Isolation and characterization of a novel thermostable non-specific lipid transfer protein-like antimicrobial protein from motherwort (Leonurus japonicus Houtt) seeds.
In screening for potent antimicrobial proteins from plant seeds, a novel heat-stable antimicrobial protein, designated LJAMP2, was purified from seeds of the motherwort (Leonurus japonicus Houtt), a medicine herb, with a procedure involving cation exchange chromatography on a CM FF column, and reverse phase HPLCs on C8 column and C18 column. LJAMP2 exhibited a molecular mass of 6.2 kDa determined. Automated Edman degradation determined the partial N-terminal sequence of LJAMP2 to be NH2-AIGCNTVASKMAPCLPYVTGKGPLGGCCGGVKGLIDAARTTPDRQAVCNCLKTLAKSYSG, which displays homology with plant non-specific lipid transfer proteins (nsLTPs). In vitro bioassays showed that LJAMP2 inhibits the growth of a variety of microbes, including filamentous fungi, bacteria and yeast. The growth of three phytopathogenic fungi, Alternaria brassicae, Botrytis maydis, and Rhizoctonia cerealis, are inhibited at 7.5 microM of LJAMP2, whereas Bacillus subtilis is about 15 microM. The IC(50) of LJAMP2 for Aspergillus niger, B. maydis, Fusarium oxysporum, Penicillium digitatum and Saccharomyces cerevisiae are 5.5, 6.1, 9.3, 40.0, and 76.0 microM, respectively.
Yang
Xingyong
X
Key Laboratory of Biotechnology and Crop Quality Improvement, Ministry of Agriculture of China and Biotechnology Research Center, Southwest University, Chongqing 400716, PR China. yanggxy@swau.edu.cn
Li
Jun
J
Li
Xianbi
X
She
Rong
R
Pei
Yan
Y
D000595:Amino Acid Sequence; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000302:isolation & purification* / Q000502:physiology*; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000502:physiology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D006801:Humans; D031332:Leonurus / Q000378:metabolism*; D008969:Molecular Sequence Data; D012639:Seeds / Q000378:metabolism*
D023181:Antimicrobial Cationic Peptides; D002352:Carrier Proteins; C030363:lipid transfer protein