Chen Gan-Hong, Hsu Ming-Pin, Tan Chi-Hsing, Sung Hsien-Yi, Kuo C George, Fan Ming-Jen, Chen Huei-Mei, Chen Shu, Chen Ching-San
Institute of Microbiology and Biochemistry and Department of Agricultural Chemistry, National Taiwan University, Taipei 106, Taiwan.
J Agric Food Chem. 2005 Feb 23;53(4):982-8. doi: 10.1021/jf0402227.
A recombinant mungbean defensin VrD1 was previously shown to exhibit antifungal and bruchid-resistant activity. To study the function and regulation of VrD1, genomic DNAs of plant defensins were isolated from Vigna radiata VC6089A and azuki bean Vigna angularis Kao Hsiung No. 6. The azuki bean defensin genomic DNA VaD1 was sequenced and converted to VaD1 cDNA. VaD1 defensin was purified from Vigna angularis Kao Hsiung No. 6 to apparent homogeneity. The complete amino acid sequence of the purified VaD1 was determined and was found to be exactly the same as the sequence deduced from VaD1 cDNA. VaD1 is a basic protein containing 46 amino acids with four conserved disulfide bonds and shares high sequence homology (78.3%) with VrD1. VaD1 inhibited the growth of Fusarium oxysporum, Fusarium oxysporum f. sp. pisi, Staphylococcus epidermidis, and Salmonella typhimurium. VaD1 also inhibited in vitro protein synthesis and bruchid larval development, but was less active than the recombinant VrD1.
先前的研究表明，重组绿豆防御素VrD1具有抗真菌和抗豆象活性。为了研究VrD1的功能和调控机制，从绿豆Vigna radiata VC6089A和赤小豆Vigna angularis高雄6号中分离出植物防御素的基因组DNA。对赤小豆防御素基因组DNA VaD1进行测序，并将其转化为VaD1 cDNA。从赤小豆高雄6号中纯化出VaD1防御素，使其达到表观均一性。测定了纯化后的VaD1的完整氨基酸序列，发现其与从VaD1 cDNA推导的序列完全相同。VaD1是一种碱性蛋白，含有46个氨基酸，有四个保守的二硫键，与VrD1具有较高的序列同源性（78.3%）。VaD1抑制尖孢镰刀菌、尖孢镰刀菌豌豆专化型、表皮葡萄球菌和鼠伤寒沙门氏菌的生长。VaD1还抑制体外蛋白质合成和豆象幼虫发育，但活性低于重组VrD1。
