Jeyanthi Venkadapathi, Anbu Periasamy, Vairamani Mariappanadar, Velusamy Palaniyandi
Department of Biotechnology, School of Bioengineering, SRM University, Kattankulathur, 603 203, Tamil Nadu, India.
Department of Biological Engineering, Inha University, Incheon, 402-751, South Korea.
Bioprocess Biosyst Eng. 2016 Mar;39(3):429-39. doi: 10.1007/s00449-015-1526-0. Epub 2015 Dec 31.
A halotolerant bacterial isolate-MHC10 with broad spectrum antibacterial activity against clinical pathogens was isolated from saltpans located in Tuticorin and Chennai (India). 16S rRNA gene analysis of MHC10 revealed close similarity to that of Bacillus methylotrophicus. The culture conditions of B. methylotrophicus MHC10 strain were optimized for antibacterial production using different carbon and nitrogen sources, as well as varying temperature, pH, sodium chloride (NaCl) concentrations and incubation periods. The maximum antibacterial activity of B. methylotrophicus MHC10 was attained when ZMB was optimized with 1 % (w/v) glucose, 0.1 % (w/v) soybean meal which corresponded to a C/N ratio of 38.83, temperature at 37 °C, pH 7.0 and 8 % NaCl. The activity remained stable between 72 and 96 h and then drastically decreased after 96 h. Solvent extraction followed by chromatographic purification steps led to the isolation of hydroquinone (benzene-1,4-diol). The structure of the purified compound was elucidated based on FTIR, (1)H NMR, and (13)C NMR spectroscopy. The compound exhibited efficient antibacterial activity against both Gram-positive and Gram-negative bacterial pathogens. The minimum inhibitory concentration (MIC) for Gram-positive pathogens ranged from 15.625 to 62.5 µg/mL(-1), while it was between 7.81 and 250 µg/mL(-1) for Gram-negative bacterial pathogens. This is the first report of hydroquinone produced by halotolerant B. methylotrophicus exhibiting promising antibacterial activity.
从位于印度杜蒂戈林和金奈的盐田中分离出一株具有耐盐性的细菌菌株MHC10,它对临床病原体具有广谱抗菌活性。对MHC10的16S rRNA基因分析表明,它与甲基营养芽孢杆菌(Bacillus methylotrophicus)具有高度相似性。利用不同的碳源、氮源、温度、pH值、氯化钠(NaCl)浓度和培养时间,对甲基营养芽孢杆菌MHC10菌株的培养条件进行了优化,以提高抗菌物质的产量。当以1%(w/v)葡萄糖、0.1%(w/v)豆粕作为优化的ZMB培养基,对应碳氮比为38.83,温度为37℃,pH值为7.0,氯化钠浓度为8%时,甲基营养芽孢杆菌MHC10的抗菌活性达到最大值。该活性在72至96小时之间保持稳定,96小时后急剧下降。通过溶剂萃取和色谱纯化步骤,分离出了对苯二酚(苯-1,4-二醇)。基于傅里叶变换红外光谱(FTIR)、氢核磁共振(¹H NMR)和碳核磁共振(¹³C NMR)光谱对纯化化合物的结构进行了阐明。该化合物对革兰氏阳性和革兰氏阴性细菌病原体均表现出高效的抗菌活性。革兰氏阳性病原体的最低抑菌浓度(MIC)范围为15.625至62.5 μg/mL⁻¹,而革兰氏阴性细菌病原体的MIC范围为7.81至250 μg/mL⁻¹。这是关于耐盐甲基营养芽孢杆菌产生的对苯二酚具有良好抗菌活性的首次报道。