Petroleum microbiology

Last updated August 30, 2023

Petroleum microbiology is a branch of microbiology that deals with the study of microorganisms that can metabolize or alter crude or refined petroleum products. These microorganisms, also called hydrocarbonoclastic microorganisms, can degrade hydrocarbons and, include a wide distribution of bacteria, methanogenic archaea, and some fungi. Not all hydrocarbonoclasic microbes depend on hydrocarbons to survive, but instead may use petroleum products as alternative carbon and energy sources. Interest in this field is growing due to the increasing use of bioremediation of oil spills.^[1]^[2]^[3]

Applications

Bioremediation

Bioremediation of oil contaminated soils, marine waters and oily sludges in situ is a feasible process as hydrocarbon degrading microorganisms are ubiquitous and are able to degrade most compounds in petroleum oil. In the simplest case, indigenous microbial communities can degrade the petroleum where the spill occurs. In more complicated cases, various methods of adding nutrients, air, or exogenous microorganisms to the contaminated site can be applied.^[4] For example, bioreactors involve the application of both natural and additional microorganisms in controlled growth conditions that yields high biodegradation rates and can be used with a wide range of media.^[4]

Crude oils are composed of an array of chemical compounds, minor constituents, and trace metals. Making up 50-98% of these petroleum products are hydrocarbons with saturated, unsaturated, or aromatic structures which influence their biodegradability by hydronocarbonclasts.^[5] The rate of uptake and biodegradation by these hydrocarbon-oxidizing microbes not only depend on the chemical structure of the substrates, but is limited by biotic and abiotic factors such as temperature, salinity, and nutrient availability in the environment.^[6]^[7]

Alcanivorax borkumensis

A model microorganism studied for its role in bioremediation of oil-spill sites and hydrocarbon catabolism is the alpha-proteobacteria Alcanivorax , which degrades aliphatic alkanes through various metabolic activities.^[6] Alcanivorax borkumensis utilizes linear hydrocarbon chains in petroleum as its primary energy source under aerobic conditions. When further supplied with sufficient limiting nutrients such as nitrogen and phosphor, it grows and produces surfactant glucolipids to help reduce surface water tension and enhance hydrocarbon uptake.[5] For this reason, nitrates and phosphates are often commercially added to oil-spill sites to engage quiescent populations of A. borkumensis, allowing them to quickly outcompete other microbial populations and become the dominant species in the oil-infested environment.^[8]^[9]

The addition of rate-limiting nutrients promotes the microbe's biodegrading pathways, including upregulation of genes encoding multiple alkane hydroxylases that oxidize various lengths of linear alkanes.^[10] These enzymes essentially remove the problematic hydrocarbon constituents of petroleum oil while A. borkumensis simultaneously increases synthesis of anionic glucoproteins, which are used to emulsify hydrocarbons in the environment and increase their bioavailability.^[10] The presence of crude oil along with appropriate levels of nitrogen and phosphor catalyzes the removal of petroleum either by mechanisms that enhance the efficiency of substrate uptake or by direct biodegradation of aliphatic chains.

Commercial applications

Two well-known oil spills exemplify large scale marine bioremediation applications:

In 1989, the Exxon Valdez ran aground, spilling 41.6 million liters of crude oil, and launching one of the first major bioremediation efforts for an oil spill. Cleanup of Alaskan shorelines relied in part on fertilizer application to augment bacterial growth.^[11]

In 2010, the BP Deepwater Horizon oil spill released 779 million liters of oil into the Gulf of Mexico. This was the largest oil spill of all time and indigenous petroleum microorganisms played a major role in petroleum degradation and cleanup.^[12]

Biosurfactants

These are microbial-synthesized surface-active substances that allow for more efficient microbial biodegradation of hydrocarbons in bioremediation processes. There are two ways by which biosurfactants are involved in bioremediation. (1) Increase the surface area of hydrophobic water-insoluble substrates. Growth of microbes on hydrocarbons can be limited by available surface area of the water-oil interface. Emulsifiers produced by microbes can break up oil into smaller droplets, effectively increasing the available surface area. (2) Increase the bioavailability of hydrophobic water-insoluble substrates. Biosurfactants can enhance the availability of bound substrates by desorbing them from surfaces (e.g. soil) or by increasing their apparent solubility. Some biosurfactants have low critical micelle concentrations (CMCs), a property which increases the apparent solubility of hydrocarbons by sequestering hydrophobic molecules into the centres of micelles.^[13]

Oil Recovery

Microbial enhanced oil recovery (MEOR) is a technology in which microbial environments are manipulated to enhance oil recovery. Nutrients are injected in situ into porous media and indigenous or added microbes promote growth and/or generate products that mobilize oil into producing wells. Alternatively, oil-mobilizing products can be produced by fermentation and injected into the reservoir. Various products and microorganisms are useful in these applications and each will yield different results. The two general strategies for enhancing oil recovery are altering the surface properties of the interface and using bioclogging to change the flow behavior.^[14] Biomass, biosurfactants, biopolymers, solvents, acids, and gases are some of the products that are added to oil reservoirs to enhance recovery.^[4] Other resources for this application:^[15]^[16]

Biosensors

Microbial biosensors identify and quantify target compounds of interest through interactions with the microbes. For example, bacteria may be used to identify a pollutant by monitoring their response to the specific chemical. The biosensor system may simply use bacterial growth as a pollutant indicator, or rely on genetic assays wherein a reporter gene is induced by the chemical.

Many analytical techniques require expensive treatment of soil samples and/or expensive equipment to detect the presence of pollutants. Bacterial biosensor systems offer the potential for cheap, robust detection systems that are selective and highly sensitive. One developed system uses Pseudomonas fluorescens HK44 to quantitatively assay for naphthalene using bioluminescence.^[17]

Challenges

Often in the process of degrading a pollutant, a microbe can create intermediates or byproducts that are also harmful, sometimes even more harmful than the original substrate. For example, some microbes produce hydrogen sulfide as a byproduct in the degradation of certain petroleum hydrocarbons and if those gases are not detoxified before escaping the system, they can be released into the atmosphere.^[18]

Biodegradation pathways

The pathways of degradation of different petroleum products vary depending on the substrate and the microorganism (i.e. aerobic/anaerobic). Specific degradation pathways of many hydrocarbon compounds can be found on the University of Minnesota Biocatalysis/Biodegradation Database.

Related Research Articles

Geomicrobiology is the scientific field at the intersection of geology and microbiology and is a major subfield of geobiology. It concerns the role of microbes on geological and geochemical processes and effects of minerals and metals to microbial growth, activity and survival. Such interactions occur in the geosphere, the atmosphere and the hydrosphere. Geomicrobiology studies microorganisms that are driving the Earth's biogeochemical cycles, mediating mineral precipitation and dissolution, and sorbing and concentrating metals. The applications include for example bioremediation, mining, climate change mitigation and public drinking water supplies.

Bioremediation broadly refers to any process wherein a biological system, living or dead, is employed for removing environmental pollutants from air, water, soil, flue gasses, industrial effluents etc., in natural or artificial settings. The natural ability of organisms to adsorb, accumulate, and degrade common and emerging pollutants has attracted the use of biological resources in treatment of contaminated environment. In comparison to conventional physicochemical treatment methods bioremediation may offer considerable advantages as it aims to be sustainable, eco-friendly, cheap, and scalable. Most bioremediation is inadvertent, involving native organisms. Research on bioremediation is heavily focused on stimulating the process by inoculation of a polluted site with organisms or supplying nutrients to promote the growth. In principle, bioremediation could be used to reduce the impact of byproducts created from anthropogenic activities, such as industrialization and agricultural processes. Bioremediation could prove less expensive and more sustainable than other remediation alternatives.

Biological augmentation is the addition of archaea or bacterial cultures required to speed up the rate of degradation of a contaminant. Organisms that originate from contaminated areas may already be able to break down waste, but perhaps inefficiently and slowly.

Geobacter is a genus of bacteria. Geobacter species are anaerobic respiration bacterial species which have capabilities that make them useful in bioremediation. Geobacter was found to be the first organism with the ability to oxidize organic compounds and metals, including iron, radioactive metals, and petroleum compounds into environmentally benign carbon dioxide while using iron oxide or other available metals as electron acceptors. Geobacter species are also found to be able to respire upon a graphite electrode. They have been found in anaerobic conditions in soils and aquatic sediment.

Biostimulation involves the modification of the environment to stimulate existing bacteria capable of bioremediation. This can be done by addition of various forms of rate limiting nutrients and electron acceptors, such as phosphorus, nitrogen, oxygen, or carbon. Alternatively, remediation of halogenated contaminants in anaerobic environments may be stimulated by adding electron donors, thus allowing indigenous microorganisms to use the halogenated contaminants as electron acceptors. EPA Anaerobic Bioremediation Technologies Additives are usually added to the subsurface through injection wells, although injection well technology for biostimulation purposes is still emerging. Removal of the contaminated material is also an option, albeit an expensive one. Biostimulation can be enhanced by bioaugmentation. This process, overall, is referred to as bioremediation and is an EPA-approved method for reversing the presence of oil or gas spills. While biostimulation is usually associated with remediation of hydrocarbon or high production volume chemical spills, it is also potentially useful for treatment of less frequently encountered contaminant spills, such as pesticides, particularly herbicides.

Mycoremediation is a form of bioremediation in which fungi-based remediation methods are used to decontaminate the environment. Fungi have been proven to be a cheap, effective and environmentally sound way for removing a wide array of contaminants from damaged environments or wastewater. These contaminants include heavy metals, organic pollutants, textile dyes, leather tanning chemicals and wastewater, petroleum fuels, polycyclic aromatic hydrocarbons, pharmaceuticals and personal care products, pesticides and herbicides in land, fresh water, and marine environments.

In biology, syntrophy, synthrophy, or cross-feeding is the phenomenon of one species feeding on the metabolic products of another species to cope up with the energy limitations by electron transfer. In this type of biological interaction, metabolite transfer happens between two or more metabolically diverse microbial species that live in close proximity to each other. The growth of one partner depends on the nutrients, growth factors, or substrates provided by the other partner. Thus, syntrophism can be considered as an obligatory interdependency and a mutualistic metabolism between two different bacterial species.

Biomining is the technique of extracting metals from ores and other solid materials typically using prokaryotes, fungi or plants. These organisms secrete different organic compounds that chelate metals from the environment and bring it back to the cell where they are typically used to coordinate electrons. It was discovered in the mid 1900s that microorganisms use metals in the cell. Some microbes can use stable metals such as iron, copper, zinc, and gold as well as unstable atoms such as uranium and thorium. Large chemostats of microbes can be grown to leach metals from their media. These vats of culture can then be transformed into many marketable metal compounds. Biomining is an environmentally friendly technique compared to typical mining. Mining releases many pollutants while the only chemicals released from biomining is any metabolites or gasses that the bacteria secrete. The same concept can be used for bioremediation models. Bacteria can be inoculated into environments contaminated with metals, oils, or other toxic compounds. The bacteria can clean the environment by absorbing these toxic compounds to create energy in the cell. Bacteria can mine for metals, clean oil spills, purify gold, and use radioactive elements for energy.

Biosurfactant usually refers to surfactants of microbial origin. Most of the biosurfactants produced by microbes are synthesized extracellularly and many microbes are known to produce biosurfactants in large relative quantities. Some are of commercial interest. As a secondary metabolite of microorganisms, biosurfactants can be processed by the cultivation of biosurfactant producing microorganisms in the stationary phase on many sorts of low-priced substrates like biochar, plant oils, carbohydrates, wastes, etc. High-level production of biosurfactants can be controlled by regulation of environmental factors and growth circumstances.

Microbial biodegradation is the use of bioremediation and biotransformation methods to harness the naturally occurring ability of microbial xenobiotic metabolism to degrade, transform or accumulate environmental pollutants, including hydrocarbons, polychlorinated biphenyls (PCBs), polyaromatic hydrocarbons (PAHs), heterocyclic compounds, pharmaceutical substances, radionuclides and metals.

<span class="mw-page-title-main">Phototrophic biofilm</span> Microbial communities including microorganisms which use light as their energy source

Phototrophic biofilms are microbial communities generally comprising both phototrophic microorganisms, which use light as their energy source, and chemoheterotrophs. Thick laminated multilayered phototrophic biofilms are usually referred to as microbial mats or phototrophic mats. These organisms, which can be prokaryotic or eukaryotic organisms like bacteria, cyanobacteria, fungi, and microalgae, make up diverse microbial communities that are affixed in a mucous matrix, or film. These biofilms occur on contact surfaces in a range of terrestrial and aquatic environments. The formation of biofilms is a complex process and is dependent upon the availability of light as well as the relationships between the microorganisms. Biofilms serve a variety of roles in aquatic, terrestrial, and extreme environments; these roles include functions which are both beneficial and detrimental to the environment. In addition to these natural roles, phototrophic biofilms have also been adapted for applications such as crop production and protection, bioremediation, and wastewater treatment.

S-200 is a bioremediation product used to clean up oil spills. It is an oleophilic nitrogen-phosphorus nutrient that promotes the growth of micro-organisms that degrade hydrocarbons. S-200 bonds to the hydrocarbon to eliminate the need to reapply in tidal or rain events. S-200 is identified as a bioremediation accelerator and as such, does not contain bacterial cultures, but rather contributes to the establishment of a robust microbial population. In the laboratory, considerable biodegradation to alkanes was seen over the course of treatment. Field trials have yielded inconsistent results.

Alcanivorax borkumensis is an alkane-degrading marine bacterium which naturally propagates and becomes predominant in crude-oil-containing seawater when nitrogen and phosphorus nutrients are supplemented.

The water associated fraction (WAF), sometimes termed the water-soluble fraction (W.S.F.), is the solution of low molecular mass hydrocarbons naturally released from petroleum hydrocarbon mixtures in contact with water. Although generally regarded as hydrophobic, many petroleum hydrocarbons are soluble in water to a limited extent. This combination often also contains less soluble, higher molecular mass components, and more soluble products of chemical and biological degradation.

Microbial enhanced oil recovery (MEOR) is a biological based technology consisting in manipulating function or structure, or both, of microbial environments existing in oil reservoirs. The ultimate aim of MEOR is to improve the recovery of oil entrapped in porous media while increasing economic profits. MEOR is a tertiary oil extraction technology allowing the partial recovery of the commonly residual two-thirds of oil, thus extending the life of mature oil reservoirs.

Bioremediation of petroleum contaminated environments is a process in which the biological pathways within microorganisms or plants are used to degrade or sequester toxic hydrocarbons, heavy metals, and other volatile organic compounds found within fossil fuels. Oil spills happen frequently at varying degrees along with all aspects of the petroleum supply chain, presenting a complex array of issues for both environmental and public health. While traditional cleanup methods such as chemical or manual containment and removal often result in rapid results, bioremediation is less labor-intensive, expensive, and averts chemical or mechanical damage. The efficiency and effectiveness of bioremediation efforts are based on maintaining ideal conditions, such as pH, RED-OX potential, temperature, moisture, oxygen abundance, nutrient availability, soil composition, and pollutant structure, for the desired organism or biological pathway to facilitate reactions. Three main types of bioremediation used for petroleum spills include microbial remediation, phytoremediation, and mycoremediation. Bioremediation has been implemented in various notable oil spills including the 1989 Exxon Valdez incident where the application of fertilizer on affected shoreline increased rates of biodegradation.

Bioremediation is the process of decontaminating polluted sites through the usage of either endogenous or external microorganism. In situ is a term utilized within a variety of fields meaning "on site" and refers to the location of an event. Within the context of bioremediation, in situ indicates that the location of the bioremediation has occurred at the site of contamination without the translocation of the polluted materials. Bioremediation is used to neutralize pollutants including Hydrocarbons, chlorinated compounds, nitrates, toxic metals and other pollutants through a variety of chemical mechanisms. Microorganism used in the process of bioremediation can either be implanted or cultivated within the site through the application of fertilizers and other nutrients. Common polluted sites targeted by bioremediation are groundwater/aquifers and polluted soils. Aquatic ecosystems affected by oil spills have also shown improvement through the application of bioremediation. The most notable cases being the Deepwater Horizon oil spill in 2010 and the Exxon Valdez oil spill in 1989. Two variations of bioremediation exist defined by the location where the process occurs. Ex situ bioremediation occurs at a location separate from the contaminated site and involves the translocation of the contaminated material. In situ occurs within the site of contamination In situ bioremediation can further be categorized by the metabolism occurring, aerobic and anaerobic, and by the level of human involvement.

Banwari Lal is an Indian environmental and industrial biotechnologist and the director of the Environmental and Industrial Biotechnology Division at The Energy and Resources Institute (TERI). Known for the development of oilzapper technology, Dr. Lal is the chief executive officer of ONGC-TERI Biotech Limited, a collaborative venture between TERI and the Oil and Natural Gas Corporation since 2008. The Department of Biotechnology of the Government of India awarded him the National Bioscience Award for Career Development, one of the highest Indian science awards, for his contributions to biosciences in 2004. He have many Indian and international joint patents with ONGC, DBT, IOCL, OIL INDIA and TERI.

Gordonia sp. nov. Q8 is a bacterium in the phylum of Actinomycetota. It was discovered in 2017 as one of eighteen new species isolated from the Jiangsu Wei5 oilfield in East China with the potential for bioremediation. Strain Q8 is rod-shaped and gram-positive with dimensions 1.0–4.0 μm × 0.5–1.2 μm and an optimal growth temperature of 40 °C. Phylogenetically, it is most closely related to Gordonia paraffinivorans and Gordonia alkaliphila, both of which are known bioremediators. Q8 was assigned as a novel species based on a <70% ratio of DNA homology with other Gordonia bacteria.

Hydrocarbonoclastic bacteria are a heterogeneous group of prokaryotes which can degrade and utilize hydrocarbon compounds as source of carbon and energy. Despite being present in most of environments around the world, several of these specialized bacteria live in the sea and have been isolated from polluted seawater.

References

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