低温萘降解菌的筛选、鉴定及降解条件优化

doi:10.13560/j.cnki.biotech.bull.1985.2019-0362

摘要/Abstract

摘要： 研究采用富集培养法从黑龙江省大庆油田地区污染土壤中筛选能以萘为唯一碳源和能源的低温菌株,采用气相色谱-质谱法（GC-MS）研究降解菌在萘-无机盐培养基中对萘的降解情况,通过单因素试验与正交试验测定降解菌的培养条件并进行优化,同时分析降解阶段其主控因素。结果表明：筛选出2株在低温条件下高效降解萘的菌株,编号为GN1和GN2。在低温条件下GN1和GN2可以快速降解萘,在对照组非生物因素影响基础上,萘（300 mg/L）的降解率在4 d内达到94.43%和95.47%,在耐受能力和降解速度方面具明显优势;经形态观察、生理生化特性和16S rDNA基因序列鉴定两株降解菌皆属于假单胞菌属（Pseudomonas）;均在萘-无机盐培养基（萘浓度300 mg/L）,培养温度15℃,初始pH 6.0,培养转数180 r/min,培养时间7 d的条件下生长最佳。2株降解菌的生长与5种环境因素均有显著关系。

关键词: 低温, 萘, 生物降解, 假单胞菌属

Abstract: The enrichment culture method was used to screen low temperature strains using naphthalene as the sole carbon source and energy source from the contaminated soil in Daqing Oilfield,Heilongjiang Province. The gas chromatography-mass spectrometry（GC-MS）was employed to study the degradation of naphthalene by degrading bacteria in naphthalene-inorganic salt medium. The single-factor test and orthogonal test were applied to determine and optimize the culture conditions of the degrading bacteria,and the main controlling factors in the degradation stage were analyzed. The results showed that 2 strains efficiently degrading naphthalene under low temperature condition were screened and named as GN1 and GN2. GN1 and GN2 rapidly degraded naphthalene under low temperature conditions. On the basis of the non-biological factors of the control group,the degradation rate of naphthalene（300 mg/L）reached 94.43% and 95.47% in 4 days,indicating that they had obvious advantages in tolerance and degradation rate. The 2 strains were identified as Pseudomonas by morphological observation,physiological and biochemical characteristics and 16S rDNA gene. The strains GN1 and GN2 grew best in naphthalene-inorganic salt medium（Naphthalene concentration 300 mg/L）,culture temperature 15℃,initial pH 6.0,culture rotation 180 r/min,and culture time 7 d. The growth of the 2 bacteria was significantly related to 5 environmental factors.

Key words: low temperature, naphthalene, biodegradation, Pseudomonas

郭亚男, 张馨予, 胥梦, 王继华. 低温萘降解菌的筛选、鉴定及降解条件优化[J]. 生物技术通报, 2019, 35(7): 100-107.

GUO Ya-nan, ZHANG Xin-yu, XU Meng, WANG Ji-hua. Screening,Identification of Naphthalene Degrading Bacteria at Low Temperature and Optimization of Their Degradation Conditions[J]. Biotechnology Bulletin, 2019, 35(7): 100-107.

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