Alkane Hydroxylase Research Articles

Whole-genome sequence-based analysis of the Paenibacillus aquistagni strain DK1, a polyethylene-degrading bacterium isolated from landfill.

Polyethylene-degrading bacteria have been emerging as a rational and safe alternative in bioremediation strategies. In this context, some Paenibacillus species produce enzymes involved in the biodegradation of pollutants. Among the enzymes involved in the biodegradation of polyethylene, the alkane hydroxylases, encoded by alkB homologous genes, play a key role in this process. Therefore, this study aimed to identify and perform a genomic investigation of the first polyethylene-degrading Paenibacillus sp. strain, named DK1. The whole-genome sequence-based analysis revealed that the DK1 strain belonged to the species Paenibacillus aquistagni and shared a total of 4327 CDSs with P. aquistagni strain 11. On the other hand, a comparison of the gene clusters showed that DK1 strain harbored a genetic context surrounding the alkB-like gene similar to that found in Pseudomonas sp. strains. The percentage of similarity ranged from 47.88 to 99.76% among all complete amino acid sequences of AlkB-like proteins analyzed. Nevertheless, the predicted amino acid sequences of AlkB-like contained typical structural motifs of alkane hydroxylases, such as His boxes and the HYG motif. These findings associated with the previously reported phenotypic results highlighted the potential of P. aquistagni strain DK1 to biodegrade polyethylene. Therefore, further studies focusing on the biochemical and structural properties of the AlkB-like protein from Paenibacillus may also contribute to the development of sustainable bioremediation strategies.

Enhanced degradation of different crude oils by defined engineered consortia of Acinetobacter venetianus RAG-1 mutants based on their alkane metabolism

Microbial consortia offer an attractive biodegradation strategy for removing hydrocarbons from oil-contaminated sites. In this study, we explored the degradation properties of Acinetobacter venetianus strain RAG-1 (RAG-1). RAG-1 effectively degrades three crude oils with excellent emulsification activity and cell surface hydrophobicity, while exhibiting broad environmental tolerance. RAG-1 accepts a range of alkane substrates (C10–C38) using three alkane hydroxylases (AlkMa, AlkMb, and AlmA). Bacterial mutant with alkMa or alkMb deletion enhanced degradation of C10–C20 or C22–C32 n-alkanes, respectively. Based on the substrate metabolism of the mutants, adjustable and targeted consortia consisting of ΔalkMa/almA and ΔalkMb were constructed, achieving enhanced degradation (10 days) of light crude oil (73.42% to 88.65%), viscous crude oil (68.40% to 90.05%), and high waxy crude oil (47.46% to 60.52%) compared with the single wild-type strain. The degradation properties of RAG-1 and the engineered consortia strategy may have potential use in microbial biodegradation applications.

Potential for and Distribution of Enzymatic Biodegradation of Polystyrene by Environmental Microorganisms.

Polystyrene (PS) is one of the main polymer types of plastic wastes and is known to be resistant to biodegradation, resulting in PS waste persistence in the environment. Although previous studies have reported that some microorganisms can degrade PS, enzymes and mechanisms of microorganism PS biodegradation are still unknown. In this study, we summarized microbial species that have been identified to degrade PS. By screening the available genome information of microorganisms that have been reported to degrade PS for enzymes with functional potential to depolymerize PS, we predicted target PS-degrading enzymes. We found that cytochrome P4500s, alkane hydroxylases and monooxygenases ranked as the top potential enzyme classes that can degrade PS since they can break C–C bonds. Ring-hydroxylating dioxygenases may be able to break the side-chain of PS and oxidize the aromatic ring compounds generated from the decomposition of PS. These target enzymes were distributed in Proteobacteria, Actinobacteria, Bacteroidetes, and Firmicutes, suggesting a broad potential for PS biodegradation in various earth environments and microbiomes. Our results provide insight into the enzymatic degradation of PS and suggestions for realizing the biodegradation of this recalcitrant plastic.

New evidence for a hydroxylation pathway for anaerobic alkane degradation supported by analyses of functional genes and signature metabolites in oil reservoirs

Microbial degradation of recalcitrant alkanes under anaerobic conditions results in the accumulation of heavy oil fraction in oil reservoirs. Hydroxylation of alkanes is an important activation mechanism under anaerobic conditions, but the diversity and distribution of the responsible microorganisms in the subsurface environment are still unclear. The lack of functional gene polymerase chain reaction (PCR) primers and commercially available intermediate degradation chemical compounds are the major obstacles for this research. In this investigation, PCR primers for the ahyA gene (encoding alkane hydroxylase) were designed, evaluated, and improved based on the nucleotide sequences available. Using microbial genomic DNA extracted from oil-contaminated soil and production water samples of oil reservoirs, ahyA gene nucleotide sequences were amplified and retrieved successfully from production water sample Z3-25 of Shengli oilfield. Additionally, the signature biomarker of 2-acetylalkanoic acid was detected in both Shengli and Jiangsu oilfields. These results demonstrate that anaerobic hydroxylation is an active mechanism used by microorganisms to degrade alkanes in oxygen-depleted oil reservoirs. This finding expands the current knowledge of biochemical reactions about alkane degradation in subsurface ecosystems. In addition, the PCR primers designed and tested in this study serve as an effective molecular tool for detecting the microorganisms responsible for anaerobic hydroxylation of alkanes in this and other ecosystems.

Microbial-induced biosurfactant-mediated biocatalytic approach for the bioremediation of simulated marine oil spill

Marine crude oil spill is the most alarming environmental issue and the dreadful instance of petroleum hydrocarbon pollution around the world. Successful remediation of these toxic pollutants demands for the ample production of biosurfactant and biocatalysts by the native hydrocarbon-degrading microbes. The main focus of this investigation is to determine the potential of a halo-tolerant and biosurfactant-producing hydrocarbonoclastic bacterium, Enterobacter hormaechei, for the effective bioremediation of marine oil spill. The strain had the ability to produce biosurfactant with excellent surface and emulsification activities along with biocatalysts. The activity of the extracellular enzymes such as lipase and laccase was found to be 160 U/ml and 38 U/ml, respectively, whereas the intracellular enzymes like alkane hydroxylase, alcohol dehydrogenase and esterase showed the corresponding activities of 48 U/ml, 86 U/ml and 102 U/ml. Overall, E. hormaechei could be able to degrade nearly 85% of petroleum hydrocarbons present in crude oil within 10 days of incubation. The biosurfactant was characterized to be an anionic, high molecular weight (48 kDa) lipoprotein-type biosurfactant. The biosurfactant was further characterized by Fourier transform infrared spectroscopy. The E. hormaechei was employed for the treatment of simulated marine oil spill and the degradation followed pseudo-second-order kinetics with rate constant k2 0.2775 and R2 0.9923. The crude oil degradation was confirmed by gas chromatography–mass spectrometry. The study suggested that the E. hormaechei is a potential biosurfactant and biocatalysts producer with the effective management of accidental marine oil spills.

Phylogeny and diversity of alkane-degrading enzyme gene variants in the laurentian great lakes and western atlantic.

Many aquatic environments are at risk for oil contamination and alkanes are one of the primary constituents of oil. The alkane hydroxylase (AlkB) is a common enzyme used by microorganisms to initiate the process of alkane-degradation. While many aspects of alkane bioremediation have been studied, the diversity and evolution of genes involved in hydrocarbon degradation from environmental settings is relatively understudied. The majority of work done to-date has focused on the marine environment. Here we sought to better understand the phylogenetic diversity of alkB genes across marine and freshwater settings using culture-independent methods. We hypothesized that there would be distinct phylogenetic diversity of alkB genes in freshwater relative to the marine environment. Our results confirm that alkB has distinct variants based on environment while our diversity analyses demonstrate that freshwater and marine alkB communities have unique responses to oil amendments. Our results also demonstrate that in the marine environment, depth is a key factor impacting diversity of alkB genes.

Bacterial bioaugmentation enhances hydrocarbon degradation, plant colonization and gene expression in diesel-contaminated soil.

Environmental contamination by hydrocarbons is a major problem, and hydrocarbon accumulation in soil poses hazardous threat to ecosystems. Phytoremediation, which involves plants, is an encouraging technique for the removal of hydrocarbons from polluted soil and water. The purpose of this investigation was to examine whether bacterial inoculation enhanced the phytoremediation of hydrocarbons in diesel-contaminated soil vegetated with maize (Zea mays L.). The two cultivars of maize, MMRI Yellow and Pearl White, were planted in diesel-polluted soil (0, 1.5, 2.5, and 3.5g diesel kg-1 soil), and inoculated with the consortium of three alkane-degrading bacterial strains, Arthrobacter oxydans ITRH49, Pseudomonas sp. ITRI73 and Pseudomonas sp. MixRI75. Bacterial inoculation enhanced plant growth and hydrocarbon degradation. Between two cultivars, MMRI Yellow showed better growth and hydrocarbon degradation in the presence and absence of bacterial inoculation. Maximum hydrocarbon degradation (80%) was observed in the soil having minimum concentration of diesel (1.5g kg-1 soil), and vegetated with bacterial inoculated MMRI Yellow maize cultivar. Furthermore, more bacterial colonization, and abundance and expression of the alkane hydroxylase gene (alkB) were observed in the root interior than in the rhizosphere and shoot interior of the plants. The bacteria-mediated phytoremediation of soil contaminated with hydrocarbons suggested that the collective use of plants and bacteria was the most beneficial approach for the reclamation of diesel-contaminated soil in comparison with vegetation alone.

Enhanced biodegradation of crude oil by constructed bacterial consortium comprising salt-tolerant petroleum degraders and biosurfactant producers

Bioremediation is an attractive strategy of utilizing bacteria to remove crude oil contaminants. In this study, two salt-tolerant crude oil-degrading and biosurfactant-producing bacteria, Dietzia sp. CN-3 and Acinetobacter sp. HC8–3S, were functionally combined to construct a bacterial consortium. The consortium achieved 95.8% degradation efficiency of crude oil in 10 days and various n-alkanes, cycloalkanes, branched alkanes and aromatic hydrocarbons were all depleted more effectively than single strains. Functional optimization of the consortium degraded crude oil efficiently in a wide range of pH (4–10) and salinity (0–120 g L−1). Furthermore, two alkane hydroxylase genes, alkB in CN-3 and alkM in HC8–3S, were cloned and their expression were examined by real-time quantitative polymerase chain reaction, indicating that alkB was more prominent in long-chain alkanes (C20, C24 and C26) utilization and alkM played crucial roles in medium- and long-chain alkanes (C14, C16, C20, C24 and C26) degradation. In soil microcosms artificially contaminated with crude oil and bioaugmented with the consortium, 58.3% of total petroleum hydrocarbons were depleted after 60 days and the degradation rate (485.8 mg kg−1 d−1) was higher than those reported in previous studies. Consequently, the consortium is a promising candidate in crude oil bioremediation.

Newly Isolated Alkane Hydroxylase and Lipase Producing Geobacillus and Anoxybacillus Species Involved in Crude Oil Degradation

Isolation and studies of novel, crude oil biodegrading thermophilic strains may provide a wider knowledge in understanding their role in petroleum degradation. In this study, the screening of ten new thermophilic strains revealed that all strains were alkane hydroxylase producers and seven of them produced lipase concurrently. Three best strains were characterized and identified through 16S rRNA sequence analysis as Geobacillus sp. D4, Geobacillus sp. D7, and Anoxybacillus geothermalis D9 with GenBank accession numbers MK615934.1, MK615935.1, and MK615936.1, respectively. Gas chromatography (GC) analysis showed that all three strains were able to breakdown various compounds in crude oil such as alkanes, toxic poly-aromatic hydrocarbons (PAHs), organosulfur, carboxylic acids, alkene, resins, organosilicon, alcohol, organochlorine, and ester. For the first time, alkane hydroxylase and lipase activity as well as crude oil degradation by A. geothermalis species were reported. Geobacillus sp. D7 is the best alkane degrader followed by A. geothermalis D9 and Geobacillus sp. D4 with 17.3%, 13.1%, and 12.1% biodegradation efficiency (BE%), respectively. The potential of thermophiles isolated can be explored further for bioremediation of sites polluted by petroleum and oil spills.

GDSL lipase occluded stomatal pore 1 is required for wax biosynthesis and stomatal cuticular ledge formation.

The plant leaf surface is coated with a waterproof cuticle layer. Cuticle facing the stomatal pore surface needs to be sculpted to form outer cuticular ledge (OCL) after stomatal maturation for efficient gas exchange. Here, we characterized the roles of Arabidopsis GDSL lipase, Occlusion of Stomatal Pore 1 (OSP1), in wax biosynthesis and stomatal OCL formation. OSP1 mutation results in significant reduction in leaf wax synthesis and occlusion of stomata, leading to increased epidermal permeability, decreased transpiration rate, and enhanced drought tolerance. We demonstrated that OSP1 activity is critical for its role in wax biosynthesis and stomatal function. In vitro enzymatic assays demonstrated that OSP1 possesses thioesterase activity, particularly on C22:0 and C26:0 acyl-CoAs. Genetic interaction analyses with CER1 (ECERIFERUM 1), CER3 (ECERIFERUM 3) and MAH1 (Mid-chain Alkane Hydroxylase 1) in wax biosynthesis and stomatal OCL formation showed that OSP1 may act upstream of CER3 in wax biosynthesis, and implicate that wax composition percentage changes and keeping ketones in a lower level play roles, at least partially, in forming stomatal ledges. Our findings provided insights into the molecular mechanism mediating wax biosynthesis and highlighted the link between wax biosynthesis and the process of stomatal OCL formation.

Isolation and characterization of hydrocarbon-degrading bacteria from gas station leaking-contaminated groundwater in the Southern Amazon, Brazil.

Twenty-three hydrocarbon-degrading bacteria strains were isolated from gas station leaking-contaminated groundwater located in the Southern Amazon, Brazil. Based on hydrocarbon (diesel, hexadecane, benzene, toluene and xylene) degradation ability, two strains were selected for further study. The amplification and sequencing of the 16S rRNA gene showed that these two strains belonged to the genus Bacillus (Bacillus sp. L26 and Bacillus sp. L30). GC-MS analysis showed that strain L30 was the most effective in degrading n-alkane (C10-C27) from diesel after 7 days of cultivation in mineral medium. Both strains produced biosurfactants and showed emulsification activity, specially the strain L30. Alkane hydroxylase gene (group III), which is important for alkane biodegradation, was present in strains. As a result, this study indicated that these bacteria could have promising applications in hydrocarbon bioremediation.

Identification of a Transcription Factor that Represses Alkane Hydroxylase Expression

Some gram negative and gram positive bacteria have evolved enzymes for metabolizing alkanes. A medium‐chain alkane hydroxylase has been found in the model organism Caulobacter crescentus, belonging to the CYP153 family of cytochrome P450 enzymes. However, the molecular mechanisms regulating expression of this enzyme are still unknown. We carried out a reverse genetics screen and RNA‐seq to identify CoxR as a transcription factor regulating this alkane hydroxylase. A deletion of coxR resulted in upregulation of both the alkane hydroxylase as well as ferredoxin and ferredoxin reductase enzymes. EMSA assays showed that CoxR bound directly to a short region of the alkane hydroxylase promoter. In summary, we have identified a novel transcription factor involved in metabolism of alkanes.

Integrated Comparative Genomic Analysis and Phenotypic Profiling of Pseudomonas aeruginosa Isolates From Crude Oil.

Pseudomonas aeruginosa is an environmental microorganism that can thrive in diverse ecological niches including plants, animals, water, soil, and crude oil. It also one of the microorganism widely used in tertiary recovery of crude oil and bioremediation. However, the genomic information regarding the mechanisms of survival and adapation of this bacterium in crude oil is still limited. In this study, three Pseudomonads strains (named as IMP66, IMP67, and IMP68) isolated from crude oil were taken for whole-genome sequencing by using a hybridized PacBio and Illumina approach. The phylogeny analysis showed that the three strains were all P. aeruginosa species and clustered in clade 1, the group with PAO1 as a representitive. Subsequent comparative genomic analysis revealed a high degree of individual genomic plasticity, with a probable alkane degradation genomic island, one type I-F CRISPR-Cas system and several prophages integrated into their genomes. Nine genes encoding alkane hydroxylases (AHs) homologs were found in each strain, which might enable these strains to degrade alkane in crude oil. P. aeruginosa can produce rhamnolipids (RLs) biosurfactant to emulsify oil, which enables their survival in crude oil enviroments. Our previous report showed that IMP67 and IMP68 were high RLs producers, while IMP66 produced little RLs. Genomic analysis suggested that their RLs yield was not likely due to differences at genetic level. We then further analyzed the quorum sensing (QS) signal molecules that regulate RLs synthesis. IMP67 and IMP68 produced more N-acyl-homoserine lactones (AHLs) signal molecules than that of PAO1 and IMP66, which could explain their high RLs yield. This study provides evidence for adaptation of P. aeruginosa in crude oil and proposes the potential application of IMP67 and IMP68 in microbial-enhanced oil recovery and bioremediation.

Draft Genome Sequence of Rhodococcus aetherivorans JCM 14343T, a Bacterium Capable of Degrading Recalcitrant Noncyclic and Cyclic Ethers.

Here, we report the draft genome sequence of Rhodococcus aetherivorans JCM 14343T, which possesses the versatile ability to degrade recalcitrant noncyclic and cyclic ether compounds. The 4.2-Mbp genome of this bacterium contains alkane hydroxylase and propane monooxygenase genes involved in the degradation of noncyclic and cyclic ethers, respectively.

Kurstakin molecules facilitate diesel oil assimilation by Acinetobacter haemolyticus strain 2SA through overexpression of alkane hydroxylase genes

ABSTRACT Biodegradation is a cost-effective process commonly used to eliminate many xenobiotic hydrocarbons such as diesel oils. However, their hydrophobic character reduces the biodegradation efficiency. In order to overcome this hurdle, kurstakins isolated from Bacillus thuringiensis strain 7SA were used as emulsifying agents. The influence of kurstakin molecules on diesel oil degradation by Acinetobacter haemolyticus strain 2SA was evaluated in the presence and absence of the aforementioned lipopeptide. The degradation rates and gene expressions of alkane hydroxylases were evaluated at days 4, 10, 14 and 21. Results showed that kurstakin molecules increased the hydrophobicity of 2SA. Moreover, diesel oil degradation activities were higher in the presence of kurstakin with 29%, 35%, 29% and 23% improvement at 4th, 10th, 14th and 21st day respectively. Statistical analysis indicated that the difference between the degradation rates in the presence and absence of kurstakin was significant with p = 0.03. The detection of three different hydroxylase genes namely alkB, almA and cyp153 in 2SA genome, might have allowed more efficient degradability of alkanes. According to the real-time PCR results, cyp153 was the most induced gene during diesel oil degradation in the presence and absence of kurstakin. Yet, the three genes demonstrated higher levels of expression in the presence of kurstakin when compared to its absence. This study showed that kurstakins enhance the diesel oil biodegradation rate by increasing the hydrophobicity of 2SA. In addition to their anti-fungal activities, kurstakins can be used as biosurfactant to increase biodegradation of diesel oil.

Elucidation of multiple alkane hydroxylase systems in biodegradation of crude oil n-alkane pollution by Pseudomonas aeruginosa DN1.

The purpose of this study was to elucidate the characteristics of multiple alkane hydroxylase systems in Pseudomonas aeruginosa DN1, including two homologues of AlkB (AlkB1 and AlkB2 ), a CYP153 homologue (P450), and two homologues of Alm-like (AlmA1 and AlmA2 ). DN1 was capable of utilizing diverse n-alkanes with chain lengths from 8 to 40 C atoms as the sole carbon source, and displayed high degradation efficiency (＞85%) of crude oil and a majority of n-alkanes using gas chromatography method. RT-qPCR analysis showed that the five enzyme genes could be induced by n-alkanes ranging from medium-chain length to long-chain length which indicated the dissimilarity of expression between those genes when grown on different n-alkanes. Notably, the expression of alkB2 gene was upregulated in the presence of all of the tested n-alkanes, particularly responded to long-chain n-alkanes like C20 and C32 . Meanwhile, long-chain n-alkanes (C20 -C36 ) significantly elevated cyp153 expression level, and the expression of two almA genes was only upregulated in the presence of n-alkanes with chain lengths of 20C's and longer. Furthermore, the disruption of those genes demonstrated that AlkB2 appeared to play a key role in the biodegradation of substrates of a broad-chain length ranges, besides other alkane hydroxylase systems ensured the utilization of n-alkanes with chain lengths of from 20 to 40 C atoms. The five functional alkane hydroxylase genes make DN1 an attractive option for its versatile alkane degradation, which is primarily dependent on the expression of alkB2 . Our findings suggest that P. aeruginosa DN1 is a predominately potential long-chain n-alkane-degrading bacterium with multiple alkane hydroxylase systems in crude oil-contaminated environment.

Low-Abundance Dietzia Inhabiting a Water-Flooding Oil Reservoir and the Application Potential for Oil Recovery.

With the development of molecular ecology, increasing low-abundance microbial populations were detected in oil reservoirs. However, our knowledge about the oil recovery potential of these populations is lacking. In this study, the oil recovery potential of low-abundance Dietzia that accounts for less than 0.5% in microbial communities of a water-flooding oil reservoir was investigated. On the one hand, Dietzia sp. strain ZQ-4 was isolated from the water-flooding reservoir, and the oil recovery potential was evaluated from the perspective of metabolisms and oil-displacing test. On the other hand, the strain has alkane hydroxylase genes alkB and P450 CYP153 and can degrade hydrocarbons and produce surfactants. The core-flooding test indicated that displacing fluid with 2% ZQ-4 fermentation broth increased 18.82% oil displacement efficiency, and in situ fermentation of ZQ-4 increased 1.97% oil displacement efficiency. Furthermore, the responses of Dietzia in the reservoir accompanied by the nutrient stimulation process was investigated and showed that Dietzia in some oil production wells significantly increased in the initial phase of nutrient injection and sharply decreased along with the continuous nutrient injection. Overall, this study indicates that Dietzia sp. strain has application potential for enhancing oil recovery through an ex situ way, yet the ability of oil recovery in situ based on nutrient injection is limited.

Functional and Genomic Characterization of a Pseudomonas aeruginosa Strain Isolated From the Southwestern Gulf of Mexico Reveals an Enhanced Adaptation for Long-Chain Alkane Degradation

Indigenous bacterial populations play an important role in the restoration of crude oil-polluted marine environments. The identification and characterization of these bacteria are key in defining bioremediation strategies for the mitigation of possible future oil spills. In this work, we characterized Pseudomonas aeruginosa strain GOM1, which was isolated from the water column in the southwestern Gulf of Mexico. Phylogenetic analysis revealed that GOM1 strain was most closely related to P. aeruginosa WC55, a strain isolated from the northern Gulf of Mexico after the Deepwater Horizon oil spill. The hydrocarbon-degrading capacity of P. aeruginosa GOM1 was investigated using various approaches. This strain degraded 96% of the aliphatic fraction (C12-C38) of crude oil during a 30-day incubation period, exhibiting a high activity on long-chain alkanes, and expressing alkane hydroxylases AlkB1, AlkB2 and AlmA. Addition of nitrogen and phosphate to seawater culture medium enhanced hexadecane degradation by GOM1. Additionally, the strain exhibited high surfactant/rhamnolipid production and emulsifying activity when grown in a complex medium in the presence of hexadecane. Comparisons of growth kinetics, hydrocarbon degradation and gene expression between GOM1 and the closely related P. aeruginosa laboratory strain PAO1 revealed that the marine isolate is better adapted to degrade alkanes. Taken together, our results place P. aeruginosa GOM1 as a potentially effective candidate to be included in a consortium for use in the bioremediation of oil-polluted sites.

Crude oil degradation and biosurfactant production abilities of isolated Agrobacterium fabrum SLAJ731

Abstract Microbial biosurfactants have a wide range of applications as surface active agents. This study aimed to explore an indigenous bacterial strain, Agrobacterium fabrum SLAJ731, isolated from a core sample of Assam oil field for the biosurfactant production and biodegradation of crude oil, which were elucidated in terms of alkane hydroxylase (AH) activity. Preliminary screening was performed using drop collapse assay and oil displacement activity and culture conditions were optimized for biosurfactant production. The biodegradation of spent crude oil was explored under the optimized conditions in the presence of glucose as a co-substrate. The strain produced 5.77 ± 0.3 g/L of biosurfactant under optimal conditions of pH 6 and 30 °C using glucose and yeast extract (C:N = 2:1) with high emulsification activity of 65 ± 0.5%. The crude biosurfactant was characterized using FTIR and 1H NMR and was found to be lipopeptide with a critical micelle concentration value of 650 mg/L and reduced surface tension to 34 ± 0.5 mN/m. It was revealed from FTIR and GC-MS analyses that bacterium was able to degrade oil more efficiently in the presence of glucose and degraded aliphatic (47%) and aromatic hydrocarbons (70%) as compared to abiotic. Both extra and intra-cellular AH activities were found to enhance the biosurfactant production. This study puts forward a distinctive mechanism involved in the biosurfactant production and oil degradation, in the presence of a co-substrate. The notable lipopeptide production and oil degradation ability of A. fabrum endorse its suitability for bioremediation and other biotechnological applications.

Biodegradation of squalene and n-hexadecane by Gordonia amicalis HS-11 with concomitant formation of biosurfactant and carotenoids

Gordonia amicalis HS-11 has been enriched from a hydrocarbon contaminated tropical soil sample. The ability of this organism to utilize a triterpenic polyunsaturated hydrocarbon, squalene (2,6,10,15,19,23-hexamethyl-6,6,10,14,18,20-tetracosahexane) and the model saturated hydrocarbon n-hexadecane is described here. The isolate degraded squalene and n-hexadecane (79 ± 3.02 and 96 ± 4.11%, respectively) after eight days of incubation. The isolate produced an extracellular biosurfactant that reduced surface tension from 69 ± 2.83 to 40 ± 1.63 and 35 ± 2.34 mN m−1 with squalene and n-hexadecane as carbon sources, respectively, after 6 days. The Actinomycete cleaved squalene to geranylacetone and farnesyl acetaldehyde that were further utilized for supporting growth. n-Hexadecane was degraded via monoterminal oxidation and activities of important enzymes (alkane hydroxylase and alcohol dehydrogenase) were highest (215 ± 8.76 and 169 ± 6.02 units mg−1 protein, respectively) after four days. Cells grown on squalene were short and with n-hexadecane there were clumps of longer cells. Squalene and n-hexadecane-grown cell surfaces were smooth possibly due to extracellular surface active compounds. While growing on hydrophobic substrates, some cells were seen adhering to droplets and others were in the free form. The culture was able to simultaneously degrade hydrocarbons and produce two commercially relevant value-added products. The yield of the extracellular biosurfactant on n-hexadecane was 480 mg l−1 and cells grown on squalene and n-hexadecane also yielded carotenoids (2.3 ± 0.14 and 2.9 ± 0.10 mg g−1 dry cell weight, respectively). This is the first report on the utilization of squalene by Gordonia.