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801.
Xiao-Qiu Shen 《Carbohydrate research》2009,344(15):2063-1796
A protected trisaccharide imidate, 2,3-di-O-acetyl-4,6-O-benzylidene-β-d-glucopyranosyl-(1→3)-2-O-chloroacetyl-3-O-benzyl-4-isobutyryl-α-l-rhamnopyranosyl-(1→4)-2-O-isobutyryl-α-l-rhamnopyranosyl trichloroacetimidate (1), has been synthesized by a block synthesis approach. Compound 1 can serve as a key intermediate in the total synthesis of resin glycoside merremoside H2. 相似文献
802.
Masayuki Okuyama Hironori Hondoh Haruhide Mori Isao Tanaka 《Journal of molecular biology》2009,392(5):1232-312
Glycoside hydrolase family 97 (GH 97) is a unique glycoside family that contains inverting and retaining glycosidases. Of these, BtGH97a (SusB) and BtGH97b (UniProtKB/TrEMBL entry Q8A6L0), derived from Bacteroides thetaiotaomicron, have been characterized as an inverting α-glucoside hydrolase and a retaining α-galactosidase, respectively. Previous studies on the three-dimensional structures of BtGH97a and site-directed mutagenesis indicated that Glu532 acts as an acid catalyst and that Glu439 and Glu508 function as the catalytic base in the inverting mechanism. However, BtGH97b lacks base catalysts but possesses a putative catalytic nucleophilic residue, Asp415. Here, we report that Asp415 in BtGH97b is the nucleophilic catalyst based on the results of crystal structure analysis and site-directed mutagenesis study. Structural comparison between BtGH97b and BtGH97a indicated that OD1 of Asp415 in BtGH97b is located at a position spatially identical with the catalytic water molecule of BtGH97a, which attacks on the anomeric carbon from the β-face (i.e., Asp415 is poised for nucleophilic attack on the anomeric carbon). Site-directed mutagenesis of Asp415 leads to inactivation of the enzyme, and the activity is rescued by an external nucleophilic azide ion. That is, Asp415 functions as a nucleophilic catalyst. The multiple amino acid sequence alignment of GH 97 members indicated that almost half of the GH 97 enzymes possess base catalyst residues at the end of β-strands 3 and 5, while the other half of the family show a conserved nucleophilic residue at the end of β-strand 4. The different positions of functional groups on the β-face of the substrate, which seem to be due to “hopping of the functional group” during evolution, have led to divergence of catalytic mechanism within the same family. 相似文献
803.
Jindou S Xu Q Kenig R Shulman M Shoham Y Bayer EA Lamed R 《FEMS microbiology letters》2006,254(2):308-316
We have sequenced a new gene, cel9B, encoding a family-9 cellulase from a cellulosome-producing bacterium, Acetivibrio cellulolyticus. The gene includes a signal peptide, a family-9 glycoside hydrolases (GH9) catalytic module, two family-3 carbohydrate-binding modules (CBM3c-CBM3b tandem dyad) and a C-terminal dockerin module. An identical modular arrangement exists in two putative GH9 genes from the draft sequence of the Clostridium thermocellum genome. The three homologous CBM3b modules from A. cellulolyticus and C. thermocellum were overexpressed, but, surprisingly, none bound cellulosic substrates. The results raise fundamental questions concerning the possible role(s) of the newly described CBMs. Phylogenetic analysis and preliminary site-directed mutagenesis studies suggest that the catalytic module and the CBM3 dyad are distinctive in their sequences and are proposed to constitute a new GH9 architectural theme. 相似文献
804.
Yang Cui Qiao Wang Xiaowei Shi Xiaowei Zhang Xiaona Sheng Lantong Zhang 《Phytochemical analysis : PCA》2010,21(3):253-260
Introduction – Forsythia suspensa is a commonly used traditional Chinese medicine including phenylethanoid glycosides, lignans, flavonoids, terpenes and volatile oils. Quantification of multi‐components is important for the quality control of Forsythia suspensa. Objective – To establish a liquid chromatography–electrospray ionisation–mass spectrometry method for simultaneous quantification of 14 bioactive constituents of Forsythia suspensa in different places of China and different parts of this herb. Methodology – The optimal chromatographic conditions were achieved on a Kromasil C18 column (150 ¥ 4.6 mm, 5 mm) with gradient elution of methanol, acetonitrile and 0.1% formic acid in 27 min. Detection was performed in negative ionisation mode by monitoring the precursor–product combination in multiple reaction monitoring (MRM) mode. The validation of the method included tests of linearity, sensitivity, precision, repeatability, stability and accuracy. Results – All calibration curves showed good linear regression (r > 0.9990) within test ranges. The established method showed good precision and accuracy with overall intra‐day and inter‐day variations of 0.7–4.3 and 1.1–3.9% respectively, and overall recoveries of 96.65–101.2% for the compounds analysed. Conclusion – The proposed method was successfully applied for the quantitative analysis of 14 constituents in 12 Forsythia suspensa samples. Copyright © 2010 John Wiley & Sons, Ltd. 相似文献
806.
Nobuya Inagaki Akinori Iguchi Takahiro Yokoyama Ken-ji Yokoi Yasushi Ono Ayanori Yamakawa Akira Taketo Ken-Ichi Kodaira 《Gene》2009
The major autolysin AcmA of Lactococcus lactis ssp. cremoris MG1363 is a modular protein consisting of an N-terminal signal sequence, a central enzymatic region (gluacma as a glucosaminidase), and a C-terminal cell-recognition domain (LysM123). gluacma (about 160 amino acids) belongs to the glycoside hydrolase (GH) 73 family, and the two acidic residues E128 and D153 have been thought to be catalytically important. In this study, amino-acid substitution analysis of AcmA was first carried out in the Escherichia coli system. Point mutations E94A, E94Q, E128A, D153A, and Y191A markedly reduced cell-lytic activity (3.8%, 1.1%, 4.2%, 4.8%, and 2.4%, respectively), whereas E128Q and D153N retained significant residual activities (32.1% and 44.0%, respectively). On the other hand, Y191F and Y191W mutations retained high activities (66.2% and 46.0%, respectively). These results showed that E94 (rather than E128 and D153) and the aromatic residue Y191 probably play important roles in catalysis of AcmA. Together with mutational analysis of another GH73 glucoaminidase Gluatlwm from the Staphylococcus warneri M autolysin AtlWM, these results suggested that the GH73 members cleave a glycosidic bond via a substrate-assisted mechanism, as postulated in the GH20 members. AcmA and Gluatlwm were purified from E. coli recombinant cells, and their enzymatic properties were studied. 相似文献
807.
甜叶菊糖苷对果蝇生长和繁殖的影响 总被引:1,自引:0,他引:1
以黑腹果蝇Drosophila melanogaster Meigen作试验材料,探讨甜叶菊糖苷(steviol glycosides)对果蝇生殖、寿命等的影响。方法:在果蝇培养基中添加甜叶菊糖苷,质量分数分别为0,1%,2%,5%,10%;培养温度为(25±1)℃;取1对未交配的雌雄果蝇放入培养基中,待出现子代成蝇时,统计成蝇数量,记录成蝇羽化的数目,连续培养4代,观察甜叶菊糖苷对果蝇生育能力的影响;随机选取100只雌雄成蝇放入培养基中,统计其寿命,结果:过高浓度的甜叶菊糖苷对果蝇有一定程度的伤害,致使果蝇寿命和生育能力下降。 相似文献
808.
Jérôme Le Nours Leonardo De Maria Ditte Welner Christel T. Jørgensen Lars L. H. Christensen Torben V. Borchert Sine Larsen Leila Lo Leggio 《Proteins》2009,75(4):977-989
Microbial β‐1,4‐galactanases are glycoside hydrolases belonging to family 53, which degrade galactan and arabinogalactan side chains in the hairy regions of pectin, a major plant cell wall component. They belong to the larger clan GH‐A of glycoside hydrolases, which cover many different poly‐ and oligosaccharidase specificities. Crystallographic complexes of Bacillus licheniformis β‐1,4‐galactanase and its inactive nucleophile mutant have been obtained with methyl‐β(1→4)‐galactotetraoside, providing, for the first time, information on substrate binding to the aglycone side of the β‐1,4‐galactanase substrate binding groove. Using the experimentally determined subsites as a starting point, a β(1→4)‐galactononaose was built into the structure and subjected to molecular dynamics simulations giving further insight into the residues involved in the binding of the polysaccharide from subsite ?4 to +5. In particular, this analysis newly identified a conserved β‐turn, which contributes to subsites ?2 to +3. This β‐turn is unique to family 53 β‐1,4‐galactanases among all clan GH‐A families that have been structurally characterized and thus might be a structural signature for endo‐β‐1,4‐galactanase specificity. Proteins 2009. © 2008 Wiley‐Liss, Inc. 相似文献
809.
从毛叶飞蛾藤(Porana racemosa Roxb.)全草的95%乙醇提取物中分离并鉴定了11个化合物,其中一新的C30甾体化合物鉴定为(22E,24ξ)-24-正丙基胆甾-7,22-二烯-3β-醇(飞蛾藤素,1).其余10个已知化合物分别为东莨菪素(2)、东莨菪苷(3)、伞形华内酯(4)、β-D-甲基吡喃果糖苷(5)、丁香脂素4-O-β-D-吡喃葡萄糖苷(6)、斛皮素-3-O-β-D-吡喃葡萄糖苷(7)、斛皮素-3-O-α-L-吡喃鼠李糖苷(8)、异泽兰黄素(9)、山奈素-3-O-β-D-吡喃葡萄糖苷(10)和(E)-N-2-(2,3-二羟基苯基)乙基肉桂酰胺(11). 相似文献
810.
植物配糖体对人肠道厌氧菌群的影响 总被引:3,自引:0,他引:3
通过从我国传统中药三七、薯蓣、青叶胆、天麻中分离到的天然植物配糖体三七皂甙R1、Dioscin、Protodiscin、Gracillin、Gastroside及Saponin D等对下沉人肠道厌氧菌群进行64h生长培养,用比浊法测定培养液中厌氧菌的数量,发现不同的植物配糖体对不同人的肠道厌氧菌群生长活性影响不同,为我国传统中药的辩证施治提供了现代的试验依据。 相似文献