玫瑰茄花色素的提取工艺研究

通过单因素和正交实验对玫瑰茄花色素提取工艺条件进行了比较筛选和优化。结果表明:玫瑰茄花色素提取的适宜溶剂为乙醇,适宜的酸碱度为p H 2.0;影响提取效果因素依次是提取时间、乙醇浓度、温度、料液比;最佳提取工艺条件为粉碎粒度40目、乙醇浓度70%(V∶V)、提取温度40℃、提取时间1.5 h、最佳料液比1∶20,在此优化条件下,玫瑰茄花色素的提取率可达0.485%。     

响应曲面法优化菌丝体中红曲色素的提取条件

以红曲菌丝体为原料,通过单因素实验及Box-Behnken设计响应曲面分析优化了红曲色素的提取工艺条件。通过实验,得到了提取率的回归方程Y=0.923-0.011X1-0.011X2-0.017X12-0.027X22-0.026X32-0.028X1X2-0.019X1X3,通过对模型解逆矩阵得优化方案:乙醇浓度X1=66%、超声功率X2=200 w、提取温度X3=52℃,模型预测结果为92.5%。在该工艺条件下进行验证实验,提取率达到92.1±0.6%(n=3)。     

苦参酮的热稳定性及其提取工艺研究

考察了苦参酮的热稳定性,并用正交实验方法优选了苦参酮的提取工艺。结果发现,苦参酮在50℃下较稳定;以苦参酮的相对提取率为考察指标,在所考察的范围内,影响苦参酮相对提取率的主要因素依次为：提取时间〉乙醇浓度〉温度〉溶剂用量,其中提取时间和乙醇浓度对苦参酮的相对提取率影响显著;苦参酮的最佳提取工艺为：提取温度50℃,提取时间120min,容积用量（mL/g）25：1,乙醇浓度（v／v）：80％,相对提取率达到99．6％。优化后的提取工艺能较高效率地提取苦参酮。     

正交试验法优化吴茱萸总碱的微波提取

以紫外-可见分光光度法为检测手段,研究了乙醇-水混合溶剂中吴茱萸总生物碱的微波提取条件,试验中分别以微波功率、萃取时间和萃取溶剂的用量、浓度及酸度等为因素,设计了五因素四水平的正交试验,考察了它们对总生物碱成分提取率的影响。结果表明,最佳提取条件为：在700W微波功率下,用浓度70％、酸度pH=2的乙醇-水混合溶剂25mL／g,提取6min,此条件下总生物碱的提取率可达0．4898％。     

红曲霉变异株A6产色素发酵条件研究

本文对红曲霉变异株Ａ６进行了产红曲色素发酵条件试验,发现在以６％饴糖为碳源,以０３％ＮａＮＯ３为氮源的培养基,初始ｐＨ４０～４５,接种量为１５％时产色素浓度最高。同时,考察了整个发酵过程中ｐＨ,残糖量与色素生成浓度的关系,认为发酵周期拟控制在８０～８５ｈｒ为宜。本试验还发现在发酵过程中补加适量饴糖有利于产红曲色素     

双水相-微波提取苎麻皮中总黄酮及抗氧化性研究

在乙醇-硫酸铵双水相体系中,以苎麻皮为原料,采用微波法提取苎麻皮中的总黄酮。通过单因素实验考察了乙醇浓度、料液比、硫酸铵用量、提取功率、提取时间对提取率的影响。用正交试验确定了最优提取条件,即在料液比1:50 g/mL、硫酸铵用量为0.2 g/mL、提取功率为200 W、提取时间为120 s,乙醇体积浓度为70%时提取率可达2.442%。采用了水杨酸法和邻苯三酚自氧化法对黄酮抗氧化性进行了研究,结果表明苎麻皮黄酮对O-·2和·OH具有较强的清除作用。     

双水相-微波提取苎麻皮中总黄酮及抗氧化性研究北大核心CSCD

在乙醇-硫酸铵双水相体系中,以苎麻皮为原料,采用微波法提取苎麻皮中的总黄酮。通过单因素实验考察了乙醇浓度、料液比、硫酸铵用量、提取功率、提取时间对提取率的影响。用正交试验确定了最优提取条件,即在料液比1:50 g/mL、硫酸铵用量为0.2 g/mL、提取功率为200 W、提取时间为120 s,乙醇体积浓度为70%时提取率可达2.442%。采用了水杨酸法和邻苯三酚自氧化法对黄酮抗氧化性进行了研究,结果表明苎麻皮黄酮对O-·2和·OH具有较强的清除作用。     

莲子心总黄酮的超声波萃取工艺研究

研究了超声波萃取法从莲子心中提取总黄酮的最佳工艺。通过单因素控制法和正交试验,以乙醇为溶剂,总黄酮提取率为考察指标,在超声波辅助下研究了乙醇浓度、提取时间、料液比和提取温度对总黄酮提取率的影响。结果表明影响莲子心中总黄酮提取率的主要因素是乙醇的浓度,其次依次为提取温度、料液比、超声波萃取时间;提取的最佳条件是:乙醇的浓度为60%,提取温度是70℃,料液比为1:24,超声波萃取时间为30 min,此条件下得到的总黄酮的提取率是10.86 mg/g。     

黑曲霉发酵花生根提取白藜芦醇的优化

本研究利用黑曲霉发酵花生根产生的纤维素酶来酶解花生根中的纤维,破坏其细胞壁,有利于自藜芦醇的提取。同时对发酵后的花生根白藜芦醇的提取进行优化。根据中心组合实验设计原理,采用四因素五水平的响应面分析法优化发酵后花生根中自藜芦醇的提取,响应面分析表明乙醇浓度、提取温度、提取时间和液固比对花生根白藜芦醇提取率有显著影响,且不是简单的线性关系。得到最佳的提取条件为乙醇浓度64％,提取温度55℃,提取时间60min,液固比8：1,在此条件下,自藜芦醇得率为0．191％。     

沙棘果渣总黄酮提取工艺优化及抗氧化活性研究

利用果胶酶协同超声波法,对沙棘果渣有效成分总黄酮的提取工艺及其抗氧化活性进行了研究。以提取率为指标,通过酶用量、液料比、乙醇浓度、提取时间、提取温度、超声功率等单因素分析,选定酶用量、液料比、超声提取时间3个因素进行响应面试验,确定提取优化工艺条件为:果胶酶用量5.1%,液料比41∶1,超声提取时间81 min,此条件下,沙棘果渣总黄酮提取率达到8.91 mg/g;以BHT为阳性对照,进行了抗氧化活性研究,得出沙棘果渣总黄酮提取液浓度为0.14 mg/mL时,对DPPH自由基的清除率达到了94.42%;提取液浓度为1.2 mg/mL时,对羟自由基和超氧阴离子的清除率分别为83.10%和43.41%。整体来看,沙棘果渣总黄酮具有较强的抗氧化能力。     

长白山红蚂蚁还原糖和粗脂肪含量的优化提取

红蚂蚁的药理和免疫功能的实验研究表明,它具有明显的抗炎、平喘、镇静、护肝等作用。本研究以长白山红蚂蚁为研究对象,通过单因素试验和响应面法进行优化设计,探索红蚂蚁中还原糖和粗脂肪提取最佳工艺条件。结果表明:提取粗脂肪的最佳工艺条件为料液比4∶25 (体积比)、浸提时间18 h、乙醇浓度72.5%,此条件下粗脂肪提取率为11.298%;提取还原糖的最佳工艺条件为DNS用量4 mL,水浴时间8 h,乙醇浓度85%,此条件下还原糖提取率为2.29%。本试验为长白山红蚂蚁的功能产品研究提供参考。     

Enhancement of pyruvate productivity in Torulopsis glabrata: Increase of NAD+ availability

This study aimed at increasing the pyruvate productivity from a multi-vitamin auxotrophic yeast Torulopsis glabrata, by increasing the availability of NAD+. We examined two strategies for increasing availability of NAD+. To supplement nicotinic acid (NA), the precursor of NAD+; and to increase the activity of alcohol dehydrogenase integrating with addition acetaldehyde as exterior electron acceptor. The addition of 8 mg l(-1) NA to the fermentation medium resulted in a significant increase in the glucose consumption rate (48.4%) and the pyruvate concentration (29%). An ethanol-utilizing mutant WSH-13 was screened and selected after nitrosoguanidine mutagenesis of the parent strain T. glabrata CCTCC M202019. Compared with the parent strain, the alcohol dehydrogenase activity of the mutant WSH-13 increased about 110% and the mutant could utilize ethanol as the sole carbon source for growth (1.8 g l(-1) dry cell weight). When growing with glucose, the addition of 4 mg l(-1) acetaldehyde to the mutant WSH-13 culture broth led to a significant increase in the glucose consumption rate (26.3%) and pyruvate production (22.5%), but the ratio of NADH/NAD+ decreased to 0.22. Acetaldehyde did not affect the glucose and energy metabolism at high dissolved oxygen (DO) concentration. However, at lower DO concentration (20%), maintaining the acetaldehyde concentration in the mutant culture broth at 4 mg l(-1) caused an increased NAD+ concentration but a decreased NADH concentration. As a consequence, the pyruvate production rate, the pyruvate yield on glucose and the pyruvate concentration were 68, 44 and 45% higher, respectively, than the corresponding values of the control (without acetaldehyde). The strategy for increasing the glycolytic flux and the pyruvate productivity in T. glabrata by increasing the availability of NAD+ may provide an alternative approach to enhance the metabolites productivity in yeast.     

Studies in gram staining.

Gram-negative bacteria stained with crystal violet are decolorized by 95% alcohol within 2 min, whereas Gram-positive bacteria require at least 3 min treatment. Aqueous solutions of safranin, neutral red, and fuschsin replace crystal violet from stained Gram-positive bacteria more quickly than alcohol alone, and alcoholic solutions of these counterstains are in most cases still more effective. Treatment of crystal violet-stained organisms with alcoholic safranin (0.25%) for 15 sec will distinguish Gram-positive bacteria (violet) from Gram-negative bacteria (pink). Alcohol containing very low concentrations of iodine generally decolorizes crystal violet-stained Gram-positive bacteria more quickly than alcohol alone. Increasing concentrations of iodine in alcohol reduce the rate of decolorization of stained bacteria, but stained Gram-negative bacteria are still readily decolorized. The addition of 0.1% iodine to alcohol increases the rate of extraction of crystal violet by alcohol from Gram-negative organisms, but delays extraction of dye from Gram-positive organisms, and this applies when counterstain is also present. A two-solution modification of Gram staining is described in which crystal violet-stained bacteria are treated with an alcoholic solution of safranin, fuchsin, and iodine.     

Studies in Gram Staining

Gram-negative bacteria stained with crystal violet are decolorized by 95% alcohol within 2 min, whereas Gram-positive bacteria require at least 3 min treatment. Aqueous solutions of safranin, neutral red, and fuchsin replace crystal violet from stained Gram-positive bacteria more quickly than alcohol alone, and alcoholic solutions of these counterstains are in most cases still more effective. Treatment of crystal viokt-stained organisms with alcoholic safranin (0.25%) for 15 scc will distinguish Gram-positive bacteria (viokt) from Gram-negative bacteria (pink).

Alcohol containing very low concentrations of iodine generally decolorizes crystal violet-stained Gram-positive bacteria more quickly than alcohol alone. Increasing concentrations of iodine in alcohol reduce the rate of decolorization of stained bacteria, but stained Gram-negative bacteria are still readily dccolorized. The addition of 0.1% iodine to alcohol increases the rate of extraction of crystal violet by alcohol from Gram-negative organisms, but delays extraction of dye from Gram-positive organisms, and this applies when counterstain is also present. A two-solution modification of Gram staining is described in which crystal violet-stained bacteria are treated with an alcoholic solution of safranin, fuchsin, and iodine.     

氟化物对酒精发酵生产的影响

ＮａＦ是ＥＭＰ途径的抑制剂,然而,目前酒精发酵生产中却用以提高出酒率,这与酒精发酵机理是矛盾的。通过实验研究,分析比较ＮａＦ对酵母菌酒精发酵,革兰氏阳性菌（乳酸杆菌）和阴性菌（醋酸杆菌）代谢活动的影响,加入ＮａＦ对发酵液中钙,镁离子浓度的改变,结果发现钙,镁离子浓度与酒精发酵密切相关,生产上,ＮａＦ之所以能改善酵母菌的酒精发酵,主要是降低了水的硬度。ＮａＦ的用量应以水的硬度而定。过量ＮａＦ对酵母菌     

Concentration effect of Riesling Icewine juice on yeast performance and wine acidity

AIMS: The objective of this study was to determine the effect of increasing juice soluble solids above 40 degrees Brix on wine yeast's ability to grow and ferment the juice, with particular focus on acetic acid production, titratable acidity (TA) changes and the maximum amount of sugar consumed by the yeast. METHODS AND RESULTS: Riesling Icewine juices at 40, 42, 44 and 46 degrees Brix were inoculated with K1- V116 at 0.5 g 1(-1) and fermented at 17 degrees C until sugar consumption ceased. Increasing soluble solids showed strong negative linear correlations with yeast growth, sugar consumption and ethanol production (r = -0.999, -0.997 and 0.984, P < 0.001, respectively). Acetic acid, glycerol and TA production normalized to sugar consumed showed strong positive correlations to the initial juice concentration (r = 0.992, 0.963, and 0.937, P < 0.001 respectively) but no correlation was found for ethanol production. The acetic acid produced as a function of sugar consumed was positively correlated to the glycerol produced (r = 0.970, P < 0.001). The final TA of the wines ranged between 11.8 and 13.7 g 1(-1) tartaric acid, increasing by 2.3-3 g 1(-1) over the starting juice. The increase in TA was positively correlated to the increase in acetic acid produced after normalizing the data to the amount of sugar consumed (r =0.975, P < 0.001). The acid equivalents resulting from the increase in acetic acid accounted for 80-100% of the TA increase when converted to units of tartaric acid. In the final Icewines, acetic acid represented 19-20% of wine TA. CONCLUSIONS: Increasing Icewine juice concentration from 40 to 46 degrees Brix increases the proportion of yeast sugar metabolism towards glycerol and acetic acid production to cope with the increased osmotic stress by decreasing yeast growth, sugar consumption rate, the total amount of sugar consumed and the total amount of ethanol produced. The high proportional contribution of acetic acid to titratable acidity in Riesling Icewine may affect acidity perception. SIGNIFICANCE AND IMPACT OF THE STUDY: We have determined that 10% v/v ethanol would not be achievable with initial juice concentrations above 42 degrees Brix and that Riesling Icewine juice above 52.5 degrees Brix would be theoretically unfermentable. The high proportional contribution of acetic acid to TA may be an important factor in the organoleptic balance of these Icewines.     

Effect of 2,3-dinitrilo-1,4-dithia-9,10-antraquinone on Ehrlich ascites carcinoma and yeast cells

The inhibition of glycolysis by 2,3-dinitrilo-1,4-dithia-9,10-antraquinone (DDA) in Ehrlich ascites carcinoma (EAC) cells as well as in the investigated respiratory and fermentative strains of yeasts was found to be the result of inactivation of thiol enzymes of this pathway. Increasing concentration of DDA caused, in EAC cells, marked inhibition of hexokinase (HK), phosphofructokinase (PFK) and practically total inhibition of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). These three enzymes, as well as alcohol dehydrogenase (ADH) were also inactivated by DDA in yeasts. DDA inhibited the biosynthetic processes as measured by following the rate of [14C]adenine and [14C)]valine incorporation into TCA-precipitable fractions proportionally to the degree of glucose consumption by EAC or the yeast cells.     

Moderate acute intake of de-alcoholized red wine, but not alcohol, is protective against radiation-induced DNA damage ex vivo -- results of a comparative in vivo intervention study in younger men

Moderate intake of wine is associated with reduced risk of cardiovascular disease and possibly cancer however it remains unclear whether the potential health benefits of wine intake are due to alcohol or the non-alcoholic fraction of wine. We therefore tested the hypothesis that the non-alcoholic fraction of wine protects against genome damage induced by oxidative stress in a crossover intervention study involving six young adult males aged 21-26 years. The participants adhered to a low plant phenolic compound diet for 48 h prior to consuming 300 mL of complete red wine, de-alcoholized red wine or ethanol on separate occasions 1 week apart. Blood samples were collected 0.5, 1.0 and 2.0 h after beverage consumption. Baseline and radiation-induced genome damage was measured using the cytokinesis-block micronucleus assay and total plasma catechin concentration was measured. Consumption of de-alcoholized red wine significantly decreased the gamma radiation-induced DNA damage at 1 and 2 h post-consumption by 20%. In contrast alcohol tended to increase radiation-induced genome damage and complete wine protected against radiation-induced genome damage relative to alcohol. The observed effects were only weakly correlated with the concentration of total plasma catechin (R=-0.23). These preliminary data suggest that only the non-alcoholic fraction of red wine protects DNA from oxidative damage but this effect cannot be explained solely by plasma catechin.     

The effect of moderate alcohol consumption on fat distribution and adipocytokines

Objective: To investigate the effect of moderate alcohol consumption on fat distribution, adipose tissue secreted proteins (adiponectin and resistin), and insulin sensitivity in healthy middle‐aged men with abdominal obesity. Research Methods and Procedures: Thirty‐four healthy men between 35 and 70 years old, with increased waist circumference (≥94 cm), participated in a randomized, controlled cross‐over design trial. They drank 450 mL of red wine (40 grams of alcohol) or 450 mL of de‐alcoholized red wine daily during 4 weeks. At the end of each treatment period, fat distribution, adipose tissue proteins, and insulin sensitivity index (ISI) were measured. Results: Subcutaneous and abdominal fat contents and body weight did not change after 4 weeks of moderate alcohol consumption. Liver fat (quip index) was slightly higher after consumption of red wine (6.8 ± 0.1) as compared with de‐alcoholized red wine (6.5 ± 0.1) but not significantly different (p = 0.09). Plasma adiponectin concentration increased (p < 0.01) to 6.0 ± 0.1 μg/mL after 28 days of moderate alcohol consumption compared with de‐alcoholized red wine (5.5 ± 0.1 μg/mL). Serum resistin concentrations and ISI were not affected by alcohol consumption. Percentage changes in serum resistin correlated significantly with changes in ISI (r = ?0.69, p < 0.01), whereas this correlation was not present between changes in plasma adiponectin and ISI (r = 0.31, p = 0.22). Discussion: Moderate alcohol consumption for 4 weeks is not associated with differences in subcutaneous and abdominal fat contents or body weight. Thus, the 10% increase in adiponectin was not associated with a change in fat distribution or body weight change.     

Efficient extraction of intracellular reduced glutathione from fermentation broth of Saccharomyces cerevisiae by ethanol

Reduced glutathione (GSH) from fermentation broth of Saccharomyces cerevisiae was extracted with ethanol without disruption of the cells. The effects of ethanol concentration, extraction temperature and extraction time were assessed by using 2(3) full factorial designs (FFD). Preliminary studies showed that ethanol concentration had the most influence on GSH yield by ethanol extraction, based on the first order regression coefficients derived using MINITAB software, and an optimal ethanol concentration (25%, v/v) was obtained. However, compared to the conventional extraction technique (hot water extraction), there was no significant advantage in yield of GSH from yeast cells using ethanol extraction under these optimized conditions. But ethanol extraction has several advantages, such as lower energy consumption and lower protein concentration of extraction broth, which may reduce the complexity and cost of the purification process. Hence, ethanol extraction which does not disrupt yeast cells could be an inexpensive, simple and efficient alternative to conventional extraction techniques in the GSH industry.