Polyphenol Oxidase from Bean Sprouts (Glycine max L.)

ABSTRACT: Polyphenol oxidase (PPO) was purified and characterized from bean sprouts by ammonium sulfate precipitation, DEAE‐Toyopearl 650M, CM‐Toyopearl 650M, SuperQ‐Toyopearl 650S and QAE‐Toyopearl 550C column chromatographies. Substrate staining of the crude extract on electrophoresis showed the presence of 2 isozymic forms of this enzyme. The molecular weight of the purified enzyme was estimated to be about 54 kDa. The optimum pH was 9.0 and optimum temperature 40 °C. Heat inactivation occurred about 30 °C. PPO showed activity to catechol, pyrogallol and dopamine. These compounds such as ascorbic acid, L‐cysteine, 2‐mercaptoethanol, and glutathione used was the effective inhibitor. Enzyme activity was maintained for 7 d at 4 °C but suddenly decreased after 8 d.     

富硒处理对绿豆GSH-Px活性及GSH含量的影响

谷胱甘肽过氧化物酶(GSH-Px),最早是在动物体内发现的一种含硒酶,近年来在许多植物中也发现了该酶的存在,并发现适当浓度的外源硒能提高GSH-Px的活性.本文以绿豆为实验材料,用NaSe2O3 作为硒源,对绿豆进行富硒处理,研究适当浓度的NaSe2O3对绿豆GSH-Px活性及绿豆中谷胱甘肽(GSH)含量的影响.结果显示:富硒处理可以促进绿豆GSH-Px的合成;无论是浸泡过程还是萌芽阶段,经富硒处理的绿豆其GSH-Px的活性都比对照样高;特别是在富硒处理5 h后,绿豆GSH-Px活性显著增加,比对照提高了59.38 %;而绿豆中GSH的含量则是先下降,后升高.     

发酵与发芽处理对绿豆淀粉黏度性质的影响

将绿豆进行发酵及发芽处理,研究不同处理对绿豆淀粉黏度的影响。采用快速黏度分析仪（RVA）测定了不同处理的绿豆淀粉糊化特性。结果表明自然发酵对绿豆淀粉黏度影响不大,只有峰值黏度略有下降：生物混合发酵可使样品的峰值黏度和热浆黏度均出现较大幅度的下降,最终黏度下降。绿豆发芽后提取的淀粉其黏度变化较大,其峰黏度上升1248．00mPa·s,热浆黏度上升698．00mPa·s,淀粉的黏度显著上升。     

Geometric and Mechanical Properties of Mung Bean (Vigna Radiata L.) Grain: Effect of Moisture

In this research, selected geometric and mechanical properties of mung bean grain were evaluated as a function of moisture content. Five levels of moisture content ranging from 7.28 to 17.77% d.b. (dry basis) were used. The average length, width, thickness, arithmetic and geometric mean diameters, sphericity, thousand grain mass and angle of repose ranged from 5.145 to 6.199 mm, 3.760 to 4.474 mm, 3.537 to 4.223 mm, 4.147 to 4.965 mm, 4.090 to 4.893 mm, 0.795 to 0.789, 52.3 to 64.6 g, and 25.87 to 29.38° as the moisture content increased from 7.28 to 17.77% d.b., respectively. The bulk density was found to be decreased from 821.3 to 745.2 kg/m³, whereas the grain volume, true density, porosity, terminal velocity, and projected area were found to be increased from 27.88 to 47.33 mm³, 1230.0 to 1456.7 kg/m³, 30.43 to 46.57%, 4.86 to 5.29 m/s, and 17.48 to 19.26 mm², respectively. There is a 43% increase in surface area from grain moisture content of 7.28 to 17.77% d.b. The static coefficient of friction on various surfaces increased linearly with the increase in moisture content. The rubber as a surface for sliding offered the maximum friction followed by galvanised iron, medium density fibreboard, stainless steel, aluminium and glass sheet. As moisture content increased from 7.28 to 17.77%, the rupture forces values ranged from 67.39 to 39.44 N; 63.86 to 42.18 N, and 53.96 to 41.79 N for thickness (Z axis), length (Y-axis) and width (X-axis), respectively.     

萌发绿豆中多酚类物质动态变化规律及其抗氧化活性的研究

以产自山西的明绿豆为实验材料,探究萌发前后绿豆中酚类化合物的动态变化规律及其抗氧化活性。采用传统方法进行绿豆的萌发,通过超声-微波协同萃取法提取绿豆及绿豆芽中的多酚类化合物,结合植物广靶代谢组学的方法对绿豆萌发前后多酚类化合物的提取液进行定性定量分析,明确酚类化合物的组成,分析萌发处理对绿豆多酚组成成分的影响及与其抗氧化活性的关系。结果表明:通过超高效液相色谱-质谱联用仪检测出46种多酚类化合物,筛选出44个差异代谢物,萌发前与萌发后绿豆芽的清除DPPH·能力分别为87.94%和96.24%,总抗氧化能力分别为28.56 U/mL和33.06 U/mL;最后通过KEGG查询得到筛选出的多酚类化合物参与了20个代谢通路。初步明确了绿豆中多酚类活性成分的物质基础,为绿豆的多酚类物质的合理利用与开发提供了参考。     

绿豆皮可溶性膳食纤维的抗氧化作用

通过体外、体内实验研究绿豆皮可溶性膳食纤维的抗氧化作用。结果表明：当质量浓度为4 mg/mL时, 绿豆皮可溶性膳食纤维的还原力为1.526,对1,1-二苯基-2-三硝基苯肼自由基和羟自由基的清除率分别为82.65% 和85.16%。动物实验结果显示,与正常对照组相比,D-半乳糖衰老模型组小鼠血清和肝组织中总抗氧化能力以 及过氧化氢酶、总超氧化物歧化酶、谷胱甘肽过氧化物酶活力均有所降低,丙二醛含量升高,且大部分差异显著 （P＜0.05,P＜0.01）,说明小鼠D-半乳糖衰老模型构建成功。与模型组相比,绿豆皮可溶性膳食纤维各剂量组小 鼠血清和肝组织中丙二醛含量大幅降低,高剂量绿豆皮可溶性膳食纤维可显著提高小鼠血清和肝组织总抗氧化能力 以及过氧化氢酶、总超氧化物歧化酶、谷胱甘肽过氧化物酶活力（P＜0.05）。因此,绿豆皮可溶性膳食纤维具有 良好的抗氧化作用。     

Comparative Studies on Physico-Chemical Properties of Starches from Jackfruit Seed and Mung Bean

The physico-chemical properties of starch from jackfruit seed and mung bean were investigated. Jackfruit seed starch had much higher resistant starch content (26.99%) than that of mung bean starch (4.04%). Furthermore, jackfruit seed starch had a higher gelatinization temperature (T_o) that required more gelatinization energy (ΔH) compared to mung bean starch. However, mung bean starch had higher amylose content and its granules were much larger than that of jackfruit seed starch. Mung bean starch had the highest peak viscosity, breakdown, and setback whereas jackfruit seed starch had the highest pasting temperature. Amylopectin chain length of mung bean starch contained higher proportion of short chains (degrees of polymerization 6–12) but lower proportion of very long chains (degrees of polymerization > 37 ) comparing with jackfruit seed starch. The X-ray diffraction patterns showed both starches to be Type-A crystallinity. In addition, both starch gels showed higher the storage modulus (G′) than the loss modulus (G?) designating as rubber like material. However, mung bean starch gel exhibited higher G’ and less tan δ than that of jackfruit seed starch indicating much stronger of gel structure.     

Effect of Several Germination Treatments on Phosphatases Activities and Degradation of Phytate in Faba Bean (Vicia faba L.) and Azuki Bean (Vigna angularis L.)

Abstract: Two assays were conducted to investigate the changes of faba bean (Vicia faba L.) and azuki bean (Vigna angularis L.) phosphatases (phytase [Phy] and acid phosphatase [AcPh]) and the degradation of its substrates (inositol phosphate esters) during seed germination. The 1st assay was to establish the optimal germination conditions of faba bean and azuki bean to improve the endogenous phosphatases and increase the hydrolysis of phytate and, in the second assay, to determine the different lower phosphate esters of myo‐inositol produced during the germination process. In the 1st assay, seeds were soaked for 12 and 24 h and germinated for 3 and 5 d with and without the addition of gibberellic acid (GA₃). In the second assay, seeds were soaked for 12 h and germinated for 1, 3, and 5 d with GA₃. Phy (up to 3625 and 1340 U/kg) and AcPh (up to 9456 and 2740 U/g) activities, and inositol hexaphosphate (IP6) (8.23 and 7.46 mg/g), inositol pentaphosphate (IP5) (0.55 and 0.82 mg/g), and inositol tetraphosphate (IP4) (0.26 and 0.01 mg/g) were detected in ungerminated faba bean and azuki bean, respectively. The germination process caused a significant increase of Phy and AcPh activities in faba bean (up to 147% and 210%) and azuki bean (up to 211% and 596%) and a reduction in the phytate phosphorus content (up to 81% and 63%, respectively). Phytate phosphorus content was affected only by soaking time in the case of faba bean. Finally, during the course of germination, IP6 and IP5 were rapidly degraded in faba bean (88% and 39%) and azuki bean (55% and 56%), and IP4 was only a short‐living intermediate, which was increased during hydrolysis and degraded to inositol triphosphate. In this manner we could obtain a low‐phytate, endogenous phosphatase‐rich ingredient for enhancing human nutrition.     

速冻建莲加工与贮藏过程中多酚氧化酶活性抑制的研究

为提高速冻建莲的品质,本文研究了不同的抑制剂组合对建莲多酚氧化酶(PPO)活性的抑制效果,并观察了速冻建莲贮藏期间PP0活性的变化。结果表明:0.025％EDTA-Na_2+0.1％抗坏血酸+0.1％柠檬酸能最大限度地抑制建莲的多酚氧化褐变,以此应用于速冻建莲的生产,能有效抑制冻藏过程中建莲的多酚氧化酶活性,褐变程度明显降低。     

不同发芽条件对蚕豆植酸酶活性的影响

以蚕豆为试材,研究发芽过程中植酸酶在蚕豆不同部位的变化和性质.发芽蚕豆不同部位植酸酶活性呈现先高后低的变化趋势,子叶植酸酶在发芽第6天达最高值.超声波处理对发芽蚕豆胚根、子叶和胚芽植酸酶活性有一定促进作用,其中以15min超声波处理对胚根和胚芽植酸酶活性影响最大,5min处理、10min处理和15min处理均对子叶植酸酶活性有显著影响.在发芽过程中不同光照对蚕豆进行处理,不论胚根、子叶还是胚芽,黑暗处理植酸酶活性高于光照处理.核酸抑制剂放线菌素(Act-D)和蛋白质抑制剂环已酰胺(CHM)对发芽蚕豆胚根、子叶和胚芽中的植酸酶活性有一定影响,其中以CHM的抑制效果最为明显.     

不同发芽条件对蚕豆植酸酶活性变化和性质的影响

以蚕豆为试材,研究了发芽过程中植酸酶在蚕豆不同部位的变化和性质。发芽蚕豆不同部位植酸酶活性呈现先高后低的变化趋势,子叶植酸酶在发芽第6天达到最高值。超声波处理对发芽蚕豆胚根和子叶植酸酶活性有一定促进作用,其中以15min超声波处理对胚根植酸酶活性影响最大,5min处理、10min处理和15min处理均对子叶植酸酶活性有显著影响。在发芽过程中不同光照对蚕豆进行处理,不论胚根、子叶还是胚芽,黑暗处理植酸酶活性高于光照处理。核酸抑制剂放线菌素(Act-D)和蛋白质抑制剂环已酰胺(CHM)对发芽蚕豆胚根、子叶和胚芽中的植酸酶活性有一定影响,其中以CHM的抑制效果最为明显。     

A Rapid High-Performance Liquid Chromatography Photodiode Array Detection Method to Determine Phenolic Compounds in Mung Bean (Vigna radiata L.)

A fast method was developed for simultaneous detection and quantification of 12 phenolic compounds in mung bean, using reversed phase high-performance liquid chromatography. The method was optimized for mobile phase combination, elution gradient, detection wavelength, and solvent extraction. All the phenolic compounds (gallic acid, neochlorogenic acid, catechin, chlorogenic acid, caffeic acid, p-coumaric acid, t-ferulic acid, vitexin, isovitexin, myricetin, quercetin, and kaempferol) were eluted for 18 min and recovered within a limit as per International Council for Harmonization guidelines. The method showed good linearity with correlation coefficient of 0.998. The limit of detection and quantification of all the compounds ranges from 0.27 ± 0.01 to 3.65 ± 0.3µg/mL and 0.91 ± 0.1 to 12.17 ± 0.9µg/mL, respectively. Vitexin (28.10 ± 0.20 to 29.60 ± 0.6 mg/100 g raw material) and isovitexin (34.09 ± 0.14 to 36.83 ± 0.82 mg/100 g raw material) were the major phenolic compounds along with other phenolic compounds found in mung beans.     

发芽处理对蚕豆主要营养成分与抗营养因子的影响

研究发芽处理对启豆2号蚕豆主要营养成分与抗营养因子含量的影响。对蚕豆发芽前后蛋白质、脂肪、淀粉、VC、氨基酸、植酸和单宁含量测定。结果表明：蚕豆在发芽1d时可溶性蛋白质含量显著增加,随后逐渐下降;粗脂肪和淀粉含量均呈下降趋势;发芽2d时VC含量显著增加,达到最大58.65mg/100g,随后逐渐下降,5d后VC含量仍高于未发芽的;发芽后1～2d蚕豆的氨基酸含量略有增加;发芽1d抗营养因子植酸含量下降了33.55%,随后平缓;发芽1d蚕豆的单宁含量增加到462.70mg/100g,2～3d增加比较平稳,5d急剧增加至1306.10mg/100g。可推知,将启豆2号蚕豆发芽1～2d的作为食用及加工的较佳时期。     

超声波对多酚氧化酶酶活力的影响及其机理

以多酚氧化酶(PPO)为对象,研究不同超声波条件对PPO 酶活力的影响及其机理。结果表明：PPO 酶活力随着超声时间增加而降低,随着超声波功率增大先略增大然后逐渐降低;在超声波作用下PPO 的稳定温度和pH值分别为40℃和6.8;经超声波处理后的PPO,其动力学参数米氏常数Km 增大、最大反应速率Vmax 减小;PPO 二级结构中β- 转角相对减少。     

Fermentation and enzyme treatment of tannin sorghum gruels: effects on phenolic compounds,phytate and in vitro accessible iron

The presence of polyphenols and phytate in cereal products has been shown to interfere with the bioavailability of minerals such as iron. In the present study, we added enzymes (wheat phytase and mushroom polyphenol oxidase) during fermentation of tannin sorghum gruels prepared from flour with or without addition of 5% flour of germinated tannin-free sorghum grains (power flour), and investigated the effects on phenolic compounds, phytate and in vitro accessible iron. Assayable phenolic compounds were significantly reduced by fermentation, with high reductions observed in gruels with added enzymes. Fermentation of the gruels with addition of enzymes reduced (on average) total phenols by 57%, catechols by 59%, galloyls by 70% and resorcinols by 73%. The phytate content was significantly reduced by fermentation (39%), with an even greater effect after addition of power flour (72%). The largest reduction of phytate (88%) was, however, obtained after addition of phytase. The in vitro accessible iron was 1.0% in the sorghum flour and it increased after fermentation with power flour and/or with enzymes. The highest in vitro accessibility of iron (3.1%) was obtained when sorghum was fermented with addition of power flour and incubated with phytase and polyphenol oxidase after the fermentation process.     

浸泡处理对红芸豆的物理性质及蒸煮品质的影响

以红芸豆为原料,探讨不同浸泡温度(25、45、65℃)对芸豆物理性质及蒸煮品质的影响。结果表明:芸豆在浸泡过程中,随着浸泡温度升高与时间延长,其吸水率、体积膨胀率、固形物溶出率等均呈增加的趋势,平衡吸水率常数则呈下降趋势;浸泡温度的提高有利于达到浸泡平衡及缩短浸泡时间。在加工性能方面,45℃下浸泡可显著缩短煮豆时间并提高煮熟红芸豆的品质。     

菜豆冷害生理及其防治措施的研究

研究了热处理与间歇升温对菜豆冷害及其生理生化的影响,结果表明：间歇升温与热处理延缓了由于冷害引起的菜豆呼吸强度的上升、电导的增加和过氧化物酶活性的增强;降低了菜豆的表面水浸凹陷与锈斑的发生。两处理相结合具有协同增效作用,既能有效抑制表面水浸凹陷,又可阻止锈斑的发生,是防止菜豆冷害发生的有效措施。     

The purification and characterisation of polyphenol oxidase from green bean (Phaseolus vulgaris L.)

Four isoforms of polyphenol oxidases (PPOs) were characterised in purified extracts of coats (PPOIa and PPOIb) and pods (PPOIIa and PPOIIb) of green bean (Phaseolus vulgaris L.). The molecular weights of four isoforms have been estimated to be from 57.5 to 39.0 kDa by SDS–PAGE. The PPOII (mixture of PPOIIa and PPOIIb) was used to characterise the PPO of green bean pods. All isoforms activities were stable between pH 6.8 and pH 7.2. PPOIa and PPOII have similar thermal inactivation profiles, and PPOIb has higher thermal stability than that of PPOIa and PPOII. PPOs showed the highest affinity to pyrogallol in all selected substrates. Although activities of PPOs were markedly inhibited by l-ascorbic acid, the activity of PPOI (mixture of PPOIa and PPOIb) was significantly activated by MnSO₄ and CaCl₂.     

发芽对绿豆皮膳食纤维结构及性质的影响

为研究发芽对绿豆皮膳食纤维结构及功能性质的影响,采用X射线衍射分析、红外光谱分析和电子显微镜扫描等方法测定其结构,并对其持水力、持油力、膨胀力、阳离子交换能力、吸附葡萄糖能力、吸附胆固醇能力和吸附NO2 － 能力等功能性质进行对比研究。结果表明：发芽处理后绿豆皮中总膳食纤维含量增加3.40%,可溶性膳食纤维增加13.62%。发芽绿豆皮膳食纤维的持水力、持油力、膨胀力明显提高,分别达到（6.97±0.32）、（4.93±0.10） g/g、（4.79±0.11） mL/g,阳离子交换能力略有降低,为（0.47±0.02） mmol/g,吸附葡萄糖能力增加,为（8.37±0.18） mmol/g,吸附胆固醇能力增加,为（2.23±0.11） mg/（mL·g）,吸附NO2 － 能力有所降低,为（3.92±0.09） mg/g。扫描电子显微镜结果表明,发芽可使绿豆皮膳食纤维表面出现更多孔隙和褶皱,有利于膳食纤维吸附能力的提高;X射线衍射结果表明,发芽没有改变绿豆皮膳食纤维的结晶度,较好地保留了膳食纤维的结晶区和非结晶区;傅里叶红外光谱分析表明,发芽没有破坏绿豆皮膳食纤维的官能团结构。绿豆经发芽处理后改善了绿豆皮膳食纤维的大部分功能性质,较好地保留了其结构,有利于绿豆副产物的开发利用。