苦荞中芦丁和槲皮素对淀粉消化酶的抑制能力

采用多种光谱技术手段研究苦荞中芦丁和槲皮素对淀粉消化酶的抑制效果和作用机制,及二者联合使用时的抑制效果。结果表明：芦丁和槲皮素对α-淀粉酶的抑制类型均为疏水相互作用力和以氢键为主要驱动力的竞争性抑制,半抑制浓度分别为0.36 mg/mL和0.22 mg/mL;对α-葡萄糖苷酶均为通过氢键结合的混合型抑制,半抑制浓度分别为1.30 mg/mL和0.362 mg/mL。同时,二者均能与α-淀粉酶和α-葡萄糖苷酶结合形成复合物从而抑制酶的活性,且只存在一个（或一类）作用位点;二者按照不同浓度比联合使用均可对α-淀粉酶和α-葡萄糖苷酶产生协同抑制的作用,当二者浓度比为7∶1和3.6∶18时,对α-淀粉酶和α-葡萄糖苷酶的联合指数分别达到0.20和0.22,苦荞中黄酮对两种消化酶达到最佳协同抑制效果。本研究为进一步阐明黄酮与淀粉消化酶相互作用机制提供一定理论基础,对指导富含黄酮食药同源植物的加工利用具有较高的指导价值,有利于促进苦荞产业的良性循环。     

芹菜素-8-C-葡萄糖苷对淀粉消化酶抑制作用机制

采用酶活动力学、荧光光谱、圆二色谱和分子对接等技术系统探究芹菜素-8-C-葡萄糖苷对α-淀粉酶和α-葡萄糖苷酶活性调控效果及机制。结果显示,芹菜素-8-C-葡萄糖苷对α-葡萄糖苷酶有良好的抑制效果,IC50值为293.5 mg/L,抑制类型为非竞争性抑制。但对α-淀粉酶无显著抑制效果。荧光光谱结果表明芹菜素-8-C-葡萄糖苷可作为猝灭剂分子与α-葡萄糖苷酶结合,发生静态猝灭,改变酶蛋白氨基酸疏水环境。圆二色谱则显示芹菜素-8-C-葡萄糖苷和α-葡萄糖苷酶之间的相互作用使酶分子的二级结构变得松散,α-螺旋和β-转角下降。分子对接结果进一步证实芹菜素-8-C-葡萄糖苷和α-葡萄糖苷酶之间作用力主要为氢键,最低结合能为-7.2 kcal/mol。本研究揭示了芹菜素-8-C-葡萄糖苷对淀粉消化酶尤其是α-葡萄糖苷酶的抑制作用机制,为未来将芹菜素-8-C-葡萄糖苷作为健康食品辅料或药物开发提供一定理论基础。     

几种原花青素的降血糖作用及与常见食品原料的结合研究

以4种不同来源的原花青素为研究对象,利用高通量淀粉浊度法测定其对α-淀粉酶和α-葡萄糖苷酶活力的抑制效果,研究其与常见食物原料之间的相互作用,为进一步开发降血糖主食提供理论依据。结果表明:葡萄籽与高粱麸皮原花青素对α-淀粉酶和α-葡萄糖苷酶活力抑制效果相对较好,蔓越莓与苹果原花青素对α-淀粉酶和α-葡萄糖苷酶活力抑制效果相对较差;食物中的主要成分与原花青素结合后会影响其对酶活力的抑制效果,常见食物中糯米、散装大米、高粱米等与葡萄籽原花青素结合明显,降低其对碳水化合物消化酶活力抑制效果,但玉米淀粉、木薯粉、马铃薯粉、生粉、米粉、小西米、小黄米、玉米片、红薯粉、黑米及黑豆与葡萄籽原花青素之间几乎不结合,是适宜发挥原花青素对碳水化合物消化酶活力抑制作用的食品原料。     

4种外源植物蛋白对小麦淀粉消化性的影响

淀粉是人类膳食的主要成分,长期大量使用含快消化淀粉多的食物易诱发糖尿病、肥胖等慢性综合征,而慢消化淀粉和抗性淀粉则有利于缓解这些症状。蛋白质能与淀粉发生相互作用并减缓淀粉的消化,进而影响餐后血糖的上升。本文以燕麦蛋白、藜麦蛋白、黑豆蛋白和扁豆蛋白4种外源植物蛋白与小麦淀粉为研究对象,研究外源植物蛋白对小麦淀粉体外消化及血糖指数的影响,进一步探讨4种外源植物蛋白对α-淀粉酶和α-葡萄糖苷酶两种淀粉消化酶的抑制作用。结果表明:4种外源植物蛋白均能显著降低小麦淀粉的水解率和血糖指数,减少快消化淀粉(RDS)含量,增加慢消化淀粉(SDS)和抗性淀粉(RS)含量。其中,燕麦蛋白和藜麦蛋白对RDS含量的影响较大,黑豆蛋白对RS含量的影响较大。另外,4种外源植物蛋白中燕麦蛋白、藜麦蛋白和黑豆蛋白能降低α-淀粉酶的活性,其中黑豆蛋白对其影响最大。黑豆蛋白和扁豆蛋白对α-葡萄糖苷酶的活性具有抑制作用,且后者的抑制效果大于前者。     

玉蜀黍不同部位提取物对α-葡萄糖苷酶和α-淀粉酶抑制作用

目的:探究玉蜀黍不同部位(须、秸秆皮、秸秆芯)提取物对α-葡萄糖苷酶和α-淀粉酶活性抑制作用.方法:采用常规理化方法测定玉蜀黍不同部位中总黄酮、总皂苷、总多糖、总蛋白质提取物的含量,酶底物反应法和3,5-二硝基水杨酸比色法测定α-葡萄糖苷酶和α-淀粉酶抑制活性,考察不同pH、温度、时间对α-葡萄糖苷酶和α-淀粉酶活性影...     

糙米多酚对淀粉消化酶的抑制作用及机理

本研究将糙米多酚(brown rice polyphenols,BRP)经过提取和C₁₈固相柱纯化后,研究了BRP对α-淀粉酶和α-葡萄糖苷酶的抑制作用及其机理。液相色谱-质谱联用技术(UPLC-DAD-ESI-Q-TOF-MS)分析结果表明,BRP中大部分多酚以糖苷的形式存在,包括1种酚醛、2种酚酸酰胺、4种酚酸糖苷、6种酚酸、6种酚酸酯和8种黄酮糖苷。BRP对α-葡萄糖苷酶显示出剂量依赖型酶抑制活性,当浓度为2 mg/mL时,抑制率最高可达到39.37%,而BRP对α-淀粉酶无明显抑制作用。此外,荧光光谱和热力学研究结果表明,BRP能与α-葡萄糖苷酶发生疏水相互作用,对α-葡萄糖苷酶的内源荧光具有动态猝灭作用。以上结果表明,BRP主要通过与α-葡萄糖苷酶发生疏水相互作用抑制α-葡萄糖苷酶的活性,从而延缓碳水化合物的水解。本文为改善餐后血糖提供一定的参考。     

栀子黄对淀粉消化酶的抑制动力学及相互作用研究

淀粉酶和α-葡萄糖苷酶是淀粉消化关键酶,也是治疗Ⅱ型糖尿病的关键酶。利用酶抑制动力学和荧光光谱技术研究栀子黄对α-淀粉酶和α-葡萄糖苷酶的抑制活性及其互作机制。结果表明,栀子黄以竞争性方式抑制α-淀粉酶和α-葡萄糖苷酶,其缔合常数Kic分别为1.47 mg/mL和0.58 mg/mL。荧光光谱表明:栀子黄对α-淀粉酶和α-葡萄糖苷酶的内源荧光有较强的猝灭能力,通过Stern-Volmer方程得到栀子黄对α-淀粉酶和α-葡萄糖苷酶的猝灭是以静态猝灭为主的混合型猝灭。焓和熵的变化表明:栀子黄与葡萄糖苷酶的结合主要由范德华力和疏水相互作用驱动,结合距离分别为4.77nm和5.19 nm。同步荧光表明,栀子黄与α-淀粉酶和α-葡萄糖苷酶的结合引起酶的重排和构象变化,从而引起酪氨酸残基或/和色氨酸残基的变化。     

水果酵素体外抗氧化及抑制α-淀粉酶和α-葡萄糖苷酶活性的研究

为明确水果酵素在自然发酵过程中体外抗氧化活性和对α-淀粉酶、α-葡萄糖苷酶的抑制作用,以DPPH自由基清除能力和ABTS自由基清除能力为指标,研究酵素的抗氧化活性;以可溶性淀粉和4-硝基苯基-α-D-吡喃葡萄糖苷(PNPG)为底物测定酵素对α-淀粉酶和α-葡萄糖苷酶的抑制作用。试验结果表明,酵素在发酵结束后DPPH自由基清除率和ABTS自由基清除率分别达到87.19%和100.00%;对α-淀粉酶和α-葡萄糖苷酶的抑制率分别达到99.57%和81.67%。酵素对DPPH自由基、ABTS自由基的清除作用和对α-淀粉酶、α-葡萄糖苷酶的抑制作用明显,表明具有较好的抗氧化活性和降血糖潜力。     

黑豆皮多酚提取物体外对α-淀粉酶和α-葡萄糖 苷酶活性的影响

目的以小麦粉、大米粉、小米粉、玉米粉4种常食用谷物和4-硝基酚-α-D-吡喃葡萄糖苷(4-Nitrophenyl-α-D-glucopyranoside,p NPG)为底物,研究在不同底物下不同浓度黑豆皮多酚提取物对α-淀粉酶和α-葡萄糖苷酶活性的影响,同时探明其对酶的抑制动力学。方法采用大孔树脂纯化黑豆皮提取物,Folin-Ciocalteu比色法测定多酚含量,分光光度法测其抑制率,多重比较分析不同底物及不同浓度提取物的差异。结果提取物溶液浓度与抑制作用呈量效关系:当抑制剂浓度为0.25、0.5、1、2 mg/mL时谷物类底物间抑制率存在显著差异(P0.05),4、6 mg/mL时差异不显著;经纯化后多酚含量(78.3%)较纯化之前(39.5%)显著提高,同时其对酶抑制作用较纯化之前显著增强;动力学分析结果表明黑豆皮多酚提取物对α-淀粉酶和α-葡萄糖苷酶两种酶的抑制类型分别为竞争性抑制和非竞争性抑制类型。结论黑豆皮多酚提取物在体外对α-淀粉酶和α-葡萄糖苷酶均具有较强的活性抑制作用。     

枸杞叶黄酮对胰脂肪酶活性的抑制作用

采用酶动力学法探讨枸杞叶黄酮对胰脂肪酶活性的抑制作用。采用紫外、荧光和红外光谱研究枸杞叶黄酮与胰脂肪酶相互作用的机理。结果表明:枸杞叶黄酮对胰脂肪酶活性有抑制作用,半抑制浓度(IC50)为(0.910±0.008) mg/mL,抑制类型为可逆非竞争抑制,抑制常数为6.04 mg/mL。紫外和荧光光谱显示:枸杞叶黄酮能使胰脂肪酶芳香氨基酸暴露,微环境和酶构象改变,内源荧光猝灭,猝灭类型为动态猝灭为主的混合型猝灭。热力学参数显示:枸杞叶黄酮与胰脂肪酶的相互作用力主要为疏水相互作用。红外光谱显示:枸杞叶黄酮对酶蛋白二级结构的影响,主要表现为复合物结构变得有序化,β-折叠向α-螺旋的转化。枸杞叶黄酮通过改变胰脂肪酶结构和微环境,进而抑制胰脂肪酶活性。     

α-Glucosidase and α-amylase inhibitors from seed oil: A review of liposoluble substance to treat diabetes

One of the effective managements of diabetes mellitus, in particular, noninsulin-dependent diabetes mellitus, is to retard the absorption of glucose by inhibition of carbohydrate hydrolyzing enzymes, such as α-glucosidase and α-amylase, in the digestive organs. Currently, there is renewed interest in plant-based medicines and functional foods modulating physiological effects in the inhibition of α-glucosidase and α-amylase. Accordingly, inhibitors of α-glucosidase or α-amylase derived from various sources have also been isolated, and majority of phenolic compounds and their effects have been investigated in animals as well. As such, when the presence of α-glucosidase inhibitor in many foodstuffs was screened for, we found that vegetable seed oil also strongly inhibited α-glucosidase and α-amylase. Seed oil is an important source of liposoluble constituents with potential for inhibition of these enzymes, hence can also be used as therapeutic or functional food sources. Therefore, this review is aimed at highlighting the main liposoluble classes of α-glucosidase and α-amylase inhibitors, but it is not intended to be an exhaustive review on the subject.     

The inhibitory effects of ethanol extracts from sorghum,foxtail millet and proso millet on α-glucosidase and α-amylase activities

Cereal crops have recently experienced increased interest due to their potential health benefits. It has been suggested that the intake of whole grain foods is beneficial to the prevention and management of diabetes mellitus. In this study, we investigated the inhibitory effect of 70% EtOH extracts from different cultivars of sorghum, foxtail millet and proso millet on α-glucosidase and α-amylase. Among the six sorghum cultivars, Mongdang-susu(SS-1), Me-susu(SS-2), Susongsaengi-susu(SS-3) and Sikyung-susu(SS-4) extracts exhibited higher inhibitory activities against α-glucosidase (IC₅₀ = 1.1–1.4 μg/ml) than acarbose, reference inhibitor (IC₅₀ = 2.1 μg/ml). In addition, these extracts strongly inhibited degradation of starch by pancreatic and salivary α-amylase, whereas extracts from foxtail and proso millets exhibited no visible or detectible inhibitory effect on α-amylase or on α-glucosidase activity. These in vitro studies indicate the potential of sorghum in the development of effective anti-diabetic agents.     

1-脱氧野尻霉素对α-葡萄糖苷酶的抑制作用机制

运用紫外光谱法、荧光光谱法和圆二色光谱法,研究桑叶提取物1-脱氧野尻霉素（1-deoxynojirimycin,DNJ）对α-葡萄糖苷酶的作用。结果发现：DNJ与α-葡萄糖苷酶反应的半抑制浓度为0.297 μg/mL,作用类型为竞争型抑制;其与α-葡萄糖苷酶主要通过静电吸引力相互作用形成基态复合物,并使α-葡萄糖苷酶的内源荧光猝灭;DNJ与α-葡萄糖苷酶相互作用形成复合物的过程是熵驱动的吸热反应,静电吸引力是两者结合反应的主要驱动力。不同温度（273、298、310 K）条件下荧光猝灭常数（Ksv）分别为1.48×104、1.29×104、1.12×104 L/mol。DNJ使α-葡萄糖苷酶的构象发生变化,且使其二级结构重新排列;诱导酶活性口袋关闭,不利于底物结合到活性位点,推测这可能是DNJ抑制α-葡萄糖苷酶活性从而降低血糖水平的机理。     

桑叶提取物、茶多酚及其复配物的抗氧化和降血糖活性

为探讨桑叶提取物(MLE)、茶多酚(TP)及其复配物的抗氧化和降血糖活性,本文考察了MLE、TP及其复配物对DPPH和ABTS+自由基的清除能力和糖尿病关键酶(猪胰腺α-淀粉酶、鼠小肠蔗糖酶和麦芽糖酶)的活性抑制能力。结果表明,TP、MLE-TP复配物以及MLE的抗氧化能力依次减弱,其中MLE-TP复配物的抗氧化能力表现为拮抗作用。MLE-TP复配物对糖尿病关键酶的抑制作用均优于单独MLE和TP。复配物MLE:TP=2:1(w/w)对α-淀粉酶的联合指数CI为0.531,复配物MLE:TP=1:1(w/w)对蔗糖酶和麦芽糖酶的CI分别为0.429和0.452。MLE-TP复配物可以提高糖尿病关键酶的抑制效果,起到协同降血糖的作用。MLE和TP的协同降糖功效,为开发安全和高效的天然降血糖功能食品和相关药物提供理论依据。     

Inhibitory properties of polyphenols in Malus “Winter Red” crabapple fruit on α-glucosidase and α-amylase using improved methods

To explore the inhibitory activity of polyphenols on α-glucosidase and α-amylase, 16 polyphenols were isolated, identified, and quantified in an edible Malus “Winter Red” crabapple fruit. The limitations of two traditional methods for α-glucosidase and α-amylase activity assay in vitro were assayed. An improved method based on an HPLC assay for α-glucosidase and a colorimetric method coupled with a custom-made mini-column for α-amylase were established. Compared with positive controls, acarbose and miglito, most polyphenols, especially the four aglycones (cyanidin, quercetin, phloretin, and 3-hydroxyphloretin) showed higher inhibition rates on α-glucosidase. None of the polyphenols showed higher inhibition rates on α-amylase than acarbose, but most, especially the four aglycones, showed higher inhibition rates on α-amylase than miglito. The Malus Winter Red fruit has great potential for postprandial blood glucose management as a potential diet therapy for diabetic patients.     

Inhibition of the in vitro activities of α-amylase, α-glucosidase and pancreatic lipase by yellow field pea (Pisum sativum L.) protein hydrolysates

The aim of this work was to produce yellow field pea protein-derived peptides as inhibitors of α-amylase, α-glucosidase and pancreatic lipase activities. A pea protein concentrate was hydrolysed with alcalase, chymotrypsin, pepsin or trypsin and the hydrolysates separated into different fractions (<1, 1–3, 3–5, 5–10 kDa) by membrane ultrafiltration. Peptide sequence analysis showed that the alcalase hydrolysate had higher levels of di- and tripeptides when compared with the chymotrypsin, pepsin and trypsin hydrolysates. The peptide fractions inhibited α-amylase and α-glucosidase activities at levels that were similar to the unfractionated hydrolysates. The peptides were more active against α-amylase (inhibition at μg level) than α-glucosidase (mg level). In contrast, the fractionated peptides had reduced ability (IC₅₀ >4.2 mg mL⁻¹) when compared with the unfractionated hydrolysate (IC₅₀ <4.2 mg mL⁻¹) to inhibit lipase activity. Enzyme kinetic studies revealed that the peptides reduced α-amylase activity through competitive inhibition. However, inhibition of α-glucosidase activity was non-competitive.     

桑叶黄酮类和多糖类化合物的提取及其降血糖作用研究

从桑叶中提取总黄酮和多糖,并测定其对α-葡萄糖苷酶和猪胰液α-淀粉酶的抑制作用以评估其降血糖功能。结果表明,桑叶中提取的黄酮类和多糖类化合物对这两种酶都有较好的抑制作用,并且总黄酮和多糖对α-葡萄糖苷酶及猪胰液α-淀粉酶的抑制存在协同作用,两者混合对两种酶的抑制作用更强。     

Metabolite profiling,antioxidant activity,and glycosidase inhibition property of the mesocarp tissue extracts of sugar date palm [Phoenix sylvestris (L.) Roxb.] fruits

Phoenix sylvestris is an underutilized seasonal fruit in West Bengal, India. Methanol extract and extracts after alkaline hydrolysis of the mesocarp tissue of full-mature edible fruits of P. sylvestris were analyzed by GC-MS following a metabolomics approach. The fractions were tested for their antioxidant and inhibitory properties against the two key enzymes involved in diabetes, α-amylase, and α-glucosidase. Total 71 metabolites belonging to organic acids, amino acids, sugars, sugar alcohols, fatty acids, and phenols were identified in the methanol extract and in fractions after saponification. All the extracts and fractions showed high antioxidant, α-glucosidase, and α-amylase inhibitory activities. Sugars like raffinose (IC₅₀ = 0.36 μM), sucrose (IC₅₀ = 0.51 μM), trehalose (IC₅₀ = 0.85 μM), and phenols like taxifolin (IC₅₀ = 0.31 μM), benzoic acid (IC₅₀ = 2.74 μM) inhibited only the enzyme α-amylase. Phenolic components which inhibited both the enzymes were caffeic acid (IC₅₀ = 1.42 μM for α-amylase and IC₅₀ = 1.8 μM for α-glucosidase), 3, 4-dihydroxy benzoic acid (IC₅₀ = 0.23 μM for α-amylase and IC₅₀ = 2.58 μM for α-glucosidase), and quinic acid (IC₅₀ = 4.91 μM for α-amylase and IC₅₀ = 4.95 μM for α-glucosidase). Ferulic acid (IC₅₀ = 0.52 μM) and 4-hydroxycinnamic acid (IC₅₀ = 0.23 μM) inhibited only α-glucosidase. This study suggested that the metabolites present in the fruit mesocarp tissue showed the potential antioxidant activity and properties to inhibit the enzymes α-amylase and α-glucosidase. Further in vivo study is to be carried out to prove the efficacy of the fruits.

Abbreviations: FM: crude methanol extract; SI: ethyl acetate extract after alkaline hydrolysis step I; SII: ethyl acetate extract after alkaline hydrolysis step II