Cervantes-González E, Rojas-Avelizapa N G, Cruz-Camarillo R, García-Mena J, Rojas-Avelizapa L I
Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas IPN. Carpio y Plan de Ayala S/N. Col. Casco de Santo Tomás. México 11340 D.F., México.
Environ Technol. 2008 Feb;29(2):171-82. doi: 10.1080/09593330802028659.
The aim of this work was to isolate oil-degrading bacteria that use chitin or keratin as carbon sources from oil contaminated soils; and additionally to study if oil removal by these bacteria is enhanced when a chitinous or a keratinous waste is added to the culture media. To isolate the above-mentioned bacteria, 12 soil samples were collected close to an oil-well. Such soils showed unsuitable nutrients content, but their counts of heterotrophic bacteria ranged within 10(5)-10(8) CFU g(-1) soil, of which 0.1-77% corresponded to oil hydrocarbon-degrading ones. By sampling on plates, 109 oil-degrading bacterial isolates were obtained. Their keratinase and chitinase activities were then screened by plate assays and spectrophotometric methods, resulting in 13 isolates that were used to integrate two mixed cultures, one keratinolytic and the other chitinolytic. These mixed cultures were grown in media with oil, or oil supplemented with chicken-feathers or shrimp wastes. The oil-hydrocarbon removal was measured by gas chromatography. Results showed that keratinolytic bacteria were better enzyme producers than the chitinolytic ones, and that oil removal in the presence of chicken-feathers was 3.8 times greater than with shrimp wastes, and almost twice, in comparison with oil-only added cultures. Identification of microorganisms from the mixed cultures by 16S rDNA, indicated the presence of seven different bacterial genera; Stenotrophomonas, Pseudomonas, Brevibacillus, Bacillus, Micrococcus, Lysobacter and Nocardiodes. These findings suggest that the isolated microorganisms and the chicken-feather wastes could be applied to the cleaning of oil-contaminated environments, whether in soil or water.
本研究旨在从受石油污染的土壤中分离出以几丁质或角蛋白为碳源的石油降解细菌;此外,研究向培养基中添加含几丁质或角蛋白的废物时,这些细菌对石油的去除效果是否会增强。为了分离上述细菌,在油井附近采集了12份土壤样本。这些土壤的养分含量不合适,但其异养细菌数量在10(5)-10(8) CFU g(-1)土壤范围内,其中0.1-77%为石油烃降解菌。通过平板取样,获得了109株石油降解细菌分离株。然后通过平板测定和分光光度法筛选它们的角蛋白酶和几丁质酶活性,得到13株分离株,用于构建两种混合培养物,一种是角蛋白分解菌混合培养物,另一种是几丁质分解菌混合培养物。这些混合培养物在含油或添加鸡毛或虾壳废物的油培养基中生长。通过气相色谱法测定石油烃的去除量。结果表明,角蛋白分解菌比几丁质分解菌产生的酶更多,并且在存在鸡毛的情况下,石油去除量比存在虾壳废物时高3.8倍,与仅添加油的培养物相比几乎高出两倍。通过16S rDNA对混合培养物中的微生物进行鉴定,表明存在七个不同的细菌属:嗜麦芽窄食单胞菌、假单胞菌、短短芽孢杆菌、芽孢杆菌、微球菌、溶杆菌和诺卡氏菌属。这些发现表明,分离出的微生物和鸡毛废物可应用于清洁受石油污染的环境,无论是土壤还是水体。