Engineering organisms for industrial fuel production

Volatile fuel costs, the need to reduce greenhouse gas emissions and fuel security concerns are driving efforts to produce sustainable renewable fuels and chemicals. Petroleum comes from sunlight, CO(2) and water converted via a biological intermediate into fuel over a several million year timescale. It stands to reason that using biology to short-circuit this time cycle offers an attractive alternative--but only with relevant products at or below market prices. The state of the art of biological engineering over the past five years has progressed to allow for market needs to drive innovation rather than trying to adapt existing approaches to the market. This report describes two innovations using synthetic biology to dis-intermediate fuel production. LS9 is developing a means to convert biological intermediates such as cellulosic hydrolysates into drop-in hydrocarbon product replacements such as diesel. Joule Unlimited is pioneering approaches to eliminate feedstock dependency by efficiently capturing sunlight, CO(2) and water to produce fuels and chemicals. The innovations behind these companies are built with the market in mind, focused on low cost biosynthesis of existing products of the petroleum industry. Through successful deployment of technologies such as those behind LS9 and Joule Unlimited, alternative sources of petroleum products will mitigate many of the issues faced with our petroleum-based economy.     

Current and Potential Biofuel Production from Plant Oils

Environmental concerns and depletion of fossil fuels along with government policies have led to the search for alternative fuels from various renewable and sustainable feedstocks. This review provides a critical overview of the chemical composition of common commercial plant oils, i.e., palm oil, olive oil, rapeseed oil, castor oil, WCO, and CTO and their recent trends toward potential biofuel production. Plant oils with a high energy content are primarily composed of triglycerides (generally >?95%), accompanied by diglycerides, monoglycerides, and free fatty acids. The heat content of plant oils is close to 90% for diesel fuels. The oxygen content is the most important difference in chemical composition between fossil oils and plant oils. Triglycerides can even be used directly in diesel engines. However, their high viscosity, low volatility, and poor cold flow properties can lead to engine problems. These problems require that plant oils need to be upgraded if they are to be used as a fuel in conventional diesel engines. Biodiesel, biooil, and renewable diesel are the three major biofuels obtained from plant oils. The main constraint associated with the production of biodiesel is the cost and sustainability of the feedstock. The renewable diesel obtained from crude tall oil is more sustainable than biofuels obtained from other feedstocks. The fuel properties of renewable diesel are similar to those of fossil fuels with reduced greenhouse gas emissions. In this review, the chemical composition of common commercial plant oils, i.e., palm oil, olive oil, rapeseed oil, castor oil, and tall oil, are presented. Both their major and minor components are discussed. Their compositions and fuel properties are compared to both fossil fuels and biofuels.     

Liquid and gaseous fuels from biotechnology: challenge and opportunities

Abstract: This paper presents challenging opportunities for production of liquid and gaseous fuels by biotechnology. From the liquid fuels, ethyl alcohol production has been widely researched and implemented. The major obstacle for large scale production of ethanol for fuel is the cost, whereby the substrate represents one of the major cost components. Various scenarios will be presented for a critical assessment of cost distribution for production of ethanol from various substrates by conventional and high rate processes. The paper also focuses on recent advances in the research and application of biotechnological processes and methods for the production of liquid transportation fuels other than ethanol (other oxygenates; diesel fuel extenders and substitutes), as well as gaseous fuels (biogas, methane, reformed syngas). Potential uses of these biofuels are described, along with environmental concerns which accompany them. Emphasis is also put on microalgal lipids as diesel substitute and biogas/methane as a renewable alternative to natural gas. The capturing and use of landfill gases is also mentioned, as well as microbial coal liquefaction. Described is also the construction and performance of microbial fuel cells for the direct high-efficiency conversion of chemical fuel energy to electricity. Bacterial carbon dioxide recovery is briefly dealt with as an environmental issue associated with the use of fossil energy.     

Biodiesel production from various feedstocks and their effects on the fuel properties

Biodiesel, which is a new, renewable and biological origin alternative diesel fuel, has been receiving more attention all over the world due to the energy needs and environmental consciousness. Biodiesel is usually produced from food-grade vegetable oils using transesterification process. Using food-grade vegetable oils is not economically feasible since they are more expensive than diesel fuel. Therefore, it is said that the main obstacle for commercialization of biodiesel is its high cost. Waste cooking oils, restaurant greases, soapstocks and animal fats are potential feedstocks for biodiesel production to lower the cost of biodiesel. However, to produce fuel-grade biodiesel, the characteristics of feedstock are very important during the initial research and production stage since the fuel properties mainly depend on the feedstock properties. This review paper presents both biodiesel productions from various feedstocks and their effects on the fuel properties. JIMB 2008: BioEnergy - Special issue.     

Recent Developments in Commercial Processes for Refining Bio-Feedstocks to Renewable Diesel

The process technologies for conversion of bio-feedstocks such as vegetable oils, animal fats, and algal oil into renewable diesel have been developed and commercialized during the last decade. The global annual production capacity of renewable diesel is approaching to 5.5 million tons per year. The refining process generally includes pretreatment of the renewable feedstock to remove impurities, hydroprocessing and isomerization to produce hydrocarbons, and distillation to produce a fuel suitable for use as diesel or jet fuel. This article reviews recent development in the commercial production of renewable diesel, pretreatment technologies, chemistry of deoxygenation and cracking of triglycerides, the effect of reaction parameters on the relative activities of different reaction pathways, catalyst development, and the technical details of commercial processes for refining bio-feedstocks.     

Combustion characteristics of a turbocharged DI compression ignition engine fueled with petroleum diesel fuels and biodiesel

In this study, the combustion characteristics and emissions of two different petroleum diesel fuels (No. 1 and No. 2) and biodiesel from soybean oil were compared. The tests were performed at steady state conditions in a four-cylinder turbocharged DI diesel engine at full load at 1400-rpm engine speed. The experimental results compared with No. 2 diesel fuel showed that biodiesel provided significant reductions in PM, CO, and unburned HC, the NO(x) increased by 11.2%. Biodiesel had a 13.8% increase in brake-specific fuel consumption due to its lower heating value. However, using No. 1 diesel fuel gave better emission results, NO(x) and brake-specific fuel consumption reduced by 16.1% and 1.2%, respectively. The values of the principal combustion characteristics of the biodiesel were obtained between two petroleum diesel fuels. The results indicated that biodiesel may be blended with No. 1 diesel fuel to be used without any modification on the engine.     

Fatty acid alkyl esters: perspectives for production of alternative biofuels

The global economy heads for a severe energy crisis: whereas the energy demand is going to rise, easily accessible sources of crude oil are expected to be depleted in only 10–20 years. Since a serious decline of oil supply and an associated collapse of the economy might be reality very soon, alternative energies and also biofuels that replace fossil fuels must be established. In addition, these alternatives should not further impair the environment and climate. About 90% of the biofuel market is currently captured by bioethanol and biodiesel. Biodiesel is composed of fatty acid alkyl esters (FAAE) and can be synthesized by chemical, enzymatic, or in vivo catalysis mainly from renewable resources. Biodiesel is already established as it is compatible with the existing fuel infrastructure, non-toxic, and has superior combustion characteristics than fossil diesel; and in 2008, the global production was 12.2 million tons. The biotechnological production of FAAE from low cost and abundant feedstocks like biomass will enable an appreciable substitution of petroleum diesel. To overcome high costs for immobilized enzymes, the in vivo synthesis of FAAE using bacteria represents a promising approach. This article points to the potential of different FAAE as alternative biofuels, e.g., by comparing their fuel properties. In addition to conventional production processes, this review presents natural and genetically engineered biological systems capable of in vivo FAAE synthesis.     

Anaerobic biodegradation of no. 2 diesel fuel in soil: a soil column study

Soil and sediments are contaminated with petroleum hydrocarbons in many parts of the world. Anaerobic degradation of petroleum hydrocarbon is very relevant in removing oil spills in the anaerobic zones of soil and sediments. This research investigates the possibility of degrading no. diesel fuel under anaerobic conditions. Anaerobic packed soil columns were used to simulate and study in situ bioremediation of soil contaminated with diesel fuel. Several anaerobic conditions were evaluated in soil columns, including sulfate reducing, nitrate reducing, methanogenic, and mixed electron acceptor conditions. The objectives were to determine the extent of diesel fuel degradation in soil columns under various anaerobic conditions and identify the best conditions for efficient removal of diesel fuel. Diesel fuels were degraded significantly under all conditions compared to no electron supplemented soil column (natural attenuation). However, the rate of diesel degradation was the highest under mixed electron acceptor conditions followed in order by sulfate reducing, nitrate reducing, and methanogenic conditions. Under mixed electron acceptor condition 81% of diesel fuel was degraded within 310 days. While under sulfate reducing condition 54.5% degradation of diesel fuel was observed for the same period. This study showed evidence for diesel fuel metabolism in a mixed microbial population system similar to any contaminated field sites, where heterogeneous microbial population exists.     

Biodiesel production—current state of the art and challenges

Biodiesel is a clean-burning fuel produced from grease, vegetable oils, or animal fats. Biodiesel is produced by transesterification of oils with short-chain alcohols or by the esterification of fatty acids. The transesterification reaction consists of transforming triglycerides into fatty acid alkyl esters, in the presence of an alcohol, such as methanol or ethanol, and a catalyst, such as an alkali or acid, with glycerol as a byproduct. Because of diminishing petroleum reserves and the deleterious environmental consequences of exhaust gases from petroleum diesel, biodiesel has attracted attention during the past few years as a renewable and environmentally friendly fuel. Since biodiesel is made entirely from vegetable oil or animal fats, it is renewable and biodegradable. The majority of biodiesel today is produced by alkali-catalyzed transesterification with methanol, which results in a relatively short reaction time. However, the vegetable oil and alcohol must be substantially anhydrous and have a low free fatty acid content, because the presence of water or free fatty acid or both promotes soap formation. In this article, we examine different biodiesel sources (edible and nonedible), virgin oil versus waste oil, algae-based biodiesel that is gaining increasing importance, role of different catalysts including enzyme catalysts, and the current state-of-the-art in biodiesel production. JIMB 2008: BioEnergy—special issue.     

Non-sterile fermentations for the economical biochemical conversion of renewable feedstocks

Heavy reliance on petroleum-based products drives continuous exploitation of fossil fuels, and results in serious environmental and climate problems. To address such an issue, there is a shift from petroleum sources to renewable ones. Biochemical conversion via fermentation is a primary platform for converting renewable sources to biofuels and bulk chemicals. In order to provide cost-competitive alternatives, it is imperative to develop efficient, cost-saving, and robust fermentation processes. Non-sterile fermentation offers several benefits compared to sterile fermentation, including elimination of sterility, reduced maintenance requirements, relatively simple bioreactor design, and simplified operation. Thus, cost effectiveness of non-sterile fermentation makes it a practical platform for low cost, large volume production of biofuels and bulk chemicals. Many approaches have been developed to conduct non-sterile fermentation without sacrificing the yields and productivities of fermentation products. This review focuses on the strategies for conducting non-sterile fermentation. The challenges facing non-sterile fermentation are also discussed.     

The effect of the bioremediation of soil contaminated with petroleum derivatives on the occurrence of epigeic and edaphic fauna

This field study investigated the colonization process of soil contaminated with different petroleum products (petrol, diesel fuel, spent engine oil; dose: 6000 mg of fuel·kg^?1 dry mass [d.m.] of soil) by epigeic and edaphic invertebrates during the progress of natural bioremediation and bioremediation enhanced using selected microorganisms (ZB-01 biopreparation). Epigeic fauna was captured using pitfall traps. Occurrence of edaphic fauna in soil samples as well as total petroleum hydrocarbon contents (TPH) were also investigated. Results showed that inoculation with ZB-01 biocenosis allowed the degradation of petroleum derivatives in the soil contaminated with diesel fuel and engine oil, with 82.3% and 75.4% efficiency, respectively. Applying bioremediation to all contaminated soils accelerated the process of recolonization by edaphic invertebrates. However, the 28-month period was too short to observe full population recovery in soils contaminated with diesel fuel and engine oil. Microbe-enhanced bioremediation accelerated recolonization by epigeic invertebrates on soil contaminated with diesel fuel, whereas it exerted inhibitory effect on recolonization of soil contaminated with engine oil (especially by Collembola). The observed discrepancies in the rates of recolonization for soils contaminated with petrol and diesel fuel that were still noted at the stage of no longer different TPH levels justify the idea to include the survey of edaphic faunal density as one of the parameters in the ecological risk assessment of various bioremediation techniques.     

Stimulation of Diesel Fuel Biodegradation by Indigenous Nitrogen Fixing Bacterial Consortia

Abstract Successful stimulation of N₂ fixation and petroleum hydrocarbon degradation in indigenous microbial consortia may decrease exogenous N requirements and reduce environmental impacts of bioremediation following petroleum pollution. This study explored the biodegradation of petroleum pollution by indigenous N₂ fixing marine microbial consortia. Particulate organic carbon (POC) in the form of ground, sterile corn-slash (post-harvest leaves and stems) was added to diesel fuel amended coastal water samples to stimulate biodegradation of petroleum hydrocarbons by native microorganisms capable of supplying a portion of their own N. It was hypothesized that addition of POC to petroleum amended water samples from N-limited coastal waters would promote the growth of N₂ fixing consortia and enhance biodegradation of petroleum. Manipulative experiments were conducted using samples from coastal waters (marinas and less polluted control site) to determine the effects of POC amendment on biodegradation of petroleum pollution by native microbial consortia. Structure and function of the microbial consortia were determined by measurement of N₂ fixation (acetylene reduction), hydrocarbon biodegradation (¹⁴C hexadecane mineralization), bacterial biomass (AODC), number of hydrocarbon degrading bacteria (MPN), and bacterial productivity (³H-thymidine incorporation). Throughout this study there was a consistent enhancement of petroleum hydrocarbon degradation in response to the addition of POC. Stimulation of diesel fuel biodegradation following the addition of POC was likely attributable to increases in bacterial N₂ fixation, diesel fuel bioavailability, bacterial biomass, and metabolic activity. Toxicity of the bulk phase water did not appear to be a factor affecting biodegradation of diesel fuel following POC addition. These results indicate that the addition of POC to diesel-fuel-polluted systems stimulated indigenous N₂ fixing microbial consortia to degrade petroleum hydrocarbons. Received: 29 December 1998; Accepted: 6 April 1999     

Improvement of engine emissions with conventional diesel fuel and diesel-biodiesel blends

In this report combustion and exhaust emissions with neat diesel fuel and diesel-biodiesel blends have been investigated. In the investigation, firstly biodiesel from non-edible neem oil has been made by esterification. Biodiesel fuel (BDF) is chemically known as mono-alkyl fatty acid ester. It is renewable in nature and is derived from plant oils including vegetable oils. BDF is non-toxic, biodegradable, recycled resource and essentially free from sulfur and carcinogenic benzene. In the second phase of this investigation, experiment has been conducted with neat diesel fuel and diesel-biodiesel blends in a four stroke naturally aspirated (NA) direct injection (DI) diesel engine. Compared with conventional diesel fuel, diesel-biodiesel blends showed lower carbon monoxide (CO), and smoke emissions but higher oxides of nitrogen (NOx) emission. However, compared with the diesel fuel, NOx emission with diesel-biodiesel blends was slightly reduced when EGR was applied.     

Impacts of diesel fuel exposure on the microphytobenthic community of an intertidal sand flat

This study employed simulated spills of weathered diesel fuel and measured the initial effects on the intertidal sand flat microphytobenthic (MPB) communities. The goals were to examine the impacts of short-term (hours) and longer-term (days) exposure to petroleum on the native sand flat MPB in coastal North Carolina and to assess recovery of the community following the exposure. We assessed changes in biomass (chlorophyll a), primary productivity (¹⁴C bicarbonate incorporation), photophysiology (P vs. I curves) and species composition (microscopy) and compared diesel exposed samples to unamended controls. We found that short-term impacts of diesel fuel pollution were confined to primary productivity and photophysiology of sand flat MPB. Short-term effects were only detected at relatively high concentrations that are not common outside of a major spill event. In the longer term, diesel fuel was again found to have effects on primary productivity, but at higher concentrations than would be likely to occur in industrialized coastal areas. However, negative impacts on photophysiology were detected at diesel fuel concentrations slightly above typical ambient conditions in coastal waters in industrialized areas. Biomass as measured by chlorophyll a was not affected by any concentration in the longer-term exposure to diesel fuel. Cell counts in the longer-term experiments found cyanobacteria had larger negative impacts from diesel fuel exposure than did diatoms. The recovery portion of this study showed the sand flat MPB communities were fairly resilient following both additions of diesel fuel. However, photophysiology and cell counts did not return to conditions equivalent to the control. Data from this study indicate that the effects of petroleum pollution on the MPB community of tidal sand flats should be considered alongside effects on other coastal microalgae in ecological and damage assessments.     

Ethanol-diesel fuel blends -- a review

Ethanol is an attractive alternative fuel because it is a renewable bio-based resource and it is oxygenated, thereby providing the potential to reduce particulate emissions in compression-ignition engines. In this review the properties and specifications of ethanol blended with diesel fuel are discussed. Special emphasis is placed on the factors critical to the potential commercial use of these blends. These factors include blend properties such as stability, viscosity and lubricity, safety and materials compatibility. The effect of the fuel on engine performance, durability and emissions is also considered. The formulation of additives to correct certain key properties and maintain blend stability is suggested as a critical factor in ensuring fuel compatibility with engines. However, maintaining vehicle safety with these blends may entail fuel tank modifications. Further work is required in specifying acceptable fuel characteristics, confirming the long-term effects on engine durability, and ensuring safety in handling and storing ethanol-diesel blends.     

Biodiesel production,properties, and feedstocks

Biodiesel, defined as the mono-alkyl esters of vegetable oils or animal fats, is an environmentally attractive alternative to conventional petroleum diesel fuel (petrodiesel). Produced by transesterification with a monohydric alcohol, usually methanol, biodiesel has many important technical advantages over petrodiesel, such as inherent lubricity, low toxicity, derivation from a renewable and domestic feedstock, superior flash point and biodegradability, negligible sulfur content, and lower exhaust emissions. Important disadvantages of biodiesel include high feedstock cost, inferior storage and oxidative stability, lower volumetric energy content, inferior low-temperature operability, and in some cases, higher NO _x exhaust emissions. This review covers the process by which biodiesel is prepared, the types of catalysts that may be used for the production of biodiesel, the influence of free fatty acids on biodiesel production, the use of different monohydric alcohols in the preparation of biodiesel, the influence of biodiesel composition on fuel properties, the influence of blending biodiesel with other fuels on fuel properties, alternative uses for biodiesel, and value-added uses of glycerol, a co-product of biodiesel production. A particular emphasis is placed on alternative feedstocks for biodiesel production. Lastly, future challenges and outlook for biodiesel are discussed. Disclaimer: Product names are necessary to report factually on available data; however, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by USDA implies no approval of the product to the exclusion of others that may also be suitable.     

Biodegradation of diesel/biodiesel blends by a consortium of hydrocarbon degraders: effect of the type of blend and the addition of biosurfactants

Biodegradation experiments for diesel/biodiesel blends in liquid cultures by-petroleum degrading microbial consortium showed that for low amendments of biodiesel (10%) the overall biodegradation efficiency of the mixture after seven days was lower than for petroleum diesel fuel. Preferential usage of methyl esters in the broad biodiesel concentration range and diminished biodegradation of petroleum hydrocarbons for 10% biodiesel blend was confirmed. Rhamnolipids improved biodegradation efficiency only for blends with low content of biodiesel. Emulsion formation experiments showed that biodiesel amendments significantly affected dispersion of fuel mixtures in water. The presence of rhamnolipids biosurfactant affected stability of such emulsions and altered cell surface properties of tested consortium.     

Influence of UV irradiation on microbiological degradation of petroleum products

Changes in the rates of microbiological degradation of kerosene, diesel fuel, and fuel oil under the effect of UV irradiation were estimated by testing the respiratory activity of microbial communities. The strongest inhibitory effect was observed upon simultaneous UV irradiation of both natural water and petroleum products. Concentrations of CO₂ in the microbial communities (microcosms) decreased from 6.7 to 3.6 vol. % upon oxidation of kerosene, from 5.9 to 0.8 vol. % upon oxidation of diesel fuel, and from 5.7 to 0.05 vol. % upon oxidation of fuel oil.     

Cashew nut shell oil as a potential feedstock for biodiesel production: An overview

Biodiesel outperforms diesel in emissions and engine performance. They burn efficiently in diesel engines and are eco-friendly. Since cashew nut shell liquid (CNSO) is waste, commercial biodiesel production from it should be profitable. CNSO is cheap and can reduce cashew processing factory waste. From cashew kernels, CNSL is extracted using various mechanical, thermal, and solvent extraction techniques. This article examines current research into using cashew nutshell liquid biodiesel (CNSLBD) in diesel engines. The work also discusses Indian biodiesel demand, availability, export information, life cycle cost analysis, cost economics of per hectare yield, Indian government initiative of CNSO. This review also evaluates the viability of this fuel as an alternative energy source. CNSLBD is a prospective alternative fuel that has the potential to benefit both the cashew nut industry and the energy industry. In addition to this, the study examines the procedures for extracting CNSO. According to the findings of the study, CNSO is a prospective alternative fuel that has the potential to benefit both the cashew nut industry and the energy industry.     

Prepared Bed Land Treatment of Soils Containing Diesel and Crude Oil Hydrocarbons

An ex situ, field-scale, prepared bed land treatment unit (LTU) was used to bio-remediate soils containing petroleum hydrocarbons. Two soils were treated in side-by-side units to compare performance: (1) a clayey silt containing crude oil hydrocarbons from releases 30 to 40 years ago and (2) a silty sand containing diesel fuel hydrocarbons from a leak about three years prior to the bioremediation. The effectiveness of the bioremediation in the LTU was evaluated over a period of 18 months. The results indicated that: (1) prepared bed bioremediation reduced the hydrocarbon concentration, mobility, and relative toxicity in the soil with the diesel fuel, and (2) chemical bioavailability appeared to limit bioremediation of the soil containing the crude oil hydrocarbons. Although the soils containing the crude oil hydrocarbons contained an average of 10,000?mg TPH/kg dry soil, these soils had limited hydrocarbon availability, nontoxic conditions, and low potential for chemical migration. For the soils containing the diesel fuel, active prepared bed bioremediation of about 15 weeks was adequate to reach an environmentally acceptable endpoint. At that time, there was little further TPH loss, no Microtox^TM toxicity, and limited hydrocarbon mobility.