DEVELOPMENT OF SUSTAINABLE MUSHROOM-BASED SEAFOOD THROUGH FLAVOR CHEMISTRY, PHYSICAL CHEMISTRY, AND SENSORY ANALYSIS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: ACTIVE
• Funding Source: AFRI COMPETITIVE GRANT
• Reporting Frequency: Annual
• Accession No.: 1028041
• Grant No.: 2022-67017-37097
• Cumulative Award Amt.: $589,250.00
• Proposal No.: 2021-09605
• Project Start Date: Aug 1, 2022
• Project End Date: Jul 31, 2026
• Grant Year: 2022
• Program Code: [A1364]- Novel Foods and Innovative Manufacturing Technologies

Recipient Organization
UNIVERSITY OF TENNESSEE
2621 MORGAN CIR
KNOXVILLE,TN 37996-4540

Performing Department
Food Science Research

Non Technical Summary
Technology-based approaches to develop alternative protein sources with great flavor and texture have enormous potential for improving the sustainability and quality of the American diet. In recent years, alternative protein sources have garnered increasing popularity in the food industry. Many different types of alternative plant-based protein offerings are commercially available to consumers (i.e., veggie burgers); however, there are less offerings that are available representing the seafood category. There is substantial interest in the incorporation of mushrooms into non-meat-based food products to help address the 2020-2025 Dietary Guidelines for Americans to lower energy density in the U.S. food supply, to increase nutrient density, and to decrease sodium consumption. The white button mushroom, Agaricus bisporus, is the most widely consumed mushroom in the United States, resulting in an enormous number of mushroom stems as a major agricultural waste stream. This project is aimed at utilizing discarded edible mushroom stems from the commercial mushroom industry as a basis for developing mushroom-based seafood alternatives with appealing seafood flavor and texture. This novel approach will combine the cross-functional disciplines of flavor chemistry, physical chemistry and consumer sensory science to achieve our goals.

Animal Health Component

33%

Research Effort Categories

Basic

67%

Applied

33%

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
502	1470	2000	75%
502	1470	3090	25%

Knowledge Area
502 - New and Improved Food Products;

Subject Of Investigation
1470 - Mushrooms and other edible fungi;

Field Of Science
3090 - Sensory science (human senses); 2000 - Chemistry;

Keywords

flavor chemistry

mushrooms

plant-based meat

seafood

sensory science

texture analysis

Goals / Objectives
Objective 1: Characterize the key odorants and aroma precursors in lobster mushroomsObjective 2:Develop a mushroom-derived seafood flavor in-situby reacting mushroom precursors under kitchen-like thermal conditions.Objective 3: Optimize the texture (seafood-like) of the mushroom-based seafood (mBSF).Objective 4: Comprehensively evaluate the sensory properties of crab cakes alternative using the mushroom-based seafood (mBSF).

Project Methods
Objective 1: Characterize the key odorants and aroma precursors in lobster mushrooms.The key odorants in lobster mushrooms will be comprehensively determined using modern flavor chemistry techniques. This includes solvent assisted flavor extraction distillation (SAFE), gas chromatography-olfactometry (GC-O), aroma extract dilution analysis (AEDA), gas chromatography-mass spectrometry (GC-MS), and stable isotope dilution assays (SIDA).Objective 2: Develop a mushroom-derived seafood flavor in-situ by reacting mushroom precursors under kitchen-like thermal conditions.Free-amino acids in both lobster and button mushroom stems will be quantitated through literature methods. Model reaction systems will then be developed to probe precursor-product relationships. Odorants will be monitored by GC-MS using SIDA.Objective 3: Optimize the texture (seafood-like) of the mushroom-based seafood (mBSF).Small-scale mushroom nanofibers with different morphologies will be prepared using a planetary ball mill. Larger-scale production of mushroom nanofibers will be conducted by a super mass collider. The nanofibers will be characterized by laser diffraction, atomic force microscopy, and the crystalline index will be determined by x-ray diffraction. Texture optimization will be guided through the analysis of mechanical properties of the samples that correlate well with the texture properties of hardness, springiness, cohesiveness, and chewiness.Objective 4: Comprehensively evaluate the sensory properties of crab cakes alternative using the mushroom-based seafood (mBSF).Standard sensory methods including descriptive sensory evaluation and consumer sensory evaluation will be used to guide recipe formulation (both flavor and texture). Attributes evaluated will include appearance, aroma, flavor, and relevant textural properties such as firmness, moisture release, and cohesiveness. An affective sensory test will be performed, and consumers will be asked for their opinion on the overall liking, texture, appearance, flavor, and aroma of the products, using three types of scaling (hedonic, intensity, and just-about-right).

Progress 08/01/24 to 07/31/25

Outputs
Target Audience:The targeted audiences for the project titled, "Development of sustainable mushroom-based seafood through flavor chemistry, physical chemistry, and sensory analysis", are 1) professionals from industry and academia, and 2) graduate students with interests in food science, flavor chemistry, natural products chemistry, and mycology. During the reporting period, two graduate students were recruited. The two graduate students are currently working on the project. In addition, two presentations were given at the American Chemical Society (ACS) Spring 2025 national meeting titled "Characterization of Odorants in Thermally Treated Lobster Mushrooms" and "Utilization of button mushroom stem waste in seafood analogs", one invited lecture titled "Characterization of odorants in dried lobster mushrooms" was given at Sonoma County Mycological Association Scholarship meeting 2025, all reaching the targeted audience: 1) professionals; both industry and academic, and 2) graduate students. Furthermore, 2 graduate students were supervised under the project and conducted the research, reaching the target audience (graduate students). Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During the reporting period, the project has provided training and professional development for two graduate students. The students gave 2 conference presentations at the ACS Spring 2025 meeting. Additionally, the students also submitted one abstract for presentation at the ACS Fall 2025 meeting. Two manuscripts on the aroma chemistry portion were submitted to ACS Journal of Agricultural of Food Chemistry. In addition, the students conducted and learned new aroma chemistry techniques including solvent-assisted flavor evaporation (SAFE), aroma extract dilution analysis (AEDA), gas chromatography - olfactometry (GC-O), gas chromatography - mass spectrometry (GC-MS), solid phase extraction (SPE), stable isotope dilution analysis (SIDA), quantitative olfactory-profile analysis. In addition, the students conducted and learned new mycology techniques including wild mushrooms collection and identification, fungi culturing and sequencing. The students were provided with training opportunities by participating in 11 wild mushroom foray events. Furthermore, the students are active members of the Asheville Mushroom Club (AMC), frequently participate in the AMC wild mushroom collection and identification events, as well as its wild mushroom identification training workshops. The fieldwork and extension services resulted in one graduate student receiving a scholarship, The Oregon Mycology Society Scholarship 2025. How have the results been disseminated to communities of interest?During the reporting period, three presentations were given, two conference presentations at the ACS Spring 2025 meeting, and one invited lecture at the Sonoma County Mycological Association Scholarship meeting 2025. In addition, 1 abstract was submitted and accepted to be presented at ACS Fall 2025. The conference presentations at ACS meetings and the invited lecture at Sonoma County Mycological Association Scholarship meeting reached both targeted audience 1 (professionals; both industry and academic) and targeted audience 2 (graduate students). In addition, two manuscripts were submitted to ACS Journal of Agricultural and Food Chemistry, reaching both targeted audience 1 (professionals; both industry and academic) and targeted audience 2 (graduate students). What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period, we plan on submitting a fourth manuscript covering the sensory consumer study of the mushroom-based crab cake model food product, publish a PhD dissertation, and award a PhD degree to the student that worked on this project. In addition, we will give 2 presentations on the aroma chemistry and sensory findings. For the sensory portion, we will conduct consumer study to evaluate the mushroom-based crab cake model food product.

Impacts
What was accomplished under these goals? Developing healthy and sustainable food that also tastes great is crucial in motivating people to adopt a nutritious and improved diets in accordance with 2020-2025 Diet Guidelines for Americans. Mushrooms can offer a promising solution in reducing energy density in the US food supply, increasing nutrient density, and lowering sodium consumption. In recent years, the plant-based meat industry has seen tremendous economic growth due to advances in food processing technology. However, seafood made from plant-based sources is still a challenge, as its flavor and texture are much harder to replicate than beef, poultry, and pork. Innovative approaches in food chemistry, including texture and in-process modification, can help develop seafood flavors, offering the opportunity to design novel mushroom-based seafood (mBSF) with enhanced flavor and texture. These technologies can meet consumer expectations for alternative seafood in a sustainable and environmentally friendly way. During this reporting period, the main chemistry focus has been the identification of the aroma-active compounds present in the rehydrated and cooked mushrooms, quantitation of the aroma-active compounds in the rehydrated and cooked mushrooms, preparation of two aroma simulation models that mimic the aroma of the rehydrated and cooked mushrooms, and sensory analysis to compare the models with the rehydrated and cooked mushrooms. The aroma chemistry research workstream resulted in one presentation at the ACS Spring 2025 Meeting and one abstract submitted and accepted to be presented at the ACS Fall 2025 Meeting. In addition, preliminary works have been done on optimizing the texture of mBSF, with a presentation given at ACS Spring 2025 Meeting. Furthermore, two manuscripts were summitted to the ACS Journal of Agricultural and Food Chemistry, one focusing on the aroma chemistry of rehydrated lobster mushrooms and the other on the cooked mushrooms. The detailed accomplishments on the objectives are as follows: Objective 1: Obtaining a reliable source of wild lobster mushrooms and aroma chemistry characterization was the primary focus of the current reporting period. Scientific research and collection permits were obtained from Tennessee Department of Environment and Conservation (TDEC) and Tennessee Wildlife Resources Agency (TWRA). Approximately 11 field trips were conducted to locate wild lobster mushroom locations. In addition, we successfully cultured the Hypomyces lactifluorum from wild lobster mushrooms collected from another location to serve as a source of material for biosynthetic pathway elucidation. The species was confirmed by ITS sequencing. This adds to the collection of totals 5 cultures of H. lactifluorum The aroma chemistry research workstream resulted in the identification of 35 aroma-active molecules in rehydrated lobster mushrooms and 48 in cooked lobster mushrooms. Stable isotope standards have been acquired, and the quantitative work is completed for cooked lobster mushrooms. Two aroma simulation model was prepared and sensorily matched with the aroma of rehydrated and cooked mushrooms, respectively. Two manuscripts were submitted to ACS Journal of Agricultural and Food Chemistry, titled "Characterization of Important Odorants in Rehydrated Lobster Mushrooms" and "Characterization of Important Odorants in Thermally Treated Lobster Mushrooms". In addition, one talk was given at the ACS Spring 2025 meeting, titled "Characterization of Odorants in Thermally Treated Lobster Mushrooms". One abstract was submitted and accepted to be presented at ACS Fall 2025 meeting, titled "Quantitation of Odorants in Thermally Treated Lobster Mushrooms". Objective 2: During this reporting period, part of the mushroom-derived seafood flavor development portion of the project was completed. The button mushroom stems were collected, freeze-dried and pulverized. A seafood analogue is developed and optimized by combining mushroom stem powder with wheat gluten. The mushroom-derived seafood flavor was developed by optimizing the cooked lobster mushrooms aroma simulation model and adjusting the application rate. In addition, the mushrooms stem powder was deodorized and recombined with undeodorized powder at an optimized ratio to reduce undesired aroma. Objective 3: During this reporting period, the mushroom-based seafood analogue texture portion of the project was completed. The button mushroom stem powder was combined with wheat gluten at different optimized ratio, determined by instrument in the previous reporting period, to create the seafood analogue. Sensory evaluation was performed on the seafood analogue to further optimize the texture by adjusting the ratio of mushroom powder and wheat gluten. The result was featured in 1 presentation at the ACS Spring 2025 meeting titled "Utilization of button mushroom stem waste in seafood analogs". Objective 4: During this reporting period, the model food product, mushroom-based crab cake, was developed by combining the mushroom-based seafood analogue with the optimized lobster mushroom flavor and other ingredients from a standard crab cake recipe. Several parameters were assessed and optimized: the amount of seafood analogue, the amount of flavor, the amount of other ingredients, and the cooking and serving procedure. The experiment for consumer sensory study was designed. The IRB for descriptive panel was obtained and the IRB for consumer study was submitted. Two graduate students participating in the project completed the required training for the IRB project. This workstream is still in progress.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2025 Citation: Ethan Darby, Thien Nguyen, and John P. Munafo. 2025. Utilization of button mushroom stem waste in seafood analogs. American Chemical Society. Spring 2025
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2025 Citation: Thien Nguyen and John P. Munafo. 2025. Quantitation of Odorants in Thermally Treated Lobster Mushrooms. American Chemical Society. Fall 2025
• Type: Peer Reviewed Journal Articles Status: Under Review Year Published: 2025 Citation: Nguyen, T.H.; Darby, E.W.; and Munafo Jr, J.P., 2025. Characterization of Important Odorants in Rehydrated Lobster Mushrooms. Journal of Agricultural and Food Chemistry.
• Type: Peer Reviewed Journal Articles Status: Under Review Year Published: 2025 Citation: Nguyen, T.H.; Darby, E.W.; and Munafo Jr, J.P., 2025. Characterization of Important Odorants in Thermally Treated Lobster Mushrooms. Journal of Agricultural and Food Chemistry.
• Type: Conference Papers and Presentations Status: Published Year Published: 2025 Citation: Thien Nguyen and John P. Munafo. 2025. Characterization of Odorants in Thermally Treated Lobster Mushrooms. American Chemical Society. Spring 2025

Progress 08/01/23 to 07/31/24

Outputs
Target Audience:The targeted audiences for the project titled, "Development of sustainable mushroom-based seafood through flavor chemistry, physical chemistry, and sensory analysis", are 1) professionals from industry and academia, and 2) graduate students with interests in food science, flavor chemistry, natural products chemistry, and mycology. During the reporting period, two Ph.D. students and one postdoctoral research associate were recruited. The two Ph.D. students are currently working on the project. In addition, four presentations were given, one at the 2024 Middle Atlantic States Mycology Conference (MASMC) at Duke University titled "Characterization of Aroma Molecules in Dried, Rehydrated, and Cooked Lobster Mushrooms", one at the USDA Project Director Meeting for Novel Foods and Innovative Manufacturing Technologies at UMass Amherst titled "Development of sustainable mushroom-based seafood through flavor chemistry, physical chemistry, and sensory analysis", two at the American Chemical Society (ACS) Fall 2024 national conference titled, "Quantitation of odorants in dried and rehydrated lobster mushrooms" and "Influence of polysaccharide addition on the cohesiveness of plant-based meat analogue", all reaching the targeted audience 1 (professionals; both industry and academic) and 2 (graduate students). Furthermore, 2 graduate students and 1 postdoctoral research associate was supervised under the project and conducted the research, reaching both targeted audience 1 (professionals; both industry and academic) and 2 (graduate students). Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During the reporting period, the project has provided training and professional development for two graduate students and one postdoctoral research associate. One graduate student gave two conference presentations, one at the 2024 Middle Atlantic States Mycology Conference (MASMC) at Duke University and the other at the American Chemical Society (ACS) Fall 2024 national conference. The students also submitted two abstracts for presentation at the ACS Spring 2025 national conference. In addition, the students conducted and learned new aroma chemistry techniques including solvent-assisted flavor evaporation (SAFE), aroma extract dilution analysis (AEDA), gas chromatography - olfactometry (GC-O), gas chromatography - mass spectrometry (GC-MS), solid phase extraction (SPE), stable isotope dilution analysis (SIDA), and quantitative olfactory-profile analysis. In addition, the students conducted and learned new mycology techniques including wild mushrooms collection and identification, fungi culturing and sequencing. The students were provided training opportunities by participating in the regional mycological events MycoBash 2024 at Black Mountain, NC, and The Mushroomers 2024 at Wildacres, NC. Furthermore, the two students are active members of the Asheville Mushroom Club (AMC), and frequently participates in the AMC wild mushroom collection and identification events, as well as its wild mushroom identification training workshops. The field works and extension services resulted in one graduate student received two awards, one is Outstanding Graduate Student Award from University of Tennessee Knoxville, the other is Sonoma County Mycological Association (SOMA) Scholarship. The postdoctoral research associate conducted food physical chemistry techniques including nanofibers preparation, seafood analogues preparation, physicochemical and morphological characterization of nanofibers, and texture properties characterization by texture analyzer. The postdoctoral research associate gave one conference presentation at the American Chemical Society (ACS) Fall 2024 national conference titled "Influence of polysaccharide addition on the cohesiveness of plant-based meat analogue". How have the results been disseminated to communities of interest?During the reporting period, four presentations were given, one at the 2024 Middle Atlantic States Mycology Conference (MASMC) at Duke University, one at the USDA Project Director Meeting for Novel Foods and Innovative Manufacturing Technologies at UMass, two at the American Chemical Society (ACS) Fall 2024 national conference. In addition, 2 abstracts were submitted to ACS Spring 2025. The USDA Project Director Meeting, The MASMC conference, and the ACS national conference presentations reached both targeted audience 1 (professionals; both industry and academic) and targeted audience 2 (graduate students). In addition, a popular press article detailing the project and requesting public assistance was published in the Sporadic News, a newsletter of a local mycological society (AMC). One manuscript was published on ACS Journal of Agricultural and Food Chemistry, reaching both targeted audience 1 (professionals; both industry and academic) and targeted audience 2 (graduate students) What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period, we plan on submitting a second manuscript and start the preparation of a third manuscript. Both manuscripts will be submitted to the Journal of Agricultural and Food Chemistry summarizing the aroma chemistry work on the lobster mushrooms. In addition, we will give 4 presentations on the aroma chemistry and physical chemistry findings. For the in-situ flavor development portion, an odorless seafood analogue will be developed and spiked with odorless precursors and thermally treated to preliminarily develop the seafood flavor.

Impacts
What was accomplished under these goals? Developing healthy and sustainable food that also tastes great is crucial in motivating people to adopt a nutritious and improved diets in accordance with 2020-2025 Diet Guidelines for Americans. Mushrooms can offer a promising solution in reducing energy density in the US food supply, increasing nutrient density, and lowering sodium consumption. In recent years, the plant-based meat industry has seen tremendous economic growth due to advances in food processing technology. However, seafood made from plant-based sources is still a challenge, as its flavor and texture are much harder to replicate than beef, poultry, and pork. Innovative approaches in food chemistry, including texture and in-process modification, can help develop seafood flavors, offering the opportunity to design novel mushroom-based seafood (mBSF) with enhanced flavor and texture. These technologies can meet consumer expectations for alternative seafood in a sustainable and environmentally friendly way. During this reporting period, the main chemistry focus has been the identification of the aroma-active compounds present in the dried, rehydrated, and cooked mushrooms, quantitation of the aroma-active compounds in the dried and rehydrated mushrooms, preparation of an aroma simulation model that mimics the aroma of the dried mushrooms, and sensory analysis to compare the model with the dried mushrooms. The aroma chemistry research workstream resulted in three presentations, one at the 2024 Middle Atlantic States Mycology Conference (MASMC) at Duke University, one at the USDA Project Director Meeting for Novel Foods and Innovative Manufacturing Technologies at UMass Amherst, one at the American Chemical Society (ACS) Fall 2024 national conference. Furthermore, one manuscript on the aroma chemistry of lobster mushrooms was published in the ACS Journal of Agricultural and Food Chemistry. In addition, preliminary works have been done on optimizing the texture of mBSF, with a presentation given at ACS Fall 2024. Two more abstracts were submitted to be presented at the ACS Spring 2025 presentation. The detailed accomplishments on the objectives are as follows: Objective 1: Obtaining a reliable source of wild lobster mushrooms and aroma chemistry characterization was the primary focus of the current reporting period. Scientific research and collection permits were obtained from Tennessee Department of Environment and Conservation (TDEC) and Tennessee Wildlife Resources Agency (TWRA). Approximately 12 field trips were conducted to locate wild lobster mushroom locations and an article was written in a local mycological society newsletter informing the public about the project and requesting assistance to find lobster mushroom locations. This resulted in a reliable source of the mushrooms. In addition, we successfully cultured the Hypomyces lactifluorum from wild lobster mushrooms collected from 4 locations to serve as a source of material for biosynthetic pathway elucidation. The species was confirmed by ITS sequencing. The aroma chemistry research workstream identified 35 aroma-active molecules in the mushrooms. Stable isotope standards have been acquired and the quantitative work is completed for dried and rehydrated mushrooms. An aroma simulation model was prepared and sensorily matched with dried mushrooms. A manuscript was published in ACS Journal of Agricultural and Food Chemistry, titled "Characterization of Important Odorants in Dried Lobster Mushrooms". In addition, three talks were given, one at the 2024 Middle Atlantic States Mycology Conference (MASMC) at Duke University titled "Characterization of Aroma Molecules in Dried, Rehydrated, and Cooked Lobster Mushrooms", one at the USDA Project Director Meeting for Novel Foods and Innovative Manufacturing Technologies at UMass Amherst titled "Development of sustainable mushroom-based seafood through flavor chemistry, physical chemistry, and sensory analysis", one at the American Chemical Society (ACS) Fall 2024 national conference titled, "Quantitation of odorants in dried and rehydrated lobster mushrooms". One abstract was submitted to be presented at ACS Spring 2025 conference, titled "Characterization of Odorants in Thermally Treated Lobster Mushrooms". Objective 2: During this reporting period, part of the in-situ mushroom-derived seafood flavor development portion of the project was completed. The button mushroom stems were collected, freeze-dried and pulverized to prepare mushroom stem nanofibers. A seafood analogue is being developed by combining mushroom stem nanofibers with wheat gluten. This seafood analogue will be spiked with the odorless precursors to develop an in-situ seafood flavor and optimized by GC-MS. This workstream is still in progress. Objective 3: During this reporting period, part of the mushroom-based seafood texture portion of the project was completed. One postdoctoral research associate was recruited to work on this portion. Mushroom cap and stem samples were collected from an industrial source. Nanofibers from the cap and stem of button mushrooms were prepared using a ball milling method. In addition, preliminary seafood analogues from plant protein with the addition of mushroom nanofibers were prepared. The texture properties including stiffness, toughness, hardness, and cohesiveness of the analogues were measured and compared to authentic crab leg and imitation crab (surimi). The results were featured in 2 presentations, one at the USDA Project Director Meeting for Novel Foods and Innovative Manufacturing Technologies at UMass Amherst and the other at the American Chemical Society (ACS) Fall 2024 national conference titled "Influence of polysaccharide addition on the cohesiveness of plant-based meat analogue". In addition, one abstract was submitted to ACS Spring 2025 national conference titled "Utilization of button mushroom stem waste in seafood analogs". This workstream is ongoing. Objective 4: During this reporting period, the preliminary lexicon for the sensory evaluation portion of the project was determined and fine-tuned. In addition, the sensory standards that represent each sensory attribute were determined. The lexicon is anticipated to be used for the formal sensory evaluation once the seafood analogs are optimized. The lexicon may require modification as the project progresses. The IRB was obtained for consumer study. Two graduate students participating in the project completed the required training for the IRB project.

Publications

• Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Nguyen, T.H. and Munafo Jr, J.P., 2024. Characterization of Important Odorants in Dried Lobster Mushrooms. Journal of Agricultural and Food Chemistry. 2024, 72, 45, 2525125260
• Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Min Li and Tao Wu. 2024. Influence of polysaccharide addition on the cohesiveness of plant-based meat analogue. American Chemical Society. Fall 2024
• Type: Conference Papers and Presentations Status: Submitted Year Published: 2025 Citation: Thien Nguyen, Ethan Darby, and John P. Munafo. 2025. Characterization of Odorants in Thermally Treated Lobster Mushrooms. American Chemical Society. Spring 2025
• Type: Conference Papers and Presentations Status: Submitted Year Published: 2025 Citation: Ethan Darby, Thien Nguyen, and John P. Munafo. 2025. Utilization of button mushroom stem waste in seafood analogs. American Chemical Society. Spring 2025
• Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Quantitation of odorants in dried and rehydrated lobster mushrooms. American Chemical Society. Fall 2024
• Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Thien Nguyen, Min Li, Tao Wu, John P. Munafo Jr. 2024. Characterization of Aroma Molecules in Dried, Rehydrated, and Cooked Lobster Mushrooms. Middle Atlantic States Mycology Conference. Spring 2024

Progress 08/01/22 to 07/31/23

Outputs
Target Audience:The targeted audiences for the project titled, "Development of sustainable mushroom-based seafood through flavor chemistry, physical chemistry, and sensory analysis", are 1) professionals from industry and academia, and 2) graduate students with interests in food science, flavor chemistry, natural products chemistry, and mycology. During the reporting period, one Ph.D. student and one postdoctoral research associate were recruited and are currently working on the project. In addition, two presentations were given, one at the USDA Project Director Meeting for Novel Foods and Innovative Manufacturing Technologies at UC Davis titled "Development of sustainable mushroom-based seafood through flavor chemistry, physical chemistry, and sensory analysis", and the other at the American Chemical Society (ACS) national conference titled, "Characterization of odorants in dried and rehydrated lobster mushrooms", both reaching the targeted audience 1 (professionals; both industry and academic). Furthermore, 1 graduate student and 1 postdoctoral research associate was supervised under the project and conducted the research, reaching both targeted audience 1 (professionals; both industry and academic) and 2 (graduate students). Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During the reporting period, the project has provided training and professional development for one graduate student and one postdoctoral research associate. The graduate student gave a presentation at the American Chemical Society (ACS) national conference titled, "Characterization of odorants in dried and rehydrated lobster mushrooms". Also, the student conducted and learned new aroma chemistry techniques including solvent-assisted flavor evaporation (SAFE), aroma extract dilution analysis (AEDA), gas chromatography - olfactometry (GC-O), gas chromatography - mass spectrometry (GC-MS), solid phase extraction (SPE), stable isotope dilution analysis (SIDA), and quantitative descriptive analysis (QDA). In addition, the student conducted and learned new mycology techniques including wild mushrooms collection and identification, fungi culturing and sequencing. The student was provided training opportunities by enrolling in the Fungi of the Southern Blue Ridge course at the Highlands Biological Station and participating in the national mycological workshop and meeting event North American Mycological Association (NAMA) Annual Forray 2023, as well as the regional mycological event The Mushroomers 2023 at Wildacres. Furthermore, the student is an active member of the Asheville Mushroom Club (AMC), and frequently participates in the AMC wild mushroom collection and identification events, as well as its wild mushroom identification training workshops. The postdoctoral research associate conducted food physical chemistry techniques including nanofibers preparation, seafood analogues preparation, physicochemical and morphological characterization of nanofibers, and texture properties characterization by texture analyzer. How have the results been disseminated to communities of interest?During the reporting period, two presentations were given, one at the USDA Project Director Meeting for Novel Foods and Innovative Manufacturing Technologies at UC Davis titled "Development of sustainable mushroom-based seafood through flavor chemistry, physical chemistry, and sensory analysis", and the other at the American Chemical Society (ACS) national conference titled, "Characterization of odorants in dried and rehydrated lobster mushrooms". The USDA Project Director Meeting and the ACS national conference presentation reached both targeted audience 1 (professionals; both industry and academic) and targeted audience 2 (graduate students). In addition, a popular press article detailing the project and requesting public assistance was published in the Sporadic News, a newsletter of a local mycological society (AMC). What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period, we plan on submitting 1 manuscript and start the preparation of a second manuscript. Both manuscripts will be submitted to the Journal of Agricultural and Food Chemistry summarizing the aroma chemistry work on the lobster mushrooms. In addition, we will give 2 presentations on the aroma chemistry findings at the American Chemical Society (ACS) national conference. For the mushroom-based seafood texture portion, the physiochemistry and morphology of mushroom nanofibers will be characterized. In addition, the texture of the seafood analogues will be characterized by a texture analyzer.

Impacts
What was accomplished under these goals? Developing healthy and sustainable food that also tastes great is crucial in motivating people to adopt a nutritious and improved diets in accordance with 2020-2025 Diet Guidelines for Americans. Mushrooms can offer a promising solution in reducing energy density in the US food supply, increasing nutrient density, and lowering sodium consumption. In recent years, the plant-based meat industry has seen tremendous economic growth due to advances in food processing technology. However, seafood made from plant-based sources is still a challenge, as its flavor and texture are much harder to replicate than beef, poultry, and pork. Innovative approaches in food chemistry, including texture and in-process modification, can help develop seafood flavors, offering the opportunity to design novel mushroom-based seafood (mBSF) with enhanced flavor and texture. These technologies can meet consumer expectations for alternative seafood in a sustainable and environmentally friendly way. Our preliminary data demonstrates the use of discarded commercial white button mushroom stems, in combination with in-process reaction flavors, to create alternative seafood similar in flavor and texture to crab legs, crab cakes, and tuna. The unique flavor of cooked mushrooms comes from a group of aroma precursors found in raw mushrooms, e.g., amino acids, peptides, and sugars. Among different edible mushrooms, the lobster mushrooms, Hypomyces lactifluorum, have a seafood-like flavor, but the main odorants responsible for their flavor have yet to be identified. By characterizing the odorants present in cooked lobster mushrooms and identifying their odorless precursors in the raw mushrooms, we propose mixing the aroma precursors (amino acid profile unique to the lobster mushroom) with the discarded button mushroom stems (containing common mushroom monosaccharides) and generating a crab-like flavor in-situ. This technology, with slight modifications, can also be applied to other mushroom-based seafood flavors such as shrimp or tuna fish flavor. The overall goal of this proposal is to develop mBSF products utilizing discarded commercial mushroom stems that mimic seafood flavor and texture. The hypothesis is that when odorless flavor precursors identified in the lobster mushroom are mixed with discarded button mushroom stems, and thermally treated under kitchen-like cooking conditions, a crab-like flavor will develop. The specific project objectives are to 1) characterize the key odorants and aroma precursors in lobster mushrooms, 2) develop a mushroom-derived seafood flavor in-situ by reacting mushroom precursors under kitchen-like thermal conditions, 3) Optimize the texture (seafood-like) of the mBSF, 4) Comprehensively evaluate the sensory properties of crab cakes alternative using the mBSF. During this reporting period, the main chemistry focus has been the identification of the aroma-active compounds present in the dried and rehydrated mushrooms. The aroma chemistry research workstream resulted in two presentations, one was given at the USDA Project Director Meeting for Novel Foods and Innovative Manufacturing Technologies at UC Davis titled "Development of sustainable mushroom-based seafood through flavor chemistry, physical chemistry, and sensory analysis", and the other at the American Chemical Society (ACS) national conference titled, "Characterization of odorants in dried and rehydrated lobster mushrooms". The detailed accomplishments on the objectives are as follows: Objective 1: Obtaining a reliable source of wild lobster mushrooms and aroma chemistry characterization was the primary focus of the current reporting period. Scientific research and collection permits were obtained from Tennessee Department of Environment and Conservation (TDEC) and Tennessee Wildlife Resources Agency (TWRA). Approximately 15 field trips were conducted to locate wild lobster mushroom locations and an article was written in a local mycological society newsletter informing the public about the project and requesting assistance to find lobster mushroom locations. This resulted in a reliable source of the mushrooms. In addition, we successfully cultured the Hypomyces in our laboratory to serve a source of material for biosynthetic pathway elucidation. The species was confirmed by ITS sequencing.The aroma chemistry research workstream resulted the identification of 35 aroma-active molecules in the mushrooms. Stable isotope standards have been acquired and the quantitative work is in progress. A manuscript is in preparation and is anticipated to be submitted to the Journal of Agricultural and Food Chemistry. In addition, two talks were presented at the USDA Project Director Meeting and the other at the American Chemical Society (ACS) national conference titled, "Characterization of odorants in dried and rehydrated lobster mushrooms." Objective 2: During this reporting period, part of the in-situ mushroom-derived seafood flavor development portion of the project was completed. The free amino acid profiles of button mushroom stems and lobster mushrooms were characterized. The results were presented at the USDA Project Director Meeting at UC Davis." This workstream is still in progress. Objective 3: During this reporting period, part of the mushroom-based seafood texture portion of the project was completed. One postdoctoral research associate was recruited to work on this portion. Mushroom capand stem samples were collected from an industrial source. Nanofibers from the cap and stem of button mushrooms were prepared using a ball milling method. In addition, preliminary seafood analogues from plant protein with and without the addition of mushroom nanofibers were prepared. This workstream is ongoing. Objective 4: During this reporting period, the preliminary lexicon for the sensory evaluation portion of the project was determined. The lexicon is anticipated to be used for the formal sensory evaluation once the seafood analogs are optimized. The lexicon may require modification as the project progresses.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Characterization of odorants in dried and rehydrated lobster mushrooms. American Chemical Society.
• Type: Other Status: Published Year Published: 2023 Citation: Mushroom of the Month: Help Study Lobster Mushroom Flavor. Sporadic News, AMC Newsletter, June 8, 2023.