Source: EXOPOLYMER, INC. submitted to NRP
PRODUCTION AND PERFORMANCE ANALYSIS OF NEXT GENERATION HYDROCOLLOIDS FOR THE HEALTHCARE, PERSONAL CARE, AND FOOD MARKETS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1022817
Grant No.
2020-33610-31998
Cumulative Award Amt.
$99,735.00
Proposal No.
2020-00544
Multistate No.
(N/A)
Project Start Date
Sep 1, 2020
Project End Date
Apr 30, 2021
Grant Year
2020
Program Code
[8.8]- Biofuels and Biobased Products
Recipient Organization
EXOPOLYMER, INC.
737 INDUSTRIAL ROAD
SAN CARLOS,CA 940703310
Performing Department
(N/A)
Non Technical Summary
The development of fermentation derived hydrocolloid ingredients began in the U.S. with the discovery of xanthan gum in the 1960s by researchers at the USDA. In recent decades, however, production of this important ingredient has largely gone offshore, as has the production of plant-based products such as guar gum and carrageenan. ExoPolymer intends to produce an entirely new portfolio of high-performance hydrocolloids that meet current and growing needs in several markets. Our technology and its use of corn-derived sugars will allow competitive manufacturing of these ingredients to return to the U.S. and open new opportunities for U.S. agricultural products and production in rural communities.Hydrocolloids are widely used ingredients found in a diverse range of products and processes. Despite their important role, there has been little development of new products over the last fifty years. Existing products, such as alginate, xanthan gum and guar gum, have limited capabilities and cannot provide needed functionality to a growing number of established and emerging markets. Examples of these performance deficits include efficacy in drug delivery and overall quality and stability of packaged food products. Businesses in the healthcare, personal care, food, and energy markets need new ingredients with performances that are not available today. During this project, we will measure multiple parameters to identify scalable fermentation, recovery, and purification strategies for different hydrocolloid products. We will also measure the novel performance properties of our products to determine which advantages they have over existing products and identify the optimal path to commercialization. This will allow us to target first generation products to specific markets and ultimately understand which properties have the greatest potential to expand current and emerging markets for hydrocolloids.ExoPolymer's technology will have broad impact in several areas. Economically, new hydrocolloids that are produced domestically will add considerable value to the U.S. agriculture industry, since new, high performance products will generate a significant financial return. The use of domestic corn as the feedstock of choice for hydrocolloid production will decrease existing import and geopolitical complexities, since industrial grade hydrocolloid products are made mostly in China and India. ExoPolymer's technology will also positively affect personal care and food market growth, which is currently being driven by consumer trends such as clean label, environmental awareness, non-GMO products, and alternative protein meat and dairy products. Final products will benefit from the unique and differential properties of ExoPolymer's natural hydrocolloid ingredients. By providing an entirely new portfolio of ingredients with novel and needed performances, ExoPolymer's hydrocolloids will enable success and expansion in several large and important markets while benefiting U.S. agricultural and rural communities.
Animal Health Component
50%
Research Effort Categories
Basic
0%
Applied
50%
Developmental
50%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
50340102020100%
Goals / Objectives
This Phase I SBIR research project has two major goals. First, working with the USDA ARS in Peoria, IL, we will explore the rheological performance of our base products, to identify specific market applications with the highest commercial potential. Second, we will define the specific fermentation conditions that allow for the optimized production of our novel hydrocolloids.Hydrocolloid ingredients are used broadly in a range of industries, but there are several unmet needs across healthcare, personal care, food, and energy markets. Incumbent hydrocolloid products, such as xanthan gum or guar gum, are effective, but were approved for use in consumer products over fifty years ago. There has been little innovation with regard to these ingredients in the past several decades. Current market trends, such as consumer demand for clean label or the shift towards alternative protein meat and dairy products, require new products with performance capabilities that are not available today. The purpose of this work is to generate foundational data on new, valuable hydrocolloid ingredients that are derived from domestic agricultural products such as corn. Milestones will include establishing robust fermentation, recovery, and purification protocols, and gathering precise rheological performance data of our products versus incumbents. This information will result directly in our ability to produce new ingredients cost effectively, and to strategically enter markets based on novel and differentiated performance of our products. GoalsIdentify market opportunities based on precise rheological performance measurement of ExoPolymer hydrocolloidsDefine scalable and precise fermentation, recovery, and purification methods and processes to optimize production of ExoPolymer hydrocolloidsObjectivesGenerate basic rheological performance metrics for ExoPolymer base hydrocolloidsCompare basic rheological behavior (shear thinning, pseudoplasticity, gel formation) of ExoPolymer hydrocolloid products to incumbent products (Xanthan, Gellan, Guar)Compare rheological behavior of ExoPolymer hydrocolloids to incumbent products at varying pH , temperature, and osmolarityExamine the ability to chemically modify and functionalize ExoPolymer hydrocolloids (in collaboration with USDA ARS Peoria)Perform structural characterization of modified hydrocolloids versus unmodified natural products (in collaboration with USDA ARS Peoria)Develop procedure for fermentation process that is representative of large scaleSurvey carbon sources to identify a sustainable, cost-effective feed stockExamine the relationship between productivity and nutrient availability (carbon, nitrogen, phosphate, etc.)Determine productivity benefits or differences between batch and fed-batch fermentation protocolsDefine precise bioreactor parameters (flow rate, agitation, etc.) for optimized, scalable, fermentation processIdentify scalable methods for product recoveryIdentify scalable methods for product purification
Project Methods
EffortsExoPolymer's microbial strains display unique properties that make them particularly well suited for large scale fermentation and production of novel hydrocolloids. To maximize these capabilities, and to ultimately enable the commercialization of our technology, we will establish a robust, scalable methodology for fermentation, recovery, and purification of our novel products. Efforts with regard to fermentation will include identifying the optimal seed train for bioreactor inoculation. Specifically, this will include the composition of growth medium and timing of inoculation for large scale production. Other specific efforts will include the determination of the optimal components in production medium, such as carbon and nitrogen sources, along with their concentrations and feeding strategies. We will also determine optimal fermentation parameters such as agitation rate, air flow/sparge rate, pH, and duration of production. For product recovery, efforts will include observation and determination of the effects of bulk recovery versus cell separation, and how product should be treated prior to purification. We will observe the effects of temperature, centrifugation, protein removal, ionic strength, and pH on recovery to determine the best strategy for product quality and market application. For the final purification step, efforts will include determining methods and steps for drying and milling. ExoPolymer's technology is based on the unique rheological performance of novel hydrocolloids, and one of the technical objectives of this project is to specifically map performance to potential market application. Our efforts to understand rheological benefits will include the study of how our hydrocolloids behave in comparison to incumbent products such as Xanthan gum or Gellan gum. Specific efforts will include measuring fundamental rheology on a TA Instruments DHR3 rheometer. We will observe the performance of our products in different formulations. Formulation variables will include types of buffers, ionic strength, pH, and salt concentration. Other variables will include temperature, humidity, and hydrocolloid concentration. We will determine the differences in rheological behavior, specifically the effect on shear stress and strain, of ExoPolymer hydrocolloids in comparison to the incumbent products listed above.EvaluationMilestone - A defined, scalable process for fermentation, recovery, and purificationDetermination of optimal growth conditions / timing of seed train (media composition, temperature, pH, inoculation concentration)Determination of optimal growth conditions / timing of fermentation (media composition, temperature, pH, total fermentation time)Precise definition of scalable fermentation conditions (aeration rate, agitation rate, titer, yield, productivity)Precise definition of scalable recovery method for individual hydrocolloid products (centrifugation, filtration, bulk recovery, treatment options)Determination of scalable purification conditions for individual hydrocolloid products (drying methods, temperature, duration, milling)Milestone - Rheological performance and mapping behavior to market opportunityDetermination of how formulation affects rheology of individual hydrocolloid products (salt concentration, buffer type, pH, ionic strength, shear stress and strain)Determination of how temperature, humidity, and concentration affect rheology of individual hydrocolloid products (shear stress and strain measurements in response to high and low temperatures, humidity, and amount of product in formulation)Generation of a dataset of how individual ExoPolymer hydrocolloids behave in comparison to incumbent productsIdentification of the optimal performance-based market opportunities for initial ExoPolymer products

Progress 09/01/20 to 04/30/21

Outputs
Target Audience:Target audiences for this effort include manufacturers of personal care, healthcare, and food products, where hydrocolloids are key ingredients that provide stability, viscosity, and other performance properties. Audiences can also include end consumers of products in these market spaces. Specifically, results from Phase I target consumers of personal care and cosmetic products with anti-aging benefits. Other interested communities are the companies and government laboratories who may become our partners. Our current outreach and partnership with government laboratories is providing new opportunities and pushing back frontiers in their corresponding work. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During the Phase I period, ExoPolymer employees received training and developed proficiency in rheology of biopolymer products, using a DHR3 rheometer from TA instruments. Rheology is a core competency for ExoPolymer and fulfilling the objectives of this project allowed for professional development in this field. How have the results been disseminated to communities of interest?Based on findings in Phase I, ExoPolymer has approached a global cosmetics company to gauge interest in our first product. The superior performance of our products generated a high level of enthusiasm among the strategic development team at the company, and we are now under NDA and in the process of generating samples to provide to them for internal testing. We are also in advanced discussions with a global food ingredient company. Further, we are speaking with several other interested strategic partners who produce polymers for the personal care, healthcare, and food markets. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Biopolymers - hydrocolloids, polysaccharides, or more generally, carbohydrates - are essential components of modern manufacturing and formulation. Uses range from personal care, pharmaceutical compounding, and food stabilization, to lubrication in devices, machines, and energy drilling. The microbial biopolymer market is dominated by two major products, xanthan gum and gellan gum. Xanthan gum, produced by fermentation of Xanthomonas campestris, was approved for use by the WHO in the early 1970s, and gellan gum, produced by fermentation of Sphingomonas elodea, was approved in 1990. ExoPolymer has evolved an agriculture-based approach, using natural microbes, that allows innovation in this commercial and scientific space. The biopolymer industry has lagged behind other industrial biotechnology areas that have benefitted from recent advances in synthetic biology. Moreover, the array of products that are currently available - xanthan gum, guar gum, carrageenan, alginate, and others - cannot meet growing, often novel performance demands in the personal care, food, healthcare, and energy industries. The time is right to apply modern genetic and molecular tools to the biopolymer industry, and ExoPolymer intends to be the first mover in this space. With early revenue from entry into the personal care market, we intend to aggressively pursue additional applications for our technology and products and to become the world leader in biopolymer design, biosynthesis, and manufacturing. ExoPolymer successfully satisfied the technical objectives of Phase I of this project. We developed commercially relevant growth conditions for the production and purification of novel biopolymers and characterized the suitability of these molecules for commercial applications. Rheological and performance capability data from Phase I resulted in the identification of three unique biopolymers with improved properties in comparison to hyaluronic acid (HA). HA is a high-value biopolymer used in a range of commercial applications including personal care and healthcare. HA is used in personal care applications for its unique ability to penetrate the skin, bind water, and reduce the appearance of wrinkles. However, it is derived from either animals or pathogenic bacteria, and there are few alternative products with performance that is similar to HA. Biopolymers identified in Phase I showed up to 3 times the water binding capacity of HA and are therefore ideal targets for commercialization in the personal care market. Based on the findings in Phase I, ExoPolymer intends to bring to market several alternative biopolymers that are natural, non-animal, non-GMO, derived from non-pathogenic microorganisms using plant-based feedstock, and most importantly, with improved performance over HA. Critical to Phase I success was the development of fermentation conditions that resulted in high yields and recovery of our biopolymers. The ability to produce target molecules at scale is a gating feature for success in industrial biotechnology. ExoPolymer thus conducted experiments to optimize growth and production on commercially relevant feedstock, and successfully scaled our process to 5-liter bioreactors. In pilot experiments, we showed that renewable corn feedstock is an ideal and cost-effective input material for strain growth and biopolymer production. We also examined rheological behavior of biopolymers to identify unique features and identify suitable market opportunities. In a partnership with a USDA research facility, we further demonstrated that our biopolymers can be derivatized to provide new functionalities and downstream applications. Major accomplishments during Phase I include: (1) Identification of optimized growth and production conditions for base strains on commercial feedstock, (2) Determination of media formulations for fermentation - identification of specific media components including nitrogen, carbon, and phosphate as well as concentrations for optimal production at bench scale, (3) Strain construction for improved productivity - identification of a set of optimal genotypes for high productivity strains, (4) Successful scale up to 2-liter flask and 5-liter fed-batch fermentation with high productivity and yield, (5) Successful baseline measurements of rheological behavior of novel biopolymers, (6) Performance characterization of biopolymers - emulsion stability and other benefits, (7) Identification of improved moisture binding capacity of novel biopolymers in comparison to hyaluronic acid (HA), and (8) Successful functionalization of base biopolymers for improved performance.

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