迷迭香中天然防腐剂的提取方法及其抑菌作用研究

本文研究了迷迭香中天然防腐剂的提取方法和抑菌作用,结果表明,用食用乙醇提取迷迭香中防腐物质的最佳提取工艺参数为:固液比1:15、提取温度80℃、提取时间为15 h。迷迭香醇提取物对实验用常见食品污染菌有较强的抑制作用,对金黄色葡萄球菌和枯草芽孢杆菌的最低抑菌质量浓度(MIC)为6.25 g.L-1,对大肠杆菌和汉逊氏酵母菌为12.5 g.L-1,对青霉和黑曲霉为25 g.L-1,对金黄色葡萄球菌、大肠杆菌和枯草芽孢杆菌的抑菌活性pH范围均为4~7,对汉逊氏酵母菌、青霉和黑曲霉为4~6。     

紫苏籽油抗菌活性研究

以革兰氏阴性菌大肠杆菌和革兰氏阳性菌枯草芽孢杆菌作为供试菌株,采用滤纸片法测定评估了紫苏籽油经过皂化提取之后其提取液抑制大肠杆菌和枯草芽孢杆菌的能力,并将所得的结果与大豆油的抑菌作用进行比较。结果表明,紫苏籽油提取液对大肠杆菌和枯草芽孢杆菌两个供试菌株的抑制效果较强,且对于枯草芽孢杆菌的抑制效果要优于大肠杆菌,且紫苏籽油对大肠杆菌和枯草芽孢杆菌的抑制效果要优于大豆油。     

樟叶乙醇提取液的抑菌作用

樟叶的乙醇提取物对金黄色葡萄球菌、大肠杆菌、枯草芽孢杆菌、汉逊氏酵母菌、青霉、毛霉有一定的抑菌作用。其最低抑菌浓度对三种细菌及酵母为 12 .5m g/ ml,对青霉、毛霉为 2 5m g/ m l     

文冠果种仁油中总甾醇的皂化法提取及组成分析北大核心CSCD

本文研究了皂化法提取文冠果种仁油中甾醇化合物的工艺条件,并在纯化的基础上利用气相色谱-质谱仪分析了提取物的基本组成。首先通过单因素试验和正交试验探讨了乙酸乙酯用量、皂化温度、料液比、皂化时间对甾醇化合物含量的影响;结果表明:乙酸乙酯用量200 m L,皂化温度67℃,料液比1∶5(g/m L),皂化时间2 h,在此条件下文冠果种仁中总甾醇含量达0.498%。粗甾醇采用溶剂结晶法进行纯化后的气质联用分析结果显示,文冠果种仁油中总甾醇中主要含有5种单体,分别是豆甾醇、豆甾-7,22-二烯-3-醇、麦角甾-7,22-二烯-3-酮、β-谷甾醇、33-降柳珊瑚甾-5,24(28)-二烯-3-醇。该研究结果为进一步探讨文冠果种仁甾醇的功能奠定了基础。     

文冠果种仁油中总甾醇的皂化法提取及组成分析

本文研究了皂化法提取文冠果种仁油中甾醇化合物的工艺条件,并在纯化的基础上利用气相色谱-质谱仪分析了提取物的基本组成。首先通过单因素试验和正交试验探讨了乙酸乙酯用量、皂化温度、料液比、皂化时间对甾醇化合物含量的影响;结果表明:乙酸乙酯用量200 m L,皂化温度67℃,料液比1∶5(g/m L),皂化时间2 h,在此条件下文冠果种仁中总甾醇含量达0.498%。粗甾醇采用溶剂结晶法进行纯化后的气质联用分析结果显示,文冠果种仁油中总甾醇中主要含有5种单体,分别是豆甾醇、豆甾-7,22-二烯-3-醇、麦角甾-7,22-二烯-3-酮、β-谷甾醇、33-降柳珊瑚甾-5,24(28)-二烯-3-醇。该研究结果为进一步探讨文冠果种仁甾醇的功能奠定了基础。     

苦参配方的醇提工艺优化及抑菌活性研究

利用响应面法设计实验优化苦参抑菌配方的回流提取工艺,以配方对金黄色葡萄球菌抑菌率作为响应值,通过考察提取工艺的单因素:浸泡时间、回流时间、水浴温度、液料比、乙醇体积分数等,选取具有显著影响的参数作为响应面实验设计变量。经过数学模型预测与验证实验确定最优提取工艺为:乙醇体积分数80. 5%、回流时间1. 6h、水浴温度95℃。在0. 003 g/m L生药给药条件下,最优工艺的醇提物的抑菌率达到94. 29%。验证结果稳定且与预测值偏差低于2%,该工艺合理可行。优化后的工艺下,所提浸膏对枯草芽孢杆菌有良好的抑制作用,对大肠杆菌抑制作用较弱。     

葛根素提取及其抑菌实验研究

以湘西葛根为实验材料,选取水、甲醇和95％的乙醇作为提取溶剂,索氏提取法提取其葛根素。以提取率为指标,综合提取工艺中加入量、提取时间的影响,采用正交实验筛选出乙醇提取工艺,最佳参数为每次加入10倍量、回流提取时间3．5h。用滤纸片法研究葛根提取液对大肠杆菌等8种常见食品腐败菌的抑菌活性,结果表明,各种菌体的抑菌效果为：大肠杆菌〉金黄色葡萄球菌〉枯草芽孢杆菌〉假丝酵母青霉,对青霉无抑菌作用。各种菌的抑菌效果与浓度的关系为,葛根素浓度升高,抑菌效果增强。     

硬枝树花中4个单体化合物的抑菌活性

研究了从硬枝树花中提取得到的4个单体化合物松萝酸(usnic acid)、去甲环萝酸(evernic acid)、巴尔巴地衣酸(barbatic acid)和水杨嗪酸(salazinic acid)对大肠杆菌、沙门氏菌、金黄色葡萄球菌、枯草芽孢杆菌4种细菌及甜瓜根腐病菌、禾谷镰刀病菌、番茄灰霉病菌3种植物病原菌的抑制能力。结果表明,水杨嗪酸对大肠杆菌的抑制能力较强,对枯草芽孢杆菌几乎无抑制作用。松萝酸对大肠杆菌和枯草芽孢杆菌基本无抑制作用。高浓度的去甲环萝酸对大肠杆菌及金黄色葡萄球菌的抑制效果较好。巴尔巴地衣酸对4种细菌的抑制作用不强。水杨嗪酸对3种植物病原菌的抑制效果明显,去甲环萝酸对3种植物病原菌的抑菌效果较好,巴尔巴地衣酸对番茄灰霉病菌的抑制能力较好,而松萝酸对植物病原菌的抑制能力较弱。     

辣椒素的提取及抑菌活性研究

以干红辣椒为原料,选取95%的乙醇和正己烷作为提取溶剂,利用索氏提取法提取其中的辣椒素。结果表明,以95%的乙醇作为提取溶剂的效果好于正己烷,辣椒素的提取率最高可达1.77%,高于正己烷的1.12%的提取率。用滤纸片法研究辣椒素对大肠杆菌等8种常见食品腐败菌的抑菌活性。结果表明:辣椒素对金黄色葡萄球菌、枯草芽孢杆菌有明显的抑菌效果,对大肠杆菌抑菌作用较弱,对黑曲霉、青霉、保加利亚乳酸杆菌和噬热链球菌没有作用。辣椒素经过高温灭菌后,抑菌作减弱,培养时间对抑菌圈的影响不大。以正己烷为助溶剂,确定辣椒素对枯草芽孢杆菌的最低抑菌浓度(MIC)为50mg/mL。     

长足大竹象雄性附腺提取物抑菌作用研究

以3种细菌(大肠杆菌Escherichia coli、金黄色葡萄球菌Bacillus subtilis、枯草芽孢杆菌Staphyloccocus aureus)为供试菌,测定了长足大竹象Cyrtotrachelus buqueti雄性附腺提取物的抑菌活性。结果表明:长足大竹象雄性附腺提取物对革兰氏阳性菌如枯草芽孢杆菌、金黄色葡萄球菌均有一定的抑菌活性。不同浓度的粗提物对枯草芽孢杆菌及金黄色葡萄球菌抑菌影响差异均具有高度统计学意义(P0.01),抑菌活性随着粗提物浓度的增加而增强。粗提物对金黄色葡萄球菌和枯草芽孢杆菌的最小抑菌浓度分别为0.2 mg·m L~(-1)、0.4 mg·m L~(-1)。不同处理温度对长足大竹象雄性附腺粗提物的抑菌作用有一定影响。     

鼻咽癌组织的显微切割及其 RNA 线性扩增

从微小体积鼻咽癌活检标本中获取纯净癌细胞一直是鼻咽癌分子生物学研究中的难题 . 为了寻找一种能从鼻咽癌活检组织中获得高纯度、高质量 RNA 来完成 cDNA 微阵列 (cDNA Microarray) 实验的简便实用方法,采用 RNAlater 技术保存鼻咽癌活检组织,显微切割技术来获得高纯度鼻咽癌细胞,利用 RNA 线性扩增技术得到 cDNA 微阵列实验所需 RNA. 结果表明：利用 RNAlater 技术可以很好地保持组织 RNA 的稳定,通过优化显微切割和 RNA 线性扩增的条件获得了 cDNA 微阵列实验所需的高纯度、高质量 RNA.     

α-亚麻酸甾醇酯的理化特性及氧化稳定性研究

本文研究了采用无溶剂直接酯化法合成得到的α-亚麻酸甾醇酯的理化特性、脂溶性、结晶特性、油脂氧化稳定性。结果表明,α-亚麻酸甾醇酯具有理想的理化特性,酸价和过氧化值分别为1.2 mg KOH/g和0.56meq/kg,反式脂肪酸含量小于0.1%,在不同植物油脂中的溶解性达到30%以上,结晶温度区间在-25.9～-29.6℃之间。α-亚麻酸甾醇酯在大豆油、油菜籽油和亚麻籽油中的浓度分别小于0.1%、0.1%和0.3%,油脂的氧化诱导时间随浓度增加而增加。因此α-亚麻酸甾醇酯良好的理化特性表明其在不同形态食品、保健品和医药产品等中将具有较广的应用范围,是一种具有较高营养价值的功能性食品添加剂。     

Genetic dissection of tocopherol and phytosterol in recombinant inbred lines of sunflower through quantitative trait locus analysis and the candidate gene approach

Sunflower (Helianthus annuus L.) contains tocopherol, a non-enzymatic antioxidant known as lipid-soluble vitamin E, and phytosterol, with interesting properties, which can result in decreased risk of chronic diseases in humans and with several beneficial effects in plants. The genetic control of tocopherol and phytosterol content in a population of 123 recombinant inbred lines of sunflower was studied through quantitative trait loci (QTL) analysis using 190 simple sequence repeats and a gene-based linkage map. Seven experiments were conducted in different environments in France and Iran during 2007 and 2008. Each experiment consisted of three replications. Means over all environments were used for QTL mapping. Five QTL for total tocopherol content on linkage groups 1, 8, 10 and 14 accounted for 45% of phenotypic variation, whereas four QTL for total phytosterol content on linkage groups 1, 2, 16 and 17 explained 27% of the phenotypic variation. GST, PAT2, SFH3 and POD genes showed co-localization with QTL for total phytosterol content. SMT2 is also mapped on linkage group 17 near the QTL of total phytosterol content. Four candidate genes, VTE4, HPPD, GST and Droug1, exhibited co-localization with QTL for total tocopherol content. The candidate genes associated with tocopherol and phytosterol, especially HPPD, VTE4 and SMT2, could be used for alternation of the tocopherol and phytosterol content of sunflower seeds through the development of functional markers.     

决明子营养价值分析及蛋白提取工艺研究

以大决明子为原材料,基于氨基酸比值系数法,对决明子的营养价值进行了分析;并以碱性缓冲液为提取剂,进行决明子蛋白提取工艺研究,探讨了缓冲液浓度及pH值、料液比、浸泡时间对蛋白提取率的影响,最后用正交试验确定大决明子蛋白的最佳提取工艺。结果表明决明子的营养价值高于大豆和紫花苜蓿,与南瓜接近略低于鸡蛋;提取工艺条件最佳为50 mmol/L、pH值=8.0的KH2PO4-NaOH缓冲液,料液比1 g：50 mL,浸泡提取时间12h。此条件下决明子蛋白的提取率为93.3%。     

提取工艺和植物生长调节剂对罗布麻愈伤组织中黄酮含量的影响

以罗布麻愈伤组织粉末为材,在单因素实验的基础上,利用响应曲面法对罗布麻愈伤组织中黄酮的提取工艺进行优化。响应曲面分析结果表明,提取试剂和提取温度对提取的黄酮含量存在显著影响。通过响应曲面分析得到罗布麻愈伤组织中黄酮提取的最佳条件为:提取试剂为70%甲醇,物料比1∶40,提取时间为4 h,提取温度为70℃。培养并比较了30种不同植物生长调节剂浓度与配比诱导100 d生长的愈伤组织中黄酮的含量,结果得出MB+KT(1.0 mg/L)+NAA(0.2 mg/L)上培养约100 d的愈伤组织中黄酮含量最高,为73.90mg/g。测定愈伤组织培养30 d内黄酮的积累动态,探明从培养的愈伤组织提取黄酮的最佳时段。通过优化提取工艺和筛选最佳植物生长调节剂浓度与配比,运用组织培养技术提高了罗布麻愈伤组织中黄酮含量。     

Enhanced Production of Androst-1,4-Diene-3,17-Dione by Mycobacterium neoaurum JC-12 Using Three-Stage Fermentation Strategy

To improve the androst-1,4-diene-3,17-dione (ADD) production from phytosterol by Mycobacterium neoaurum JC-12, fructose was firstly found favorable as the initial carbon source to increase the biomass and eliminate the lag phase of M. neoaurum JC-12 in the phytosterol transformation process. Based on this phenomenon, two-stage fermentation by using fructose as the initial carbon source and feeding glucose to maintain strain metabolism was designed. By applying this strategy, the fermentation duration was decreased from 168 h to 120 h with the ADD productivity increased from 0.071 g/(L·h) to 0.108 g/(L·h). Further, three-stage fermentation by adding phytosterol to improve ADD production at the end of the two-stage fermentation was carried out and the final ADD production reached 18.6 g/L, which is the highest reported ADD production using phytosterol as substrate. Thus, this strategy provides a possible way in enhancing the ADD production in pharmaceutical industry.     

Production of testosterone from phytosterol using a single-step microbial transformation by a mutant of Mycobacterium sp.

A testosterone (TS)-producing mutant, ST2, was derived from a phytosterol-assimilating and androst-4-ene-3,17-dione (AD)-producing bacterium, Mycobacterium sp. B-3805S, using nitrosoguanidine (NTG) mutagenesis. Production of TS from phytosterol using a single-step microbial transformation process by ST2 was investigated in a 5-l surface-aeration microprocessor-controlled fermentor loaded with a synthetic medium supplemented with 0.1% phytosterol, 2% glucose and 1% peptone at 30°C. An increase in dissolved oxygen at the initial stage of fermentation favored the side-chain degradation of phytosterol to AD. Later in the fermentation, a decrease in the dissolved oxygen to zero resulted in a decrease in pH to 6.0 as well as the reduction of AD to TS. Under optimal fermentation conditions, the maximum conversion ratio of phytosterol to TS was 31% after 120 h cultivation. It was concluded that the control of dissolved oxygen in the fermentation culture is the most important parameter for production of TS from phytosterol via AD. TS was isolated from the fermentation culture by addition of Amberlite XAD-7 resin and was further purified by flash chromatography on a silica gel column. After crystallization, TS was obtained as needle crystals with the correct melting point. Journal of Industrial Microbiology & Biotechnology (2002) 28, 280–283 DOI: 10.1038/sj/jim/7000243 Received 20 September 2001/ Accepted in revised form 16 January 2002     

Phytosterols and lecithin do not have an additive effect in lowering plasma and hepatic cholesterol levels in diet-induced hypercholesterolemic rats

Both plant sterols and lecithin are used as dietary supplements for lowering blood cholesterol in Western countries. This study evaluated the possibility of an additive effect of these ingredients on the regulation of lipid concentrations and cholesterol metabolism. Male Sprague-Dawley rats were randomly divided into three groups, and fed one of the following diets for 5 weeks; high cholesterol diet (HCD), phytosterol mixture-supplemented diet (PD, HCD+0.25% phytosterols), or phytosterol mixture and lecithin-supplemented diet (PLD, PD+0.15% lecithin). Feeding the PD for 5 weeks resulted in a 34% and 41% decrease in plasma total- and VLDL+LDL-cholesterol levels, respectively, and a 23% decrease in hepatic cholesterol content compared to those for the HCD rats (p < 0.05). These cholesterol-lowering properties of the phytosterol mixture were also associated with the down-regulation of hepatic acyl CoA:cholesterol acytransferase (ACAT) activity (p < 0.05). Addition of lecithin plus phytosterol mixture to the hypercholesterolemic diet did not significantly affect blood and hepatic lipid concentrations (with the exception of 36% decrease in hepatic triglyceride level, p < 0.05) as well as hepatic ACAT activity compared to feeding the hypercholesterolemic diet supplemented with phytosterol alone. These results indicate that combining lecithin, at a 0.15% level, with a phytosterol mixture-supplemented diet does not exhibit an additive effect in regulating hepatic ACAT activity or lowering blood cholesterol in hypercholesterolemic rats.     

Oral absorption of phytosterols and emulsified phytosterols by Sprague-Dawley rats

Clinical studies have demonstrated that consumption of phytosterol esters in lipid-based foods decreases serum concentrations of total and LDL cholesterol. These substances represent minimal potential for adverse effects when consumed orally because of their low bioavailability. However, some studies have reported estrogenic and other effects in laboratory animals treated parenterally with phytosterols, demonstrating that these substances may have the potential to cause adverse effects if absorbed. Water-soluble phytosterols have been prepared by formulation with emulsifiers to expand delivery options to include non-lipid-based foods. However, emulsifiers are used as excipients in the formulation of lipophilic pharmaceuticals to increase solubility, thereby increasing their absorption. Therefore, oral consumption of emulsified water-soluble phytosterols could potentially increase their absorption. In the current study, absorption of phytosterols prepared as water-soluble emulsified micelles with two different food-grade emulsifiers was evaluated in Sprague-Dawley rats and compared with absorption of non-micellar free phytosterols and esterified phytosterol mixtures dissolved in a lipophilic vehicle (soybean oil). Rats were dosed via gavage with 42 mg/kg of formulated phytosterol preparations. Blood was collected at 8, 16, 24, and 32 hours, extracted with hexane, derivatized with benzoyl chloride, and analyzed by high-performance liquid chromatography to determine concentrations of beta-sitosterol, and campesterol. Plasma concentrations and AUC(0-32 hours) [microg/mL/h] of beta-sitosterol and campesterol were lower in plasma obtained from rats treated with emulsified phytosterol preparations than in animals treated with free phytosterols dissolved in soybean oil. Because the pharmacokinetic profile of water-soluble phytosterols is similar to that of phytosterols administered in a lipid vehicle, the safety profile is likely to be the same as that of phytosterols and phytosterol esters in currently used applications.     

化学衍生-溶剂结晶法从植物甾醇混合组分中分离豆甾醇

采用化学衍生和溶剂结晶法,建立起一条从植物甾醇混合组分分离豆甾醇的技术途径。通过对植物甾醇的乙酰化,溴化-脱溴和皂化反应,并结合相应试样的重结晶操作以分离豆甾醇。利用此方法从植物甾醇混合组分分离豆甾醇,其产率为6.9%,回收率45.7%,具有生物工业应用潜力。