| NSF Org: |
MCB Div Of Molecular and Cellular Bioscience |
| Recipient: |
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| Initial Amendment Date: | September 3, 2020 |
| Latest Amendment Date: | October 14, 2020 |
| Award Number: | 2036768 |
| Award Instrument: | Standard Grant |
| Program Manager: |
David Rockcliffe
drockcli@nsf.gov (703)292-7123 MCB Div Of Molecular and Cellular Bioscience BIO Direct For Biological Sciences |
| Start Date: | September 1, 2020 |
| End Date: | August 31, 2022 (Estimated) |
| Total Intended Award Amount: | $499,999.00 |
| Total Awarded Amount to Date: | $499,999.00 |
| Funds Obligated to Date: |
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| History of Investigator: |
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| Recipient Sponsored Research Office: |
110 8TH ST TROY NY US 12180-3590 (518)276-6000 |
| Sponsor Congressional District: |
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| Primary Place of Performance: |
NY US 12180-3522 |
| Primary Place of Performance Congressional District: |
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| Unique Entity Identifier (UEI): |
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| Parent UEI: |
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| NSF Program(s): |
NSF 2026 Fund, FM-Future Manufacturing |
| Primary Program Source: |
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| Program Reference Code(s): |
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| Program Element Code(s): |
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| Award Agency Code: | 4900 |
| Fund Agency Code: | 4900 |
| Assistance Listing Number(s): | 47.074 |
ABSTRACT
The sustainability of mankind?s endeavors will require future manufacturing of materials with closed-loop lifecycles, rather than the traditional linear extract-process-consumer-dispose paradigm. In this project, investigators will develop new technologies and conceptual frameworks towards enabling a regenerative circular bioeconomy for the plastics industry. The project proposes to develop new engineered microbes that will be able to grow on a plastic produced from a petrochemical feedstock, and produce protein polymers that have properties that are appropriate for use in a wide range of consumer products. The investigators at Rensselear Polytechnic Institute will partner with Conagen, Inc. on this project. The project will generate new biotechnologies and elucidate fundamental concepts that enable upcycling of nonbiodegradable plastic waste as well as design and synthesize protein-based polymers as a sustainable solution to petroleum-based plastics. Furthermore, the investigators will establish a "Biomanufacturing of Sustainable Materials" certificate program for undergraduate students as part of the Arch Program at Rensselaer Polytechnic Institute (RPI), which will provide hands-on training in the multidisciplinary skillsets required to engage in future manufacturing for a circular economy. This training program will also connect students with off-campus work experiences which will further provide career opportunities and workforce development.
With support from 1) the Future Manufacturing Program in the Division of Molecular and Cellular Biosciences, and 2) the NSF 2026 Fund Program in the NSF Office of Integrated Activities, the team of researchers from RPI will bioengineer bacterial strains that upcycle petrochemical plastic waste into de novo designed recombinant silk fibroins (RSFs), a class of structural proteins that can potentially exhibit material properties resembling a wide range of commonly used plastics. In parallel, using experimental and computational approaches, the investigators will examine the relationships between molecular/supramolecular structure, processing methods and conditions, and thermomechanical properties in these RSFs, with particular emphasis on identifying primary sequences that are suitable for melt processing into common thermoplastic products. With their industry partner, Conagen Inc., they will explore RSF bioproduction with the engineered microbes at pilot scales to provide a key link between lab-scale research efforts and commercial-scale biomanufacturing. These combined efforts represent a multidisciplinary approach towards solving grand standing challenges in the biomanufacturing of protein-based polymers for use as sustainable plastics in a circular economy. This project addresses priorities of the Future Manufacturing Program. The project also directly addresses several of the NSF2026 Idea Machine winning entries: "a world without waste", "designing ecosystems for the future", and "repurposing, recycling, renewable energy".
This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
PUBLICATIONS PRODUCED AS A RESULT OF THIS RESEARCH
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PROJECT OUTCOMES REPORT
Disclaimer
This Project Outcomes Report for the General Public is displayed verbatim as submitted by the Principal Investigator (PI) for this award. Any opinions, findings, and conclusions or recommendations expressed in this Report are those of the PI and do not necessarily reflect the views of the National Science Foundation; NSF has not approved or endorsed its content.
For the health of Earth's natural environment and the sustainability of mankind's endeavors, future manufacturing will need to shift away from a linear extract-process-consume-dispose paradigm to a regenerative circular economy, where the aim is to keep materials and products at their highest utility and value at all times. The benefits of a truly closed loop lifecycle include conservation of finite resources and prevention of waste accumulation on land and in water. A manufacturing workflow in which petroleum-based plastic waste is upcycled into renewable protein-based polymers by genetically engineered microbes can play a key role in enabling a circular economy for the plastics industry.
The goal of this Future Manufacturing Seed Grant project was to explore the synthesis of renewable protein-based biomaterials using engineered microbes. Specifically, this project aimed to develop bacteria that can produce bespoke silk-mimetic proteins while metabolizing molecules derived from waste plastic as a nutrient source. These bacterial strains can effectively valorize waste plastic, which end up as recalcitrant environmental pollutants in our current linear plastics economy. Moreover, this project aimed to study the bottlenecks in cellular machinery that negatively impact production yield for recombinant silk proteins and to develop strategies for substantially increasing production yield. Lastly, this project also aimed to develop computational tools to enable rational design of silk-mimetic biomaterials with targeted properties. These in silico tools can facilitate rapid development of renewable biobased materials with optimized properties for commercial applications.
Significant project outcomes include:
1) The first proof-of-concept demonstration of producing genetically encoded recombinant proteins using engineered Pseudomonas bacteria that metabolize plastic-derived hydrocarbons. This engineered bacterial strain is able to convert these hydrocarbons, which can potentially be obtained from waste plastic through pyrolysis or chemical degradation, into high value protein products, such as protein fluorophores or artificial silk.
2) The discovery of cellular toxicity and metabolic burden as a reason for low production yields for silk-mimetic proteins in bacterial hosts. The production of disordered proteins, such as silk proteins, in bacteria negatively impacts cell growth and viability and is thus a potentially limiting factor for achieving high yields in these hosts. Changing the recombinant protein sequence to decreased disorder was shown to be a viable strategy for increasing production yield.
3) The development of a novel Escherichia coli strain optimized for synthesizing conventionally "difficult" to produce protein-based polymers, such as recombinant silk, in high yields.
4) The development of engineered Bacillus bacteria that produce and extracellularly secrete recombinant silk protein, which substantially simplifies purification processes for the artificial silk product.
5) The development of computational simulations that can predict the tensile properties of silk-mimetic polymers based on their known chemical structure.
Additionally, this project supported the development of an educational training program for the next generation of sustainability-focused scientists and engineers. The Biomanufacturing of Sustainable Plastics summer training program was offered in 2021 and 2022 to undergraduate students at Rensselaer Polytechnic Institute. This training program provided students with a unique perspective and skillset to pursue future opportunities in the field of sustainable materials. Students gained a conceptual understanding of sustainable plastics through lecture, discussion sessions, and readings. Additionally, students received hands-on training in research techniques relevant to biomanufacturing and characterizing sustainable plastics. Students in the program were further given the opportunity to participate in an undergraduate research project associated with the training program. This project led to the creation of a searchable property database of commercially available biobased plastics, which can help manufacturers and designers with selecting appropriate renewable plastics for their consumer products. This database is publicly available free of charge at https://sustainablebioplas.wixsite.com/sustainability
Last Modified: 12/30/2022
Modified by: Runye H Zha
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