Seven new triterpene glycosides from the pericarps of Stryphnodendron fissuratum

Seven new triterpene glycosides on the basis of the lupane skeleton (1–7) were isolated from the pericarps of Stryphnodendron fissuratum (Leguminosae). The structures of 1–7 were determined on the basis of extensive spectroscopic analysis, including two-dimensional NMR data, and the results of hydrolytic cleavage.     

无患子果实生长发育及其内含物的变化特征北大核心CSCD

以福建建宁县无患子为材料,观测果实生长动态,并采用Logistic方程对生长指标进行曲线拟合,观察生长过程中果皮显微结构变化,确定无患子果实生长发育的重要时期,为无患子的高效栽培管理和果实采收策略的制定提供科学依据。结果表明:(1)无患子果实生长、果皮总皂苷和种子油脂的积累规律相似,总体上呈Logistic增长模型的单“S”型曲线;果皮总皂苷和种子油脂的主要积累时期分别为45~90 DAP(花授粉后天数)和75~105 DAP。(2)无患子果皮除含有角质层、表皮细胞、厚角细胞、薄壁细胞、维管束、石细胞等基本结构外,还含有溶生式分泌腔和草酸钙簇晶等特征性结构;随着果实生长,分泌腔体积逐渐变大。(3)根据果实生长变化规律和Logistic方程拟合结果,可将无患子果实生长发育进程划分为生长初期(0~30 DAP)、速生期(30~90 DAP)、生长后期(90~120 DAP)和果实成熟期(120~150 DAP)4个阶段。在生产实践中果实成熟期内均适合果实采收,其中135和150 DAP分别为果实皂用和油用采收的最佳时期,此外还应根据每年天气状况对果实采收时间进行适当调整。     

New triterpene oligoglycosides from the Caribbean sponge Erylus formosus

Seven new triterpene glycosides, erylosides R₁ (1), T₁ (3), T₂ (4), T₃ (5), T₄ (6), T₅ (7), and T₆ (8) along with the known formoside (2) were isolated from the sponge Erylus formosus collected along the Caribbean coast of Mexico. Glycoside 1 was determined as a trisaccharide, glycoside 2 as a tetrasaccharide while glycosides 3–8 were hexasaccharide. Their carbohydrate chains were unprecedented and have never been found in oligosaccharides from other biological sources, except Erylus spp. Three carbohydrate chains in the glycosides 3 and 6, 4 and 7, 5 and 8 correspondingly are new. The glycosides 1–5 have penasterol as aglycone while glycosides 6–8 proved to be glycoconjugates of 24-methylene-14-carboxy-lanost-8(9)-en-3β-ol.     

雷州半岛风水林中无患子的空间分布格局与空间关联性

为探讨无患子(Sapindus saponaria)在生态修复和城市园林中的应用,在广东湛江雷州市龙门镇足荣村无患子风水林设置1 hm2样地,调查群落的物种组成,并采用点格局分析法,结合完全随机零模型和异质泊松零模型,分析无患子的空间分布格局与关联性.结果表明,样地中胸径≥1 cm的乔木共计28科60属73种3585株...     

Triterpenoid saponins from the fruits and galls of Sapindus mukorossi

Six saponins, sapinmusaponin K (1) [hederagenin-3-O-(3-O-acetyl-alpha-L-arabinopyranosyl)-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside], sapinmusaponin L (2) [hederagenin-3-O-(4-O-acetyl-alpha-L-arabinopyranosyl)-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabino-pyranoside], sapinmusaponin M (3) [hederagenin-3-O-(2,3-O-diacetyl-beta-D-xylopyranosyl)-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside], sapinmusaponin N (4) [hederagenin-3-O-(2,4-O-diacetyl-beta-D-xylopyranosyl)-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside], sapinmusaponin O (5) [3,7,20(S)-trihydroxydammar-24-ene-3-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranoside], and sapinmusaponin P (6) [3,7,20(R)-trihydroxydammar-24-ene-3-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-d-glucopyranoside], along with seven known saponins (7-13), were isolated from fruits and the galls of Sapindus mukorossi. Their structures were elucidated by 1D and 2D NMR spectroscopic techniques and acid hydrolysis. Biological evaluation indicated that saponins 1-4 and 7-13 showed moderate cytotoxicity against several human tumor cell lines.     

西南丘陵地区紫色土酸性对无患子幼树生长和光合特性的影响

探讨无患子(Sapindus mukorossi Gaertn.)生长和光合作用对酸性土壤胁迫的响应机制,为其高效种植提供理论依据。采用随机区组实验设计,以重庆市紫色土不同程度酸性区(pH分别为7.56、5.65、4.41)3年生无患子幼树为研究对象,分析紫色土酸性对无患子幼树生长、叶性状指标和光合作用的影响。结果表明:中性区土壤速效氮(AN)、速效磷(AP)含量极显著低于酸性区,中性区叶片全氮(TN)、全钾(TK)含量极显著高于酸性区(P<0.01)。与中性区相比,酸性区幼树的株高、净光合速率(P_n)、总叶绿素(Chls)、叶绿素a(Chl a)、类胡萝卜素(Cars)含量、叶绿素a/b(Chl a/b)比值极显著减小(P<0.01),最大光化学效率(F_v/F_m)和非光化学淬灭系数(qN)显著减少(P<0.05),而叶绿素b(Chl b)含量和叶绿素/类胡萝卜素(Chls/Cars)比值极显著增加(P<0.01),冠幅、电子传递效率(ETR)、光化学淬灭系数(qP)、实际原初光能捕获效率(Y(Ⅱ...     

Sesquiterpenes and triterpenoids from the rhizomes of Alisma orientalis and their pancreatic lipase inhibitory activities

Five new terpenoids, including three sesquiterpenes 11-hydroxy-8-ox-alismoxide (1), 11-oxo-13-nor-alismol (2), and 1β,11-dihydroxy-β-cyperone (3), and two protostane-type triterpenoids 16β-acetoxy alisol B (4) and 16α-acetoxy alisol B (5) were isolated from the rhizomes of Alisma orientalis together with 11 known compounds. Their structures were established using 1D and 2D NMR and HRESIMS spectroscopic analyses. The isolated compounds were assayed for their inhibitory activities against pancreatic lipase. Compounds 6 and 13 showed inhibitory effects with IC₅₀ values of 64.4 and 45.5 μM, respectively.     

无患子光合生理日变化及其与生理生态因子的关系

以8年生无患子植株为试验材料,采用便携式Li-6400光合测定仪,分别于花期和果期测定了其净光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)、胞间CO2浓度(Ci)、空气温度(Ta)、叶片光合有效辐射(PAR)、空气相对湿度(RH)和空气CO2浓度(Ca)等参数的日变化,以揭示无患子光合特征及其与主要环境因子的关系。结果显示:(1)无患子Pn和Tr在花期和果期日变化均呈"M"型双峰曲线,并存在"光合午休"现象,第1和第2峰值分别出现在10:00和14:00左右。(2)无患子花期Pn、Tr、Gs、水分利用效率(WUE)日均值高于果期,而果期的Ci日均值高于花期。(3)在8:00~14:00,无患子花期光能利用效率(LUE)大于果期,并在10:00左右出现峰值,而在14:00~18:00其果期LUE大于花期。(4)无患子叶片Pn与Tr、Gs、PAR具有极显著正相关关系,且与Gs关系最为密切,与Ci呈极显著负相关关系,与Ca则表现为显著负相关关系。(5)影响无患子叶片Pn变化的决定生理因子是Gs和Tr,主要限制生理因子是Ci;主要决定生态因子是RH,主要限制生态因子是Ta,且Gs是影响无患子Pn的最重要的生理生态因子。研究表明,无患子对光强有较强的适应能力,是喜光植物,这为其人工林定向培育,以及花期和果期的管理提供了一定理论依据。     

有机栽培龙脑百里香精油对白色念珠菌体外抑菌活性的研究

【背景】白色念珠菌(Candidaalbicans)属于条件致病性真菌,可引起严重的黏膜真菌感染及全身系统性真菌感染,是导致患者高发病率和高死亡率的主要菌群之一。【目的】探究百里香精油对白色念珠菌的抑菌活性及抑制机理。【方法】测定5种百里香精油对白色念珠菌的抑菌圈直径,分析具有高抑菌活性的精油成分。在此基础上,通过扫描电子显微镜(scanning electron microscope, SEM)观察精油对白色念珠菌菌体细胞形态的影响。测定碱性磷酸酶(alkaline phosphatase, AKP)含量、胞外溶液电导率并进行碘化丙啶(propidium iodide, PI)染色分析,探究精油对白色念珠菌生物膜的形成与黏附及磷脂酶活性的影响,并通过实时荧光定量PCR法分析与白色念珠菌生物膜形成相关基因(凝集素样序列基因ALS4,从酵母型向菌丝型细胞的形态转变基因HWP1、磷脂酶基因PLB1)的表达水平,探究该精油对白色念珠菌的抑菌机制。【结果】筛选出了对白色念珠菌高度敏感的有机栽培龙脑百里香精油(Thymus vulgaris CT borneol essential oil, T...     

三孢布拉霉crgA启动子的克隆和活性分析

【背景】CrgA是三孢布拉霉(Blakesleatrispora,Bt)中调控类胡萝卜素合成的关键负调控因子,其表达水平会影响类胡萝卜素的合成。【目的】克隆三孢布拉霉crgA启动子并分析其活性,为进一步解析CrgA表达调控机制奠定基础。【方法】通过综合微生物基因组(integrated microbial genomes, IMG)数据库提供的基因组序列,克隆crgA翻译起始位点上游2 000 bp序列,分析其顺式调控元件和转录起始区域预测,通过RT-qPCR分析不同光照时间对三孢布拉霉crgA相对转录水平的影响;构建4个不同长度的crgA启动子截短序列驱动的GUS-mGFP5重组表达载体p1303-procrgAF、F1、F2和F3,利用农杆菌侵染整合到三孢布拉霉基因组中,在黑暗和光照条件下测定β-D-葡萄糖苷酸酶(β-D-glucuronidase,GUS)酶活性并观察荧光信号。【结果】crgA启动子不仅包含基础的TATA-box和CAAT-box元件,还包括多个与光响应相关的元件。观察荧光结果显示CaMV35S和构建的4个突变启动子均能在三孢布拉霉体内驱动下游基因表达,检测GUS...     

Halimodendrin I, a new acylated triterpene glycoside from Halimodendron halodendron (Fabaceae)

Halimodendrin I, a new acylated triterpene glycoside (1), was isolated and chemically characterized as 3β-O-palmitoyl-28-[3′-palmitoyl-β-d-glucopyranosyl]-olean-12-en-28-oic acid from the aerial part of Halimodendron halodendron (Fabaceae) by IR, 1D and 2D NMR, HR-ESI-MS and LR-ESI-MS experiments. In addition, seven known compounds were isolated and identified as: palmitic acid, glycerol-2-linoleneate, glycerol-1,3-dilinoleneate, ferulic acid, 3-O-methylquercetin, β-sitosterol, and β-sitosterol-3-O-β-d-glucopyranoside. Nine fatty acids were identified and quantified in the saponifiable matter of the hexane extract. These fatty acids are: myristic, n-pentadecanoic, palmitoleic, palmitic, linoleic, oleic, stearic, arachidic, and behenic acids. The volatile oil was isolated by hydrodistillation (0.013%, w/w) with unpleasant smell. Twenty-seven components were identified in the oil by GC/MS.     

基于空腔填充效应构建伯克霍尔德菌脂肪酶LipA的热稳定性突变体

【目的】中温伯克霍尔德菌胞外脂肪酶LipA在工业领域具有重要的应用价值。利用蛋白质工程技术来提高其热稳定性,对开发脂肪酶LipA酶制剂及提高其应用范围及应用效果,具有重要的意义。【方法】利用生物信息学软件Castp、Voronoia和Cave分析LipA分子中存在的空腔及其组成氨基酸残基;利用FoldX软件构建上述氨基酸残基的突变体电子文库,并基于空腔效应(体积变小)、自由能变化值(降低)和空间结构特点等对前述突变体电子文库进行筛选。从突变体电子文库中选择具有代表性的突变体,通过基因工程技术,引入突变。经诱导表达后,实验验证并筛选出热稳定性的突变体。【结果】构建了一个由58个突变体组成的电子文库;并对其中17个代表性的突变体进行了实验验证;筛选到2个热稳定性有明显提高的突变体LipA-His15Pro和LipA-Ala210Val;其叠加突变体LipA-His~(15)Pro/Ala~(210)Val的T50~(12)较野生型LipA提高了8°C,在55°C下的半衰期较野生型脂肪酶LipA提高了23.1倍。【结论】基于空腔填充技术构建热稳定性伯克霍尔德菌胞外脂肪酶LipA突变体,是一种行之有效的策略。     

Molecular modeling of substrate selectivity of Candida antarctica lipase B and Candida rugosa lipase towards c9, t11- and t10, c12-conjugated linoleic acid

Molecular modeling was used to clarify the mechanism of the selectivity of Candida antarctica lipase B and Candida rugosa lipase towards cis9, trans11 (c9, t11-) and trans10, cis12 (t10, c12-) conjugated linoleic acid. Hydrogen bonds network, substrate conformation, binding affinity and water molecules in the binding site were analyzed. Substrate conformation and binding affinity were not correlated with the experimental results of the substrate selectivity. On the contrary, better enzyme preference towards a substrate was correlated with two stronger hydrogen bonds (His-NH-O_a and His-NH-Ser-O_γ) and less water molecules between the substrate the binding pocket. Possible explanation of these was discussed.     

马醉木二萜成分及其乙酰胆碱酯酶抑制活性研究

为研究马醉木叶中的二萜成分及其乙酰胆碱酯酶抑制活性,该研究根据薄层色谱显色特征,使用硅胶、MCI和半制备高效液相等色谱技术对其进行分离与纯化,并通过波谱数据(NMR和MS)分析且结合文献报道数据进行对比,鉴定了所得化合物的结构,同时采用Ellman法首次对其乙酰胆碱酯酶抑制活性进行评价。结果表明：从马醉木叶中分离并鉴定了8个二萜化合物,分别是pierisformoside F (1)、3-epi-grayanotoxinⅩⅧ(2)、3-epi-grayanotoxin B (3)、asebotoxin-X (4)、pierisformosin B (5)、asebotoxinⅢ(6)、rhodojaponinⅢ(7)和pierisformosin C (8)。其中,化合物1为首次从该植物中分离得到,化合物8表现出乙酰胆碱酯酶抑制活性。综上表明,马醉木中含有丰富的二萜成分和活性成分。该研究结果丰富了马醉木的化学成分多样性,为其后续综合开发和利用提供了一定的理论依据,也为寻求更多的活性成分提供了借鉴。     

Efficient, galactose-free production of Candida antarctica lipase B by GAL10 promoter in Δgal80 mutant of Saccharomyces cerevisiae

An efficient yeast gene expression system with GAL10 promoter that does not require galactose as an inducer was developed using Δgal80 mutant strain of Saccharomyces cerevisiae. We constructed several combinations of gal mutations (Δgal1, Δgal80, Δmig1, Δmig2, and Δgal6) of S. cerevisiae and tested for their effect on efficiency of recombinant protein production by GAL10 promoter using a lipase, Candida antarctica lipase B (CalB), as a reporter. While the use of Δgal1 mutant strain required the addition of a certain amount of galactose to the medium, Δgal80 mutant strain did not require galactose. Furthermore, it was found that the recombinant CalB could be produced more efficiently (1.6-fold at 5 L-scale fermentation) in Δgal80 mutant strain than in the Δgal1 mutant. The Δgal80 mutant strain showed glucose repressible mode of expression of GAL10 promoter. Using Δgal80 mutant strain of S. cerevisiae, CalB was efficiently produced in a glucose-only fermentation at volumes up to 500 L.     

Immobilization of Candida rugosa lipase on sporopollenin from Lycopodium clavatum

Sporopollenin is a natural polymer obtained from Lycopodium clavatum, which is highly stable with constant chemical structure and has high resistant capacity to chemical attack. In this study, immobilization of lipase from Candida rugosa (CRL) on sporopollenin by adsorption method is reported for the first time. Besides this, the enzyme adsorption capacity, activity and thermal stability of immobilized enzyme have also been investigated. It has been observed that under the optimum conditions (Spo-E_(0.3)), the specific activity of the immobilized lipase on the sporopollenin by adsorption was 16.3 U/mg protein, which is 0.46 times less than that of the free lipase (35.6 U/mg protein). The pH and temperature of immobilized enzyme were optimized, which were 6.0 and 40 °C respectively. Kinetic parameters V_max and K_m were also determined for the immobilized lipase. It was observed that there is an increase of the K_m value (7.54 mM) and a decrease of the V_max value (145.0 U/mg-protein) comparing with that of the free lipase.     

浙江低山地区多用途植物无患子的开花物候特征

无患子(Sapindus mukorossi Gaertn.)是我国长江以南地区传统的重要绿化树种,其果皮富含皂苷,种仁富含油脂,是国家林业局审定的新型木本油料树种之一。为研究多用途树种无患子在浙江低山地区的开花特征,2012年和2013年连续两年对位于浙江省天台县9年生无患子人工林在群体、个体、花序和单花水平进行开花物候观测和比较,并运用开花日期、相对开花强度和同步性等指数研究了无患子开花物候特征。观察结果显示:无患子花呈浅黄白色,花的类型有雄花和两性花,没有发现雌花。雄花较两性花大(花径分别为5.09 mm和3.72 mm),雄蕊多为8枚,个别7或9、10枚;雌蕊退化仅留下浅绿色凸起。两性花花萼稍抱拢,花药藏于花被片下,雄蕊大多8枚,极少数7枚或9枚,柱头高于花药并伸出花苞,子房一般具3室,极少数4室或仅2室。无患子2012年的开花进程略早于2013年,其花期集中在在5月中旬至6月上旬,单花从花蕾膨大到花朵凋谢一般为8—9 d。在2012年和2013年,无患子在群体、个体和花序水平的花期约为30 d、20 d、11 d和28 d、19 d、13 d。个体水平的开花振幅均呈单峰曲线,年际间相似性较高;开花同步性在个体水平同步性较高(同步指数为0.868),表现出一种大量、集中的开花式样;相对开花强度在单株间分布范围相对宽泛,但主要分布在30%—40%,在年际间和年际内均呈现极显著差异。花期同步指数在两年的变异范围分别是0.81—0.97和0.70—0.98,不同单株开花同步性在年际内差异极显著,但在年际间差异性则不显著。由此可见,无患子的生殖资源分配存在明显的时空差异,较长的花期可以减少非法花粉的干扰、保持其种群基因多样性,遗传因子是决定无患子开花物候的主要因素,生态环境对无患子开花物候的影响还需进一步研究,本研究以期为探索无患子开花的主要限制因子奠定基础。     

Chaperone-mediated activation in vivo of a Pseudomonas cepacia lipase

An extracellular Pseudomonas cepacia lipase, LipA, is inactive when expressed in the absence of the product of the limA gene. Evidence has been presented that LimA is a molecular chaperone. The lipA and limA genes have been cloned in separate and independently inducible expression systems in Escherichia coli. These systems were used to test the molecular chaperone hypothesis by investigating whether LimA could activate presynthesized prelipase and whether presynthesized LimA could activate newly synthesized prelipase. The results show that LimA cannot activate presynthesized prelipase and that presynthesized LimA can activate only a limited number of de novo synthesized prelipase molecules. Co-immunoprecipitation of prelipase/lipase with LimA generated a 1:1 complex of prelipase/lipase and LimA. The results suggest that a 1:1 complex of LipA and LimA is required for prelipase processing and secretion of active lipase.     

Xanthones from the stems of Cudrania tricuspidata and their inhibitory effects on pancreatic lipase and fat accumulation

Nine new xanthones, cudracuspixanthones I - Q (12–14, 25, 32–36), and 30 known xanthones (1–11, 15–24, 26–31, 37–39) were isolated from the stems of Cudrania tricuspidata (Moraceae). The structures of isolated compounds were established by using 1D and 2D NMR in combination with HR-TOF-MS. Xanthones from the stems of C. tricuspidata exerted pancreatic lipase inhibitory activity. In addition, cudracuspixanthone P (35), a new xanthone, reduced the fat accumulation in liver cells stimulated with fatty acids. Therefore, these compounds might be beneficial in the treatment of metabolic diseases.