Effect of rice bran protein extract on enzymatic browning inhibition in potato puree

Effect of rice bran protein extract (RBPE) on enzymatic browning inhibition in potato puree was studied by colour measurement and polyphenol oxidase (PPO) inhibition. RBPE inhibited browning in potato puree and showed a higher per cent potato PPO inhibition at pH 4.0 and 5.0 than those at pH 3.0, 6.0 and 7.0 (P ≤ 0.05). RBPE heated to 40 and 60 °C had browning inhibition in potato puree and per cent potato PPO inhibition at a similar extent to unheated RBPE (P > 0.05). Browning inhibition and per cent potato PPO inhibition of RBPE were decreased when it was heated to 80 °C (P ≤ 0.05). RBPE inhibited browning in potato puree higher than 5, 10 and 20 mm ascorbic acid and 5 and 10 mm citric acid (P ≤ 0.05). Regarding the kinetics study, RBPE exhibited a mixed‐type inhibition for potato PPO. Therefore, RBPE has a potential to be used as a natural antibrowning agent in the potato industry.     

美拉德反应产物抑制冷冻甘薯片的褐变

氨基酸与葡萄糖形成的美拉德反应产物（MRPs）对甘薯中的多酚氧化酶（PPO）以及冷冻甘薯片的褐变均具有一定的抑制作用.氨基酸与葡萄糖在不同的反应条件下（氨基酸的种类、pH值、反应时间以及氨基酸与葡萄糖之比）所形成的美拉德反应产物对甘薯中多酚氧化酶的抑制作用不同.其最佳的美拉德反应条件是反应pH值为3.0,L-半胱氨酸,氨基酸与葡萄糖的质量比为1：2,反应时间为5 h.     

双孢蘑菇酶促褐变特性及褐变的控制

为有效控制双孢蘑菇褐变,研究pH值、温度、底物浓度以及抑制剂对双孢蘑菇的多酚氧化酶(polyphenol oxidase,PPO)活性的影响。结果表明：以邻苯二酚为底物,双孢蘑菇PPO最适反应pH6.8,最适反应温度20℃,最适反应底物浓度为0.06mol/L,米氏常数Km为0.1072mol/L。采用计算机图像处理技术对双孢蘑菇褐变程度进行定量检测,得出对双孢蘑菇贮藏保鲜效果较好的褐变抑制剂及其最佳配比为0.30mmol/L半胱氨酸、0.06％抗坏血酸、0.05%乙酸。     

芒果多酚氧化酶的特性及抑制研究

以邻苯二酚为底物,采用分光光度法在420nm处测定芒果多酚氧化酶(PPO)的活性,研究了温度、pH值、底物浓度对其活性的影响,并建立了酶促褐变反应动力学方程,探讨了L-半胱氨酸、抗坏血酸、亚硫酸氢钠、柠檬酸和醋酸五种抑制剂对芒果酶促褐变的抑制效果。结果表明,芒果多酚氧化酶的最适温度为50℃,最适pH值为6.86;酶促褐变反应动力学符合米氏方程所描述的单底物酶促反应动力学,以邻苯二酚为底物的最大反应速度Vmax=192(U/min·g),Km=0.0173mol/L,相应的动力学方程为v=192[S]/(0.0173+[S]);五种物质对该酶均表现出不同的抑制作用,抑制效果为:抗坏血酸>L-半胱氨酸>亚硫酸氢钠>柠檬酸>醋酸。     

酶促诱发儿茶素对模拟苹果汁非酶褐变特性的影响

以苹果中含量较多的儿茶素为研究对象,以磷酸缓冲液为苹果汁褐变模拟体系,在多酚氧化酶（polyphenol oxidase,PPO）酶促诱发儿茶素后,钝化PPO,通过色差仪连续测定模拟体系亮度（L*）值、红度（a*）值、黄度（b*）值及总色差（ΔE*）值的变化趋势,研究酶促诱发条件对模拟果汁体系非酶褐变特性的影响。结果表明：酶促诱发时间及诱发体系温度对儿茶素参与的非酶褐变无显著性影响（P＞0.05）,而酶促诱发体系pH值以及儿茶素浓度对模拟体系褐变程度影响显著（P＜0.05）,pH 4.0时,模拟果汁体系褐变程度较大,褐变程度随儿茶素浓度的增大而增强;本实验同时发现,PPO诱发儿茶素发生非酶聚合,并与聚合产物相互作用而沉淀,导致果汁产生后浑浊。     

Effect of nitric oxide on pericarp browning of harvested longan fruit in relation to phenolic metabolism

The effects of nitric oxide (NO) on enzymatic browning of harvested longan fruit in relation to phenolic metabolisms were investigated. Fruits were dipped for 5 min in 1 mM sodium nitroprusside (SNP), a nitric oxide donor, then packed in 0.03 mm thick polyethylene bags, and finally stored for 6 days at 28 °C. Changes in pericarp browning and pulp breakdown were evaluated, while total phenol content, activities of phenolic-associated enzymes, polyphenol oxidase (PPO), peroxidase (POD) and phenylalanine ammonia lyase (PAL), and concentrations of total soluble solids, titratable acidity and ascorbic acid were measured. SNP treatment delayed pericarp browning, inhibited activities of PPO, POD and PAL and maintained a high total phenol content of longan fruit during storage. Furthermore, NO showed a significant inhibition of the in vitro activities of PPO and POD, indicating that the beneficial effect of NO was direct. Moreover, application of NO resulted in a lower pulp breakdown and maintained relatively high levels of total soluble solids and ascorbic acid.     

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%。     

The Effect of Antibrowning Agents on Inhibition of Potato Browning,Volatile Organic Compound Profile,and Microbial Inhibition

Abstract: Burbank and Norkotah potato slices were dipped into 3% sodium acid sulfate (SAS), citric acid (CA), sodium erythorbate (SE), malic acid (MA), sodium acid pyrophosphate (SAPP), or a combination of SAS‐CA‐SE. Browning by polyphenol oxidase (PPO) obtained from potato extract with 0.04 to 0.016 g/mL of antibrowning solutions at pH 2.0 to 6.9 were measured by UV‐Vis spectroscopy. The color of slices dipped in antibrowning solutions at pHs 2 to 7 and stored at 4 °C for 15 d was measured every 5 d by colorimeter. Headspace analysis of volatiles in raw and cooked potato samples was performed by selected ion flow tube mass spectrometer (SIFT‐MS) and soft independent modelling by class analogy (SIMCA) analysis of the calculated odor activity values (OAV) determined interclass distances. Microbial growth was measured at 15 d. At unadjusted pHs (1.1 to 7.1), the PPO browning of the control and samples with SAPP was not significantly different, SAS, CA, and MA produced some inhibition and SE and SAS‐CA‐SE prevented browning. At pH 5 to 7, only SE and SAS‐CA‐SE were effective browning inhibitors. Based on the color of potato slices, SE was the most effective at pH 2 to 7, but SAS was most effective at unadjusted pH. Cooking increased volatile levels in the treated potatoes and decreased differences between volatile profiles. Differences between cooked samples may not be noticeable by the consumer because volatiles with high discriminating powers have low OAVs. SAS, CA, and SAS‐CA‐SE treatments inhibited microbial growth but SAPP, control, and SE did not, most likely due to pH. Practical Application: Antibrowning agents inhibit polyphenol oxidase, increasing shelf life and consumer acceptability of processed raw potato products by preserving the color. Their effectiveness was shown to be mainly due to a pH effect, except SE, which was not pH dependent. MA, CA, and SAS‐CA‐SE are better acidulants for inhibition of color change as well as growth of spoilage bacteria, yeast, and mold than SAPP, the industry standard.     

Role of endogenous and exogenous phenolics in litchi anthocyanin degradation caused by polyphenol oxidase

The degradation of anthocyanins and/or the oxidation of phenolics caused by polyphenol oxidase (PPO) results in an enzymatic browning reaction of fruits and vegetables. This work was conducted with a view to explaining the unexpected observation that litchi (Litchi chinensis Sonn.) PPO did not directly oxidise litchi anthocyanins. PPO and anthocyanin from litchi fruit pericarp were extracted and purified, respectively, and then the anthocyanin degradation by PPO in the presence of (−)-epicatechin (endogenous PPO substrate), and catechol and gallic acid (exogenous PPO substrates) were analysed comparatively. The results showed that catechol was the most effective in litchi anthocyanin degradation, followed by (−)-epicatechin and gallic acid, but no significant differences existed between catechol and (−)-epicatechin. The study suggested that litchi PPO directly oxidised (−)-epicatechin; then oxidative products of (−)-epicatechin in turn catalysed litchi anthocyanin degradation, and finally resulted in the browning reaction, which can account for pericarp browning of postharvest litchi fruit.     

抗坏血酸与壳聚糖处理对鲜切香芋贮藏效果的研究

以湖南香芋头为试材,研究不同浓度抗坏血酸与壳聚糖处理对鲜切香芋的贮藏效果及生理生化变化的影响。结果表明,在15℃贮藏温度下,与对照相比,0、1.0、5.0、10.0g/L 抗坏血酸(VC)分别与15.0g/L 壳聚糖配合处理较好地保持了鲜切香芋的外观和营养,减少鲜切香芋的失重率和褐变度,并抑制多酚氧化酶(PPO)、过氧化物酶(POD)和淀粉酶的活性。其中,5.0g/L 抗坏血酸与壳聚糖配合处理的保鲜效果最明显。     

Purification and enzymatic characteristics of a novel polyphenol oxidase from lotus seed (Nelumbo nucifera Gaertn.)

A polyphenol oxidase (PPO) from lotus seed was purified by the procedures including ammonium sulphate precipitation and affinity chromatography. The apparent molecular mass was 38.6 kDa by SDS‐PAGE. Kinetic studies showed that the K_m and V_max values for catechol were 6.04 mm and 416.67 U, respectively. The PPO performed optimal activity in 20 °C and pH 7.0. The enzymatic activity could be mainly maintained up to 50 °C and pH 4.0–7.0. The activity could be inhibited by various inhibitors including thiourea, urea, sodium hydrogen sulphite, EDTA·2Na, SDS, citric acid, guanidine hydrochloride, ascorbic acid, sodium sulphite and sodium thiosulphate. The metal ions Ba²⁺, Mg²⁺, Ca²⁺, Mn²⁺, Co²⁺ and Zn²⁺ could inhibit the activity of PPO, while Cu²⁺ performed obvious enhancement. The enzymatic properties of PPO could probably provide practical application in inhibiting the PPO activity and preventing enzymatic browning in the process of picking, transportation, processing and storage of fresh lotus seeds.     

Polyphenol oxidase activity,phenolic acid composition and browning in cashew apple (Anacardium occidentale, L.) after processing

This study describes the extraction and characterisation of cashew apple polyphenol oxidase (PPO) and the effect of wounding on cashew apple phenolic acid composition, PPO activity and fruit browning. Purification factor was 59 at 95% (NH₄)₂SO₄ saturation. For PPO activity, the optimal substrate was catechol and the optimum pH was 6.5. PPO K_m and V_max values were 18.8 mM and 13.6 U min⁻¹ ml⁻¹, respectively. Ascorbic acid, citric acid, sodium sulphite and sodium metabisulphite decreased PPO activity, while sodium chloride increased PPO activity. Wounding at 2 °C and 27 °C for 24 h increased PPO activity but storage at 40 °C reduced PPO activity. Gallic acid, protocatechuic acid and cinnamic acid (free and conjugate) were identified in cashew apple juice. Cutting and subsequent storage at 40 °C hydrolysed cinnamic acid. 5-Hydroxymethylfurfural content in cashew apple juice increased after injury and storage at higher temperatures, indicating non-enzymatic browning.     

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.     

Purification and characterisation of polyphenol oxidase from leaves of Cleome gynandra L.

A polyphenol oxidase was purified and characterised from leaves of the common spiderflower. Purification sequentially with ammonium sulphate, dialysis, DEAE-Sepharose ion-exchange chromatography and Sephadex G-75 gel filtration chromatography resulted in 37.8-fold enrichment in the specific activity and 44.3% recovery of the total activity. Purified PPO is a monomeric protein of 52.6 kDa revealed by Coomassie and active staining and Western blot. It was optimally active at pH 8.0 and 60 °C, and stable from pH 3.0 to 9.0 and below 60 °C. It displayed enzymatic activity towards monophenols, diphenols and triphenols, especially towards diphenols, and substrate specificity towards methylated and methoxylated substrates. Its activity was slightly increased by 0.1% SDS, heavily inhibited by Hg²⁺ and Pb²⁺, and completely inhibited by 1.0 mM of ascorbic acid, l-cysteine, β-mercaptoethanol, sodium diethyldithiocarbamate and thiourea, and by 10 mM of dithioerythritol, sodium metabisulphite and sodium sulphite.     

Inhibition of enzymatic browning in actual food systems by the Maillard reaction products

BACKGROUND: The Maillard reaction occurring between amino acids and sugars produces neo‐formed compounds having certain levels of antioxidant activity depending on the reaction conditions and the type of reactants. The objective of this study was to investigate enzymatic browning inhibition capacity of Maillard reaction products (MRPs) formed from different amino acids including arginine (Arg), histidine (His), lysine (Lys) and proline (Pro). RESULTS: The inhibitory effects of the MRPs on polyphenol oxidase (PPO) were determined. The total antioxidant capacity (TAC) of MRPs derived from different amino acids were in the order Arg > His > Lys > Pro. The TAC and PPO inhibition of MRPs were evaluated as a function of temperature (80–120 °C), time (1–6 h) and pH (2–12). Arg‐Glc and His‐Glc MRPs exhibited strong TAC and PPO inhibition. Increasing temperature (up to 100 °C) and time also increased TAC and PPO inhibition. Kinetics analysis indicated a mixed type inhibition of PPO by MRPs. CONCLUSION: The results indicate that the MRPs derived from Arg and His under certain reaction conditions significantly prevent enzymatic browning in actual food systems. The intermediate compounds capable of preventing enzymatic browning are reductones and dehydroreductones, as confirmed by liquid chromatographic–mass spectrometric analyses. Copyright © 2010 Society of Chemical Industry     

果蔬酶促褐变机理的研究进展

酶促褐变对果蔬在加工保鲜过程中的品质有重大影响，有关酶促褐变机理的相关研究也越来越多。本文整理归纳了国内外围绕褐变机理进行的多酚氧化酶和酚类底物的相关研究，包括对多酚氧化酶种类、催化位点、提取、分离、纯化、活性测定方法的介绍；酚类化合物种类、含量变化及其变化原因的阐述；酚类酶促氧化反应、醌的生成途径、次级代谢过程、氧化终产物特性以及抗褐变剂抑制酶促褐变机理的报道，以期为酶促褐变机理的系统深入研究提供借鉴与参考。     

荔枝与荔枝酒褐变控制

以荔枝及荔枝酒为试料,研究荔枝多酚氧化酶(PPO)的酶学特性及荔枝酒发酵过程中的褐变机理及控制措施。结果表明：荔枝PPO存在同功酶,荔枝PPO最适底物为没食子酸;以没食子酸为底物,荔枝PPO最适反应pH值为8.0,最适反应温度50℃,荔枝PPO的动力学参数为：米氏参数(Km)为26.033mmol/L,最大反应速度(Vmax)为34.816g/(L·min);荔枝PPO对酸碱、对热稳定性较差;荔枝酒发酵过程中褐变的主要原因是其PPO引起的酶促褐变。因此,低温贮藏和高温热处理可有效抑制荔枝及荔枝酒在贮藏和加工中酶促褐变反应。     

Suppression of internal browning and maintenance of antioxidants in beeswax plus salicylic acid coated pear fruit during different storage conditions

Postharvest application of edible waxes blended with antioxidants is an effective strategy to regulate postharvest quality and antioxidant losses in fruits. Effect of beeswax coatings (BW 1.0% and 2.0%) alone or enriched with salicylic acid (SA) on antioxidant properties of pear fruit was studied during 67 days of cold storage and 20 days of supermarket storage conditions respectively. Results showed that under both the storage conditions fruits coated with BW + SA maintained higher total phenolic content (TPC), antioxidant activity and ascorbic acid content (AsA) as compared to the control. Furthermore, combined coatings significantly reduced polyphenol oxidase (PPO) activities and inhibited internal browning (IB) in pears throughout cold and supermarket storage. Overall, the BW 2.0% + SA 2.0 mM exhibited the most competitive effect in terms of preserving the antioxidant properties and preventing internal browning in pears during storage.     

Effect of whey protein concentrate on phenolic profile and browning of fresh-cut lettuce (Lactuca Sativa)

The in vitro kinetics of lettuce PPO with respect to dissolved oxygen using catechin, chlorogenic acid, caffeic acid and gallic acid has been examined. In-vitro lettuce polyphenol oxidase (PPO) activity was determined by measuring the consumption of oxygen during the oxidation reaction. The effect of whey protein concentrate (WPC) was tested on the inhibition of lettuce PPO comparing with ascorbic acid (AA) and cysteine. A competitive model that considered inhibitors was the most appropriate model to explain reaction kinetics. Browning of lettuce was also monitored during storage for 24 h. Addition of WPC prevented loss of lightness in lettuce. Loss of identified phenolic compounds in lettuce was measured during the enzymatic browning process by high-performance liquid chromatography. Degradation of identified phenolic compounds followed first order kinetics during storage. Combination of WPC with cysteine was proposed for the protection of phenolics compounds against PPO-catalysed oxidation.     

In silico guided pre-treatment of sugarcane juice with natural inhibitors of polyphenol oxidase (PPO) is a new and effective strategy for development of spray-dried formulation

Sugarcane juice is a popular beverage and is also processed to produce sugar. The polyphenol oxidase (PPO) in sugarcane juice causes enzymatic browning and makes the process of sugar production complex and cumbersome. Storage of sugarcane juice is also hampered by the high sugar content and rapid microbial fermentation. The present research assessed the potential of lemon juice (LJ) and ginger extract (GE) as natural inhibitors of PPO. Enzyme kinetics and the mechanism of inhibition of LJ and GE were studied. Primary investigation was carried out using molecular docking approach to assess the inhibitory potential of LJ and GE and to determine the nature of interaction between the enzyme and inhibitors. Extracts were used as inhibitors and studies revealed that both reduced the PPO activity. Subsequently, pure bioactive inhibitors such as ascorbic acid, citric acid, and 6-shogaol present in these natural extracts were used to study the mode of inhibition of PPO. Citric acid decreased PPO activity by lowering pH, while ascorbic acid was found to be a competitive inhibitor of PPO with a K_i of 75.69 µM. The proportion of LJ and GE required in sugarcane juice was optimized on the basis of browning index and sensory acceptance. Further, the sugarcane cane juice after inhibition of PPO under optimized conditions was spray dried and evaluated for reconstitution properties. The product formulated in the present study is a new and effective approach to address quality-compromising issues associated with long-term storage of cane juice.