DBIA Research Program

This project addresses the challenge of increased acid whey (AW) production driven by the rising demand for Greek yogurt and acid-coagulated cheese. This collaborative endeavor involves Galasys (Madison, WI), a startup company, and research groups led by Professor George Huber and Professor Scott Rankin at UW-Madison.

Despite various attempts, the food industry has yet to find a comprehensive solution for AW disposal beyond land application and animal feed. To confront this issue, our team has pioneered a groundbreaking process that utilizes acid catalysts to transform Greek yogurt AW into valuable products, such as glucose-galactose syrup (GGS) and milk minerals.

Unfortunately, the GGS currently possesses a brown coloration and contains 5-Hydroxymethylfurfural (HMF), limiting its application in food and beverages. In this project, our team will work on enhancing the quality of GGS by refining the process and incorporating a downstream adsorbent bed.

Our primary objectives encompass achieving high GGS yields from various whey sources on a 100-liter batch scale, improving GGS quality by reducing HMF concentration and coloration, and conducting a techno-economic analysis to evaluate the feasibility of large-scale production based on our pilot plant results. We believe that these innovative efforts have the potential to revolutionize the dairy industry by effectively addressing the AW challenge and creating new opportunities within the sweetener market.

The goal of our project is to sustainably produce bioplastic polymers from lactose and lactate-rich dairy industry residues. Our strategy is a bioconversion approach where we will engineer bacteria to consume components of large-volume dairy industry residue streams including acid whey and permeate. Acid whey and permeate are contain high concentrations of lactose and lactate which the engineered bacteria can convert into biopolymers. These biopolymers can be harvested and manufactured into biodegradable plastics.

Our work on this specific project involves improving the performance of the engineered bacteria and the biopolymer yields on acid whey and permeate. Then later milestones will test initial scalability of the process.

The processing and manufacturing of milk, cheeses, and yogurts generates several coproducts with low economic value (e.g., milk permeates and whey permeates). One promising option to add value to this coproducts is the production of the lactic acid enantiomers, D-lactic acid and L-lactic acid, which could enter the market as green chemicals for the manufacturing of biodegradable plastics such as polylactic acid (PLA).

This project will evaluate the use of genetically engineering Lactobacillus plantarum strains to produce each lactic acid enantiomer with high purity. We will first investigate, in 500-mL bench-scale bioreactors, the yields, rates, and titers of D-lactic acid production when an engineered L. plantarum strain is fed different lactose-rich dairy coproducts.

Based on these experiments we will select a coproduct (or coproduct mixture) that could be provided by an industrial partner, and operational conditions to evaluate performance at the pilot scale (e.g., 400-liter bioreactor). Data from experiments at this scale will help establish mass and energy balances, evaluate lactic acid extraction procedures, and other logistical items that need to be considered when assessing the technoeconomic viability of valorizing dairy coproducts by fermentation.

This project led by Prof. Xuejun Pan is to synthesize prebiotic galacto-oligosaccharide (GOS) from an underutilized dairy industry waste (byproduct), whey permeate. The major component of the whey permeate is lactose (~80 % of the total solids). The annual cheese whey production is around 57 billion pounds in the United States. At present, the protein in whey has been well marketed as the whey protein concentrate (WPC), while lactose in whey permeate is underutilized. Therefore, exploring new and enduring uses of whey permeate (lactose) is of great interest and importance to the dairy industry.

Prebiotic GOS has the potential to develop beneficial microorganisms in the gut environment, which can result in the reduction of many inflammatory diseases in the intestine. Pan’s Lab has developed and patented a facile synthesis of GOS from lactose with a very high yield (> 90 %) under mild conditions (70 °C and 20 min). The synthesized GOS has demonstrated prebiotic potential both in in vitro and in vivo conditions. In this project, we will scale up the process using the facilities and equipment available at the Center of Dairy Research (CDR). Specifically, we will establish process mass balance, and conduct a preliminary techno-economic analysis (TEA) to evaluate the economic feasibility of the technology. We will develop potential prebiotic applications of the synthesized GOS in dairy products and animal feeds.

The success of the project would be beneficial to the dairy industry by valorizing waste (lactose) into high-value prebiotic products. Further, supplementing the synthesized GOS into dairy products (yogurts, fermented milk, ice creams, and cheeses) and animal feeds can potentially increase health benefits in both humans and animals.

Oleochemicals are a class of important commodity chemicals used as fuels, solvents, lubricants, and as part of specialty materials. Oleochemicals are currently derived from either unsustainable plant oils or low-yielding chemical syntheses. The Pfleger lab has developed a family of engineered microorganisms capable of converting sugars and organic acids to model oleochemical products. In order to be economically viable, processes using these organisms must be based on low-cost feedstocks because oleochemical prices are relatively low. D

airy wastes such as acid whey and milk permeate contain sugars and amino acids that are readily used by microorganisms and could prove to be an attractive feedstock for oleochemical synthesis. This project will demonstrate conversion of model dairy wastes to model oleochemicals at laboratory scale, conduct technoeconomic analysis of the process, and ultimately demonstrate an optimized process at pilot scale.

Whey permeate is an underutilized dairy residue that holds significant promise as a feedstock for large scale production of renewable chemicals. Therefore, this project will combine fermentation science and microbial engineering to improve butanol production with whey permeate as feedstock. Butanol is an important industrial chemical used in the production of paint, detergent, plastics and synthetic rubber, among other applications. Additionally, butanol has tremendous promise as a renewable aviation fuel. We unmasked genetic modifications that led to the production of 45% greater butanol by Clostridium beijerinckii with whey permeate as feedstock.

In this project, we will engineer a strain of C. beijerinckii with enhanced capacity to produce butanol in whey permeate, based on the aforementioned genetic modifications. We expect the resulting strain to exhibit enhanced butanol production, potentially reaching 16 – 20 g/L or more. Subsequently, we will scale up the whey permeate-to-butanol fermentation to 80-liter bioreactor.

Upon completion, we expect to demonstrate the feasibility of bio-converting whey permeate to butanol at laboratory (12.5-liters) and pilot scales (80-liters), thereby, establishing a basis for future commercialization efforts.

Research

Conduct research into the conversion of dairy by-products or isolation of value-added components from these coproducts. Research proposals should advance existing capabilities or investigate novel approaches in the conversion of dairy waste streams. Examples could include:

• Enhancing bioconversion rates using micro-organisms (e.g., bio fermentation) or by enzymatic processes
• Developing alternative (or enhanced) separation processes to isolate chemicals of interest generated in these streams

Possible targets could include:

• Organic acids (e.g., lactic acid)
• Fatty acids (e.g., medium chain fatty acids)
• Biodegradable bioplastic biopolymers (e.g., polylactic acid, poly-3-hydroxybutyrate: PHB)

• Replacement of petrochemically derived (fossil fuel) platform chemicals (e.g., 1, 3-propanediol)

• Natural food ingredients or supplements (e.g., colors, flavors, etc.)
• Other options could include bioalcohols and high-value biofuels

Development

Each proposal must contain a development section or portion of the project that would evaluate the viability of bringing the research to market. This can be done in a number of ways, such as:

• Carrying out scale-up trials to demonstrate process viability, efficiency and/or generate product samples for evaluation or mass balance determinations (discuss with CDR their pilot scale-up capabilities for your process)
• Performing a technoeconomic analysis (TEA) that illustrates the economic benefits underpinning the technology proposed in comparison to any existing or alternative processes

A partnership with another institution, as outlined below, may support this section of the project.

• Projects should be comprised of both research and development activities
• Project budget can be up to a maximum of $80,000 per year
• Project proposals for a maximum of 3 years
• No overhead charges (indirects) are permitted as they have already been accounted for in the initial award to the DBIA (from USDA)

Applications may be in support of a Masters or Ph.D. student or postdoc. The student does not need to be identified at the time of submission.

Approved applications will receive funding for the first year up-front. Funding for subsequent years will be dependent upon the approved completion of required interim reports (every 6 months) and a final report. For projects requesting three years of funding, satisfactory progress towards milestones will have to be demonstrated by end of year two.

• Proposals must be submitted by UW-System Faculty & Staff with PI status (limited or permanent)
• Applications may include a co-PI from another academic institution (outside the UW-System) provided:
  • The Institution is from within the 11-state region covered by the DBIA
  • Funding is matched 1:1 by the partner institution if research will be conducted at that institution
• Applications may also involve a partnership between two or more entities, including:
  • Other educational institutions or non-profit organizations
  • Private companies, for-profit businesses and start-ups

It is important for the results of this work to be made available to the dairy industry. Successful applicants will be required to present their work at the CDR industry research forums. Estimated travel costs to Madison should be included in the budget proposal.

Results may be published and presented, with acknowledgements made to the funding provided by the DBIA, using the following language:

Funding for the DBIA was made possible by the U.S. Department of Agriculture’s (USDA) Agricultural Marketing Service through grant AM190100XXXXG079. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the USDA.

Interim reports must be completed every 6 months. Funding for year two (and three if applicable) will be dependent upon the approved completion of required interim reports. For projects requesting three years of funding, satisfactory progress towards the milestones will have to be demonstrated by end of year two. A final report will be requested from all awardees.

All submissions must be received via the DBIA online application system: DBIA Application Portal

Full instructions for registering an account and submitting the application can be found here: Applicant Tutorial

Confirmation e-mails will be sent to all applicants after the submission has been received. All applicants will be notified of their status.

The application includes:

1. Application Details

• Full, descriptive title to convey the importance of the project
• List the main participants and their profiles – PI, co-PIs, students (if known)
• A 250-word, non-technical summary of the project that will be used to explain to a wider audience, including:
  • Importance of the project (current status of the research topic)
  • Major objectives
  • Research basis
  • Development plans

2. Narrative

Formatting requirements: Word or PDF document; 5 page maximum, single spaced, size 12-point, Calibri font, 1-inch margins. Written in clear language that conveys the scientific and commercial details to reviewers familiar with the dairy and food industry. Please include the section headers outlined below in your narrative document (Background and Justification, Methodology and Timeline, Impact, and Future Research Goals).

Background and Justification

What is the scientific merit behind the research underpinning the proposal? This may include references, prior results, etc. What is the current state of this topic and the gap that is proposed to be addressed in the project? What are the alternative approaches currently used?

Do you have supporting evidence for the potential commercial feasibility of the proposal? This may take the form of projected market analyses, support from industry partners (letters will not count towards the 5-page maximum), current technoeconomic analysis, yield calculations, mass balances, etc.

Describe why this DBIA funding is necessary to advance this research area for your program.

Methodology and Timeline

List the key milestones of the proposal for both research and development activities.

Also include:

• Key methods and approaches to be used (recommended to include a potential process flow diagram for the development phase, if relevant)
• How will each milestone be achieved?
• Where will the work take place?
• Proposed scale of bio-conversion process (e.g. size of bio-reactor)
• Challenges likely to be faced and describe how they will be tackled (contingencies)

Impact

Discuss the likely impact on the dairy industry if successful.

Future Research Goals

Provide insight into the potential for further research & development work that may result from the efforts outlined in the proposal.

3. Budget

Please upload the completed budget template (TEMPLATE). No other version of a budget will be accepted.

The budget may include up to $80k per annum, for a maximum of three years, is available (with the stipulations regarding satisfactory progress after year two). You should include the following details on an annual basis (see provided template). You must include the following categories:

• Salary & Fringe
• Travel expenses
• Supplies
• Details of matching funding (if any)

Successful applicants will be required to present their work at the CDR industry research forums. Estimated travel costs to Madison should be included in the budget proposal.

If funding is used for work carried out at a partner’s location (i.e., non-UW system location), please note that no overhead charges as outlined above applies to any research location. Please also note the requirement for 1:1 match for all funds requested for work carried out at that location.

4. Curriculum Vitae
Please upload the CV of each key stakeholder (PI, Co-PI, Student (if known))

5. References

Please upload one document containing references. This should include supporting literature for your proposal. There is no maximum page limit.

Proposals will be reviewed and scored by a review committee made up of the DBIA management team, a representative from the Midwest Dairy Association and a representative from Dairy Management Inc.

Proposals will be scored as follows:

Background and Justification (25%)

• Is the current state of the science in this area adequately described (e.g., current processes, gaps, opportunities, etc.)?
• Is a TEA planned for a new process or an existing one provided?
• Is there an industry partner or entrepreneurial interest? Are there letters of support?
• Was the need for DBIA funding adequately described?

Methodology and Timeline (40%)

• Does the proposal significantly advance the technology?
• Were challenges adequately described and addressed?
• Is the approach novel or incremental?
• Are the development plans feasible and detailed?

Impact and Future Research (25%)

• What is the likelihood to successfully generate viable value-added products?
• If successful, how large an impact would this research make on the dairy industry?

Budget (10%)

• Is the budget adequate for the goals and objectives?