[Objective] Toisolate and characterize bacteria that can be used todevelop microbial biosensor for methanol (MeOH) determination.[Methods] We used selective medium and streak plate to isolate bacteria.Morphological,physiological characteristics and 16S rDNA sequence analysis were used to identify the strain.AnMeOHbiosensorwas then developed by immobilizing M211 along with dissolved oxygen (O2) sensor.[Result] AnMeOH utilizing bacteriumwas isolated from biogas-producing tank using methanol as the sole carbon source,and identified as Methylobacteriumorganophilium.Decrease of O2 concentration is linearly related to the MeOH concentration in the range from 0.02% to 1%,with the MeOH detection limit of O.27mg/L.The response time of the biosensor is within 20 min.Furthermore,the result of interference test and the detection of methanol sample are both satisfactory.[Conclusion] Good results are obtainedin interference test and the detection of methanol sample.The proposed methodseems very attractive in monitoring methanol.%[目的]筛选能够利用甲醇的微生物菌株,并将其应用于甲醇测定的初步研究中.[方法]采用富集培养及固体平板分离法分离菌株;通过生理生化特征及16S rDNA序列分析进行菌株鉴定;以固定化细胞和溶氧仪组合成测定系统,测定了不同含量甲醇的响应时间以及溶氧变化与甲醇含量的关系.[结果]从山西清徐农用沼气池中分离得到一株能以甲醇为唯一碳源和能源生长的菌株M211,通过生理生化特征及16SrDNA序列分析,该菌株被鉴定为嗜有机甲基杆菌(Methylobacterium organophilium).传感系统在0.02%-1％ (V/V)甲醇含量范围内测定,响应时间小于20 min,检出限为0.27 mg/L,溶氧消耗量同甲醇含量呈线性关系,拟合系数(R2)为0.9986.该测定体系不易受其它干扰物质影响,检测结果与实际样品浓度一致.[结论]该测定体系具有较强的选择性,及良好的重现性和稳定性,具有较好的应用前景.
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