50%.Lygos technology and business directly supports government wide and USDA SBIR program priorities and societal challenge areas. Benefits established from Lygos processes for production of renewable chemicals include increasing product diversification from US agricultural feedstocks, increasing energy efficiency of manufacturing processes, decreasing carbon dioxide emissions, and increasing US energy independence. This small business innovation research phase I project will support Lygos development of a biocatalyst for conversion of sugar feedstocks into fine and commodity chemicals to expand their existing markets. With regard to societal impact, Lygos microbial processes are based on renewable feedstocks and eliminate use of environmentally hazardous materials.' />
Source: Lygos Inc. submitted to NRP
PHOSPHATASE ENGINEERING FOR MICROBIAL PRODUCTION OF A HIGH-VALUE ORGANIC ACID
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1015996
Grant No.
2018-33610-28258
Cumulative Award Amt.
$100,000.00
Proposal No.
2018-00477
Multistate No.
(N/A)
Project Start Date
Aug 1, 2018
Project End Date
Mar 31, 2020
Grant Year
2018
Program Code
[8.8]- Biofuels and Biobased Products
Recipient Organization
Lygos Inc.
636 San Pablo Ave
Albany,CA 94706
Performing Department
(N/A)
Non Technical Summary
Lygos is developing novel bio-manufacturing processes to valuable chemicals. Our targeted chemicals have existing, large markets of >$250MM per year. Current petroleum-based production of these chemicals has a number of pitfalls. Current chemical manufacturing requires petroleum feedstocks, utilize environmentally damaging processes, achieve low-yields, and have high production costs. In contrast, Lygos' process converts domestic, renewable, agricultural sugars, including corn, corn-stover, woody-biomass, and other energy crops into chemicals using fermentation; our process is high-yielding, is performed at ambient temperatures, and could decrease production costs by >50%.Lygos technology and business directly supports government wide and USDA SBIR program priorities and societal challenge areas. Benefits established from Lygos processes for production of renewable chemicals include increasing product diversification from US agricultural feedstocks, increasing energy efficiency of manufacturing processes, decreasing carbon dioxide emissions, and increasing US energy independence. This small business innovation research phase I project will support Lygos development of a biocatalyst for conversion of sugar feedstocks into fine and commodity chemicals to expand their existing markets. With regard to societal impact, Lygos microbial processes are based on renewable feedstocks and eliminate use of environmentally hazardous materials.
Animal Health Component
75%
Research Effort Categories
Basic
5%
Applied
75%
Developmental
20%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
51140991060100%
Goals / Objectives
Lygos is targeting compounds that are currently derived from petroleum using expensive, inefficient and ecologically hazardous chemical processes which has hampered their market expansion. Lygos is developing robust high-yielding microbial catalysts that will decrease production costs for these select compounds. The goal of this Phase I SBIR proposal is to engineer a new class of enzymes catalyzing specific reactions for conversion of sugar feedstocks into high-value organic acids. Lygos' microbial processes are designed to be compatible with a range of domestic agricultural inputs, including traditional, corn-derived sugars as well as sugars derived from non-food biomass and waste agricultural and forestry materials. Thus, if successful, the proposed technology will provide new manufacturing routes to domestically produce high-value chemicals, providing an additional revenue source to relevant agricultural stakeholders.
Project Methods
Lygos employs state-of-the-art synthetic biology techniques to rapidly prototype and optimize metabolic pathways. These methods include development of product screening tools, rapid biocatalyst construction, protein engineering tools, and identification and deployment of novel metabolic pathways in industrial micro-organisms. The general workflows and methods Lygos is developing for synthetic biology can drastically decrease the cost and time required for commercialization of biocatalysts. The scientific results from this effort will be a greater understanding of how to engineer a new class of enzymes.

Progress 08/01/18 to 03/31/19

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Lygos developed an alternate methodology for screening 3-PG phosphatase activity in vivo in engineered yeast strains and identified active 3-PG phosphatase enzymes. The best strains produced a yield of glyceric acid approaching 20%, but the glyceric acid production rate is slow, likely due to limiting 3-PG phosphatase activity, and must be still improved. Thus, we devised more sophisticated computational modeling approaches to generate rational and semi-rational libraries of variants of the best 3-PG phosphatases.We also devised a selection strategy for improved 3-PG phosphatase activity, which should allow exploration of a very large sequence space and allow discovery of mutations improving the catalytic rate of the phosphatase that may be difficult to predict rationally. Lygos strain engineering capabilities allow for the generation of libraries of tens of thousands of pooled transformants and, coupled with conventional PCR-based mutagenesis methods, are ideally suited to generate the initial diversity for a selection. In absence of a convenient system for in vitro characterization of our engineered enzymes, we are confident that the alternate approaches developed here will produce 3-PG phosphatases fulfilling the requirements for industrial production of glycolic acid.

Publications