Characterization of dioxygenases and biosurfactants produced by crude oil degrading soil bacteria

ABSTRACT Role of microbes in bioremediation of oil spills has become inevitable owing to their eco friendly nature. This study focused on the isolation and characterization of bacterial strains with superior oil degrading potential from crude-oil contaminated soil. Three such bacterial strains were selected and subsequently identified by 16S rRNA gene sequence analysis as Corynebacterium aurimucosum, Acinetobacter baumannii and Microbacterium hydrocarbonoxydans respectively. The specific activity of catechol 1,2 dioxygenase (C12O) and catechol 2,3 dioxygenase (C23O) was determined in these three strains wherein the activity of C12O was more than that of C23O. Among the three strains, Microbacterium hydrocarbonoxydans exhibited superior crude oil degrading ability as evidenced by its superior growth rate in crude oil enriched medium and enhanced activity of dioxygenases. Also degradation of total petroleum hydrocarbon (TPH) in crude oil was higher with Microbacterium hydrocarbonoxydans. The three strains also produced biosurfactants of glycolipid nature as indicated d by biochemical, FTIR and GCMS analysis. These findings emphasize that such bacterial strains with superior oil degrading capacity may find their potential application in bioremediation of oil spills and conservation of marine and soil ecosystem.

Evaluation of bacterial diversity recovered from petroleum samples using different physical matrices

ABSTRACT Unraveling the microbial diversity and its complexity in petroleum reservoir environments has been a challenge throughout the years. Despite the techniques developed in order to improve methodologies involving DNA extraction from crude oil, microbial enrichments using different culture conditions can be applied as a way to increase the recovery of DNA from environments with low cellular density for further microbiological analyses. This work aimed at the evaluation of different matrices (arenite, shale and polyurethane foam) as support materials for microbial growth and biofilm formation in enrichments using a biodegraded petroleum sample as inoculum in sulfate reducing condition. Subsequent microbial diversity characterization was carried out using Scanning Electronic Microscopy (SEM), Denaturing Gradient Gel Electrophoresis (DGGE) and 16S rRNA gene libraries in order to compare the microbial biomass yield, DNA recovery efficiency and diversity among the enrichments. The DNA from microbial communities in petroleum enrichments was purified according to a protocol established in this work and used for 16S rRNA amplification with bacterial generic primers. The PCR products were cloned, and positive clones were screened by Amplified Ribosomal DNA Restriction Analysis (ARDRA). Sequencing and phylogenetic analyses revealed that the bacterial community was mostly represented by members of the genera Petrotoga, Bacillus, Pseudomonas, Geobacillus and Rahnella. The use of different support materials in the enrichments yielded an increase in microbial biomass and biofilm formation, indicating that these materials may be employed for efficient biomass recovery from petroleum reservoir samples. Nonetheless, the most diverse microbiota were recovered from the biodegraded petroleum sample using polyurethane foam cubes as support material.

Microbial demetallization of crude oil using Aspergilus sp.: vanadium oxide octaethyl porphyrin [VOOEP] as a model of metallic petroporphyrins

An isolate from polluted soil identified as Aspergillus sp. MS-100 was able to consume vanadium oxide octaethyl porphyrin as a model for protoporphyrins in crude oil. The isolate degrades about 55% of vanadium oxide octaethyl porphyrin [VOOEP] under optimum conditions during 7 days. The release of more than 0.96 mgL-1 of free vanadium into the aqueous phase was confirmed using atomic absorption. By using the Taguchi experimental design method, the optimum values of pH, temperature and initial concentration of VOOEP were determined as 5.5, 30C, and 20 mg/l, respectively. The reduction of VOOEP in the culture medium was accelerated by Ag+ and inhibited by Zn2+ and EDTA. The Sn2+ and Pb2+ ions showed a stimulatory effect at 0.1 mM and an inhibitory effect at 1 mM

Biodegradabilidad de componentes de mezclas naturales de hidrocarburos previamente sometidas a landfarming / Components biodegradability of natural hydrocarbon mixtures previously subjected to landfarming

La compleja composición del petróleo crudo, derivados y deshechos de las diferentes etapas de la industria petrolera hacen de este producto una mezcla que presenta diferentes dificultades para su eliminación por métodos biológicos. El objetivo de este trabajo fue mostrar el potencial de biodegradación de comunidades bacterianas autóctonas frente a hidrocarburos obtenidos de cuatro sitios contaminados y sometidos a biorremediación, en un sistema de Landfarming por casi un década. Los resultados mostraron una marcada diferencia de biodegradabilidad de las tres principales fracciones alifáticas, aromáticas, y polares, obtenidas por cromatografía en columna. Si bien todas las fracciones fueron utilizadas como fuente de carbono y energía, existieron variaciones importantes en la producción de biomasa entre ellas, como asimismo en la cinética de biodegradación, según la composición de cada fracción.

Effect aeration on biodegradation of petroleum waste

Large amaounts of oily sludge are generated as residues by the oil industry, representing a real problem for refineries. This work studied the technical viability of treating oily aludge biologically, through stimulation of native microorganisms, at bench scale. Such microorganisms were able to grow in a medium containing oily sludge as the only carbon and energy source. Two oily sludge concentrations were studied, 5(per cent) (v/v) and 10 (per cent) (v/v), with a C:N ratio of 100:1. Higher microbial populations were observed in the first case. Substrate inhibition and/or toxic effect took place in the second case. The importance of aeration on the microbial activity and on the biodegradation of the residue was ascertained. In terms of n-paraffins, pristane and phytane consumption, maximum global efficiency of the genus "Pseudomonas sp." predominated. Two yeast species were also identified and two filamentous fungi were isolated.

Some petroleum-utilizing bacterial isolates from Saudi Arabian soil

Microbial degradation of oil remains the best candidate for rapid and efficient removal of accidental pollution of the environment. Several bacterial species were isolated from urbian soil, obtained from different geographical areas in the kingdom, which could grow on synthetic medium containing whole petroleum as a carbon source. All isolates grew on upto 4% of the oil and on three types of oils from different sources. Most isolates belonged to the actinomycetes including Coryneform group and streptomyces. Gas chromatographic analysis of growth-exhausted oils showed the ability of some strains to utilize many components of these oils, While others could utilize low molecular weight components only