黑茶曲霉属和散囊菌属真菌产胞外水解酶能力的比较分析

【目的】曲霉属和散囊菌属真菌是黑茶渥堆中的优势微生物,对形成黑茶风味、品质和生理功能 起关键作用。但在茶叶发酵过程中这些真菌水解酶的差异尚不清楚。【方法】采用鉴别培养基平板检测和酶活 性检测相结合,分析曲霉属和散囊菌属不同菌株在合成培养基上和茶叶发酵条件下胞外水解酶活性的差异。【结 果】鉴别培养基平板检测结果显示：供试的 9 个菌株均不同程度呈现纤维素酶、木聚糖酶和单宁酶活性,7 个 菌株显示淀粉酶活性,2 个菌株显示果胶酶活性,1 个菌株显示蛋白酶活性。同一菌株可以同时分泌多种胞外 水解酶,多个菌株也可以表现同一水解酶的活性,不同菌株产生胞外水解酶能力存在显著差异。在茶叶发酵过 程中,黑曲霉菌株 PE-3 水解大分子碳水化合物酶活性显著高于冠突散囊菌菌株 PE-1。其中,淀粉酶活力达到 33.81 U/mL。而菌株 PE-1 的单宁酶和蛋白酶活性则显著高于菌株 PE-3。其中,单宁酶活力达到 132.09 U/mL。【结 论】在茶叶发酵过程中,曲霉属和散囊菌属的菌株可能具有对不同茶叶成分的选择性,水解酶的协同作用影响 菌株对茶叶成分的利用效率。     

高效液相色谱法测定新促孕液中淫羊藿甙的含量

通过流动相筛选和线性关系考察、精密度试验、稳定性试验、重复性试验、加样回收试验和空白试验建立了淫羊藿甙的高效液相色谱法,并测定新促孕液中淫羊藿甙的含量.结果表明,甲醇-水为流动相在6:4时分离度最好,在色谱柱为Kromasil C 18、柱温为室温、流速1 ml/min、检测波长270 nm、进样量20μl的色谱条件下,测定方法稳定可行、精密度较高、线性关系和重复性良好;4批样品中淫羊藿甙的含量在0.106～0.124 mg/ml之间,因而可将含量下限定为0.1 mg/ml,作为控制新促孕液质量的指标之一.     

Synthesis of isoacteoside, a dihydroxyphenylethyl glycoside

The total chemical synthesis of isoacteoside (1), 2-(3,4-dihydroxyphenyl)ethyl 6-O-caffeoyl-3-O-(-l-rhamnopyranosyl)--d-glucopyranoside, is described. An acteoside acetate with benzyl groups at the catechols (3: 2-(3,4-dibenzyloxyphenyl)ethyl 2,6-di-O-acetyl-4-O-[3,4-bis(O-benzyl)caffeoyl]-3-O-(-l-rhamnopyranosyl)--d-glucopyranoside) was treated with a solution of methy-lamine in methanol (MeNH₂ in MeOH) to perform both deacetylation and caffeoyl migration, affording an isoacteoside derivative with benzyl groups at the catechols4b: 2-(3,4-dibenzyloxyphenyl)ethyl 6-O-[3,4-bis(O-benzyl) caffeoyl] -3-O-(-l-rhamnopyranosyl)--d-glucopyranoside —in 34% yield. Debenzylation of4b was successfully accomplished by catalytic transfer hydrogenation using 1,4-cyclohexadiene to give the target compound isoacteoside (1) in 54% yield.¹H and¹³C nuclear magnetic resonance spectral data of the synthesized isoacteoside (1) were identical with those of the natural isoacteoside isolated fromPaulownia tomentosa (Thumb.) Steud.Part of this research was presented at the 51st Annual Meeting of the Japan Wood Research Society, Tokyo, April 2001     

Oceanalin B,a Hybrid α,ω-Bifunctionalized Sphingoid Tetrahydroisoquinoline β-Glycoside from the Marine Sponge Oceanapia sp.

Oceanalin B (1), an α,ω-bipolar natural product belonging to a rare family of sphingoid tetrahydoisoquinoline β-glycosides, was isolated from the EtOH extract of the lyophilized marine sponge Oceanapia sp. as the second member of the series after oceanalin A (2) from the same animal. The compounds are of particular interest due to their biogenetically unexpected structures as well as their biological activities. The structure and absolute stereochemistry of 1 as a α,ω-bifunctionalized sphingoid tetrahydroisoquinoline β-glycoside was elucidated using NMR, CD and MS spectral analysis and chemical degradation. Oceanalin B exhibited in vitro antifungal activity against Candida glabrata with a MIC of 25 μg/mL.     

Effect of host plant alternation on some adaptive enzymes of the bird cherry–oat aphid, <Emphasis Type="Italic">Rhopalosiphum padi</Emphasis> (L.)

Changes in the activity of some adaptive enzymes of the bird cherry–oat aphid R. padi after transfer from primary (bird cherry) to secondary (triticales) host plants were assessed. The following groups of enzymes were studied: (1) transferases—glutathione S-transferase (GST) and UDP-glucosyltransferase (UDPGT); (2) antioxidant enzymes—superoxide dismutase (SOD) and catalase (CAT); (3) oxidoreductases—polyphenol oxidase (PPO) and peroxidase (PX); and (4) glucoside hydrolases—α- and β-glucosidase. The activity of the transferases and the antioxidant enzymes increased after transfer to the secondary host, but the level of activity was closely associated with feeding duration on the secondary host. The strongest induction was noted for SOD, the activity of which was more than three times greater on the secondary than on the primary host. In contrast, transfer of the bird cherry–oat aphid was accompanied by a decline in activity of PPO, PX, and β-glucosidase; PPO and PX activity was 50% less in aphids fed on the secondary host rather than on the primary host. Activity of α-glucosidase increased after prolonged feeding on the secondary host. The results indicated that the adaptive enzymes of the bird cherry–oat aphid enable it to feed on distantly related host plants.     

一个棉花β-1,3-葡聚糖酶基因全长cDNA的克隆与特征分析

利用本实验室分离的防卫基因类似物片段,通过5’和3’RACE,从海岛棉海7124中克隆了一个β-1,3-葡聚糖酶基因全长cDNA,命名为GbGLU。其全长1 266 bp,含一个编码361个氨基酸残基的开放阅读框。该基因编码产物理论分子量和等电点为MW = 39.66 kD,pI = 9.15,含有糖基水解酶家族17的保守结构域,N端有26个氨基酸残基组成的信号肽。Southern结果显示,GbGLU在棉花基因组中以单拷贝形式存在,研究发现,GbGLU的表达受黄萎病菌的诱导。聚类分析表明,棉花葡聚糖酶基因可分为3类,分别含有糖基水解酶家族3、9、17的保守结构域。无论棉花中的还是本研究中参考的其他作物中,受病菌诱导的葡聚糖酶基因都含有糖基水解酶家族17的保守结构域,故推断其为此类基因参与抗病反应的功能域。     

酶学分析在昆虫抗有机农药研究中的应用

概述了抗有机农药的不同昆虫的酶类的活性变化。抗性品系的水解酶类和氧化酶类均有明显的变化,即对水解酶类而言,酯酶活力尤其是羟酸酯酶活力增加,但乙酰胆碱酯酶则变化不明显或敏感性下降;对氧化酶类而言,则多功能氧化酶的活力升高。     

黑布林籽β-葡萄糖苷酶聚合物的制备与稳定性研究

为探究β-葡萄糖苷酶聚合物的制备方法及其稳定性,以黑布林籽β-葡萄糖苷酶为原料制备交联酶聚合物(CLEAs),优化了制备过程中沉降剂的种类及比例、戊二醛交联剂的浓度等工艺参数,并采用扫描电镜分析其表面结构特征;在此基础上,以红景天苷的酶促合成为模型反应,研究CLEAs在有机溶剂中的热稳定性和操作稳定性。结果表明,制备β-葡萄糖苷酶CLEAs的最佳沉降剂为95%乙醇,其适宜添加量为酶液体积的3倍;戊二醛交联剂的最适浓度为30 mmol·L^-1,交联时间为1.0 h,酶活回收率高达91.4%。扫描电镜结果表明,在此条件下制备的CLEAs有较大的比表面积和致密的多孔道结构。稳定性结果表明,将β-葡萄糖苷酶CLEAs在60℃下保温2.0 h,其酶活回收率为75.1%,是游离酶的6.1倍;重复使用8次后,仍保持73.5%的初始酶活性。综上,黑布林籽β-葡萄糖苷酶CLEAs具有良好的生物催化活性、热稳定性和操作稳定性,可用于糖苷类化合物的高效合成。本研究为β-葡萄糖苷酶CLEAs的研究提供了理论依据,对工业化生产具有现实意义。     

双水相–SPE–TFA–GC/MS测定烟草中糖苷类潜香物质方法的建立

建立了双水相–固相萃取(SPE)–三氟乙酰衍生化(TFA)–GC/MS测定烟草中糖苷类潜香物质的方法:烟草样品通过微波双水相提取、0.45μm膜过滤后得到的糖苷粗提物,过HP–20 SPE小柱,再依次经过正戊烷–二氯甲烷和甲醇洗脱,洗脱液经浓缩至干燥后得到的糖苷精提物,通过三氟乙酰胺(MBTFA)衍生化,采用葡萄糖苷标准物及NIST谱库比照定性,并与内标2–硝基苯–β–D–葡萄糖苷的衍生化产物面积比进行定量分析,可测定烟草中18种糖苷类潜香物质。利用该方法测定烤烟K326、白肋烟Kenek、香料烟Basma的糖苷类潜香物质,共测得18种糖苷类香味成分,糖苷包括14种葡萄糖苷、1种芸香糖苷和3种巢菜糖苷,苷元包括脂肪醇、芳香族化合物、降异戊二烯类、倍半萜类多酚类化合物;烤烟中糖苷类潜香物含量最高,香料烟中含量最低。     

Yield,Nutrient Uptake,and Quality of Stevia as Affected by Organic Sources of Nutrient

A field experiment was conducted during 2008 and 2009 at the Council of Scientific and Industrial Research-Institute of Himalayan Bioresource Technology, Palampur, India, to study the effect of organic sources of nutrient on yield, nutrient uptake, fertility status of soil, and quality of stevia crop in the western Himalayan region. The experiment comprised eight different combinations of organic manure [farmyard manure (FYM), vermicompost (VC), and apple pomace manure (AP)]. Total leaf dry biomass increased by 149% over the control with application of VC 1.5 t ha^?1 + AP 5 t ha^?1. Application of organic manures enhanced organic carbon and available nutrient status of soil more than the control. Nitrogen (N) and phosphorus (P) content in stem were significantly affected by the application of organic manures over the control. Stevia plants supplied with FYM 10 t ha^?1 + AP 2.5 t ha^?1 recorded more total glycoside than other treatments. Stevioside yield (kg ha^?1) was greater with application of FYM 10 t ha^?1 + AP 2.5 t ha^?1.