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991.
New oligosaccharyltransferase assay method 总被引:2,自引:0,他引:2
We developed a new in vitro assay for oligosaccharyltransferase (OST), which catalyzes the transfer of preassembled oligosaccharides on lipid carriers onto asparagine residues in polypeptide chains. The asparagine residues reside in the sequon, Asn-X-Thr/Ser, where X can be any amino acid residue except Pro. We demonstrate the potency of our assay using the OST from yeast. In our method, polyacrylamide gel electrophoresis is used to separate the glycopeptide products from the peptide substrates. The substrate peptide is fluorescently labeled and the formation of glycopeptides is analyzed by fluorescence gel imaging. Two in vitro OST assay methods are now widely used, but both the methods depend on previous knowledge of the oligosaccharide moiety: One method uses lectin binding as the separation mechanism and the other method uses biosynthetically or chemoenzymatically synthesized lipid-linked oligosaccharides as donors. N-linked protein glycosylation is found in all three domains of life, but little is known about the N-glycosylation in Archaea. Thus, our new assay, which does not require a priori knowledge of the oligosaccharides, will be useful in such cases. Indeed, we have detected the OST activity in the membrane fraction from a hyperthermophilic archaeon, Pyrococcus furiosus. 相似文献
992.
993.
模拟大气CO2浓度和温度升高对亚高山冷杉(Abies faxoniana)林土壤酶活性的影响 总被引:1,自引:0,他引:1
采用控制环境生长室研究了川西亚高山森林生态系统中与C、N、P循环有关的土壤转化酶、脲酶、硝酸还原酶和酸性磷酸酶活性的月动态及其对模拟大气CO2浓度增加、温度升高以及交互作用的动态响应。在一个生长季节内,土壤有机层和矿质土壤层的转化酶、脲酶、硝酸还原酶和酸性磷酸酶的活性高峰均出现在温度较高的夏季。其中,土壤有机层的转化酶活性高峰出现在6月份,但土壤矿质层的转化酶活性高峰出现在7月份,土壤有机层和矿质土壤层的脲酶和酸性磷酸酶的活性高峰均出现在7月份,而硝酸还原酶的活性高峰均出现在8月份。升高大气CO2浓度处理(EC)对土壤有机层和矿质土壤层的转化酶、脲酶、硝酸还原酶和酸性磷酸酶活性没有显著影响。升高温度处理(ET)显著增加了土壤有机层和矿质土壤层的酶活性,并且土壤有机层的转化酶、硝酸还原酶和脲酶活性增加更显著。大气CO2浓度增加和温度升高之间的交互作用(ECT)对土壤有机层和矿质土壤层酶活性的影响主要是温度升高引起的。 相似文献
994.
侵蚀退化红壤自然恢复下土壤生物学质量演变特征 总被引:14,自引:1,他引:13
为了探讨严重侵蚀退化红壤区自然植被恢复过程中土壤生物学质量演变特征,对南方严重侵蚀退化红壤自然植被恢复的4个演替阶段(裸地,地衣地,苔藓地和草地),以及该地区人工马尾松林地的土壤微生物量、酶活性和线虫数量进行了比较研究。结果表明:在侵蚀红壤自然恢复过程中土壤生物学性质演变特征明显。在恢复初期,地衣和苔藓对土壤生物学性质的改善主要体现在土壤表层。在0~2cm土层地衣地土壤微生物量碳、氮、蔗糖酶和脲酶活性高于裸地,但差异不显著;苔藓地表层微生物量氮、脲酶和酸性磷酸酶活与人工马尾松林地已无显著差异,表明苔藓地是严重侵蚀退化红壤自然恢复过程中土壤质量改善的重要阶段。裸地、地衣地和苔藓地土壤线虫恢复程度低于微生物量和酶活性。草地土壤微生物量碳、氮和3种酶活性以及线虫数量则显著高于自然恢复初期各阶段。与人工恢复林相比较,自然恢复草地表层土壤生物学质量优于人工马尾松林地,但对深层土壤的改善效果不如林地。相关分析表明自然恢复过程中土壤微生物与酶活性的改善程度比较一致,而土壤线虫对自然植被恢复响应与微生物和酶活性不尽相同。 相似文献
995.
The study focused on the dynamics of Malondialdehyde (MDA) contents and the activities of protective enzymes in the leaves of alfalfa varieties with various resistances to Aphis medicaginis Koch. The results showed that susceptible varieties always had higher MDA contents than resistant varieties, and the MDA contents tended to rise in both susceptible and resistant varieties in period of the varieties were pierced and sucked by aphids. Superoxide dismutase (SOD), peroxidase (POD) and phenylalanine ammonia-lyase (PAL) activities in susceptible varieties were lower than those in resistant varieties, and in both susceptible and resistant varieties the SOD and POD activities tended to rise at first and then decline, and the PAL activities rose to their peaks and then tended to remain stable. In the susceptible and resistant varieties the catalase (CAT) activities appeared to rise and decline alternatively; the PPO activities in resistant varieties were lower than those in susceptible varieties in early growth, but higher than those in susceptible varieties in later growth. It follows that infested by aphids, susceptible and resistant varieties had the MDA contents, variations of SOD, POD, PAL and PPO activities were closely correlated with their aphid resistances, hence these indexes could be used as physiological indexes for testing aphid resistance of alfalfa, whereas the relations of their CAT activities to their resistances needed to be further studied. 相似文献
996.
The landward changes of soil enzyme activities and physico-chemical properties of the surface sediment in Chongming Dongtan of the Yangtze River Estuary, were studied. Along the elevation gradient or succession series, the contents of total phosphorus (TP), total nitrogen (TN) and organic matter (OM) in the sediment increased, but the average grain size (AGS) of the sediment and the content of the dissolved inorganic phosphorus (DIP) decreased. The activity of alkaline phosphatase increased gradually along the elevation gradient, and was positively correlated with the values of TP, TN and OM (P<0.05), but negatively to AGS and DIP (P<0.05). It was correlated with a mechanism of substrate inductivity and product inhibition. Catalase activity had the similar trend of gradual increase along the elevation gradient, enhancing the fertility of the soil and the oxidative process of OM in the sediment. Along the succession series, from the tidal flat to the bulrush (Scirpus mariqueter) zone, and then to the reed (Phragmites australis) zone, the activity of sucrase only changed insignificantly, but there was a higher activity in the bulrush zone than in other zones. The activity of proteinase decreased from the tidal flat to the reed zone, and the activity was negatively correlated with OM and TN (P<0.05), but positively with DIP (P<0.05). Through the succession zones a decrease in the number of diatoms resulted in a decline in the concentration of protein, which influenced the proteinase activity, suggesting that the proteinase in the sediment was produced by diatoms. 相似文献
997.
验证了荧光假单胞菌(Pseudomonas fluorescensATCC13525)香兰素脱氢酶基因(vanillin dehydrogenasegene,vdh)的功能。基因vdh表达产物(Vdh)的活性测定结果显示Vdh具有很高的活性,而且不经IPTG诱导的Vdh也具有同样高的活性。经过4 h的体外酶促反应,重组蛋白Vdh能把95%以上的香兰素转化为香兰素酸,从而验证了vdh基因的表达产物具有香兰素脱氢酶的功能。同时发现NAD 是从香兰素到香兰素酸体外转化必不可少的因素。 相似文献
998.
999.
1000.
Yuasa HJ Hasegawa T Nakamura T Suzuki T 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》2007,146(4):461-469
The indoleamine 2,3-dioxygenase (IDO)-like myoglobin (Mb) is a unique type of Mb isolated from the buccal mass of several archgastropod species. Here, we expressed Sulculus diversicolor IDO-like Mb as a GST-fusion protein in bacteria. The visible spectrum of GST-fusion IDO-like Mb shows characteristic α- and β-peaks, indicating that it binds oxygen. To identify residues important in heme and oxygen binding, we constructed site-directed mutants. We initially replaced each of the 7 histidines of S. diversicolor IDO-like Mb with alanine. The spectra of three mutants (H74A, H288A, and H332A) revealed a remarkable loss of absorbance around 414 nm, indicating that they cannot bind heme. His74, His288, and His332 were also replaced by arginine or tyrosine. Neither H332R nor H332Y contains heme, suggesting that His332 is the proximal ligand of IDO-like Mb. In contrast, both H74R and H288Y mutants were isolated in the heme-binding oxy-form. The autoxidation rates of these two mutants showed that they can bind oxygen as stably as wild-type. His74 and His288 might be partially associated with heme-binding, but do not act as the distal ligand. The S. diversicolor IDO-like Mb seems to stably bind oxygen in a different manner from normal myoglobins. 相似文献