Mechanism of Browning in Fresh Highbush Blueberry Fruit (Vaccinium corymbosum L). Role of Blueberry Polyphenol Oxidase,Chlorogenic Acid and Anthocyanins

Browning reactions which occurred after crushing fresh blueberries (Vaccinium corymbosum L) were followed by determination of chlorogenic acid colour density, polymer colour and the level of polymers. A polyphenol oxidase (PPO) activity plays a dominant role in enzymatic browning. Peroxidase (POD) does not contribute to brown colour formation. The loss of 29% of the colour in a model system containing PPO and blueberry anthocyanins indicated that PPO could act directly on these pigments. However, the rate of anthocyanin degradation was stimulated by the addition of chlorogenic acid. © 1997 SCI.     

高灌蓝越橘多酚氧化酶的提取和纯化

采用丙酮粉沉淀抽滤法、硫酸铵分级沉淀法及苯基琼脂糖疏水性层析法对高灌蓝越橘多酚氧化酶进行提取和纯化。在提取和纯化中，用加入不同的酚吸收剂和抑制剂并比较酶活性的高低来确认最佳方法。结果表明在粗提中加入酚吸收剂、增溶剂和抑制剂提取效果较好，苯基琼脂糖疏水性层析法对此类酶纯化效果较佳并为后来的辅酶的分离提供了较高的纯化倍数。     

Changes in Anthocyanins and Polyphenolics During Juice Processing of Highbush Blueberries (Vaccinium corymbosum L.)

Frozen blueberries ( Vaccinium corymbosum L.) were processed into juice and concentrate, and the changes in anthocyanin pigments and polyphenolics (cinnamates, procyanidins, flavonol glycosides) were monitored. While juice yield was 83%, only 32% of the anthocyanins were recovered in single-strength juice. Flavonol, procyanidin and chlorogenic acid recoveries in juice were 35%, 43%, and 53%, respectively. The proportion of polyphenolics remaining in the press-cake residue ranged from 1% (chlorogenic acid) to 18% (anthocyanins). Pronounced losses of anthocyanins and polyphenolics during milling and depectinization are believed to be due to native polyphenol oxidase. Losses during concentration ranged from 1.5% (anthocyanins) to 20% (procyanidins). Striking changes occurred in the anthocyanin profile with malvidin glycosides being most stable and delphinidin glycosides the least.     

杨桃多酚氧化酶的部分纯化及其特性研究

杨桃多酚氧化酶粗酶液经过DEAE-Toyopearl650M离子交换柱层析,Butyl-Toyopearl650M疏水柱层析后,被纯化了21.6倍,回收率为45.2%。该酶能迅速地催化焦性没食子酸的酶促氧化反应,而对邻苯二酚、间苯二酚、对苯二酚和绿原酸则完全无催化活性。该酶对焦性没食子酸的Km值为7.92mmol/L,其最适pH为8.0,pH稳定性范围在pH4.0-11.0,最适温度为60℃,热稳定性相对较高,在≥90℃加热30min后仍残留约9%的酶活性。该酶的最佳抑制剂是抗坏血酸,其次是NaHSO3和盐酸-L-半胱氨酸,Cu2+、Zn2+、Ca2+等金属离子对该酶也有一定的抑制作用。     

番石榴多酚氧化酶的部分纯化及其特性研究

番石榴PPO的粗提取液经过80%硫酸铵盐析和DEAE-Toyopearl 650M、CM-Sephadex C-50离子交换柱层析分离后,被纯化了约126倍,回收率为16.13%。该酶迅速地催化焦性没食子酸的酶促氧化反应,而对对苯二酚和绿原酸则完全无催化活性。该酶对焦性没食子酸的Km值为4.6 mmol/L,其最适pH为7.5,pH稳定性范围在pH4.0~11.0,最适温度为50℃,热稳定性相对较高,在≥90℃加热10 min后仍残留约15%的酶活性。该酶的最佳抑制剂是抗坏血酸和NaHSO3,其次是植酸、盐酸-L-半胱氨酸、柠檬酸,Ca2+、Mg2+等金属离子对该PPO也有较强的抑制作用。     

树脂法提取高灌蓝莓果色素的研究

以高灌蓝莓果为原料,研究不同树脂对蓝莓果色素的吸附率和解吸效果及不同洗脱剂的洗脱效果,确定用树脂法提取高灌蓝莓果色素的工艺,并对用所得的蓝莓果色素的性质进行检测。结果表明,D101-A树脂对高灌蓝莓果色素的吸附和解吸效果较好,用60％乙醇洗脱100min得到的产品质量较好,色价达42,产品收率为0．256％;且D101—A树脂重复使用20次后吸附率仅降低1．24％。在该提取条件下所提色素水溶性好,在酸性条件（pH≤4）下具有较好的稳定性,有一定的耐光性,在80℃以下热稳定性较好,且对低浓度的常用食品添加剂比较稳定。     

莲藕多酚氧化酶的纯化方法研究

研究了莲藕多酚氧化酶(PPO)的提取纯化。结果表明：莲藕多酚氧化酶采用30% 硫酸铵盐析除杂、80%硫酸铵沉淀、DEAE Sephadex A-50 柱层析、Sephadex G-75 柱层析四步进行分离纯化,所得纯化物经SDS-PAGE检验为均一单带,其分子量约为21kD,其纯化倍数和产率分别为103 倍和0.38%。     

涩柿贮藏过程中PPO及褐变控制

实验以新鲜的柿子为主要原料,采用测定吸光值的变化,研究了柿果中多酚氧化酶(PPO)活性的最适条件和抑制剂、烫漂温度和时间对柿果中多酚氧化酶(PPO)活性及褐变的影响。结果表明:柿子中的多酚氧化酶的最适温度是19℃,最适pH为6．5,最适底物浓度为12mmol/L。用0．1%亚硫酸钠+0．3%植酸和100℃+120s处理能有效的抑制柿果中PPO的活性,褐变速度明显减慢。     

雪梨多酚氧化酶酶学特性及其果汁褐变控制技术研究

雪梨在加工过程中极易发生酶褐变,为抑制雪梨汁的酶褐变,试验以雪梨为原料制备多酚氧化酶(Polyphenol oxidase,PPO)粗酶提取液,加入邻苯二酚为底物,采用分光光度法对其PPO的酶学特性及不同护色剂对PPO活性的影响进行了研究;并选择L-半胱氨酸、D-异抗坏血酸钠、抗坏血酸3种添加剂对雪梨汁进行护色,通过响应面优化试验确定雪梨汁最优护色组合。结果表明:雪梨PPO的最适pH为4.5,最适温度为30 ℃;雪梨PPO具有一定的热稳定性,随着温度的提高,抑制PPO活性所需要的时间逐渐减少;雪梨PPO催化底物邻苯二酚的酶促反应动力学与米氏方程高度符合,最大反应速率Vmax=217.39 U/min,米氏常数Km=0.152 mol/L。雪梨汁加工的最优护色剂组合为:6.56 mmol/L的L-半胱氨酸、4.58 mmol/L的D-异抗坏血酸钠和6.18 mmol/L的抗坏血酸,在此条件下对雪梨汁褐变的抑制率可达90.82%。     

香菜叶多酚氧化酶的分离纯化与部分酶学性质研究

新鲜香菜叶经匀浆、缓冲液提取、硫酸铵分级沉淀、DEAE-Sepharose离子交换层析、Superdex-200凝胶过滤层析,获得电泳纯的多酚氧化酶。该酶比活力达到5 622.95 U/mg,酶活回收率为3.90%,纯化倍数为126.08;全酶分子质量为111.10 kD,亚基分子质量为55.60 kD;最适温度为37 ℃,最适pH值为6.5;在25～45 ℃及pH 6.0～7.0范围内有较好的稳定性;在最适条件下测得其Km值为4.04×10－2 mol/L;甲醇、乙醇、异丙醇、氯仿及柠檬酸、抗坏血酸、Ca2＋、Hg2＋、Ba2＋对其有抑制作用,Co2＋、Pb2＋对其具有一定的激活作用。     

丝瓜多酚氧化酶的分离纯化及酶学性质

从丝瓜中分离纯化出多酚氧化酶（polyphenol oxidase,PPO）,并对其部分酶学性质进行研究。采用硫酸铵分级盐析、透析、DEAE-Cellulose DE-52离子交换层析和Sephadex G-75分子筛凝胶过滤层析分离纯化丝瓜PPO。纯化所得酶的比活力为957.9 U/mg,纯化倍数为28.3,酶活力回收率为18.5%;十二烷基磺酸钠聚丙烯酰胺凝胶电泳（sodium dodecyl sulfate polyacrylamide gel electrophoresis,SDS-PAGE）及非变性聚丙烯酰胺凝胶电泳（Native-PAGE）检测结果显示该酶蛋白呈单一条带,为单亚基蛋白,其分子质量约为67.8 kD,且无同工酶;该酶最适pH6.0,最适温度30 ℃,以左旋多巴（L-dopa）为底物,其米氏常数（Km）为1.32 mmol/L,最大反应速率（Vmax）为0.22 OD475 nm/min。     

薯蓣多酚氧化酶特性及褐变控制

从薯蓣中提取多酚氧化酶，采用分光光度法对其动力学特性进行研究。结果表明，该酶最适作用pn为5．8，最适作用温度为20℃，Km=1．82mol/L，Vmax=1．11(△OD/min)。本文同时考察了抑制剂对薯蓣多酚氧化酶活性的影响。     

芋头多酚氧化酶的分离纯化与酶学特性

以鲜芋头为原料,通过提取、30％～80％饱和度硫酸铵沉淀、透析、超滤,以及柱层析等步骤对芋头多酚氧化酶(polyphenol oxidase,PPO)进行逐级分离纯化,并探讨其酶学特性及抑制剂对其作用规律.结果 表明:经DEAE-Sepharose、Superdex G-75色谱柱纯化后,得到了电泳纯PPO,比活力为2...     

紫薯中多酚氧化酶活性的研究及褐变控制

研究紫薯组织中多酚氧化酶(PPO)的活性,最适pH值、最适温度等特性,并在此基础上,研究加热温度、加热时间、抗坏血酸和柠檬酸添加量对PPO褐变的抑制效果。结果表明：紫薯PPO最适pH值范围为5.5～7.5,最适温度为30℃。在较好的分离淀粉的前提下,最佳抑制褐变条件是：加热温度80℃、加热时间10min、抗坏血酸添加量0.9‰、柠檬酸添加量1.2g/L。     

甜柿多酚氧化酶特性的研究及褐变控制

本文研究了从罗田甜柿中提取的多酚氧化酶的酶学特性。以邻苯二酚为底物时它的最适温度为20℃，最适pH值为6．6，Km=9．617mmol／L Vmax=0．0495A420／min。四种抑制剂的效果由强到弱依次是亚硫酸氢钠、L-半胱氨酸、抗坏血酸、柠檬酸。在温度为90℃时加热3min，酶活性被完全抑制。     

Purification and Characterization of Polyphenol Oxidase from Akko XIII Loquat (Eriobotrya japonica cv Akko XIII)

Akko XIII is an important loquat variety grown in Turkey. As with many fruits and vegetables, enzymatic browning catalyzed by polyphenol oxidase (PPO) also occurs in loquats. PPO from Akko XIII loquat was extracted and purified through (NH₄)₂SO₄ precipitation, dialysis and ion exchange chromatography. The enzyme showed several peaks with PPO activity on DEAE-Toyopearl 650 M column, of which only two (isoenzyme A and isoenzyme B) were characterized. Assay of activity of the isoenzymes between pH 3.04 and 7.80 using catechol as substrate showed two activity peaks, one at acidic pH and the other at neutral pH. pH optima of isoenzyme A and B were found to be at 7.4 and 4.98, respectively. The Km values of isoenzyme A and B using catechol as substrate were found to be 152.3 mM and 5.4 mM, respectively. They both displayed maximal activity at 30^oC. The two isoenzymes displayed different heat resistance and sensitivity towards various inhibitors.     

Polyphenol Oxidase from Apple (Malus domestica Borkh. cv Bramley's Seedling): Purification Strategies and Characterization

ABSTRACT Polyphenol oxidase (PPO) was isolated from Bramley's Seedling apples with 75.7‐fold purification and 26.5% recovery by ammonium sulfate precipitation, phenyl sepharose chromatography, ion exchange chromatography, and hydroxyapatite chromatography. Molecular weight was estimated to be about 45 kDa by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS PAGE). Optimum PPO activity was at pH 6.5 and greater than 50% activity was retained during storage for 72 h at pH 5.5 to 6.5. Optimum temperature for activity was 30 °C and the enzyme had residual activity of greater than 50% during storage for 72 h at 20 °C to 30 °C and for 24 h at 40 °C to 50 °C. Of the substrates tested, activity was greatest with 4‐methylcatechol followed by catechol, pyrogallol, and (?)epicatechin. The most effective inhibitors tested were sodium metabisulfite and ascorbic acid.     

The Influence of Selected Osmotic Dehydration and Pretreatment Parameters on Dry Matter and Polyphenol Content in Highbush Blueberry (Vaccinium corymbosum L.) Fruits

The paper presents an assessment of the influence of selected highbush blueberry pretreatment methods and parameters on the process of osmotic dehydration conducted in 65 °Brix sucrose solution for 5 to 240 min at 30–70 °C. The pretreatment methods used included: fruit immersion in boiling water (15 s) and in 0.5 % NaOH solution (15 s at 95 °C), exposure to ultrasound at atmospheric pressure (vibration frequency of 35?±?5 kHz, 500 W, for 15 min.) and at low pressure (0.92 kg?cm^?1), and enzymatic processing; pectinase (enzyme activity of 46,000 PGU/mL; 0.6 mL/90 g of fruits; 30 min at approx. 22 °C) and lipase (enzyme activity of 750 PGU/mL; 0.7 mL/90 g of fruits; 30 min at approx. 22 °C) were used. Dehydration was also conducted in the presence of pectinolytic enzymes. The dehydrated material was analyzed in terms of the content of dry matter, total polyphenols, and particular polyphenols using high performance liquid chromatography. It was observed that dehydration was much more intensive at 60 and 70 °C, but such temperatures led to substantial losses of phenolic compounds (by 15–30 % after 2-h dehydration) and unfavorable changes in the texture of the final product. A promising method of pretreatment is fruit immersion in solutions containing pectinolytic and lipolytic enzymes, which increase dry matter content by 26 % (after 1 h of dehydration at 30 °C) with a low loss of phenolic compounds (4 %). Among the identified anthocyanins, the greatest retention during dehydration at various temperatures was displayed by petunidin-3-galactoside (over 80 % after 1 h of dehydration) and petunidin-3-glucoside (over 78 %).     

桑叶多酚氧化酶分离及活性研究

采用分光光度法和扫描电子显微镜对桑叶多酚氧化酶(PPO)的结构、活性以及不同抑制剂的抑制效果进行研究。结果表明:桑叶多酚氧化酶为纤维状蛋白;其最适pH值为7.0,最适温度为40℃;在相同抑制剂浓度下,总体抑制效果为:抗坏血酸>亚硫酸氢钠>柠檬酸>氯化镁>氯化钠。