(C6H6O2)量为0.3~0.9ml时,毛蟹、铁观音、黄旦、肉桂酶活力随底物浓度的增加呈曲线增长,含量太高会产生抑制作用,当底物含量∶酶含量=1.5∶1时,酶活力表现最高;毛蟹、铁观音、黄旦、肉桂酶活力在45~65℃范围内均呈ω型变化趋势,依次在45℃、55℃、65℃时出现高峰。在50~90℃下水浴处理1h,毛蟹、铁观音、黄旦、肉桂酶活力分别丧失34.66%~55.91%,22.68%~50.89%,23.14%~53.57%,31.58%~57.89%;酶活力的适宜pH相对偏碱,在pH8.6时表现为一个活力高峰。相同浓度的五种抑制剂对酶均有一定的抑制效果,其中NaHSO3对酶活力的抑制作用最强,L-半胱氨酸其次,NaCl的抑制效果最差。     

椰甲截脉姬小蜂寄生对椰心叶甲幼虫体内几种酶活性的影响

采用分光光度法测定被椰甲截脉姬小蜂寄生0.5～4 d内的椰心叶甲3龄和4龄幼虫体内几种酶活性的变化.结果表明:寄生后,寄主幼虫体内过氧化氢酶(CAT)和过氧化物酶(POD)活性均高于对照;多酚氧化酶(PPO)活性受抑制;酯酶(EST)活性先升高后降低.结果揭示了椰心叶甲幼虫对寄生产生的生理防御反应.