耐盐石油烃降解菌的筛选鉴定及其特性研究

为得到高效耐盐石油烃降解菌,从黄河三角洲石油污染盐渍化土壤中分离出39株细菌,经液体培养初筛和土壤培养复筛实验,得到1株高效耐盐石油烃降解菌BM38.通过形态特征、生理生化特征和16S rDNA序列分析,确定该菌为恶臭假单胞菌（Pseudomonas putida）.通过液体培养实验,研究了BM38的耐盐和产生物表面活性剂特性以及对不同烃的利用能力.结果表明,在含0.5%～6.0%NaCl液体培养基中BM38生长良好,属中度耐盐菌.在高盐环境下BM38具有较强的分解石油烃能力,其中在含1.0%NaCl液体培养基中,降解7 d后,原油降解率达到73.5%;在含盐量0.22%和0.61%土壤中添加BM38,降解40 d后,土壤总石油烃降解率达到40%以上.BM38能产生一种生物乳化剂,盐浓度对这种乳化剂的乳化能力影响较大,当NaCl浓度增加到1.0%,乳化值（EI24）开始迅速降低,但在NaCl浓度为2.0%时,EI24仍达到61.0%.BM38能够利用环己烷、甲苯、异辛烷、菲和正十六烷为唯一碳源生长,其中对正构烷烃和芳烃具有较强的利用能力.

Isolation, Identification and Characterization of Halotolerant Petroleum-degrading Bacteria

To obtain efficient halotolerant petroleum-degrading bacteria, 39 bacteria strains were isolated from 30 petroleum contaminated saline soil samples in Yellow River Delta, an important base of petroleum production in China. One bacterium (strain BM38) was found to efficiently degrade crude oil in highly saline environments based on a series of liquid and soil incubation experiments. According to its morphology, physiochemical characteristics and 16S rDNA sequence analysis, this strain was identified as Pseudomonas putida. Moreover, a series of liquid incubation experiments were conducted to investigate its characteristics such as halotolerance, biosurfactants production and degrading efficiency for various hydrocarbons. The salt resistance test demonstrated that strain BM38 grew well at NaCl concentrations ranging from 0.5% to 6.0%. Petroleum degradation experiments showed that strain BM38 could degrade 73.5% crude oil after 7 days in a liquid culture medium containing 1.0% NaCl and remove more than 40% of total petroleum hydrocarbons after 40 days in the soil with 0.22% and 0.61% of salinity, these results proved that the strain was effective in removing petroleum hydrocarbons. Strain BM38 could produce a bioemulsifier in a liquid culture medium. The NaCl concentration had the significant effect on the EI24 of fermentation broth, which decreased sharply if the NaCl concentration was greater than 1.0%. However, the EI24 of BM38 was still quite high in the presence of 2.0% of NaCl, and the value was 61.0%. Furthermore, this strain was also able to grow in mineral liquid media amended with hexadecane, toluene, phenanthrene, isooctane and cyclohexane as the sole carbon sources. Among these hydracarbons, strain BM38 showed relatively high ability in degrading n-alkanes and aromatic hydracarbons. The results indicated that strain BM38 had potential for application in bioremediation of petroleum-contaminated saline soil.