Activity based metaproteomic analysis of nitrous oxide reductase in soil
Project/Area Number |
24658063
|
Research Category |
Grant-in-Aid for Challenging Exploratory Research
|
Allocation Type | Multi-year Fund |
Research Field |
Plant nutrition/Soil science
|
Research Institution | Yamaguchi University |
Principal Investigator |
|
Project Period (FY) |
2012-04-01 – 2015-03-31
|
Project Status |
Completed (Fiscal Year 2014)
|
Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
|
Keywords | 一酸化二窒素還元酵素 / 二次元電気泳動 / 活性染色 / 土壌 / 脱窒菌 / 一酸化二窒素 / 亜酸化窒素還元酵素 / メタボローム / 亜酸化窒素 / 二次元ネイティブ電気泳動 / 多様性解析 |
Outline of Final Research Achievements |
This study attempted to extract active nitrous oxide reductases (N2ORs) of denitrifying bacteria in soil and to analyze their diversity by detecting them based on the activity-dependent staing on a two-dimensional native PAGE gel. It was not successfule to avoid isoelectric precipitation of the proteins during the isoelectric electrophoresis, the first step of the 2-D native PAGE. The activity of N2OR preparations was lost inreversively. The extraction of active N2ORs from soil was conducted by indirect methods where bacterials cells were separated from soil particles by density gradient centrifugation. The amount of proteins thus extracted was not sufficient to create a Coomacy Brilliant Blue-positive spots on the native PAGE gel.
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Report
(4 results)
Research Products
(2 results)