Advanced biofuel production in microbes

The cost-effective production of biofuels from renewable materials will begin to address energy security and climate change concerns. Ethanol, naturally produced by microorganisms, is currently the major biofuel in the transportation sector. However, its low energy content and incompatibility with existing fuel distribution and storage infrastructure limits its economic use in the future. Advanced biofuels, such as long chain alcohols and isoprenoid- and fatty acid-based biofuels, have physical properties that more closely resemble petroleum-derived fuels, and as such are an attractive alternative for the future supplementation or replacement of petroleum-derived fuels. Here, we review recent developments in the engineering of metabolic pathways for the production of known and potential advanced biofuels by microorganisms. We concentrate on the metabolic engineering of genetically tractable organisms such as Escherichia coli and Saccharomyces cerevisiae for the production of these advanced biofuels.     

Engineering biofuel tolerance in non-native producing microorganisms

Large-scale production of renewable biofuels through microbiological processes has drawn significant attention in recent years, mostly due to the increasing concerns on the petroleum fuel shortages and the environmental consequences of the over-utilization of petroleum-based fuels. In addition to native biofuel-producing microbes that have been employed for biofuel production for decades, recent advances in metabolic engineering and synthetic biology have made it possible to produce biofuels in several non-native biofuel-producing microorganisms. Compared to native producers, these non-native systems carry the advantages of fast growth, simple nutrient requirements, readiness for genetic modifications, and even the capability to assimilate CO₂ and solar energy, making them competitive alternative systems to further decrease the biofuel production cost. However, the tolerance of these non-native microorganisms to toxic biofuels is naturally low, which has restricted the potentials of their application for high-efficiency biofuel production. To address the issues, researches have been recently conducted to explore the biofuel tolerance mechanisms and to construct robust high-tolerance strains for non-native biofuel-producing microorganisms. In this review, we critically summarize the recent progress in this area, focusing on three popular non-native biofuel-producing systems, i.e. Escherichia coli, Lactobacillus and photosynthetic cyanobacteria.     

Metabolic engineering of microbial pathways for advanced biofuels production

Production of biofuels from renewable resources such as cellulosic biomass provides a source of liquid transportation fuel to replace petroleum-based fuels. This endeavor requires the conversion of cellulosic biomass into simple sugars, and the conversion of simple sugars into biofuels. Recently, microorganisms have been engineered to convert simple sugars into several types of biofuels, such as alcohols, fatty acid alkyl esters, alkanes, and terpenes, with high titers and yields. Here, we review recently engineered biosynthetic pathways from the well-characterized microorganisms Escherichia coli and Saccharomyces cerevisiae for the production of several advanced biofuels.     

Biofuel production from microalgae as feedstock: current status and potential

Algal biofuel has become an attractive alternative of petroleum-based fuels in the past decade. Microalgae have been proposed as a feedstock to produce biodiesel, since they are capable of mitigating CO₂ emission and accumulating lipids with high productivity. This article is an overview of the updated status of biofuels, especially biodiesel production from microalgae including fundamental research, culture selection and engineering process development; it summarizes research on mathematical and life cycle modeling on algae growth and biomass production; and it updates global efforts of research and development and commercialization attempts. The major challenges are also discussed.     

Metabolic engineering for advanced biofuels production from Escherichia coli

Global energy and environmental problems have stimulated increasing efforts toward synthesizing liquid biofuels as transportation energy. Compared to the traditional biofuel, ethanol, advanced biofuels should offer advantages such as higher energy density, lower hygroscopicity, lower vapor pressure, and compatibility with existing transportation infrastructure. However, these fuels are not synthesized economically using native organisms. Metabolic engineering offers an alternative approach in which synthetic pathways are engineered into user-friendly hosts for the production of these fuel molecules. These hosts could be readily manipulated to improve the production efficiency. This review summarizes recent progress in the engineering of Escherichia coli to produce advanced biofuels.     

Metabolic engineering strategies toward production of biofuels

Exacerbation of climate change and air pollution around the world have emphasized the necessity of replacing fossil fuels with clean and sustainable energy. Metabolic engineering has provided strategies to engineer diverse organisms for the production of biofuels from renewable carbon sources. Although some of the processes are commercialized, there has been continued effort to produce advanced biofuels with higher efficiencies. In this article, metabolic engineering strategies recently exploited to enhance biofuel production and facilitate utilization of non-edible low-value carbon sources are reviewed. The strategies include engineering enzymes, exploiting new pathways, and systematically optimizing metabolism and fermentation processes, among others. In addition, metabolic and bioprocess engineering strategies to achieve competitiveness of current biofuel production systems compared with fossil fuels are discussed.     

Biofuel production in Escherichia coli: the role of metabolic engineering and synthetic biology

The microbial production of biofuels is a promising avenue for the development of viable processes for the generation of fuels from sustainable resources. In order to become cost and energy effective, these processes must utilize organisms that can be optimized to efficiently produce candidate fuels from a variety of feedstocks. Escherichia coli has become a promising host organism for the microbial production of biofuels in part due to the ease at which this organism can be manipulated. Advancements in metabolic engineering and synthetic biology have led to the ability to efficiently engineer E. coli as a biocatalyst for the production of a wide variety of potential biofuels from several biomass constituents. This review focuses on recent efforts devoted to engineering E. coli for the production of biofuels, with emphasis on the key aspects of both the utilization of a variety of substrates as well as the synthesis of several promising biofuels. Strategies for the efficient utilization of carbohydrates, carbohydrate mixtures, and noncarbohydrate carbon sources will be discussed along with engineering efforts for the exploitation of both fermentative and nonfermentative pathways for the production of candidate biofuels such as alcohols and higher carbon biofuels derived from fatty acid and isoprenoid pathways. Continued advancements in metabolic engineering and synthetic biology will help improve not only the titers, yields, and productivities of biofuels discussed herein, but also increase the potential range of compounds that can be produced.     

Synthetic biology for biofuels: Building designer microbes from the scratch

The ultimate goal in the production of biofuels is to produce fuels identical or similar to petroleum-derived transportation fuels more efficiently and in commercial quantities. Synthetic biologists have been engineering microbes to synthesize biofuels, such as butanol and fatty acid- or isoprenoid-based fuels, which are nearly identical to gasoline and diesel. One of the most urgent demands along this direction is to attain a solid framework for characterizing and standardizing the biological parts and devices. It seems quite promising because biotechnologies specially based on miniaturizations have been making a big contribution to this work. Therefore, in this review, recent advances and difficulties in the biofuel field are discussed, along with the advances of synthetic biology, which will make it possible to create designer microorganisms that produce economically viable next generation biofuels, aside from bioethanol, from corn or sugar cane, and biodiesel from plant or animal oils.     

Methods of downstream processing for the production of biodiesel from microalgae

Despite receiving increasing attention during the last few decades, the production of microalgal biofuels is not yet sufficiently cost-effective to compete with that of petroleum-based conventional fuels. Among the steps required for the production of microalgal biofuels, the harvest of the microalgal biomass and the extraction of lipids from microalgae are two of the most expensive. In this review article, we surveyed a substantial amount of previous work in microalgal harvesting and lipid extraction to highlight recent progress in these areas. We also discuss new developments in the biodiesel conversion technology due to the importance of the connectivity of this step with the lipid extraction process. Furthermore, we propose possible future directions for technological or process improvements that will directly affect the final production costs of microalgal biomass-based biofuels.     

生物柴油研究进展

介绍了国内外生物柴油的发展现状,探讨了我国发展生物柴油的原料来源途径,包括木本油料植物、转基因油料作物、废弃油脂、微生物油脂和微藻油脂等,综述了制备生物柴油的化学法、酶法、超临界法等生产技术及其进展,概括了当前生物柴油主要的品质问题与改性对策,分析了生物柴油副产物的高值化利用策略,指出了我国生物柴油产业化面临的原料、技术和生物炼制方面的主要问题。     

Plants to power: bioenergy to fuel the future

Bioenergy should play an essential part in reaching targets to replace petroleum-based transportation fuels with a viable alternative, and in reducing long-term carbon dioxide emissions, if environmental and economic sustainability are considered carefully. Here, we review different platforms, crops, and biotechnology-based improvements for sustainable bioenergy. Among the different platforms, there are two obvious advantages to using lignocellulosic biomass for ethanol production: higher net energy gain and lower production costs. However, the use of lignocellulosic ethanol as a viable alternative to petroleum-based transportation fuels largely depends on plant biotechnology breakthroughs. We examine how biotechnology, such as lignin modification, abiotic stress resistance, nutrition usage, in planta expression of cell wall digestion enzymes, biomass production, feedstock establishment, biocontainment of transgenes, metabolic engineering, and basic research, can be used to address the challenges faced by bioenergy crop production.     

The path to next generation biofuels: successes and challenges in the era of synthetic biology

Volatility of oil prices along with major concerns about climate change, oil supply security and depleting reserves have sparked renewed interest in the production of fuels from renewable resources. Recent advances in synthetic biology provide new tools for metabolic engineers to direct their strategies and construct optimal biocatalysts for the sustainable production of biofuels. Metabolic engineering and synthetic biology efforts entailing the engineering of native and de novo pathways for conversion of biomass constituents to short-chain alcohols and advanced biofuels are herewith reviewed. In the foreseeable future, formal integration of functional genomics and systems biology with synthetic biology and metabolic engineering will undoubtedly support the discovery, characterization, and engineering of new metabolic routes and more efficient microbial systems for the production of biofuels.     

Designing the perfect plant feedstock for biofuel production: using the whole buffalo to diversify fuels and products

Petroleum-derived liquid fuels and commodities play a part in nearly every aspect of modern daily life. However, dependence on this one natural resource to maintain modern amenities has caused negative environmental and geopolitical ramifications. In an effort to replace petroleum, technologies to synthesize liquid fuels and other commodities from renewable biomass are being developed. Current technologies, however, only use a portion of plant biomass feedstocks for fuel and useful products. "Using the whole feedstock buffalo" or optimally using all portions and biochemicals present in renewable biomass will enhance the economic and environmental feasibility of biofuels and coproducts. To accomplish this optimization, greater understanding of the relationship between liquid fuel and bioproduct properties and plant chemistries is needed. Liquid fuel properties and how they relate to biochemistry and petrochemistry are discussed. Enhanced biofuel yields and high-value commodities from biomass are needed to sustainably replace petroleum-based products. Several metabolic engineering strategies are discussed. We will describe paths of possible fuel and product diversification using dedicated lignocellulosic biomass (e.g., switchgrass).     

Anaerobic fermentation of glycerol: a path to economic viability for the biofuels industry

Although biofuels such as biodiesel and bioethanol represent a secure, renewable and environmentally safe alternative to fossil fuels, their economic viability is a major concern. The implementation of biorefineries that co-produce higher value products along with biofuels has been proposed as a solution to this problem. The biorefinery model would be especially advantageous if the conversion of byproducts or waste streams generated during biofuel production were considered. Glycerol-rich streams generated in large amounts by the biofuels industry, especially during the production of biodiesel, present an excellent opportunity to establish biorefineries. Once considered a valuable 'co-product', crude glycerol is rapidly becoming a 'waste product' with a disposal cost attributed to it. Given the highly reduced nature of carbon in glycerol and the cost advantage of anaerobic processes, fermentative metabolism of glycerol is of special interest. This review covers the anaerobic fermentation of glycerol in microbes and the harnessing of this metabolic process to convert abundant and low-priced glycerol streams into higher value products, thus creating a path to viability for the biofuels industry. Special attention is given to products whose synthesis from glycerol would be advantageous when compared with their production from common sugars.     

Production of fuels and chemicals from renewable resources using engineered Escherichia coli

Biotechnological production of fuels and chemicals from renewable resources is an appealing way to move from the current petroleum-based economy to a biomass-based green economy. Recently, the feedstocks that can be used for bioconversion or fermentation have been expanded to plant biomass, microbial biomass, and industrial waste. Several microbes have been engineered to produce chemicals from renewable resources, among which Escherichia coli is one of the best studied. Much effort has been made to engineer E. coli to produce fuels and chemicals from different renewable resources. In this paper, we focused on E. coli and systematically reviewed a range of fuels and chemicals that can be produced from renewable resources by engineered E. coli. Moreover, we proposed how can we further improve the efficiency for utilizing renewable resources by engineered E. coli, and how can we engineer E. coli for utilizing alternative renewable feedstocks. e.g. C1 gases and methanol. This review will help the readers better understand the current progress in this field and provide insights for further metabolic engineering efforts in E. coli.     

Anaerobic digestion of microalgal biomass for bioenergy production,removal of nutrients and microcystin: current status

Using renewable microalgal biomass as active feedstocks for biofuels and bioproducts is explored to substitute petroleum-based fuels and chemicals. In the last few years, the importance of microalgae biomass has been realized as a renewable feedstock due to several positive attributes associated with it. Biorefinery via anaerobic digestion (AD) of microalgal biomass is a promising and sustainable method to produce value-added chemicals, edible products and biofuels. Microalgal biomass pretreatment is a significant process to enhance methane production by AD. Findings on the AD microbial community’s variety and organization can give novel in turn on digester steadiness and presentation. This review presents a vital study of the existing facts on the AD microbial community and AD production. Co-digestion of microalgal biomass with different co-substrates was used in AD to enhance biogas production, and the process was economically viable with improved biodegradability. Microcystins, which are produced by toxic cyanobacterial blooms, create a severe hazard to environmental health. Anaerobic biodegradation is an effective method to degrade the microcystins and convert into nontoxic products. However, for the cost-effective conversion of biomass to energy and other beneficial byproducts, additional highly developed research is still required for large-scale AD of microalgal biomass.     

丁醇在大肠杆菌中的生物合成研究进展*

生物丁醇作为一种重要的化学品和石油基燃料的替代品引起了人们的广泛关注。大肠杆菌(Escherichia coli)是生物合成化学品的优良底盘菌株,已在其体内构建了丁醇的生物合成途径。但大肠杆菌合成丁醇存在:(1)代谢通量非最优;(2)辅因子和氧化还原不平衡;(3)丁醇产量和产率低等问题,为此,从高效酶选择、碳代谢流调控、辅因子调控、丁醇生产和工艺等方面已经对丁醇合成途径和丁醇发酵进行了优化。从该角度出发阐述近几年来大肠杆菌生物合成丁醇的研究进展,并展望了利用工程大肠杆菌生产丁醇的研究方向,旨在为应用其进行高效的丁醇生产提供参考。     

Combinatorial genetic perturbation to refine metabolic circuits for producing biofuels and biochemicals

Recent advances in metabolic engineering have enabled microbial factories to compete with conventional processes for producing fuels and chemicals. Both rational and combinatorial approaches coupled with synthetic and systematic tools play central roles in metabolic engineering to create and improve a selected microbial phenotype. Compared to knowledge-based rational approaches, combinatorial approaches exploiting biological diversity and high-throughput screening have been demonstrated as more effective tools for improving various phenotypes of interest. In particular, identification of unprecedented targets to rewire metabolic circuits for maximizing yield and productivity of a target chemical has been made possible. This review highlights general principles and the features of the combinatorial approaches using various libraries to implement desired phenotypes for strain improvement. In addition, recent applications that harnessed the combinatorial approaches to produce biofuels and biochemicals will be discussed.     

The role of biochemical engineering in the production of biofuels from microalgae

Environmental changes that have occurred due to the use of fossil fuels have driven the search for alternative sources that have a lower environmental impact. First-generation biofuels were derived from crops such as sugar cane, corn and soybean, which contribute to water scarcity and deforestation. Second-generation biofuels originated from lignocellulose agriculture and forest residues, however these needed large areas of land that could be used for food production. Based on technology projections, the third generation of biofuels will be derived from microalgae. Microalgae are considered to be an alternative energy source without the drawbacks of the first- and second-generation biofuels. Depending upon the growing conditions, microalgae can produce biocompounds that are easily converted into biofuels. The biofuels from microalgae are an alternative that can keep the development of human activity in harmony with the environment. This study aimed to present the main biofuels that can be derived from microalgae.     

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.