文献类型：期刊文献

英文题名：Exploration of the antibacterial proteins in pearl oyster Pinctada fucata induced by bacterial inoculation

作者：Lin, Haisheng[1,2];Ishizaki, Shoichiro[1];Nagashima, Yuji[1];Nagai, Kiyohito[3];Maeyama, Kaoru[4];Watabe, Shugo[5]

机构：[1]Tokyo Univ Marine Sci & Technol, Grad Sch Marine Sci & Technol, Minato Ku, 4-5-7 Konan, Tokyo 1088477, Japan;[2]Guangdong Ocean Univ, Coll Food Sci & Technol, Haida Rd 1, Mazhang 524088, Zhanjiang, Peoples R China;[3]Mikimoto Co Ltd, Pearl Res Lab, Shima 5170403, Japan;[4]Mikimoto Pharmaceut Co Ltd, Ise 5168581, Japan;[5]Kitasato Univ, Sch Marine Biosci, Sagamihara, Kanagawa 2520373, Japan

年份：2017

卷号：83

期号：3

起止页码：489

外文期刊名：FISHERIES SCIENCE

收录：SCI-EXPANDED(收录号：WOS:000403085500015)、、WOS

基金：This study was partially supported by the Sasakawa Scientific Research Grant from the Japan Science Society (28-317).

语种：英文

外文关键词：Innate immunity; Bacterial invasion; Gill antibacterial protein; High-performance liquid chromatography

外文摘要：The aim of this research was to characterize immune-related antibacterial substances from pearl oyster Pinctada fucata induced by bacterial invasion. Bacteria inoculation was performed by injecting 0.1 ml of 1.0 x 10(12) colony-forming units/ml Vibrio parahaemolyticus into adductor muscle. Acidic extracts were prepared with 0.1% trifluoroacetic acid from different tissues after 8 h of injection, and antibacterial activity against V. parahaemolyticus was determined via the microdilution broth method. The acidic extracts from gills of inoculated oysters (AEg) showed stronger antibacterial activity than those from non-inoculated ones. Based on this result, antibacterial proteins were purified from AEg via two-step gel filtration chromatography, followed by high-performance liquid chromatography using a TSkgel G3000 column. Protein components were analyzed by both sodium dodecyl sulfate and native polyacrylamide gel electrophoresis. As a result, two antibacterial proteins, APg-1 (with a molecular mass of approximately 210 kDa) and APg-2 (of approximately 30 kDa), were obtained from AEg. Matrix-assisted laser desorption/ionization time of flight mass spectrometry analysis and partial amino acid sequences revealed that these proteins might be novel antibacterial proteins. These results indicate that antibacterial proteins are potentially upregulated in the gill of pearl oysters or released there-from for defense against bacterial invasion.