Development of scalable bio-platforms for production of...
2
Development of scalable bio-platforms for production of novel products for industrial, fuel, and pharmaceutical applications Mark Running ([email protected]), David Schultz ([email protected]) Industrial manufacturing, like many other sectors of the economy, relies heavily on inexpensive and abundant petroleum and other fossil fuels for energy and raw materials input. While advanced extraction technologies have recently provided supply stability, we are still on track to largely deplete petroleum reserves in a few generations, with price instability likely to become a recurring issue, as it has many times in the past. A great deal of interest has focused on photosynthetic organisms as an alternative, renewable source for liquid fuels, as well as a source of substitutions for petroleum inputs in the manufacturing of plastics, lubricants, surfactants, and pharmaceuticals. The aim of our proposed contribution to the Kentucky RII Track I application is the development of novel bio-based production platforms for large scale production of raw materials suitable for industrial, fuel, and medicinal applications. In this way we hope to contribute to the EPSCoR central theme’s broad category of inputs, including development of new materials using synthetic biology approaches, ultimately resulting in key applications that enhance Kentucky’s competitiveness in agricultural, transportation (fuel) and health (biopharmaceutical production) industries. Cross-disciplinary collaboration statewide with faculty and industry partners will play a key role in the success of this project. Project: engineering of plant biosynthetic pathways to optimize production of products for downstream applications. Plants produce an enormous variety of chemical structures, more than 50,000 distinct compounds. However, these plant specialized metabolites are typically underutilized in industrial applications, in large part because these products may only be found in crops not compatible with large-scale agricultural production, are found in low quantities, or are too expensive to purify from plant tissues. Biotechnology provides a mechanism to produce selected compounds in crops that are already optimized for agricultural production. Working with EPSCoR faculty and industrial collaborators, we will target enhanced value compounds for production in crop plants widely grown in Kentucky, and in the moss system described below. Two example areas of interest are production of unusual monoenoic fatty acids (with uses as low temperature lubricants, feedstock for polymer production, etc.) and anacardic acids (with uses as resins, coatings, rubber compounds, medicinal applications, etc.) through heterologous expression of target genes. Project: development and scaling of novel plant-based systems for large-scale production. We have identified an altered strain of the moss Physcomitrella patens that grows as single, rapidly dividing cells that do not differentiate and do not senesce (Figure 1). These plants, like the green algae progenitors of land plants, also accumulate high levels of oils, as indicated by structural studies (Figure 2) and metabolite measurements. Preliminary investigations indicate that these cells are more photosynthetically active, as indicated by higher levels of expression of photosynthesis genes and greater numbers of photosynthetic structures. The enhanced ability of this moss strain to manufacture and accumulate lipids, along with its ease of growth, ease of transformation, and scalability, makes this a potentially extremely useful system for industrial oil and biofuel production. Once compounds targeted for large-scale production are identified (in consultation with statewide academic and industrial collaborators), we will introduce the genes necessary for synthesis of these compounds into this moss strain, and implement and optimize technologies that allow for high-level production of the compound of interest.
Transcript of Development of scalable bio-platforms for production of...
Development of scalable bio-platforms for production of novel products for industrial, fuel, and pharmaceutical applications
Mark Running ([email protected]), David Schultz ([email protected])
Industrial manufacturing, like many other sectors of the economy, relies heavily on inexpensive and abundant petroleum and other fossil fuels for energy and raw materials input. While advanced extraction technologies have recently provided supply stability, we are still on track to largely deplete petroleum reserves in a few generations, with price instability likely to become a recurring issue, as it has many times in the past. A great deal of interest has focused on photosynthetic organisms as an alternative, renewable source for liquid fuels, as well as a source of substitutions for petroleum inputs in the manufacturing of plastics, lubricants, surfactants, and pharmaceuticals. The aim of our proposed contribution to the Kentucky RII Track I application is the development of novel bio-based production platforms for large scale production of raw materials suitable for industrial, fuel, and medicinal applications. In this way we hope to contribute to the EPSCoR central theme’s broad category of inputs, including development of new materials using synthetic biology approaches, ultimately resulting in key applications that enhance Kentucky’s competitiveness in agricultural, transportation (fuel) and health (biopharmaceutical production) industries. Cross-disciplinary collaboration statewide with faculty and industry partners will play a key role in the success of this project. Project: engineering of plant biosynthetic pathways to optimize production of products for downstream applications. Plants produce an enormous variety of chemical structures, more than 50,000 distinct compounds. However, these plant specialized metabolites are typically underutilized in industrial applications, in large part because these products may only be found in crops not compatible with large-scale agricultural production, are found in low quantities, or are too expensive to purify from plant tissues. Biotechnology provides a mechanism to produce selected compounds in crops that are already optimized for agricultural production. Working with EPSCoR faculty and industrial collaborators, we will target enhanced value compounds for production in crop plants widely grown in Kentucky, and in the moss system described below. Two example areas of interest are production of unusual monoenoic fatty acids (with uses as low temperature lubricants, feedstock for polymer production, etc.) and anacardic acids (with uses as resins, coatings, rubber compounds, medicinal applications, etc.) through heterologous expression of target genes. Project: development and scaling of novel plant-based systems for large-scale production. We have identified an altered strain of the moss Physcomitrella patens that grows as single, rapidly dividing cells that do not differentiate and do not senesce (Figure 1). These plants, like the green algae progenitors of land plants, also accumulate high levels of oils, as indicated by structural studies (Figure 2) and metabolite measurements. Preliminary investigations indicate that these cells are more photosynthetically active, as indicated by higher levels of expression of photosynthesis genes and greater numbers of photosynthetic structures. The enhanced ability of this moss strain to manufacture and accumulate lipids, along with its ease of growth, ease of transformation, and scalability, makes this a potentially extremely useful system for industrial oil and biofuel production. Once compounds targeted for large-scale production are identified (in consultation with statewide academic and industrial collaborators), we will introduce the genes necessary for synthesis of these compounds into this moss strain, and implement and optimize technologies that allow for high-level production of the compound of interest.
Figure 1. (Left) Wild type cells of the moss Physcomitrella patens grows as filamentous structures. (Right) The ggb strain of Physcomitrella patens grows as individual cells that rapidly divide and do not differentiate, making them suitable for bio-based production. Figure 2. (Left) Wild type Physcomitrella patens cells show little oil body accumulation. (Right) The ggb strain shows large, visible oil bodies even at an early stage of development.
