作者机构：
[杨文敏; 谢方平*; 刘科; 孙松林; 李仲恺; 叶强] College of Engineering, Hunan Agriculture University, Changsha 410128, China;[谢方平*; 杨文敏; 孙松林] Hunan Provincial Engineering Technology Research Center for Modern Agricultural Equipment, Changsha 410128, China;[杨国顺] College of Horticulture and Landscape, Hunan Agriculture University, Changsha 410128, China
通讯机构：
College of Engineering, Hunan Agriculture University, China
作者机构：
[衡周] Horticulture and Landscape College, Hunan Agricultural University, Changsha 410128, China;[熊兴耀] The Institute of Vegetables and Flowers Chinese Academy of Agricultural Sciences, Beijing 100081, China;[周江] The Library of Hunan Agricultural University, Changsha 410128, China;[苏小军; 熊兴耀; 胡秋龙; 衡周] Hunan Engineering Laboratory for Alcohol Fuels from Biomass, Changsha 410128, China
通讯机构：
Hunan Engineering Laboratory for Alcohol Fuels from Biomass, China
作者机构：
[周鸣谦] School of Marine Science and Technology, Huaihai Institute of Technology, Lianyungang 222005, Jiangsu, China;[周鸣谦; 王仁才] College of Landscape and Horticulture, Hunan Agriculture University, Changsha 410128, China
通讯机构：
School of Marine Science and Technology, Huaihai Institute of Technology, China
作者机构：
[唐鑫; 吴灿; 夏延斌] College of Food Science Technology, Hunan Agricultural University, National R and D Center For Vegetable Processing, Changsha 410128, China
通讯机构：
College of Food Science Technology, Hunan Agricultural University, National R and D Center For Vegetable Processing, China
摘要：
High-speed counter-current chromatography was successfully applied to separate and purify bioactive ingredients from sweet potato leaves. Four caffeoylquinic acid derivatives and a mixture of two flavonoids were successfully obtained in one step. The caffeoylquinic acid derivatives were 3-O-caffeoylquinic acid (1), 4,5-di-O-caffeoylquinic acid (4), 3,5-di-O-caffeoylquinic acid (5), and 3,4-di-O-caffeoylquinic acid (6). The two-phase solvent system was composed of n-hexane\ethyl acetate\ethanol\water\acetic acid (1:5:2:4:0.1, v/v). The upper layer was used as the stationary phase, and the lower layer was used as the mobile phase. The flow rate was 1.5 mL/min, the revolution speed was 850 rpm, and the injection volume was 280 mg. The mixture of flavonoids was separated by preparative high-performance liquid chromatography into quercetin-3-O-β-D-galactopyranoside (2) and quercetin-3-O-β-D-glucoside (3). The purities of compounds 1–6 were 95.8% (5.4 mg), 99.5% (6.1 mg), 98.7% (15.1 mg), 97.8% (14.5 mg), 96.2% (10.3 mg), and 96.8% (7.8 mg), respectively, as determined by HPLC with a pulsed amperometric detector. The chemical structures were identified by electrospray ionization-tandem mass spectrometry and nuclear magnetic resonance imaging. The results showed the efficiency of the method in purifying bioactive compounds from sweet potato leaves, and compound 2 was separated from sweet potato for the first time.
通讯机构：
[He, Li Bo] H;Hunan Agr Univ, Coll Hort & Landscape Architecture, Changsha 410128, Hunan, Peoples R China.
会议名称：
International Conference on Civil, Architectural and Hydraulic Engineering (ICCAHE 2012)
会议时间：
AUG 10-12, 2012
会议地点：
Zhangjiajie, PEOPLES R CHINA
会议主办单位：
[He, Li Bo;Xiong, Xing Yao] Hunan Agr Univ, Coll Hort & Landscape Architecture, Changsha 410128, Hunan, Peoples R China.
会议论文集名称：
Applied Mechanics and Materials
关键词：
Eight scenes of yuelu academy;Landscape restoration;Yuelu academy
摘要：
Yuelu Academy is one of the four most prestigious academies in the history, its architecture part had been reconstructed in 1980's, but the garden landscape lacked unified design. Nowadays, the garden landscape of academy is losing its poetic imagery gradually. Under the principle of respecting history and spreading garden tradition, the conception of improving landscape axis for the academy and restoring Eight Scenes of Yuelu Academy is proposed for the overall restoration of the academy landscape. It is meaningful for setting a good example for the Chinese classical academy's garden and replenishing the traditional garden art.
摘要：
Although much effort has been made in the field of membrane proteomics, the analysis of membrane proteins particularly integral membrane proteins with poor water solubility still presents a great challenge. In this paper, 2% SDS was used to extract membrane proteins and experimental conditions for the application of acetone precipitation method to the cleanup of SDS-solubilized membrane protein sample were optimized. For improving the re-dissolution and trypsinolysis of acetone-precipitated proteins, several commonly used additives, urea, methanol and sodium deoxycholate (SDC), were employed and compared. The results showed that, when the pre-cooled acetone-to-sample ratio was 6:1 (v/v) with one additional washing step, residual SDS in the protein sample could be lowered to below 0.01% and more than 90% of the proteins were precipitated and therefore recovered. 1% SDC-containing buffer could improve the re-dissolution and digestion of the acetone precipitated proteins more efficiently than the others. Using the combinative sample preparation strategy developed, 398 proteins were identified from the rat liver membrane-enriched fraction, including 188 membrane proteins. Compared with other three representative solution-based sample preparation methods commonly used in membrane proteomics, the newly developed combinative strategy increased the number of identified total proteins and membrane proteins on average by 29.2% and 28.5%, respectively. This combinative strategy was demonstrated to be easily operated at low cost and suitable for the analysis of membrane proteins varying in type and sample volume, etc. (C) 2012 Published by Elsevier B.V.
摘要：
In order to discover the formation mechanism of carotenoid derived aroma, which has been wildly used on protection of crop against insect attacks, the full-length cDNA of an Artemisia annua carotenoid cleavage dioxygenase (AaCCD1) was cloned by rapid amplification of cDNA ends. The function of AaCCD1 was characterized by expression of AaCCD1 in a strain of E. coli accumulating carotenoids and enzyme assay in vitro. The completed open read frame of AaCCD1 was 1629 bp and it encoded a 542-amino acid protein with a 77% amino acid identity to Arabidopsis thaliana CCD1, a predicted molecular mass of 61.04 kDa and a pI of 5.8. AaCCD1 efficiently cleaves carotenoids and regulate the formation of terpenoid compounds. This is the first time to report the cloning and identification of carotenoid cleavage dioxygenase from Atemisia annua, which will play a great role on understanding the regulation of volatile compounds.
作者机构：
[胡新丽; 黄磊; 邹宗兴; 徐伟; 倪卫达; 刘晓] Faculty of Engineering, China University of Geosciences, Wuhan, Hubei 430074, China;[徐伟] Chengdu Institute of Geology and Mineral Resources, Chengdu, Sichuan 610081, China
通讯机构：
Faculty of Engineering, China University of Geosciences, China
作者机构：
[戴思慧; 孙小武; 熊兴耀] College of Horticulture and Landscape, Changsha 410128, China;[姚珺; 李明] College of Engineering, Hunan Agricultural University, Changsha 410128, China;[戴思慧; 孙小武; 熊兴耀] Hunan Provincial Key Laboratory for Germplasm Innovation and Utilization of Crop, Changsha 410128, China
通讯机构：
College of Horticulture and Landscape, China
摘要：
Analysis of the membrane proteins, particularly the integral membrane proteins, is limited by the inherent membrane hydrophobicity. Sodium dodecyl sulfate (SDS) is one of the most efficient reagents used for the extraction of membrane proteins, but its presence in samples interferes with LC-MS-based proteomic analyses because it affects RP-LC separations and electrospray ionization. In this paper, we present an improved sample preparation strategy based on SOS-assisted digestion and peptide-level SDS-removal using an optimized potassium dodecyl sulfate (KDS) precipitation method (SSDP method) for shotgun analysis of the membrane proteome. This method utilizes a high concentration of SDS (1.0%) to lyse the membranes and to solubilize the hydrophobic membrane proteins, resulting in a more complete protein digestion in the diluted SDS buffer (0.1% SOS), and a high efficiency of SOS removal and peptide recovery by the optimized KDS precipitation for protein identification. The SSDP method provides evidence that proteins can be efficiently digested, and the SDS can be decreased to <0.01% allowing >95% peptide recovery. Compared to other sample preparation methods commonly used in shotgun membrane proteomics, the newly developed method not only increased the identified number of the total proteins, membrane proteins and integral membrane proteins by an average of 33.1%, 37.2% and 40.5%, respectively, but also leading to the identification of highest number of matching peptides. All the results showed that the method yielded better recovery and reliability in the identification of the proteins especially the highly hydrophobic integral membrane proteins, and thus providing a promising tool for the shotgun analysis of membrane proteome. (C) 2012 Elsevier B.V. All rights reserved.
摘要：
In-gel digestion is an attractive route in mass spectrometry-based proteomic analysis, which, however, often suffers from a certain amount of sample loss mainly due to insufficient protein digestion and peptide extraction. To address this, herein we establish a partially degradable gel-assisted protein digestion and peptide recovery method by means of a simple replacement of bis-acrylamide (BA) with bis-acrylylcystamine (BAC). Concretely, the protein sample solubilized using high concentrations of sodium dodecyl sulfate (SOS) and urea were directly entrapped and immobilized into BAC-crosslinked gel by vacuum-dried gel absorption followed by fixation treatment. After removal of SDS and urea by repeated washing, the proteins were subjected to in-gel digestion and the gel was reductively treated. The tryptic peptides were recovered from the partial degradation of the gel and analyzed afterwards by capillary liquid chromatography coupled with tandem mass spectrometry (CapLC-MS/MS). Compared with conventional BA-crosslinked gel method, this new method increased the numbers of identified proteins and unique peptides by 20.2% and 20.4%, respectively. The further statistical analysis demonstrated that the method improved the recovery of tryptic peptides particularly larger and/or hydrophobic peptides, thereby significantly facilitating protein identification. Thus, the newly developed method is a promising alternative for BA-crosslinked gel-based shotgun workflows and has potential application in the related fields of protein chemistry and proteomics. (C) 2011 Elsevier B.V. All rights reserved.
