大蒜素和有机溶剂对蒜氨酸酶活性及大蒜油组成的影响

大蒜油具有重要的生物活性和调味功能。本文以化学合成的蒜氨酸为原料在蒜酶的催化下制备大蒜油,研究中间产物大蒜素、萃取溶剂对蒜酶活性的影响,以及溶剂对大蒜油组成成分的影响。结果显示:大蒜素在一定程度上会抑制蒜酶的活性,但不会导致蒜酶的完全失活;有机溶剂对蒜酶的影响比较复杂,其中氯仿、二氯甲烷使蒜酶活性完全丧失,而正己烷、正戊烷对蒜酶的活性影响则不明显。采用气质联用、核磁共振二维谱表征了不同溶剂下大蒜油的组成。用氯仿、正己烷等非极性溶剂萃取大蒜素,得到的大蒜油以3-乙烯基-1,2-二硫环己-4-烯(4X)和3-乙烯基-1,2-二硫环己-5-烯(5X)为主;而用乙酸等质子溶剂萃取,则大蒜油成分以二烯丙基三硫醚(DATS)、二烯丙基二硫醚(DADS)为主。     

Deodorization of Garlic Breath by Foods,and the Role of Polyphenol Oxidase and Phenolic Compounds

Garlic causes a strong garlic breath that may persist for almost a day. Therefore, it is important to study deodorization techniques for garlic breath. The volatiles responsible for garlic breath include diallyl disulfide, allyl mercaptan, allyl methyl disulfide, and allyl methyl sulfide. After eating garlic, water (control), raw, juiced or heated apple, raw or heated lettuce, raw or juiced mint leaves, or green tea were consumed immediately. The levels of the garlic volatiles on the breath were analyzed from 1 to 60 min by selected ion flow tube mass spectrometry (SIFT‐MS). Garlic was also blended with water (control), polyphenol oxidase (PPO), rosemarinic acid, quercetin or catechin, and the volatiles in the headspace analyzed from 3 to 40 min by SIFT‐MS. Raw apple, raw lettuce, and mint leaves significantly decreased all of the garlic breath volatiles in vivo. The proposed mechanism is enzymatic deodorization where volatiles react with phenolic compounds. Apple juice and mint juice also had a deodorizing effect on most of the garlic volatiles but were generally not as effective as the raw food, probably because the juice had enzymatic activity but the phenolic compounds had already polymerized. Both heated apple and heated lettuce produced a significant reduction of diallyl disulfide and allyl mercaptan. The presence of phenolic compounds that react with the volatile compounds even in the absence of enzymes is the most likely mechanism. Green tea had no deodorizing effect on the garlic volatile compounds. Rosmarinic acid, catechin, quercetin, and PPO significantly decreased all garlic breath volatiles in vitro. Rosmarinic acid was the most effective at deodorization.     

大蒜油的抗凝血作用

为研究大蒜油的抗凝血作用,本实验从大蒜中分离提取大蒜油,经气相色谱-质谱联用仪鉴定该大蒜油中主要成分为二烯丙基硫醚（allyl sulfide,DAS）、二烯丙基二硫醚（diallyl disulfide,DADS）和二烯丙基三硫醚（diallyl trisulfide,DATS）。体外实验结果表明大蒜油、DAS、DADS、DATS均具有部分抑制凝血酶活性功能,且抑制作用由强到弱依次为DATS＞大蒜油＞DADS＞DAS。利用Lineweaver-Burk双倒数作图法进行酶动力学分析,结果表明大蒜油、DAS、DADS、DATS对凝血酶的抑制作用属于混合性抑制（竞争性和非竞争性抑制）。分子荧光光谱分析结果表明大蒜油、DAS、DADS、DATS能与凝血酶蛋白中的发色基团相互作用导致荧光猝灭。三维分子对接分析结果显示出DATS与凝血酶结合性能良好;二维分子对接分析结果显示出DATS与凝血酶蛋白活性位点处Asp189、Gly219和Ala190形成氢键,与酶活性位点的Gly216、Ser195、Trp215形成疏水作用力,体现竞争性抑制作用,与非活性位点Gly226、Asp221A、Tyr225、Glu217、Cys191、Val213形成疏水作用,体现非竞争性抑制作用。体内实验结果表明,大蒜油、DAS、DADS、DATS分别通过正向调控大鼠凝血四项指标而直接展现出抗凝血功能,且抗凝血能力依次为DATS＞DADS＞DAS＞大蒜油。大鼠体内抗氧化实验结果表明大蒜油、DAS、DADS、DATS均只有微弱抗氧化能力,大蒜油、DAS、DADS、DATS具有的抗凝血功能应与其抗氧化能力不相关。     

基于主成分分析的大蒜药用质量评价

通过比较分析大蒜的药用品质,建立大蒜药用质量评价体系。以水分、灰分、水溶性浸出物、大蒜素含量、蒜氨酸含量、大蒜辣素含量和蒜酶活力为指标,分析甘肃民乐、江苏邳州、山东金乡、河南郑州、重庆巫溪和新疆且末、拜城、种马场、虎头镇、大有镇、新地乡等11个产地大蒜的药用品质特征及差异,并通过相关性分析、主成分分析和聚类分析对大蒜质量进行综合评价。结果表明,不同产地大蒜的上述指标都具有显著性差异。相关性分析表明,蒜酶活力与水分呈极显著正相关性（P<0.01）,蒜氨酸含量与灰分和大蒜辣素含量呈显著正相关性（P<0.05）。利用主成分分析可筛选出3个累计贡献率达到81.134%的主成分,在贡献率最大的第1主成分中,蒜氨酸对大蒜药用品质影响最大,其次为大蒜辣素和蒜酶活力。对11个产地大蒜进行综合评价,甘肃民乐大蒜的主成分综合评分得分最高为1.44,其余4个得分大于零的皆为新疆大蒜。系统聚类分析可将大蒜聚为四类,其中郑州大蒜单独为一类,金乡、巫溪和种马场大蒜聚为一类,新地、大有和民乐大蒜聚为一类,其余三地聚为一类。蒜氨酸、大蒜辣素和蒜酶活力可用于体现不同产地大蒜药用品质间的差距,利用综合评分可筛...     

Volatile distribution in garlic (Allium sativum L.) by solid phase microextraction (SPME) with different processing conditions

Treatments of autoclaving, high temperature aging (aged-black garlic), crushing, and roasting at 100, 150, and 200°C were applied to alter the volatile profiles of garlic (Allium sativum L.). Headspace volatiles in samples were analyzed by a solid phase microextraction (SPME)-GC/MS. Total peak areas of crushed-raw garlic were the highest and those of aged-black garlic clove were the lowest. Crushing effects were clearly observed in raw garlic, aged-black garlic, and roasted garlic at 200°C for 60 min. Sulfur-containing volatiles including diallyl disulfide and diallyl trisulfide were major volatiles. Generally, peak areas of diallyl disulfide decreased when garlic received autoclaving and roasting treatment while diallyl trisulfide and allyl methyl trisulfide increased during heat treatment compared to raw garlic. Roasting at 200°C for 60 min caused the formation of pyrazines greatly in garlic. Principal component analysis (PCA) for the volatile profiles by SPME-GC/MS could discriminate types of processed garlic successfully.     

Antiyeast Potency of Heated Garlic in Relation to the Content of Allyl Alcohol Thermally Generated from Alliin

ABSTRACT:  The appearance of antiyeast activity in heated garlic extract was influenced by the temperature, heating time, and pH. The maximum antiyeast activity was attained at the temperature range between 110 and 130 °C for the heating time range of 90 and 30 min, respectively. Reduced antiyeast activity occurred above 130 °C. More potent antiyeast activity was generated when the pH of the garlic extract was adjusted to and below 4.0. The sensitivity to the antiyeast activity of heated garlic or heated alliin was quite different depending on the yeast strain. Species belonging to Zygosaccharomyces were the least sensitive, while Candida utilis and Pichia membranaefaciens were among the most sensitive. The alliin content of the garlic was in the range of 0.5% to 1.32%. When garlic extracts were heated at 121 °C for 120 min, the content of allyl alcohol generated from alliin through thermal degradation was in the range of 0.08% to 0.16%. The allyl alcohol produced upon heating was positively proportional to the content of alliin in garlic. The antiyeast potency expressed as the minimum inhibitory concentration (MIC) was negatively proportional to the content of allyl alcohol and thus to the content of alliin.     

不同品种和加工工艺制得黑蒜的成分分析

目的比较不同品种和加工工艺制得黑蒜的成分。方法以独头蒜和多瓣蒜为试验材料,采用湿热加工法制备黑蒜,与相应鲜蒜和干热加工法生产的黑蒜(市售)比较,分析其主要成分的变化。采用苯酚硫酸法测定总糖含量,福林酚法测定多酚样物质含量,DPPH、ABTS、超氧阴离子自由基清除率以及FRAP法测定抗氧化性,硫酸钡吸光比浊法测定总硫含量,高效液相法测定含硫化合物含量。结果独头蒜与多瓣蒜相比,其总糖含量较高,但多酚样物质含量无明显差异;其DPPH自由基清除率较低,但ABTS和超氧阴离子自由基清除率以及总抗氧化能力无明显差异;除蒜氨酸含量稍高外,总硫、二烯丙基二硫(diallyl disulfide,DADS)和二烯丙基三硫(diallyltrisulfide,DATS)含量均无明显差异。无论独头蒜还是多瓣蒜,湿热加工成黑蒜后,总糖含量和多酚样物质均含量增高;ABTS、DPPH、超氧阴离子自由基清除率和总抗氧化能力均显著增高;蒜氨酸、DADS和DATS含量均降低,但S-烯丙基半胱氨酸(S-allyl cysteine,S-AC)含量增高,总硫含量无明显变化。湿热加工法制备的黑蒜与市售黑蒜相比,湿法加工黑蒜其多酚样物质含量较高,而市售黑蒜总糖含量较高;其ABTS、DPPH、超氧阴离子自由基清除率和总抗氧化能力均较高;蒜氨酸、DADS和DATS含量均较高,而S-AC含量较低。结论独头蒜与多瓣蒜制备而得的黑蒜的主要成分无明显差异。湿热加工法制备的黑蒜与市售黑蒜相比,抗氧化性较强,但S-AC含量较低。不同大蒜品种和加工工艺制备黑蒜可以为黑蒜生产和功能特性探究提供依据。     

Odor Components of Human Breath After the Ingestion of Grated Raw Garlic

The major odor components of finely grated, raw garlic in the air of the human mouth have been studied. Immediately after garlic ingestion, tests of the mouth air of six subjects revealed two major peaks that were positively identified as being allyl mercaptan and diallyl disulfide; these findings are based on an evaluation of the fragment patterns of a GC-MS analysis and from the actual retention times of these authentic compounds. The allyl mercaptan level was higher than the level of diallyl disulfide, although both compounds smell like garlic. Our results suggest that after garlic ingestion, allyl mercaptan is the major garlic-smelling compound in human mouth air, and diallyl disulfide is secondary.     

Deodorization of Garlic Breath Volatiles by Food and Food Components

The ability of foods and beverages to reduce allyl methyl disulfide, diallyl disulfide, allyl mercaptan, and allyl methyl sulfide on human breath after consumption of raw garlic was examined. The treatments were consumed immediately following raw garlic consumption for breath measurements, or were blended with garlic prior to headspace measurements. Measurements were done using a selected ion flow tube‐mass spectrometer. Chlorophyllin treatment demonstrated no deodorization in comparison to the control. Successful treatments may be due to enzymatic, polyphenolic, or acid deodorization. Enzymatic deodorization involved oxidation of polyphenolic compounds by enzymes, with the oxidized polyphenols causing deodorization. This was the probable mechanism in raw apple, parsley, spinach, and mint treatments. Polyphenolic deodorization involved deodorization by polyphenolic compounds without enzymatic activity. This probably occurred for microwaved apple, green tea, and lemon juice treatments. When pH is below 3.6, the enzyme alliinase is inactivated, which causes a reduction in volatile formation. This was demonstrated in pH‐adjusted headspace measurements. However, the mechanism for volatile reduction on human breath (after volatile formation) is unclear, and may have occurred in soft drink and lemon juice breath treatments. Whey protein was not an effective garlic breath deodorant and had no enzymatic activity, polyphenolic compounds, or acidity. Headspace concentrations did not correlate well to breath treatments.     

Antiyeast Activity of Heated Garlic in the Absence of Alliinase Enzyme Action

Garlic heated to 121 °C was found to strongly inhibit the growth of yeasts, but not that of bacteria. The potency and stability of the antiyeast activity of heated garlic were compared with those of fresh garlic, garlic oil, and allyl isothiocyanate. The inhibitory activity of heated garlic was stable, and the minimum inhibitory concentration did not change for up to 30 d at 37 °C. The antiyeast activity of heated garlic was not influenced by pH. Alliin heated in distilled water showed an antiyeast activity pattern similar to that of heated garlic, suggesting that the compound(s) thermally generated from alliin are the principal antiyeast compound(s) of heated garlic. The antiyeast activity was increased as time of heating increased up to 45 min at 121 °C, and the activity did not change when garlic was further heated for up to 120 min.     

High hydrostatic pressure treatment for manufacturing of garlic powder with improved microbial safety and antioxidant activity

The traditional method of manufacturing garlic powder (GP) that includes simple grinding of air-dried garlic slices has problems of microbial safety and a pungent flavour for this product. Microbiologically safe GP with a less pungent flavour and better antioxidant activities was manufactured using high hydrostatic pressure (HHP), wet grinding and freeze-drying process. The numbers of total aerobic bacteria and yeasts and molds in untreated (without HHP) GP were 3.64 and 2.47 log CFU/g respectively. Garlic powder treated with 600 MPa HHP for 5 min exhibited a total aerobes count of 1.62 CFU/g and a yeasts and molds count of 1.43 log CFU/g. The diallyl disulfide content, which is responsible for the pungent odour of garlic, was also significantly reduced by HHP due to a decrease in the alliinase activity. Hence, a novel process using HHP can help to produce GP with improved microbial safety, flavour and nutrition.     

Effects of pH Adjustment and Subsequent Heat Treatment on the Formation of Volatile Compounds of Garlic

Effects of pH adjustment during the blending of garlic cloves and subsequence heat treatment on the formation of volatile compounds of garlic were studied by means of gas chromatography (GC). The formation of the two isomeric cyclic compounds 3-vinyl-[4H]-l,2-dithiin and 2-vinyl-[4H]-l,3-dithiin, which were artifacts from allicin, was favored around pH 5.5. Formation of diallyl trisulfide, methyl allyl trisulfide, cis-1-propenyl allyl disulfide, isobutyl isothiocyanate, 2,4-dimethylfuran, 1,3-dithiane, aniline, methyl allyl sulfide, dimethyl disulfide and 3-methyl-2-cyclopentene-l-thione were favored in neutral or weak acidic conditions, whereas formation of diallyl disulfide, diallyl sulfide, methyl allyl disulfide, propenylthiol, propyl allyl disulfide and 1,2-epithiopropane were favored around pH 9.0.     

金属离子对蒜氨酸酶性质的影响

以新鲜大蒜为原料,采用硫酸铵分级沉淀法、PEG6000沉淀法、葡聚糖G-200柱层析法提取分离蒜氨酸酶,获得适宜蒜氨酸酶粗分离的条件为：4℃、pH6.5磷酸盐缓冲液浸提,20%的PGE6000盐析,2000×g离心30min。再采用SDS-PAGE电泳法鉴定其纯度,发现纯度是粗酶液的16.85倍,回收率为41.26%,蒜氨酸酶亚基的分子质量约为54.7kD。利用正交试验优化金属离子交互作用对蒜氨酸酶活性的影响,研究金属离子对蒜氨酸酶反应动力学及紫外-可见光谱的影响。结果表明：不同浓度的离子组合对蒜氨酸酶活性有显著的影响,Mn2+是影响蒜氨酸酶活性最重要的因素;不同离子交互作用最佳条件为：Mn2+ 浓度10mmol/L、Fe3+浓度10mmol/L、Ca2+浓度5mmol/L、辅助因子蔗糖浓度5mmol/L,此条件下蒜氨酸酶相对酶活性为155.59%。而10mmol/L的Cu2+能够显著抑制蒜氨酸酶的活性;以蒜氨酸为底物,测得蒜氨酸酶Km值为0.25mmol/L,Vmax为1.29μmol/min;紫外-可见光谱证实了辅基磷酸吡哆醛的存在,金属离子对蒜氨酸酶的激活与抑制机制可能是因为激活剂和抑制剂影响了蒜氨酸酶辅助因子5,-磷酸吡哆醛的结构以及其与蒜氨酸酶主链的结合,从而影响了蒜氨酸酶的活性。     

不同切割伤害程度对大蒜次生代谢物质合成的影响

为研究不同切割伤害程度对大蒜次生代谢物质合成的影响,应用气相色谱法对大蒜切割伤害后的呼吸强度进行测定,应用丙酮酸法对大蒜切割伤害后催化合成大蒜素的蒜氨酸酶活力进行测定,在感官评定的同时应用静态顶空-气相色谱-质谱联用法对大蒜挥发性成分进行测定。结果表明:大蒜受伤后进行跃变型呼吸,切割伤害程度越大,大蒜呼吸强度越高,蒜氨酸酶活力越高。同时,切割伤害程度越大,大蒜风味强度明显越强、风味物质种类越多。大蒜受到切割伤害后的风味物质主要由二烯丙基二硫化物、甲基-2-丙烯基二硫化物等硫化物以及硫醚类物质组成,在6 h蒜泥中检测出种类最多的风味物质。     

超临界CO2-分子蒸馏纯化蒜油与水蒸气蒸馏法制取蒜油挥发性成分分析

采用分子蒸馏技术(MD)将超临界CO2(SFE)萃取的蒜油进行纯化,以顶空固相微萃取气质联用技术(HSSPME-GC-MS)分析蒜油中的挥发性成分,确定最佳工艺：进样速度1 滴/s,蒸馏温度60℃,刮膜转速250r/min。纯化的蒜油挥发性成分仅有4 种,二烯丙基二硫醚和二烯丙基三硫醚、2- 乙烯基-1,3- 二硫杂-4- 环己烯和3- 乙烯基-1,2- 二硫杂-5- 环己烯;与水蒸气蒸馏法制得的新鲜蒜油(F-GO)比较,SFE 法是一种理想的制取蒜油的方法;蒜油放置30d(S-GO)后有效成分含量下降,导致品质下降。     

Inhibitory activity of essential oils of garlic and onion against bacteria and yeasts

The essential oils of garlic and onion and their constituent sulfides with three or more sulfur atoms were potent inhibitors of yeast growth. The minimum inhibitory concentrations of garlic oil, onion oil, diallyl trisulfide, diallyl tetrasulfide, and dimethyl trisulfide for all the yeasts tested ranged between 2 and 45 ppm. The oils and their constituent sulfides, however, were only very weakly antibacterial, showing minimum inhibitory concentrations of greater than 300 ppm for most of the bacteria tested. The antiyeast activity of garlic oil and onion oil was storage stable and was not influenced by pH. Film formation on soy sauce by Zygosaccharomyces rouxii SS1 was completely prevented for 30 days by the addition of 30 and 40 ppm of garlic oil and onion oil, respectively.     

Stabilization of sunflower oil by garlic extract during accelerated storage

Efficacy of garlic extract in stabilizing sunflower oil during accelerated storage has been studied. Extracts of garlic were prepared in different solvents; extract yield was in the range of 6.24–23.2% and antioxidant activity range in the linoleic acid system was 14.1–93.2%. Being highest in yield and antioxidant potential, methanolic extract was thermally evaluated by heating the extract at 185 °C for different intervals, i.e. 0–80 min and evaluating antioxidant activity of the heated extract in the linoleic acid system (71.6% inhibition). Methanolic extract of garlic at three different concentrations, i.e. 250 (SFO-250), 500 (SFO-500) and 1000 ppm (SFO-1000) were added to preheated RBD sunflower oil. BHA (SFO-BHA) and BHT (SFO-BHT) at 200 ppm served as standards besides the control. Weight gain (WG), antioxidant activity index (AAI), free fatty acid (FFA) content, peroxide value (PV), conjugated dienes (CD), conjugated trienes (CT) and thiobarbituric acid-reactive substances (TBARS) were taken as parameters for evaluation of effectiveness of garlic in stabilization of sunflower oil. Results from different parameters were in agreement with each other, suggesting the highest efficiency of SFO-1000, followed by SFO-BHT, SFO-BHA, SFO-500, SFO-250 and Ctrl. Results reveal garlic to be a potent antioxidant for stabilization of sunflower oil.     

大蒜中硫代亚磺酸酯提取工艺及其稳定性研究

采用乙醇溶液提取大蒜中的硫代亚磺酸酯,研究了乙醇浓度、提取温度、提取时间、料液比对硫代亚磺酸酯提取量的影响,在单因素实验的基础上采用响应面法确定了最佳提取条件:乙醇浓度为64.89%,提取温度为19.06℃,提取时间为1.48 h,料液比为1∶4.48(g∶mL),硫代亚磺酸酯含量可达4.17 mmol/100 g。对硫代亚磺酸酯的稳定性研究表明:随着温度的升高,硫代亚磺酸酯稳定性下降,提取液在贮存过程中,随着硫代亚磺酸酯含量的降低,二烯丙基二硫醚(DADS)与二烯丙基三硫醚(DATS)的含量会上升。     

Inhibition of heterocyclic aromatic amine formation in fried ground beef patties by garlic and selected garlic-related sulfur compounds

The effects of garlic and selected organosulfur compounds (diallyl disulfide, dipropyl disulfide, diallyl sulfide, allyl methyl sulfide, allyl mercaptan, cysteine, and cystine) on the formation of heterocyclic aromatic amines (HAAs) in fried ground beef patties were evaluated. Minced garlic cloves (ca. 4.8 to 16.7%, wt/wt) or organosulfur compounds (0.67 mmol) were added directly to ground beef. Patties (100 g) were fried at 225 degrees C (surface temperature) for 10 min per side. Two patties were fried for each replication, and five replicates were analyzed for each treatment. For each replicate, four subsamples were analyzed (two unspiked subsamples for concentration and two spiked subsamples for the recovery of HAA standards). The volatile sulfur compounds significantly (P < 0.05) reduced concentrations of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine by reductions of 46 to 81%, while average reductions of 35, 22, and 71%, were achieved with cystine, cysteine, and whole garlic, respectively. The volatile sulfur compounds reduced concentrations of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline by 34 to 67%, while reductions of 25, 19, and 63% (P < 0.05) were achieved with cystine, cysteine, and whole garlic, respectively. These studies confirm that garlic and some organosulfur compounds have the potential to reduce HAA formation incooked beef patties.     

Comparison of encapsulation properties of major garlic oil components by hydroxypropyl β-cyclodextrin

The major garlic oil (GO) components were encapsulated by hydroxypropyl β-cyclodextrin (HP-β-CD), and their encapsulation properties were compared in this study. Our results showed that the optimal conditions for the GO encapsulation were as follows: weigh ratio at 12:88 (GO: HP-β-CD), pH at 5.0, and inclusion time at 4 h. Furthermore, results from the phase solubility curves indicated that two components of diallyl disulfide (DADS) and diallyl trisulfide (DATS) had higher water solubility caused by HP-β-CD and that DATS-HP-β-CD complex had higher stability constant (K _1:1 = 6.702 × 10⁵) and more significant change in free energy ( \Updelta G_\textCOMP^* \Updelta G_{\text{COMP}}^{*}  = −3.496 × 10⁴) than DADS-HP-β-CD. These findings indicated that DATS was better suited to be encapsulated by HP-β-CD compared to DADS. The result finally obtained from gas chromatography–flame ionization detector (GC–FID) confirmed the easier inclusion properties of DATS by HP-β-CD.