Synthetic biology has been essential in engineering the LS9 microbial catalysts. The biosynthetic pathways to produce finished fuel products do not exist in the native E. coli host, and prior to our efforts alkane biosynthetic genes were unknown. LS9 designed the pathways, synthesized the genes encoding each enzyme in the pathway, and constructed multigene biosynthetic operons enabling production. To improve yield, productivity, and titer – the drivers of process economic efficiency – the biosynthetic pathways and host metabolism have required significant genetic optimization. LS9 developed capabilities for the computational design and automated parallel construction of gene, operon, and recombinant cell libraries that have enabled the rapid construction and evaluation of thousands of rationally engineered microorganisms. This capability in combination with state of the art screening, process development, and analytical methodologies has enabled LS9 in only a few years to advance from concept to a process slated for commercial-scale demonstration.
This same technology platform has been leveraged for the production of surfactants for use in consumer products in collaboration with Procter & Gamble. The ability to exchange biosynthetic parts and leverage the core host “chassis” has enabled the development of this chemical product line much faster, achieving in months what had taken years for the earlier products.
LS9 intends to continue to leverage the power of synthetic biology to further advance these and future products as quickly and cost effectively as possible. We feel strongly these technologies are essential to the goal of weaning our dependence on fossil feedstocks and the further development of a world leading industrial biotechnology industry.
Increasing Rates of Natural Fermentation for Polymers
Metabolix is bringing new, clean solutions to the plastics, chemicals and energy industries based on highly differentiated technology. For 20 years, Metabolix has focused on advancing its foundation in polyhydroxyalkanoates (PHA), a broad family of biopolymers. Through a microbial fermentation process, the base polymer PHA is produced within microbial cells and then harvested. Development work by Metabolix has led to industrial strains of the cells, which can efficiently transform natural sugars into PHA. The recovered polymer is made into pellets to produce Mirel™ Bioplastics by Telles products.
Conventional plastics materials like polyvinyl chloride (PVC), polyethylene teraphthalate (PET), and polypropylene (PP) are made from petroleum or fossil carbon. The PHA in Mirel bioplastics is made through the fermentation of sugar and can be biodegraded by the microbes present in natural soil or water environments. Although PHAs are produced naturally in many microorganisms, the cost and range of compositions required for successful commercialization dictated that PHA pathways had to be assembled in a robust industrial organism that does not naturally produce the product.
Metabolic pathway engineering was used to accomplish this task, relying on modern tools of biotechnology. These include DNA sequencing and synthetic construction of genes encoding the same amino acid sequence as in the donor strain, but optimized for expression in the engineered industrial host. These technologies provided rapid development and optimization of robust industrial production strains that would not have been feasible using classical techniques relying on isolation and transfer of DNA from one species to the other.
This has allowed Metabolix to successfully commercialize Mirel bioplastics. More than 50 years after it was first considered as a potentially useful new material and following several efforts by leading chemical companies to commercialize PHAs based on natural production hosts, Metabolix has made these products available at a commercial scale.
Increasing Efficiency in Bioprocessing of Pharmaceuticals
Sitagliptin, Merck’s first-in-class dipeptidyl peptidase-4 inhibitor, is marketed under the trade name Januvia® as a treatment for type II diabetes. The chemical manufacturing route to Sitagliptin developed by Merck won a Presidential Green Chemistry Challenge Award in 2006, but there were still several opportunities for improvement. Codexis and Merck collaborated to develop a novel, environmentally benign alternative manufacturing route. Using synthetic biology and its directed evolution technologies, Codexis discovered and developed a transaminase capable of enabling the new biocatalytic route, which is currently in scale-up towards commercial manufacture.
One common definition of “synthetic biology” is “the design and construction of new biological entities that do not exist in the natural world.” In this instance, there was no known enzyme that could perform the reaction required to enable the biocatalytic route. By designing and generating new enzyme variants, Codexis was able to identify a novel enzyme that provided detectable initial activity. This enzyme was then improved greater than 25,000-fold in order to generate the highly active, stable, enantioselective and practical enzyme from a starting activity that did not previously exist in the natural world. This work was awarded with the Presidential Green Chemistry Challenge Award in June, 2010.
Producing a Building Block for Everyday Products