构建酿酒酵母工程菌合成香紫苏醇

香紫苏醇是一种来源于植物的双环二萜醇,常用于香味成分且具有重要生物学活性。为实现香紫苏醇的微生物生产,以酿酒酵母为宿主,表达焦磷酸赖百当烯二醇酯合酶和香紫苏醇合酶,构建香紫苏醇的人工生物合成途径。发现过表达前体代谢关键酶、蛋白质融合增强底物通道效应及去除异源蛋白信号肽等,有利于香紫苏醇合成。在摇瓶培养条件下,组合优化得到的工程菌株S6的香紫苏醇产量达到8.96 mg/L。研究结果对其他萜类化合物的异源生物合成具有参考价值。     

酿酒酵母工程菌高密度培养生产香紫苏醇

为提高酿酒酵母工程菌S7香紫苏醇产量,采用摇瓶培养,研究了其生长和代谢特点,发现产物合成与菌体生长密切关联。在3 L发酵罐中通过补料-溶氧联动控制的方式,以葡萄糖、乙醇和葡萄糖/乙醇混合物为碳源进行高密度培养,香紫苏醇产量分别达到253 mg/L、386 mg/L和408 mg/L,最高产量是摇瓶培养的27倍。说明添加乙醇作为碳源有助于香紫苏醇合成。研究结果对优化酿酒酵母细胞工厂,高效生产萜类化合物具有重要参考价值。     

香紫苏开花期蒸腾和光合作用日变化特征及其影响因子研究

对开花期香紫苏功能叶片的蒸腾作用、光合作用日变化规律以及二者与环境因子之间的相互作用进行了研究.结果表明:处于花期的香紫苏蒸腾速率和气孔导度的日变化总体趋势是先升高后降低;不同花期净光合速率的日变化之间则存在明显的差别,初花期的净光合速率明显高于盛花期和终花期;湿度和光合有效辐射是影响香紫苏蒸腾作用的最主要因素,而光合有效辐射和CO2浓度变化则对香紫苏的净光合速率影响最大.     

大肠杆菌通过甲羟戊酸途径合成紫苏醇

紫苏醇,即[4-异丙烯基-1-环己烯]甲醇,是一种具有类似芳樟醇和松油醇特殊气味的单环单萜烯醇。在医药、食品和化妆品等行业具有广阔市场空间和研究价值。文中研究了以工程大肠杆菌通过甲羟戊酸途径合成紫苏醇的方法。首先在大肠杆菌中构建来源于粪肠球菌的MVA代谢途径合成柠檬烯,随后柠檬烯通过细胞色素P450烷烃羟化酶的羟基化转化为紫苏醇。然后将构建的紫苏醇合成菌株在摇瓶发酵条件下进行优化,研究发现工程大肠杆菌以葡萄糖为原料,通过MVA代谢途径可合成约50.12 mg/L的紫苏醇。本研究构建合成紫苏醇的MVA代谢途径也可用于其他萜类化合物的合成,为今后生物法合成萜类化合物提供了理论依据和技术支持。     

香紫苏及其栽培技术

香紫苏,学名salvia sclarea L,别名莲座鼠尾草,南欧丹参。香紫苏是一种高产量的农作物,其鲜花和种子的深加工产品,具有较离的经济价值。香紫苏鲜花精油主要用于配制日用化妆品香精,也用于食品和制酒工业。种子油脂用于陶器和瓷器生产,花是很好的蜜源。香紫苏亩产鲜花450至600公斤,鲜花含油量因年份而不同。种植第一年、第三年较高,第二年略低。亩产精油量:第一年1.31至1.74公斤,第二年0.7至0.95公斤,第三年0.99至1.32公斤。亩产种子,直播四26.6公斤,移载田26.4公斤。香紫苏精油理化性质,色泽淡黄色或橙黄笆,相对密度(25℃)0.906～0.925,折见指     

干旱胁迫对香紫苏生理特性和光合特性的影响研究

目的:研究土壤干旱胁迫对香紫苏生理特性和光合特性的影响.方法:以香紫苏(Salvia sclarea L)的功能叶为研究对象,分实验组和对照组进行盆栽实验,在随后的5天中,分别测定两组香紫苏叶片中脯氨酸、丙二醛和叶绿素的含量,同时测定并分析蒸腾速率、气孔导度、净光合速率、细胞间隙CO2浓度的变化情况.结果:随着干旱胁迫程度的加重,香紫苏叶片内脯氨酸的含量总体趋势增加,与干旱胁迫的时间正相关;而叶片中丙二醛的含量变化不明显;干旱胁迫时,叶片内叶绿素的含量升高,在第四天时达到最大值,随干旱胁迫的加重,叶绿素含量又呈下降的趋势.其蒸腾速率变化曲线呈双峰型,随干旱时间的延长,蒸腾速率明显下降;香紫苏气孔导度变化曲线为单峰型,当干旱胁迫加重时,香紫苏气孔导度下降明显,气孔导度与胞间CO2浓度呈负相关,净光合速率变化与气孔导度变化曲线较为一致.结论:香紫苏生理因子和光合作用对干旱胁迫有一定的适应能力,但重度胁迫会对其造成严重影响.     

水稻害虫生态调控系统中推-拉策略的初步应用

本文在江门市新会区水稻试验田,构建利用"推-拉"策略的生态调控系统,以紫苏、烟草和香根草分别作为"推"和"拉"的成分,并对紫苏、烟草和香根草的生态功能进行了评价。Y型嗅觉仪测定结果表明,白背飞虱和褐飞虱对"紫苏+水稻"组选择率分别为38.70%和33.87%;对"烟草+水稻"组的选择率分别为30.12%和39.65%;通过笼罩选择性测定,白背飞虱和褐飞虱对"紫苏+水稻"组选择率分别为33.90%和38.10%;对"烟草+水稻"组的选择率分别为34.74%和39.00%,均显著低于对"水稻"对照组的选择率(60%-70%)。通过Y型嗅觉仪和笼罩选择性测定,拟水狼蛛对"香根草+水稻"组的选择率分别为67.55%和66.07%,显著高于对"水稻"的选择率32.45%和33.93%。进一步在田间试验区调查,结果表明,在"推-拉"处理区白背飞虱和褐飞虱的种群数量分别为563.4头/百丛和490.7头/百丛,显著低于对照田(819.6头/百丛和763.8头/百丛);在水稻分蘖后期处理区蜘蛛数量(213.33头/百丛)极显著高于对照田(80.00头/百丛),同时在稻田旁的香根草上能发现栖息的蜘蛛和卵块。由此得出,稻田边种植紫苏和烟草对稻飞虱有明显的驱避作用,而稻田边种植香根草对蜘蛛具有引诱作用,形成了紫苏、烟草和香根草的"推-拉"生态调控系统,既减少了害虫稻飞虱的数量,又增加了天敌蜘蛛数量,其结果可为生态防治害虫及科学保护天敌提供科学依据。     

尖尾芋石油醚部位化学成分及其乳腺癌细胞毒活性研究

研究天南星科海芋属植物尖尾芋Alocasia cucullata(Lour.) Schott石油醚部位化学成分。采用硅胶、Sephadex LH-20等色谱方法进行分离纯化,运用波谱学技术进行结构鉴定。采用CCK-8法测试化合物对人乳腺癌MDA-MB-231、MCF-7细胞及小鼠乳腺癌4T1细胞的体外细胞毒活性。从尖尾芋根茎石油醚部位中分离得到15个化合物,分别为对羟基苯甲醛(1)、苯甲酸(2)、香草酸乙酯(3)、没食子酸乙酯(4)、7-羟基香豆素(5)、肉豆蔻酸甲酯(6)、角鲨烯(7)、齐墩果酸(8)、熊果酸(9)、香紫苏醇(10)、香紫苏内酯(11)、原儿茶醛(12)、原儿茶酸(13)、二十八烷醇(14)和没食子酸(15),化合物2～15为海芋属首次分离得到,所有化合物均为尖尾芋首次分离得到。乳腺癌细胞毒活性研究显示：化合物4、8和9对3种乳腺癌细胞均有一定的抑制作用。     

烟草与紫苏科间远缘杂种后代化学成分及医药成分的研究

为利用药用植物资源以缓解吸烟与健康的矛盾,通过创新育种的方法将普通烟草品种78-04与紫苏进行科间远缘杂交,对选育出的新型烟草稳定品系—紫苏烟的内在化学成分及医药成分进行分析。酯酶与过氧化物酶同工酶分析证实,通过药用植物与烟草基因间的渐渗、交流以及重组,紫苏烟中转入了父本紫苏的遗传物质。烟叶主要化学成分分析显示,紫苏烟具有低糖、中高烟碱的特点,芸香苷的含量比亲本普通烟草品种78-04提高了70.4%。气相色谱-质谱联用技术初步分析表明,紫苏烟中含有α-丁香烯、β-丁香烯、芳樟醇和薄荷醇等紫苏挥发油的成分,与紫苏比较其薄荷醇的含量提高了2倍多;并产生了α-桉醇、香叶烯、桉油精、愈创木奥、α-郁金烯和γ-榄香烯等亲本所没有的医药成分和香料成分。药用植物与普通烟草科间远缘杂交能创造变异、产生烟草新类型,对降低卷烟产品的危害性具有一定作用,紫苏烟在医药和食品等领域也具有很高的综合利用价值。     

山茱萸抗过敏活性组分的筛选及其对RBL-2H3细胞脱颗粒的作用CSCD

为研究山茱萸中抗过敏活性成分,采用透明质酸酶抑制实验对山茱萸中活性组分进行初步筛选;采用细胞脱颗粒抑制实验对透明质酸酶抑制活性较高的组分进一步评估;采用气相色谱-质谱联用法(GC-MS)进行成分分析。结果表明山茱萸石油醚部位(petroleum ether extract of Corni Fructus,COPE)具有显著透明质酸酶抑制活性,其IC50值为3.11 mg/mL;经硅胶柱层析分离得到4个透明质酸酶抑制活性更高的组分,其中组分COPE-8对RBL-2H3细胞脱颗粒抑制作用更显著,呈剂量依赖形式且具低细胞毒性。经20μg/mL COPE-8处理后,RBL-2H3细胞脱颗粒释放的组胺和β-氨基己糖苷酶分别减少46%和39%;经GC-MS分析,COPE-8中主要活性物质为萜类物质,包括α-香树脂醇、β-香树脂醇和香紫苏醇。结果表明COPE-8具有显著的抗过敏活性,在抗过敏活性物开发方面具有一定的应用价值。     

Terpenoids from Salvia palaestina

Six known terpenoids: vergatic acid, ursolic acid, crataegolic acid, lupane-3β-,11α, 20-triol, sclareol and sitosteryl 3β-glucoside were isolated from the leaves of Salvia palaestina and were identified by spectral data. Among the compounds, sclareol showed high activity against Staphylococcus aureus, S. epidermis, Escherichia coli, Proteus vulgaris and Pseudomonas aeruginosa, while the triterpenoids were not tested due to solubility problems.     

Enzymatic biotransformation of terpenes as bioactive agents

The plant-derived terpenoids are considered to be the most potent anticancer, anti-inflammatory and anticarcinogenic compounds known. Enzymatic biotransformation is a very useful approach to expand the chemical diversity of natural products. Recent enzymatic biotransformation studies on terpenoids have resulted in the isolation of novel compounds. 14-hydroxy methyl caryophyllene oxide produced from caryophyllene oxide showed a potent inhibitory activity against the butyryl cholinesterase enzyme, and was found to be more potent than parent caryophyllene oxide. The metabolites 3β,7β-dihydroxy-11-oxo-olean-12-en-30-oic acid, betulin, betulonic acid, argentatin A, incanilin, 18β glycyrrhetinic acid, 3,11-dioxo-olean-12-en-30-oic acid produced from 18β glycyrrhetinic acid were screened against the enzyme lipoxygenase. 3,11-Dioxo-olean-12-en-30-oic acid, was found to be more active than the parent compound. The metabolites 3β-hydroxy sclareol 18α-hydroxy sclareol, 6α,18α-dihydroxy sclareol, 11S,18α-dihydroxy sclareol, and 1β-hydroxy sclareol and 11S,18α-dihydroxy sclareol produced from sclareol were screened for antibacterial activity. 1β-Hydroxy sclareol was found to be more active than parent sclareol. There are several reports on natural product enzymatic biotransformation, but few have been conducted on terpenes. This review summarizes the classification, advantages and agents of enzymatic transformation and examines the potential role of new enzymatically transformed terpenoids and their derivatives in the chemoprevention and treatment of other diseases.     

The role of acid catalysis in the genesis of amber

Analysis of the hydrocarbon fraction from baltic amber is described. Transformations which have occurred in resins during the formation of amber are discussed on the grounds of acid-catalysed reactions undergone by 7,13-abietadiene and sclareol.     

Mitochondria-targeted liposomes improve the apoptotic and cytotoxic action of sclareol

Current efforts toward improving the effectiveness of drug therapy are increasingly relying on drug-targeting strategies to effectively deliver bioactive molecules to their molecular targets. Pharmaceutical nanocarriers represent a major tool toward this aim, and our efforts have been directed toward achieving nanocarrier-mediated subcellular delivery of drug molecules with mitochondria as the primary subcellular target. Meeting the need for specific subcellular delivery is essential to realizing the full potential of many poorly soluble anticancer drugs. In this article, we report that mitochondria-targeted liposomes significantly improve the apoptotic and cytotoxic action of sclareol, a poorly soluble potential anticancer drug. The results support the broad applicability of our nanocarrier-mediated subcellular targeting approach as a means to improve the effectiveness of certain anticancer therapeutics.     

Application of Heavy Metal Rich Tannery Sludge on Sustainable Growth,Yield and Metal Accumulation by Clarysage (Salvia sclarea L.)

A field experiment was conducted to evaluate the effective utilization of tannery sludge for cultivation of clarysage (Salvia sclarea) at CIMAP research farm, Lucknow, India during the year 2012–2013. Six doses (0, 20, 40, 60, 80, 100 tha^?1) of processed tannery sludge were tested in randomised block design with four replications. Results revealed that maximum shoot, root, dry matter and oil yield were obtained with application of 80 tha^?1of tannery sludge and these were 94, 113 and 61% higher respectively, over control. Accumulation of heavy metals (Cr, Ni, Fe, Pb) were relatively high in shoot portion of the plant than root. Among heavy metals, magnitude of chromium accumulation was higher than nickel, iron and lead in shoot as well as in root. Linalool, linalyl acetate and sclareol content in oil increased by 13,8 and 27% respectively over control, with tannery sludge application at 80 tha^?1. Heavy metals such as chromium, cadmium and lead content reduced in postharvest soil when compared to initial status. Results indicated that clarysage (Salvia sclarea) can be grown in soil amended with 80 tha^?1sludge and this can be a suitable accumulator of heavy metals for phytoremediation of metal polluted soils.     

A comparative study of the effects of cholesterol and sclareol, a bioactive labdane type diterpene, on phospholipid bilayers

Sclareol (labd-14-ene-8,13-diol) is a highly water-insoluble molecule that belongs to the labdane type diterpenes and is characterized as a biologically active molecule, due to its cytotoxic and cytostatic effects against human leukemic cell lines. A superimposition study between sclareol and cholesterol, based on their corresponding hydrophobic and polar molecular segments calculated from their lipophilic profiles, revealed their spatial similarities. This structural similarity between the two molecules prompted us to compare their effects on the structure and stability of phospholipid dipalmitoylphosphatidylcholine (DPPC) membranes. Differential scanning calorimetry (DSC) was applied to compare the thermal changes caused by either cholesterol or sclareol when are incorporated in DPPC bilayers. The results showed that sclareol is incorporated into phospholipid model membranes and mimics the thermal effects of cholesterol especially at concentrations up to X(sclareol)=9.1 mol%. These effects can be summarized as the abolition of pre-transition, lowering of the main phase transition and reduction of the enthalpy change (DeltaH) of the gel to liquid-crystalline phase transition of DPPC bilayers. At concentrations X> or =16.7 mol%, sclareol and cholesterol caused different heterogeneity in lipid bilayers or a reversible transition from a vesicular suspension to an extended peak bilayer network. This different fluidization, exerted by the two molecules at high concentration, may be related to their different stability and the z-average mean diameter of the liposomes they form. Small unilamellar vesicles, prepared by the thin film hydration method showed that DPPC bilayers containing a high concentration of sclareol in equimolar ratio sclareol:cholesterol were unstable, in contrast to the ones containing only cholesterol.     

Sclareol induces apoptosis in human HCT116 colon cancer cells <Emphasis Type="Italic">in vitro</Emphasis> and suppression of HCT116 tumor growth in immunodeficient mice

Labd-14-ene-8, 13-diol (sclareol) is a labdane-type diterpene, which has demonstrated significant cytotoxic activity against human leukemic cell lines, but its effect on solid tumor-derived cells is uknown. Here, we demonstrate that addition of sclareol to cultures of human colon cancer HCT116 cells results in inhibition of DNA synthesis, arrest of cells at the G₁ phase of the cell cycle, activation of caspases-8, -9, PARP degradation, and DNA fragmentation, events characteristic of induction of apoptosis. Intraperitoneal (ip) administration of sclareol alone, at the maximum tolerated dose, was unable to induce suppression of growth of HCT116 tumors established as xenografts in immunodeficient SCID mice. In contrast, ip administration of liposome-encapsulated sclareol, following a specific schedule, induced suppression of tumor growth by arresting tumor cell proliferation as assessed by detecting the presence of the cell proliferation-associated nuclear protein, Ki67, in thin tumor sections. These findings suggest that sclareol incorporated into liposomes may possess chemotherapeutic potential for the treatment of colorectal and other types of human cancer.