EVOLUGATE

index_page

administrateur — 2012-02-16T21:38:18Z

The Science & Technology section of our website is under renovation. Please contact us at info@evolugate if you any question. - Science & Technology

Army Reseach Lab

Ziad — 2011-09-01T19:44:05Z

Improving the Conversion Algal Oil to Bio Jet-fuel

On March 14, 2011, Evolugate was awarded a phase 1 SBIR grant from the Army Research Laboratory to increase the maximal growth temperature of a strain of heterotrophic algae.

Increasing the maximal growth temperature will reduce the ratio of unsaturated fatty acids in the algal oil, and make it more suitable for conversion to bio jet-fuel. Using its proprietary technology the Evolugator™, Evolugate will develop the new strain by successively adapting the alga to steadily increasing temperatures until the ratio of unsaturated fatty acids in the oil it produces is significantly reduced for several levels of growth temperatures.

There is tremendous interest in algal oil because many experts are convinced that converting algal oil to biofuels is the most viable method by which enough transportation fuel can be produced to replace current diesel consumption worldwide. Algae, especially the heterotrophic ones, have a lot of advantages as a raw material for biofuel production : They have a faster growth rate than any terrestrial crop ; they have a yield of oil per unit area higher than any other oil crop ; they can grow without competing for resources (land, feedstock, water) with food crops.

Algal oil is rich in fatty acids, which makes it suitable for conversion to biodiesel. However, biodiesel has a number of characteristics that make it inappropriate for airplanes. For example, it has poor cold flow properties, a lower energy density than fossil diesel, and higher kinetic viscosity. Reducing the ratio of unsaturated fatty acids in algal oil will make it a proper raw material for subsequent conversion into a biofuel that has the advantages of fossil jet fuel and none of its inconveniences (i.e. carbon print, oil prices, dependence on foreign oil).

The Army Research Laboratory provides innovative science, technology and analyses to enable a full spectrum of military operations. It serves as the bridge between the scientific and technical communities and the Army while being the lead in providing innovative solutions for current and future soldiers.

FAQs

Marc — 2010-07-15T21:54:13Z

How does continuously culturing microbes facilitate evolution ? In the real world, microbes have spent millennia naturally evolving to their environment. Wild microbes are supremely adapted for their native environment, which is usually subject to continuous, and often extreme, fluctuations in variables such as light intensity, temperature, pressure, moisture, etc. and the microbes must be prepared for and be able to thrive under these fluctuations. These fluctuations, and the need for microbes to carry the required genetic material to adapt to them, slows down the rate of natural evolution. After all, microbes in a fluctuating environment spend less time trying to adapt to each variable they encounter and, therefore, face decreased selective pressure to adapt to any one particular variable. On the other hand, when one or many variables are kept constant, microbes evolve more rapidly. This is the benefit of experimental evolution via continuous cultures—the environmental conditions and be strictly controlled and either kept constant or changed very slowly to facilitate the emergence of adaptive mutations. As continuous cultures are periodically, grown, diluted and re-grown, the fittest variants and fastest growers under these culture conditions take over the population. The result is a strain that has been growth optimized for your conditions of interest. When using microbes for industrial purposes, environmental conditions must often be as uniform as possible to optimize biocatalysis and it is essential for the economic viability of a particular industrial process that the microbial biocatalyst be able can optimally under these steady conditions. Thus, experimental evolution via continuous culture is an ideal way of optimizing wild microbes for the steady conditions found in industry.

What is done with spent culture ? Spent microbial cultures are simply destroyed according to the appropriate waste material regulations, with respect to each related material.

Continuous culture is not new and has been known for decades to be an ineffective way of changing the properties of microorganisms. What would make it work now ? The use of serial or continuous culture to experimentally evolve microorganisms is not a new idea. Indeed, the first example of this approach was published in 1878 and continuous culture had its heyday in the middle of the 1900's, culminating in the invention of the chemostat by Szilard and Monod. Unfortunately, continuous culture never reached its potential and has failed to deliver effective microbes for industrial applications. Both serial cultures and chemostats are limited by technical difficulties that either slow down the rate at which experimental evolution occurs (as in serial transfer) or prevents it completely (as in chemostats). Thus, companies that wish to develop industrial strains through experimental evolution are limited to methods that are largely ineffective. Not surprisingly, interest in experimental evolution has steadily waned in the last 30 years or so. We have developed new culture technology that circumvents the traditional problems associated with continuously culturing microbes allowing for rapid and robust experimental evolution. Improving natural micro-organisms through experimental evolution is now practical and our technology has reopened a wide field of applications for green chemistry.

Will the strains you adapt revert over time ? Clearly, microbes have spent millions of years adapting to be the way they are. If selective pressure to change is removed from a microbe we have adapted for a particular set of conditions and the microbe is returned to its original environment, it will experience pressure to revert back to its original state. However, long term experimental evolution results in the accumulation of many different genetic changes and their rapid reversion is highly improbable. Thus, it will take time for our microbes to alter phenotype once they are removed from the environment to which we have adapted them. That said, it is important for us to continuously apply selective pressure on our microbes, otherwise we run the risk of undoing the good work we have done.

When you adapt a microbe to a different optimal growth temperature, does it lose its ability to grow at its initial optimal growth temperature ? Does it narrow its range of temperature ? In evolutionary biology, there is a concept called antagonistic pleiotropy which postulates that the longer a microbe spends adapting to a certain set of conditions, the less robust it will be under other conditions. So the short answer is yes, if the difference between starting and final growth temperature is large enough and if the microbe spends enough time learning to be a specialist at a particular temperature, it is possible that a microbe will lose the ability to grow at the initial temperature. We have not pushed the limits of thermal adaptation and the strains we have produced are still capable of growing at both temperatures, although we do see a drop in robustness at the initial temperature.

Is it possible to evolve consortia of microbes in a same experiment ? If the microbes in this consortium are interdependent for growth, that is to say that the growth of one is required for the growth of another, then it is fairly straightforward. Selecting for growth rate improves the growth of the rate-limiting microbe. If the microbes are not interdependent, then this is one of the most difficult configurations to work on. In this latter case, the microbes that are to be cultured together must either have compatible replication rates or we will systematically wash out the slowest growing species. Under these circumstances, it is still possible to evolve consortia, however, it is important to run the experiment in what we call "chemostat" mode where the microbes are cultured for as long as possible prior to dilution. Under these conditions, different microbes will assume different niches in the culture chamber (e.g. eating different food sources) and become adapted to the new conditions this way.

Is it possible to predict the time it takes to achieve some specific goal ? Unlike some traditional engineering, IT, or building developments, the accumulation of biological mutations is a stochastic process. There is no way to predict when an adaptive mutation will occur, nor is it possible to predict what type of adaptive mutation might occur. Evolution is generally characterized by a series of high-probability mutational events that usually have only modest adaptive effects. However, it is our experience that the adaptation process also occasionally hits "metabolic walls" where it takes a significantly longer time for an adaptive variant to sweep through the population. At these "walls" it is likely that low-probability mutagenic events are required to continue the adaptation process. It is impossible to predict where these "walls" might occur or how long it might take to break through them. Progressing in the field of experimental evolution is like exploring a wild country beyond a new frontier.

What makes your technique better than genetic engineering ? The Evolugate technology and its ability to facilitate robust experimental evolution confers several competitive advantages over genetic engineering : • First, microorganisms have spent millions of years evolving exquisitely complex metabolic pathways, making it exceedingly difficult to properly re-engineer new functionality. We simply do not know enough about some characteristics to be able to predict what types of changes need to engineered. Moreover, even high-throughput recombinant methods of genetic engineering, such as genome shuffling, have difficulty altering complex traits, which may require the acquisition of multiple low probability mutagenic events. Experimental evolution is blind to these concerns. With enough selective pressure and time, experimental evolution can easily access the genetic diversity needed to alter complex and poorly understood traits. • Second, genetic engineering requires the ability to insert or remove genetic material. For many microbes, the tools required to make this work simply do not yet exist, requiring significant investment in the development of the appropriate molecular biology methods. There is no such limitation for experimental evolution, which only requires the ability to culture cells. • Third, tinkering with one trait via genetic engineering often comes at the expense of other traits, such as growth rate and even successful examples of genetic engineering tend to produce strains that are less robust than the parent strain. To make an analogy, it is a bit like adding or deleting some lines in a piece of software when you did not write the program and are not a master of all the code. Thus, engineered microbes may be able to perform a certain industrial task, but do so too slowly to be an effective biocatalyst from an economic standpoint. Strains produced by experimental evolution are simultaneously optimized for both the desired trait as well as growth rate. • Fourth, strains produced by experimental evolution are naturally occurring genetic variants of the original parent strain—no foreign DNA has been inserted and no endogenous DNA has been deliberately removed. As such, these strains are not considered "genetically-modified organisms" (GMOs). This is an important consideration for certain environmentally sensitive projects where the resulting microbe may be released into the biosphere. This said, our technique can be highly complementary with genetic engineering.

How is your technology integrated in the industry ? Our purpose is to produce variants of industrially important microbes that can grow robustly under the conditions that prevail in industry. To achieve this result our growth conditions are designed to be as close as possible to the actual conditions a microbe would experience in the real world. As a result, we are not making microbes that work in a lab, we will be able to more easily incorporate them into a functioning biorefinery.

When you evolve microbes, does it produce GMOs ? Strains produced by experimental evolution are naturally occurring genetic variants of the original parent strain—no foreign DNA has been inserted and no endogenous DNA has been deliberately removed. As such, these strains are not considered "genetically-modified organisms" (GMOs). This is an important consideration for certain environmentally sensitive projects where the resulting microbe may be released into the biosphere.

Is your technology harmless for the environment ? Many of the microbes we develop are destined for use in confined fermentors for bioproduct production at industrial scale, thus there risk to the environment is minimal. However, when the industry requires that a particular microbial biocatalyst be released into the biosphere, there is always a risk, however small, that that microbe can have a deleterious effect on the environment. There are two ways in which our technology minimizes this risk and makes our microbes the greenest solution to this problem. First, through experimental evolution, microbes become hyper-specialists at thriving under the conditions to which we adapted them. They do so at the expense of being able to thrive when those conditions change. Thus, once they are out in the wild they are at a distinct disadvantage once the conditions change from what they were designed for. This means that the most likely fate for our microbes after they "escape" is extinction. When they have nothing left to eat they just starve and die, resulting in microbial biomass that is highly and quickly biodegradable.

History

Marc — 2010-07-15T20:59:07Z

this relates to the first discoeries of biology and culturing microbes - Optimized Evolution

June 2010

Marc — 2010-06-03T14:01:13Z

Evolugate Launches Bioremediation Project to Mitigate the Gulf Oil Spill

As BP scientists struggle to cap the well that is leaking vast quantities of oil into the Gulf of Mexico, attention is turning to the daunting task of cleaning up the oil that is slowly creeping ashore and affecting sensitive marine ecosystems. Bioremediation is emerging as the only viable means to this end, however, significant obstacles prevent current methods of bioremediation from being effective. EVOLUGATE, LLC of Gainesville, FL has developed an innovative solution to the problems that restrain the full potential of bioremediation and has committed its resources to providing an environmentally friendly long-term microbial solution for effective and rapid bioremediation of the Gulf oil spill.

Evolugate announces the initiation of this project with scientists taking actual samples of oil from contaminated sites in the Gulf and beginning to culture microbial communities uniquely adapted for biodegrading these samples.

What is bioremediation and what are its limitations ? Oil-eating microbes are found naturally throughout the ocean and bioremediation is the process of allowing these microbes to naturally break down oil of anthropogenic origin. The problem with simply allowing these microbes to do their work is that they do so too slowly to prevent oil from doing significant environmental and economic damage. Sometimes this is because the natural flora is not accustomed to certain components of a particular spill and sometimes it is because the environment lacks other nutrients like oxygen. To solve these problems, researchers have turned to augmenting the native flora with microbes that can degrade components refractory to natural biodegradation or that can work efficiently without oxygen. However, this approach has been largely ineffective mainly because these non-native microbes are not adapted for the specific environmental conditions of a particular spill, such as water temperature, salinity, pH or oxygen concentration. EVOLUGATE is taking a unique and groundbreaking approach to this problem by pre-adapting communities of petroleum-degrading microbes to both actual oil samples taken from contaminated sites in the Gulf and the specific environmental conditions where those samples were found. The result is a catalog of microbial communities perfectly adapted for the rapid and efficient biodegradation of specific contaminated sites.

How does EVOLUGATE produce these oil-eating microbes ? The key to effective bioremediation of the Gulf oil spill is the abandonment of the "one-size-fits-all" approach to the problem. In essence, there is no magic bullet microbe capable of rapidly degrading all oil spills in any environment. Rather, effective microbes must be tailored for the unique crude oil composition and environmental conditions of affected areas of the Gulf. Only EVOLUGATE with its patented methodology for adapting microbes to new environmental conditions and food sources, can produce "designer" strains custom-made for remediating sensitive areas of the Gulf. More importantly, the EVOLUGATE method produces variants of naturally occurring marine microbes so the strains released into the environment are not GMOs. In summary, EVOLUGATE is capable of quickly procuring "designer" microbial communities that are both highly capable of degrading a real-world oil spill and of doing so as rapidly as possible and with minimal lingering impact on the environment.

Illustrated Press Release June 3rd

In the Media

Marc — 2010-02-16T16:37:37Z

August 2010

July 1, 2010

June 17, 2010

June 14, 2010

June 9, 2010

June 7, 2010

June 4, 2010

April 2010

March 2010

February 12, 2010

February 9, 2010

January 1, 2010

September 9, 2009

August 26, 2009

February 2010

Marc — 2010-02-10T20:04:41Z

Nominations. Two experienced CEOs join the Advisory Board of Evolugate, LLC. The company is proud to announce the appointment of Russell J. Howard and Terrance J. Bruggeman to its Advisory Board.

Dr. Howard currently serves as CEO of a new Cleantech company, Oakbio Inc., as well as on the boards of several leading companies and foundations in the biotechnology industry. He brings significant scientific and operational expertise having served as CEO of Maxygen, Inc., a publicly traded company, for the past eleven years. Dr. Howard has published over 140 peer-reviewed publications and received numerous awards and honors.

Mr. Bruggeman's extensive experience managing biotechnology companies, including Diversa, Inc. (now Verenium, Inc.) will be an asset to Evolugate. Mr. Bruggeman currently serves as Executive Chairman of BioTork, the spin-off company recently created from Evolugate to capitalize on the Biofuels opportunity. He also serves on a number of industry, corporate and non-profit boards.

"Evolugate has assembled the scientific and technology expertise to exploit the breakthrough new technology of “microbial evolution on demand” developed by Evolugate. I am excited to advise the team as they seek to capitalize on their unique technology capabilities to develop products in a diversity of business areas”, said Russell Howard.

"Evolugate's technology creates a myriad of commercial opportunities,” said Terry Bruggeman. “I look forward to helping guide the company to commercial success.”

Evolugate's technology exploits the power of Darwinian evolution and natural selection for tuning and optimizing the phenotype of any cell to provide it with superior commercial relevance.

Evolugate's technology can be used to rapidly produce optimally adapted and efficient strains for virtually any application, including the conversion of biomass to biofuels, biopesticides, and the improvement of any industrial bioprocess.

index_page

administrateur — 2007-10-13T10:41:35Z

Evolugate provides access to the Evolugator technology for both industrial organizations and academic community.

Commercial entities

Evolugate mainly collaborates with industry to improve any bioprocesses, in every field of biochemistry including biofuels, bioremediation, and bio-insecticides and has the capability to collaborate in other fields like biochemicals, bio-nutrition, bio-cosmetics and bio-pharmaceuticals.

Developing microbial strains to perform specific industrial tasks is a quite long and elaborated process. In order to make it as comprehensible as possible for potential partners, Evolugate has broken it into several fundamental steps : Initial Consultation, Conceptual Study, Technical Assessment, Feasibility Study, Evolution Process, Scale Up and Microbial Supply.

Our “Project Development Process” document provides information about each one of these steps such as general description, prerequisites, timing and deliverables. By understanding the Evolugate development process, our partners will be able to better plan and manage our collaborative projects, and avoid wastes of energy, time and other valuable resources. Contact us to get the “Project Development Process” document.

Our collaborations with industry are subject to a Research Service and Licensing Agreement in connection with the Work Plan elaborated jointly.

Academic institutions

Evolugate's Academic Collaboration Program provides academic research laboratories access to our technology and methods, allowing them to improve their microbial strains for a variety of non-commercial, research applications.

Our first of such collaborations was with the University of Florida. Details can be found in the collaborations section.

For more information about academic collaboration conditions, please contact us or e-mail at info@evolugate.com.

index_page

administrateur — 2007-10-13T10:40:08Z

Evolugate is a biotechnology and engineering company, operating in the nascent field of experimental evolution and metabolic engineering of micro-organisms for industrial and academic purposes.

We offer the possibility to improve existing bio-processes as well as to create new ones through the utilization of our reliable and proprietary continuous culture technology, known as the EvolugatorTM, which improves microorganisms by natural evolution and selection.

It is operated with the help of an interdisciplinary team that offers a unique blend of expertise in both science and business, federating people coming from research and industry as well.

Evolugate's interdisciplinary team offers a unique blend of expertise that enables us to support industrial organizations. Our biological staff includes experts combining cutting-edge knowledge in biocatalysis, genomics, evolution, and metabolic engineering. Our expertise will allow us to optimize both the initial engineering of target micro-organisms and their subsequent improvement through evolution in the Evolugator. Our engineering staff, which has designed and manufactured the EvolugatorTM machine, has the requisite manufacturing and engineering expertise to enable us to adapt the Evolugator for specific applications as they arise.

index_page

administrateur — 2007-10-13T10:30:57Z

For inquiry about :

Academic Collaboration Program : info@evolugate.com

Business Development : see subsidiaries

	• Biofuels
	• Biodegradation
	• Bioinsecticides
	• Other markets

Other purposes : info@evolugate.com

To contact us by mail :

	Evolugate LLC
	2153 SE Hawthorne Rd, #15
	Gainesville, FL 32641
	USA

To contact us by phone :

(352)505-8646