SBIR-STTR Award

Novel Proteolysis-based Tools for Metabolic Engineering
Award last edited on: 3/30/2022

Sponsored Program
SBIR
Awarding Agency
NSF
Total Award Amount
$1,464,964
Award Phase
2
Solicitation Topic Code
BC
Principal Investigator
Jason Kelly

Company Information

Ginkgo Bioworks Inc (AKA: Primus)

27 Drydock Avenue Floor 8
Boston, MA 02210
   (814) 422-5362
   info@ginkgobioworks.com
   www.ginkgobioworks.com
Location: Single
Congr. District: 08
County: Suffolk

Phase I

Contract Number: ----------
Start Date: ----    Completed: ----
Phase I year
2011
Phase I Amount
$150,000
This Small Business Innovation Research (SBIR) Phase I project aims to engineer microbes for the cost-effective production of the amino acid, L-Threonine. Currently, engineered microbes bear mutations that increase the production of Threonine of interest by inhibiting the cell?s ability to produce other amino acids. These mutations are critical as they effectively channel the cell?s metabolic flux toward Threonine, thereby boosting production efficiency and easing downstream purification. Unfortunately, these mutations also decrease cellular fitness and, thus, the growth media must be supplemented with costly nutrients. Technical research herein will assess the feasibility of applying novel regulated proteolysis technology to direct metabolic flux toward Threonine production in the absence of costly media supplementation. The project has 3 key objectives: 1) generate E. coli strains containing off-pathway metabolic enzymes tagged for degradation by a growth-phase dependent proteolysis system, 2) test the ability of these strains to grow on supplement-free media, and 3) assay for production of Threonine by these engineered strains. We anticipate that our engineered strains will grow robustly on minimal, un-supplemented media. Upon induction of our proteolysis system, we expect our strains to specifically eliminate off-target metabolic pathways, leading to a substantial increase in production of our target product, Threonine. This Small Business Innovation Research (SBIR) Phase I project aims to engineer microbes for the cost-effective production of the amino acid, L-Threonine. Currently, engineered microbes bear mutations that increase the production of Threonine of interest by inhibiting the cell?s ability to produce other amino acids. These mutations are critical as they effectively channel the cell?s metabolic flux toward Threonine, thereby boosting production efficiency and easing downstream purification. Unfortunately, these mutations also decrease cellular fitness and, thus, the growth media must be supplemented with costly nutrients. Technical research herein will assess the feasibility of applying novel regulated proteolysis technology to direct metabolic flux toward Threonine production in the absence of costly media supplementation. The project has 3 key objectives: 1) generate E. coli strains containing off-pathway metabolic enzymes tagged for degradation by a growth-phase dependent proteolysis system, 2) test the ability of these strains to grow on supplement-free media, and 3) assay for production of Threonine by these engineered strains. We anticipate that our engineered strains will grow robustly on minimal, un-supplemented media. Upon induction of our proteolysis system, we expect our strains to specifically eliminate off-target metabolic pathways, leading to a substantial increase in production of our target product, Threonine. The broader impact/commercial potential of this project is the generation of more cost-efficient L-Threonine producing microbial strains. Purified amino acids are estimated to constitute a U.S. market of $1.30 billion by 2013. These chemicals are used as animal feedstock supplements, precursors in production of the artificial sweetener aspartame, and have potential as biofuel precursors. Currently, key amino acids are produced commercially using highly engineered microbes that convert low-cost sugar sources (e.g. glucose) to the final amino acid product. To improve the conversion efficiency and ease downstream purification, the microbe?s ability to synthesize other, off-pathway amino acids is often eliminated. However, because these other amino acids are critical for bacterial growth, they must be added as a supplement to the growth medium, substantially increasing production costs. The technology proposed here would allow for efficient, robust production of easily purified amino acids without the need for media supplementation, dramatically reducing production costs. Moreover, the regulated degradation technology developed herein will provide next-generation regulatory tools for other industrial metabolic engineering applications.

Phase II

Contract Number: ----------
Start Date: ----    Completed: ----
Phase II year
2013
(last award dollars: 2016)
Phase II Amount
$1,314,964

This Small Business Innovation Research (SBIR) project aims to engineer microbes for the cost-effective production of specialty chemicals. Currently, engineered microbial strains bear mutations that increase the production of chemicals of interest by inhibiting the cell's ability to produce off pathway chemicals. These "loss-of-function" mutations are critical as they effectively channel the cell's metabolic flux toward the product of interest. This both boosts the production efficiency and eases downstream purification by eliminating the accumulation of undesirable but chemically-similar contaminants. Unfortunately, these mutations may also decrease the fitness of the cells and, as a result, the growth media must be supplemented with costly nutrients. Technical research herein will assess the feasibility of applying novel regulated proteolysis technology to simultaneously direct maximal metabolic flux toward the target chemical of interest while avoiding the need to supplement the growth media. If successful, this technology would provide a great cost savings and enable fermentative production to be applied more broadly in the production of specialty chemicals. The broader impact/commercial potential of this project is to provide a stable and cost-effective fermentative production route to a specialty chemical. Fermentative production of chemicals offers many advantages over traditional petrochemical or extraction-based production processes. Petrochemical production maintains the nation?s reliance on an unsustainable feedstock (oil) and also leads to national security issues as the US is largely dependent on foreign oil sources. Chemical production via extraction from plant materials also has ecological challenges. The process often uses toxic solvents, and may rely on unsustainable farming practices for many plants that are not traditional food crops. Engineered microbes fermented on sugar feedstock produced using high-efficiency agricultural practices offer a stable alternative for producing specialty chemicals, both in terms of supply and price