Aunpad Ratchaneewan, Na-Bangchang Keasara
Graduate Program in Biomedical Sciences, Faculty of Allied Health Sciences, Thammasat University, Klongluang District, Pathumthani, Thailand.
Curr Microbiol. 2007 Oct;55(4):308-13. doi: 10.1007/s00284-006-0632-2. Epub 2007 Aug 13.
A total of 34 bacterial strains with anti-methicillin-resistant Staphylococcus aureus (MRSA) activity were isolated from 69 soil and water samples collected from four areas of Thailand. One strain, WAPB4 identified as Bacillus pumilus, showed remarkable antibacterial activity against MRSA, vancomycin-resistant Enterococcus faecalis (VRE), and several Gram-positive test bacteria. Bacteriocin produced by WAPB4 was designated as pumilicin 4. It was heat stable up to 121 degrees C, 15 min and active within the pH range of 3-9. Its activity disappeared when treated with pronase E, chymotrypsin, and trypsin, demonstrating its proteinaceous nature. At high dosage (80 AU mL(-1)), the effect of pumilicin 4 was bactericidal to both MRSA and VRE. Bacteriostasis was observed for a low dose of bacteriocin (20 AU mL(-1)). Purification of pumilicin 4 was performed by a three-step procedure, i.e., solvent extraction, solid phase extraction, and reversed-phase chromatography. The molecular mass of purified pumilicin 4 as determined by mass spectrometry was 1994.62 Dalton. This present study is the first report of a novel bacteriocin, pumilicin 4, produced by B. pumilus that has potential for use as an alternative antibacterial agent for the treatment of infection with MRSA and VRE.
从泰国四个地区采集的69份土壤和水样中，共分离出34株具有抗耐甲氧西林金黄色葡萄球菌（MRSA）活性的细菌菌株。其中一株名为短小芽孢杆菌的WAPB4菌株，对MRSA、耐万古霉素粪肠球菌（VRE）以及几种革兰氏阳性测试菌表现出显著的抗菌活性。WAPB4产生的细菌素被命名为短小菌素4。它在121摄氏度下加热15分钟仍保持稳定，在pH值3 - 9的范围内具有活性。用链霉蛋白酶E、胰凝乳蛋白酶和胰蛋白酶处理后，其活性消失，表明它具有蛋白质性质。在高剂量（80 AU mL(-1)）下，短小菌素4对MRSA和VRE均有杀菌作用。低剂量细菌素（20 AU mL(-1)）时观察到抑菌作用。短小菌素4通过三步程序进行纯化，即溶剂萃取、固相萃取和反相色谱法。通过质谱测定，纯化后的短小菌素4的分子量为1994.62道尔顿。本研究首次报道了由短小芽孢杆菌产生的一种新型细菌素——短小菌素4，它有潜力作为治疗MRSA和VRE感染的替代抗菌剂。
