Zhang Beibei, Xie Chengjian, Yang Xingyong
Key Laboratory of Biotechnology and Crop Quality Improvement of Ministry of Agriculture of China and Biotechnology Research Center, Southwest University, Chongqing, China.
Peptides. 2008 Mar;29(3):350-5. doi: 10.1016/j.peptides.2007.11.024. Epub 2007 Dec 17.
A novel small antifungal peptide produced by a Bacillus strain B-TL2 isolated from tobacco stems was purified. The purification procedure consisted of ammonium sulfate precipitation, cation exchange chromatography on CM-Sepharose Fast Flow column and reverse-phase HPLC on SOURCE 5RPC column. After the final isolation step, one peptide with antifungal activity, designated as BTL, was obtained. The molecular mass of the purified BTL was determined as 2500 Da and 2237.7 Da by SDS-PAGE and matrix-assisted laser desorption/ionization time of flight mass spectrometry, respectively. The N-amino acid sequence of BTL was determined to be NH(2)-KQQLATEAESAGPIL, which shows relatively low identity to other antimicrobial peptides from bacteria. The peptide exhibited strong inhibitory activity against mycelial growth of Bipolaris maydis, Alternaria brassicae, Aspergillus niger, Cercospora personata. The purified BTL displayed thermostability, almost retaining 100% activity at 100 degrees C for 15 min.
从烟草茎中分离出的一株芽孢杆菌B-TL2产生的一种新型小抗真菌肽被纯化出来。纯化过程包括硫酸铵沉淀、在CM-Sepharose Fast Flow柱上进行阳离子交换色谱以及在SOURCE 5RPC柱上进行反相高效液相色谱。经过最后的分离步骤,获得了一种具有抗真菌活性的肽,命名为BTL。通过SDS-PAGE和基质辅助激光解吸/电离飞行时间质谱分别测定纯化后的BTL的分子量为2500 Da和2237.7 Da。BTL的N-氨基酸序列被确定为NH(2)-KQQLATEAESAGPIL,与其他细菌来源的抗菌肽相比具有相对较低的同源性。该肽对玉米小斑病菌、芸苔链格孢、黑曲霉、花生尾孢菌的菌丝生长表现出强烈的抑制活性。纯化后的BTL具有热稳定性,在100℃下15分钟几乎保留100%的活性。