The authors wish to thank the analytical laboratory of the CanmetENERGY Technology Centre for its technical support, and Suncor Energy Inc. for supplying the synthetic crude oil. Partial funding for this study was provided by Natural Resources Canada and government of Canada&#39;s interdepartmental Program of Energy Research and Development (PERD) with project ID A22.015. Yi Zhang would like to acknowledge his Natural Sciences and Engineering Research Council (NSERC) of Canada Visiting Fellowship from January 2015 to January 2016.

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The authors declare that they have no competing financial interests.

The protocol described here utilizes cyclic operation of a single reactor containing a batch of fluidized catalyst particles to simulate feed oil cracking and catalyst regeneration. The oil to be cracked is preheated and fed from the top through an injector tube with its tip close to the bottom of the fluid bed. The vapor generated after catalytic cracking is condensed and collected in a receiver, and the liquid product collected is subsequently analyzed for simulated distillation to determine yields of fractions in diff...

There is strong global interest in both the private and public sectors to find efficient and economic means to produce transportation fuels from biomass-derived feedstocks. This interest is driven by a general concern over the substantial contribution of burning petroleum fossil fuels to greenhouse gas (GHG) emissions and its associated contribution to global warming. Also, there is strong political will in North America and Europe to displace foreign-produced petroleum with renewable domestic liquid fuels. In 2008, biofuels provided 1.8% of the world&#39;s transportation fuels1. In many developed countries, it is required that biofuels replace from 6% to 10% of petroleum fuels in the near future2. In Canada, regulations require an average renewable fuel content of 5% in gasoline starting December 15, 20103. The Renewable Energy Directive (RED) in Europe has also mandated a 10% renewable energy target for the European Union transport sector by 20204.
The challenge has been to develop and demonstrate a viable economic pathway to produce fungible transportation fuels from biomass. Biological sources include triglyceride-based biomass such as vegetable oils and animal fats, as well as waste cooking oil and cellulosic biomass such as wood chips, forest wastes, and agriculture residues. Over the past two decades, research has focused on the evaluation of biomass-derived oil processing using conventional fluid catalytic cracking (FCC)5-12, a technology responsible for producing most of the gasoline in a petroleum refinery. Our novel approach in this study is to co-process canola oil mixed with oil sands bitumen-derived feedstock. Normally, bitumen must be upgraded prior to refining, producing refinery feedstocks such as synthetic crude oil (SCO)&#8212;this processing route is particularly energy intensive, accounting for 68-78% of the GHG emissions from the SCO production13 and, in 2011, constituting 2.6% of Canada&#39;s total GHG emissions14. Replacing a portion of upgraded HGO with biofeed would reduce GHG emissions, since biofuel production involves a much smaller carbon footprint. Canola oil is chosen in this work because it is abundant in Canada and the US. This feedstock possesses a density and viscosity similar to those of HGOs while the contents of sulfur, nitrogen, and metals that could affect FCC performance or product quality are negligible. Moreover, this co-processing option offers significant technological and economic advantages as it would allow utilization of the existing refinery infrastructure and, hence, would require little additional hardware or modification of the refinery. In addition, there may be potential synergy that could result in product quality improvement when co-processing a highly aromatic bitumen feed with its straight-chain biomass counterpart. However, co-processing involves important technical challenges. These include the unique physical and chemical characteristics of bio-feeds: high oxygen content, paraffinic-rich composition, compatibility with petroleum feedstocks, fouling potential, etc.
This study provides a detailed protocol for the production of biofuels at laboratory scale from canola oil through catalytic cracking. A fully automated reaction system&#160;&#8211; referred to in this work as the laboratory test unit (LTU)15 &#8211; is used for this work. Figure 1 shows schematically how this unit operates. This LTU has become the industry standard for laboratory FCC studies. The objective of this study is to test the suitability of the LTU for cracking canola oil to produce fuels and chemicals with the goal of mitigating GHG emissions.
<img alt="Figure 1" src="/files/ftp_upload/54390/54390fig1.jpg" /> 
Figure 1: Conceptual illustration of the reactor. Illustration showing flow lines of the catalyst, feed, product, and diluent. <a href="https://www.jove.com/files/ftp_upload/54390/54390fig1large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>

<table><tbody><tr><td>Advanced Cracking Evaluation (ACE) Unit</td><td>Kayser Technology Inc.</td><td>ACE R+ 46</td><td>Assembled by Zeton Inc. SN:505-46; consisting of (1) a reactor; (2) catalyst addition system; (3) feed delivery system; (4) liquid collection system; (5) gas collection system; (6) gas analyzing system; (7) catalyst regeneration system; (8) CO catalytic convertor; (9) coke analyzing system</td></tr><tr><td>Reactor (ACE)</td><td>Kayser Technology Inc.</td><td>V-105</td><td>A 1.6 cm ID stainless steel tube having a tapered conical bottom and with a diluent (nitrogen) flowing from the bottom to fluidize the catalyst and also serve as the stripping gas at the end of the run</td></tr><tr><td>Catalyst Addition System (ACE)</td><td>Kayser Technology Inc.</td><td></td><td>Six hoppers (V-120F, with respective valves) for addition of catalyst for up to 6 runs</td></tr><tr><td>Feed Delivery System (ACE)</td><td>Kayser Technology Inc.</td><td></td><td>Consisting of feed bottle (V-100), syringe (FS-115), pump (P-100), and injector (with 1.125 inch injector height, &lt;em&gt;i.e.&lt;/em&gt;, the distance from the lowest point of the conical reactor bottom to the bottom end of the feed injector)</td></tr><tr><td>Liquid Collection System (ACE)</td><td>Kayser Technology Inc.</td><td></td><td>Six liquid receivers (V-110F) immersed in a common coolant bath (Ethylene glycol/water mixture in 50:50 mass ratio) at about &amp;ndash;15 &amp;deg;C in a large tank (V-145)</td></tr><tr><td>Gas Collection System (ACE)</td><td>Kayser Technology Inc.</td><td></td><td>Based on water displacement principle; consisting of gas collection vessel (V-150) with a motor-driven stirrer (MTR-100), and a weight scale (WT-100) for weighing the displaced water collected in a beaker (V-160).</td></tr><tr><td>Gas Analyzing System (ACE)</td><td>Kayser Technology Inc.</td><td></td><td>Key element being Agilent micro GC (model 3000A) with four capillary columns equipped with respective thermal conductivity detectors (TCDs)&amp;nbsp;</td></tr><tr><td>Catalyst Regeneration System (ACE)</td><td>Kayser Technology Inc.</td><td>V-105</td><td>Spent catalyst in reactor being burned &lt;em&gt;in situ&lt;/em&gt; in air at +700 &amp;deg;C to ensure complete removal of carbon deposited on the catalyst</td></tr><tr><td>CO Catalytic Convertor&amp;nbsp; (ACE)</td><td>Kayser Technology Inc.</td><td></td><td>A reactor (V-140) with CuO as catalyst to oxidize any CO and hydrocarbons in exhausted flue gas to CO&lt;sub&gt;2&lt;/sub&gt; (to be analyzed by IR gas analyzer) and H&lt;sub&gt;2&lt;/sub&gt;O (to be absorbed by a dryer)</td></tr><tr><td>Coke Analyzing System (ACE)</td><td>Kayser Technology Inc.</td><td></td><td>Servomex (Model 1440C) IR analyzer for measuring CO&lt;sub&gt;2&lt;/sub&gt; in exhausted flue gas</td></tr><tr><td>R+MM Software Suite</td><td>Kayser Technology Inc.</td><td></td><td>Including iFIX 3.5&amp;nbsp;</td></tr><tr><td>Agilent Micro GC</td><td>Agilent Technologies</td><td>3000A</td><td>For gas analysis after cracking</td></tr><tr><td>Cerity Networked Data System</td><td>Agilent Technologies</td><td></td><td>Software for Agilent Micro GC</td></tr><tr><td>CO&lt;sub&gt;2&lt;/sub&gt; Gas Analyser</td><td>Servomex Inc.</td><td>1440C</td><td>SN: 01440C1C02/2900</td></tr><tr><td>NESLAB Refrigerated Bath</td><td>Themo Electron Corporation</td><td>RTE 740</td><td>SN: 104300061</td></tr><tr><td>Orion&amp;nbsp; Sage Syringe Pump</td><td>Themo Electron Corporation</td><td>M362</td><td>For delivering feed oil to injector tube</td></tr><tr><td>Synthetic Crude Oil (SCO)&amp;nbsp;</td><td>Suncor Energy Inc.</td><td></td><td>Identified as Suncor OSA 10-4.1</td></tr><tr><td>Catalyst P</td><td>Petro-Canada Refinery</td><td></td><td>Equilibrium catalyst</td></tr><tr><td>Balance</td><td>Mettler Toledo</td><td>AB304-S</td><td>For weighing liquid product receivers</td></tr><tr><td>Balance</td><td>Mettler Toledo</td><td>XS8001S</td><td>For weighing water displaced by gas product</td></tr><tr><td>Ethylene Glycol</td><td>Fisher Scientifc Inc.</td><td>CAS 107-21-1</td><td>Mixed with distilled water as coolant (50 v% )</td></tr><tr><td>Drierite</td><td>W.A. Hammond Drierite Co. Ltd.</td><td>24001</td><td>For water absorption after CO catalytic converter</td></tr><tr><td>Copper Oxide</td><td>LECO Corporation</td><td>501-170</td><td>Catalyst for conversion of CO to CO&lt;sub&gt;2&lt;/sub&gt;</td></tr><tr><td>Toluene</td><td>Fisher Scientific Co.&amp;nbsp;</td><td>CAS 108-88-3</td><td>For cleaning liquid receivers</td></tr><tr><td>Acetone</td><td>Fisher Scientific Co.&amp;nbsp;</td><td>CAS 67-64-1</td><td>For cleaning liquid receivers</td></tr><tr><td>Micro GC Calibration Gas</td><td>Air Liquid Canada Inc.</td><td>SPG-25MX0015306</td><td>Multicomponent standard gas</td></tr><tr><td>19.8% CO&lt;sub&gt;2&lt;/sub&gt; Standard Gas</td><td>BOC Canada Ltd.</td><td>24069890</td><td>For calibration of IR analyzer</td></tr><tr><td>Argon Gas</td><td>Linde Canada ltd.</td><td>24001306</td><td>Grade 5.0 Purity</td></tr><tr><td>Helium Gas</td><td>Linde Canada ltd.</td><td>24001333</td><td>Grade 5.0 Purity</td></tr><tr><td>Gas analyzer GC Module</td><td>Inficon</td><td>GCMOD-15</td><td>Channel A</td></tr><tr><td>Gas analyzer GC Module</td><td>Inficon</td><td>GCMOD-03</td><td>Channel B</td></tr><tr><td>Gas analyzer GC Module</td><td>Inficon</td><td>GCMOD-04</td><td>Channel C</td></tr><tr><td>Gas analyzer GC Module</td><td>Inficon</td><td>GCMOD-73</td><td>Channel D</td></tr><tr><td>HP 6890 GC</td><td>Hewlett-Packard Co.&amp;nbsp;</td><td>G1530A</td><td>For simulated distillation</td></tr><tr><td>ASTM 2887 Standard Sample</td><td>PAC L.P.</td><td>26650.150</td><td>For quality control in simulated distillation</td></tr><tr><td>ASTM 2887 Standard Sample</td><td>PAC L.P.</td><td>25950.200</td><td>For calibration in simulated distillation</td></tr><tr><td>Column for GC 6890 (simulated distillation)</td><td>Agilent Technologies</td><td>CP7562</td><td>10 m x 0.53 mm x 1.2 &amp;micro;m, HP 6890 GC column</td></tr><tr><td>Liquid Nitrogen</td><td>Air Liquid Canada Inc.</td><td>SPG-NIT1AC240LC</td><td>For use in simulated distillation&amp;nbsp;</td></tr><tr><td>Nitrogen</td><td>Air Liquid Canada Inc.</td><td>Bulk (building N2)</td><td>For use in ACE unit operation</td></tr><tr><td>Isotemp Programmable Furnace</td><td>Thermo Fisher Scientifc Inc.</td><td>10-750-126</td><td>For calcination of catalyst</td></tr><tr><td>GC Vials, Crimp Top</td><td>Chromatograghic Specialties Inc</td><td>C223682C</td><td>2 ml, for liquid product</td></tr><tr><td>Seals, Crimp Top</td><td>Chromatograghic Specialties Inc</td><td>C221150</td><td>11 mm, for use with GC vials</td></tr><tr><td>4 oz clear Boston round bottles</td><td>Fisher Scientific Co.&amp;nbsp;</td><td>02-911-784</td><td>With PE cone lined caps, for use in feed system</td></tr><tr><td>Sieve</td><td>Endecotts Ltd.</td><td>6140269</td><td>Aperture 38 micron</td></tr><tr><td>Sieve</td><td>Endecotts Ltd.</td><td>6146265</td><td>Aperture 250 &amp;mu;m</td></tr><tr><td>Shaker</td><td>Endecotts Ltd.</td><td>MIN 2737-11</td><td>Minor-Meinzer 2 Sieve Shaker for catalyst screening</td></tr><tr><td>V20 Volumetric KF Titrator</td><td>Mettler Toledo</td><td>5131025056</td><td>For water content analysis of the liquid product</td></tr><tr><td>Hydranal Composite 5</td><td>Sigma-Aldrich</td><td>34805-1L-R</td><td>Reagent for Karl Fischer titration</td></tr><tr><td>Methanol (extremely low water grade)</td><td>Fisher Scientific Co.&amp;nbsp;</td><td>A413-4</td><td>Mixed with toluene (40:60 w/w) for KF titration: also used to recover water in receiver</td></tr><tr><td>Glass Wool</td><td>Fisher Scientific Co.&amp;nbsp;</td><td>11-388</td><td>Placed inside the top of receiver outlet arm&amp;nbsp;</td></tr></tbody></table>

laboratory production of biofuels and biochemicals from a rapeseed oil through catalytic cracking conversion

The work is based on a reported study which investigates the processability of canola oil (bio-feed) in the presence of bitumen-derived heavy gas oil (HGO) for production of transportation fuels through a fluid catalytic cracking (FCC) route. Cracking experiments are performed with a fully automated reaction unit at a fixed weight hourly space velocity (WHSV) of 8 hr-1, 490-530 &#176;C, and catalyst/oil ratios of 4-12 g/g. When a feed is in contact with catalyst in the fluid-bed reactor, cracking takes place generating gaseous, liquid, and solid products. The vapor produced is condensed and collected in a liquid receiver at -15 &#176;C. The non-condensable effluent is first directed to a vessel and is sent, after homogenization, to an on-line gas chromatograph (GC) for refinery gas analysis. The coke deposited on the catalyst is determined in situ by burning the spent catalyst in air at high temperatures. Levels of CO2 are measured quantitatively via an infrared (IR) cell, and are converted to coke yield. Liquid samples in the receivers are analyzed by GC for simulated distillation to determine the amounts in different boiling ranges, i.e., IBP-221 &#176;C (gasoline), 221-343 &#176;C (light cycle oil), and 343 &#176;C+ (heavy cycle oil). Cracking of a feed containing canola oil generates water, which appears at the bottom of a liquid receiver and on its inner wall. Recovery of water on the wall is achieved through washing with methanol followed by Karl Fischer titration for water content. Basic results reported include conversion (the portion of the feed converted to gas and liquid product with a boiling point below 221 &#176;C, coke, and water, if present) and yields of dry gas (H2-C2's, CO, and CO2), liquefied petroleum gas (C3-C4), gasoline, light cycle oil, heavy cycle oil, coke, and water, if present.

The established protocol has been successfully applied to an oil blend of 15:85 volume ratio (i.e., 14.73:85.27 mass ratio) between canola oil and an SCO-derived HGO20. For practical reasons (cost, availability of canola oil, and possible challenges in commercial operation), the study was focused on feedstock containing 15 v% canola oil addition, although feeds with higher concentrations were also tried. The blend was catalytically cracked at 490-530 &#176;C and 8.0 hr...

Watch this Scientific Journal Video about Laboratory Production of Biofuels and Biochemicals from a Rapeseed Oil through Catalytic Cracking Conversion at JoVE.com

Laboratory Production of Biofuels and Biochemicals from a Rapeseed Oil through Catalytic Cracking Conversion

This paper presents an experimental method to produce biofuels and biochemicals from canola oil mixed with a fossil-based feed in the presence of a catalyst at mild temperatures. Gaseous, liquid, and solid products from a reaction unit are quantified and characterized. Conversion and individual product yields are calculated and reported.

The work is based on a reported study which investigates the processability of canola oil (bio-feed) in the presence of bitumen-derived heavy gas oil ...

laboratory-production-biofuels-biochemicals-from-rapeseed-oil-through

Research

JoVE Journal

Chemistry

13.7K Views.  Natural Resources Canada. This paper presents an experimental method to produce biofuels and biochemicals from canola oil mixed with a fossil-based feed in the presence of a catalyst at mild temperatures. Gaseous, liquid, and solid products from a reaction unit are quantified and characterized. Conversion and individual product yields are calculated and reported.

Video: Laboratory Production of Biofuels and Biochemicals from a Rapeseed Oil through Catalytic Cracking Conversion

Biomass Conversion to Produce Hydrocarbon Liquid Fuel Via Hot-vapor Filtered Fast Pyrolysis and Catalytic Hydrotreating

Lignocellulosic biomass conversion to produce biofuels has received significant attention because of the quest for a replacement for fossil fuels. Among the various thermochemical and biochemical routes, fast pyrolysis followed by catalytic hydrotreating is considered to be a promising near-term opportunity. This paper reports on experimental methods used 1) at the National Renewable Energy Laboratory (NREL) for fast pyrolysis of lignocellulosic biomass to produce bio-oils in a fluidized-bed reactor and 2) at Pacific Northwest National Laboratory (PNNL) for catalytic hydrotreating of bio-oils in a two-stage, fixed-bed, continuous-flow catalytic reactor. The configurations of the reactor systems, the operating procedures, and the processing and analysis of feedstocks, bio-oils, and biofuels are described in detail in this paper. We also demonstrate hot-vapor filtration during fast pyrolysis to remove fine char particles and inorganic contaminants from bio-oil. Representative results showed successful conversion of biomass feedstocks to fuel-range hydrocarbon biofuels and, specifically, the effect of hot-vapor filtration on bio-oil production and upgrading. The protocols provided in this report could help to generate rigorous and reliable data for biomass pyrolysis and bio-oil hydrotreating research.

Experimental methods for fast pyrolysis of lignocellulosic biomass to produce bio-oils and for the catalytic hydrotreating of bio-oils to produce fuel range hydrocarbons are presented. Hot-vapor filtration during fast pyrolysis to remove fine char particles and inorganic contaminants from bio-oil was also assessed.

Lignocellulosic biomass conversion to produce biofuels has received significant attention because of the quest for a replacement for fossil fuels. Among ...

Biochemistry

Preparation of Silica Nanoparticles Through Microwave-assisted Acid-catalysis

Microwave-assisted synthetic techniques were used to quickly and reproducibly produce silica nanoparticle sols using an acid catalyst with nanoparticle diameters ranging from 30-250 nm by varying the reaction conditions. Through the selection of a microwave compatible solvent, silicic acid precursor, catalyst, and microwave irradiation time, these microwave-assisted methods were capable of overcoming the previously reported shortcomings associated with synthesis of silica nanoparticles using microwave reactors. The siloxane precursor was hydrolyzed using the acid catalyst, HCl. Acetone, a low-tan &#948; solvent, mediates the condensation reactions and has minimal interaction with the electromagnetic field. Condensation reactions begin when the silicic acid precursor couples with the microwave radiation, leading to silica nanoparticle sol formation. The silica nanoparticles were characterized by dynamic light scattering data and scanning electron microscopy, which show the materials&#39; morphology and size to be dependent on the reaction conditions. Microwave-assisted reactions produce silica nanoparticles with roughened textured surfaces that are atypical for silica sols produced by St&#246;ber&#39;s methods, which have smooth surfaces.

Silica nanoparticles were prepared using acid-catalysis of a siloxane precursor and microwave-assisted synthetic techniques resulting in the controlled growth of nanomaterials ranging from 30-250 nm in diameter. The growth dynamics can be controlled by varying the initial silicic acid concentration, time of the reaction, and temperature of reaction.

Microwave-assisted synthetic techniques were used to quickly and reproducibly produce silica nanoparticle sols using an acid catalyst with nanoparticle ...

Thin-layer Chromatographic (TLC) Separations and Bioassays of Plant Extracts to Identify Antimicrobial Compounds

A common screen for plant antimicrobial compounds consists of separating plant extracts by paper or thin-layer chromatography (PC or TLC), exposing the chromatograms to microbial suspensions (e.g. fungi or bacteria in broth or agar), allowing time for the microbes to grow in a humid environment, and visualizing zones with no microbial growth.&#160;The effectiveness of this screening method, known as bioautography, depends on both the quality of the chromatographic separation and the care taken with microbial culture conditions.&#160;This paper describes standard protocols for TLC and contact bioautography&#160;with a novel application to amino acid-fermenting bacteria. The extract is separated on flexible (aluminum-backed) silica TLC plates, and bands are visualized under ultraviolet (UV) light.&#160;Zones are cut out and incubated face down onto agar inoculated with the test microorganism.&#160;Inhibitory bands are visualized by staining the agar plates with tetrazolium red.&#160;The method is applied to the separation of red clover (Trifolium pratense cv. Kenland) phenolic compounds and their screening for activity against Clostridium sticklandii, a hyper ammonia-producing bacterium (HAB) that is native to the bovine rumen.&#160;The TLC methods apply to many types of plant extracts&#160;and other bacterial species (aerobic or anaerobic), as well as fungi, can be used as test organisms if culture conditions are modified to fit the growth requirements of the species.

Thin-layer Chromatographic TLC Separations and Bioassays of Plant Extracts to Identify Antimicrobial Compounds

Methods are described for thin-layer chromatographic (TLC) separation of plant extracts and contact bioautography to identify antibacterial metabolites. The methods are applied to the screening of red clover phenolic compounds inhibiting hyper&#160;ammonia-producing bacteria (HAB) native to the bovine rumen.

A common screen for plant antimicrobial compounds consists of separating plant extracts by paper or thin-layer chromatography (PC or TLC), exposing the ...

Experimental Protocol for Biodiesel Production with Isolation of Alkenones as Coproducts from Commercial Isochrysis Algal Biomass

The need to replace petroleum fuels with alternatives from renewable and more environmentally sustainable sources is of growing importance. Biomass-derived biofuels have gained considerable attention in this regard, however first generation biofuels from edible crops like corn ethanol or soybean biodiesel have generally fallen out of favor. There is thus great interest in the development of methods for the production of liquid fuels from domestic and superior non-edible sources. Here we describe a detailed procedure for the production of a purified biodiesel from the marine microalgae Isochrysis. Additionally, a unique suite of lipids known as polyunsaturated long-chain alkenones are isolated in parallel as potentially valuable coproducts to offset the cost of biodiesel production. Multi-kilogram quantities of Isochrysis are purchased from two commercial sources, one as a wet paste (80% water) that is first dried prior to processing, and the other a dry milled powder (95% dry). Lipids are extracted with hexanes in a Soxhlet apparatus to produce an algal oil (&#34;hexane algal oil&#34;) containing both traditional fats (i.e., triglycerides, 46-60% w/w) and alkenones (16-25% w/w). Saponification of the triglycerides in the algal oil allows for separation of the resulting free fatty acids (FFAs) from alkenone-containing neutral lipids. FFAs are then converted to biodiesel (i.e., fatty acid methyl esters, FAMEs) by acid-catalyzed esterification while alkenones are isolated and purified from the neutral lipids by crystallization. We demonstrate that biodiesel from both commercial Isochrysis biomasses have similar but not identical FAME profiles, characterized by elevated polyunsaturated fatty acid contents (approximately 40% w/w). Yields of biodiesel were consistently higher when starting from the Isochrysis wet paste (12% w/w vs. 7% w/w), which can be traced to lower amounts of hexane algal oil obtained from the powdered Isochrysis product.

Experimental Protocol for Biodiesel Production with Isolation of Alkenones as Coproducts from Commercial Isochrysis Algal Biomass

Detailed methods are presented for the production of biodiesel along with the co-isolation of alkenones as valuable coproducts from commercial Isochrysis microalgae.

The need to replace petroleum fuels with alternatives from renewable and more environmentally sustainable sources is of growing importance. ...

Environment

A Toolkit to Enable Hydrocarbon Conversion in Aqueous Environments

This work puts forward a toolkit that enables the conversion of alkanes by Escherichia coli and presents a proof of principle of its applicability. The toolkit consists of multiple standard interchangeable parts (BioBricks)9 addressing the conversion of alkanes, regulation of gene expression and survival in toxic hydrocarbon-rich environments.
A three-step pathway for alkane degradation was implemented in E. coli to enable the conversion of medium- and long-chain alkanes to their respective alkanols, alkanals and ultimately alkanoic-acids. The latter were metabolized via the native &beta;-oxidation pathway. To facilitate the oxidation of medium-chain alkanes (C5-C13) and cycloalkanes (C5-C8), four genes (alkB2, rubA3, rubA4and rubB) of the alkane hydroxylase system from Gordonia sp. TF68,21 were transformed into E. coli. For the conversion of long-chain alkanes (C15-C36), theladA gene from Geobacillus thermodenitrificans was implemented. For the required further steps of the degradation process, ADH and ALDH (originating from G. thermodenitrificans) were introduced10,11. The activity was measured by resting cell assays. For each oxidative step, enzyme activity was observed.
To optimize the process efficiency, the expression was only induced under low glucose conditions: a substrate-regulated promoter, pCaiF, was used. pCaiF is present in E. coli K12 and regulates the expression of the genes involved in the degradation of non-glucose carbon sources.
The last part of the toolkit - targeting survival - was implemented using solvent tolerance genes, PhPFD&alpha; and &beta;, both from Pyrococcus horikoshii OT3. Organic solvents can induce cell stress and decreased survivability by negatively affecting protein folding. As chaperones, PhPFD&alpha; and &beta; improve the protein folding process e.g. under the presence of alkanes. The expression of these genes led to an improved hydrocarbon tolerance shown by an increased growth rate (up to 50%) in the presences of 10% n-hexane in the culture medium were observed.
Summarizing, the results indicate that the toolkit enables E. coli to convert and tolerate hydrocarbons in aqueous environments. As such, it represents an initial step towards a sustainable solution for oil-remediation using a synthetic biology approach.

A sustainable auto regulating bacterial system for the remediation of oil pollutions was designed using standard interchangeable DNA parts (BioBricks). An engineered E. coli strain was used to degrade alkanes via &beta;-oxidation in toxic aqueous environments. The respective enzymes from different species showed alkane degradation activity. Additionally, an increased tolerance to n-hexane was achieved by introducing genes from alkane-tolerant bacteria.

This work puts forward a toolkit that enables the conversion of alkanes by Escherichia coli and presents a proof of principle of its applicability. The ...

Bioengineering

Qualitative Characterization of the Aqueous Fraction from Hydrothermal Liquefaction of Algae Using 2D Gas Chromatography with Time-of-flight Mass Spectrometry

Two-dimensional gas chromatography coupled with time-of-flight mass spectrometry is a powerful tool for identifying and quantifying chemical components in complex mixtures. It is often used to analyze gasoline, jet fuel, diesel, bio-diesel and the organic fraction of bio-crude/bio-oil. In most of those analyses, the first dimension of separation is non-polar, followed by a polar separation. The aqueous fractions of bio-crude and other aqueous samples from biofuels production have been examined with similar column combinations. However, sample preparation techniques such as derivatization, solvent extraction, and solid-phase extraction were necessary prior to analysis. In this study, aqueous fractions obtained from the hydrothermal liquefaction of algae were characterized by two-dimensional gas chromatography coupled with time-of-flight mass spectrometry without prior sample preparation techniques using a polar separation in the first dimension followed by a non-polar separation in the second. Two-dimensional plots from this analysis were compared with those obtained from the more traditional column configuration. Results from qualitative characterization of the aqueous fractions of algal bio-crude are discussed in detail. The advantages of using a polar separation followed by a non-polar separation for characterization of organics in aqueous samples by two-dimensional gas chromatography coupled with time-of-flight mass spectrometry are highlighted.

A two-dimensional gas chromatography-time-of-flight mass spectrometry method is described for characterization of the aqueous fraction of bio-crude produced from hydrothermal liquefaction of algae. This protocol can also be employed to analyze the aqueous fraction of liquid products from fast pyrolysis, catalytic fast pyrolysis, catalytic deoxygenation and hydro-treating.

Two-dimensional gas chromatography coupled with time-of-flight mass spectrometry is a powerful tool for identifying and quantifying chemical components in ...

Fast Pyrolysis of Biomass Residues in a Twin-screw Mixing Reactor

Fast pyrolysis is being increasingly applied in commercial plants worldwide. They run exclusively on woody biomass, which has favorable properties for conversion with fast pyrolysis. In order to increase the synergies of food production and the energetic and/or material use of biomass, it is desirable to utilize residues from agricultural production, e.g., straw. The presented method is suitable for converting such a material on an industrial scale. The main features are presented and an example of mass balances from the conversion of several biomass residues is given. After conversion, fractionated condensation is applied in order to retrieve two condensates &#8212; an organic-rich and an aqueous-rich one. This design prevents the production of fast pyrolysis bio-oil that exhibits phase separation. A two phase bio-oil is to be expected because of the typically high ash content of straw biomass, which promotes the production of water of reaction during conversion.
Both fractionated condensation and the use of biomass with high ash content demand a careful approach for establishing balances. Not all kind of balances are both meaningful and comparable to other results from the literature. Different balancing methods are presented, and the information that can be derived from them is discussed.

A procedure for thermochemical conversion of biomass residues is presented that aims at maximizing the yield of liquid products (fast pyrolysis). It is based on a technology proven on an industrial scale and especially suitable for treating a straw type of biomass.

Fast pyrolysis is being increasingly applied in commercial plants worldwide. They run exclusively on woody biomass, which has favorable properties for ...

Construction and Setup of a Bench-scale Algal Photosynthetic Bioreactor with Temperature, Light, and pH Monitoring for Kinetic Growth Tests

The optimal design and operation of photosynthetic bioreactors (PBRs) for microalgal cultivation is essential for improving the environmental and economic performance of microalgae-based biofuel production. Models that estimate microalgal growth under different conditions can help to optimize PBR design and operation. To be effective, the growth parameters used in these models must be accurately determined. Algal growth experiments are often constrained by the dynamic nature of the culture environment, and control systems are needed to accurately determine the kinetic parameters. The first step in setting up a controlled batch experiment is live data acquisition and monitoring. This protocol outlines a process for the assembly and operation of a bench-scale photosynthetic bioreactor that can be used to conduct microalgal growth experiments. This protocol describes how to size and assemble a flat-plate, bench-scale PBR from acrylic. It also details how to configure a PBR with continuous pH, light, and temperature monitoring using a data acquisition and control unit, analog sensors, and open-source data acquisition software.

This paper describes the assembly process and operation of a bench-scale photosynthetic bioreactor that can be used, in conjunction with other methods, to estimate pertinent kinetic growth parameters. This system continuously monitors the pH, light, and temperature using sensors, a data acquisition and control unit, and open-source data acquisition software.

The optimal design and operation of photosynthetic bioreactors (PBRs) for microalgal cultivation is essential for improving the environmental and economic ...

Biodiesel Synthesis by Ultrasonic-Assisted Transesterification of Vegetable Oils

Ultrasonic-Assisted Preparation of Biodiesel Products from Vegetable Oils

Utilizing vegetable oil as a sustainable feedstock, this study presents an innovative approach to ultrasonic-assisted transesterification for biodiesel synthesis. This alkaline-catalyzed procedure harnesses ultrasound as a potent energy input, facilitating the rapid conversion of extra virgin olive oil into biodiesel. In this demonstration, the reaction is run in an ultrasonic bath under ambient conditions for 15 min, requiring a 1:6 molar ratio of extra virgin olive oil to methanol and a minimum amount of KOH as the catalyst. The physiochemical properties of biodiesel are also reported. Emphasizing the remarkable advantages of ultrasonic-assisted transesterification, this method demonstrates notable reductions in reaction and separation times, achieving near-perfect purity (~100%), high yields, and negligible waste generation. Importantly, these benefits are achieved within a framework that prioritizes safety and environmental sustainability. These compelling findings underscore the effectiveness of this approach in converting vegetable oil into biodiesel, positioning it as a viable option for both research and practical applications.

Author Spotlight: Employing Green-Chemistry Principles for Safe and Sustainable Synthesis of Biodiesels

A safe ultrasonic-assisted transesterification method for vegetable oils using an alkaline catalyst is presented here. The method is rapid and efficient for preparing pure biodiesel products.

Utilizing vegetable oil as a sustainable feedstock, this study presents an innovative approach to ultrasonic-assisted transesterification for biodiesel ...

Biofuels: Producing Ethanol from Cellulosic Material

Source: Laboratories of Margaret Workman and Kimberly Frye - Depaul University
In this experiment, cellulosic material (such as corn stalks, leaves, grasses, etc.) will be used as a feedstock for the production of ethanol.&#160;The cellulosic material is first pretreated (ground and heated), digested with enzymes, and then fermented with yeast.&#160;Ethanol production is monitored using an ethanol probe.&#160;The experiment can be extended to optimize ethanol production by varying the feedstock used, pretreatment conditions, enzyme variation, yeast variation, etc.&#160;An alternative method of monitoring the reaction is to measure the carbon dioxide produced (using a gas sensor) instead of the ethanol.&#160;As a low-tech alternative, glucose meters (found in any drug store) can be used to monitor the glucose during the process, if an ethanol probe or carbon dioxide gas sensor is not available.
With an increased emphasis on &#8216;inquiry-based learning&#8221;, scientific probes are becoming more popular.&#160;Handheld devices like the Vernier Lab Quest used in conjunction with a variety of probes (such as those for conductivity, dissolved oxygen, voltage, and more) allow for less focus on collecting data and/or making graphs and more on analyzing the data and making predictions.&#160;Another advantage is that these are small and lightweight and can be taken into the field for measurements.

Source: Laboratories of Margaret Workman and Kimberly Frye - Depaul University
In this experiment, cellulosic material (such as corn stalks, leaves, ...

Laboratory Production of Biofuels and Biochemicals from a Rapeseed Oil through Catalytic Cracking Conversion

Summary

Explore More Videos

Chapters in this video

Biomass Conversion to Produce Hydrocarbon Liquid Fuel Via Hot-vapor Filtered Fast Pyrolysis and Catalytic Hydrotreating

Preparation of Silica Nanoparticles Through Microwave-assisted Acid-catalysis

Thin-layer Chromatographic (TLC) Separations and Bioassays of Plant Extracts to Identify Antimicrobial Compounds

Experimental Protocol for Biodiesel Production with Isolation of Alkenones as Coproducts from Commercial Isochrysis Algal Biomass

A Toolkit to Enable Hydrocarbon Conversion in Aqueous Environments

Qualitative Characterization of the Aqueous Fraction from Hydrothermal Liquefaction of Algae Using 2D Gas Chromatography with Time-of-flight Mass Spectrometry

Fast Pyrolysis of Biomass Residues in a Twin-screw Mixing Reactor

Construction and Setup of a Bench-scale Algal Photosynthetic Bioreactor with Temperature, Light, and pH Monitoring for Kinetic Growth Tests

Ultrasonic-Assisted Preparation of Biodiesel Products from Vegetable Oils

Biofuels: Producing Ethanol from Cellulosic Material