Source: PENNSYLVANIA STATE UNIVERSITY submitted to
BIOINSPIRED ENZYMATIC MANUFACTURING OF PULSE PROTEIN HYDROGELS AS COLD/HEAT-TOLERANT FAT TEXTURIZING AGENTS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
Reporting Frequency
Annual
Accession No.
1030226
Grant No.
2023-67017-40167
Cumulative Award Amt.
$599,000.00
Proposal No.
2022-09146
Multistate No.
(N/A)
Project Start Date
Aug 1, 2023
Project End Date
Jul 31, 2027
Grant Year
2023
Program Code
[A1364]- Novel Foods and Innovative Manufacturing Technologies
Project Director
Zhang, Y.
Recipient Organization
PENNSYLVANIA STATE UNIVERSITY
408 Old Main
UNIVERSITY PARK,PA 16802-1505
Performing Department
(N/A)
Non Technical Summary
A high intake of fats, particularly saturated fatty acids, is closely associated with health risks such as obesity, type 2 diabetes, certain cancers, and cardiovascular diseases. Recent national studies on US adult consumers showed that the intake of saturated fatty acids remained above 10% of energy, which is higher than the recommended level by WHO and FAO. It is essential to provide low-fat or fat-free food products to keep consumers healthy. Fat texturizing agents can act like fat but supply fewer calories and pose no or little health risks. In the food industry, fat texturizing agents are added to processed food products, including processed meat, meat analogues, fried foods, baked foods, dairy products, frozen and refrigerated foods, and snacks to provide consumers with the desired mouthfeel. Traditional fat texturizing agents receive an increasing expression of concerns about nutrition, health, and food sustainability as they are produced using carbohydrates, fatty acids, or vegetable oils. Protein-based fat texturizing agents is the new trend, but they also carry drawbacks that limit their development and applications. The primary disadvantage is that plant-based fat texturizers cannot tolerate cold-temperature and high-temperature processing. The second drawback is their low flexibility in the complex food matrix where polar and non-polar components co-exist. There are also limited techniques for producing high-quality protein-based fat texturizing agents, considering that the current techniques rely heavily on mechanical processing. Unlike mechanical processing, enzymes catalyze chemical reactions with high specificity under mild reaction conditions with low energy consumption. Thus, we foresee the urgent need to discover novel protein-based fat texturizing agents with superior qualities via novel and clean enzymatic manufacturing techniques.We propose to use proteins from pulse crops, e.g., dry peas, lentils, and chickpeas, to create novel fat texturizing agents. Pulses are massively produced in the US and are expected to play a vital role in addressing food security, nutrition, health, and sustainability in the immediate and foreseeable future. Pulse proteins will be modified by protease enzymes to improve their functionalities to enable them to function as fat texturizing agents. To improve their tolerance to cold-temperature and high-temperature processing, we are inspired by nature, where naturally occurring transglutaminase enzymes can act to combine different proteins in living organisms for a plethora of biological functions, and the cross-linked proteins show improved stability. We, therefore, will further modify the proteins via transglutaminase catalysis. The novel pulse protein-based hydrogels will be investigated for their cold-/heat-tolerance as fat texturizing agents in a case study in meat analogue products. Overall, this project is bioinspired by biocatalysis in nature, aiming to discover novel protein-based hydrogels as new fat texturizing agents for healthy processed foods. The pulse protein hydrogels discovered in this project as new texturizing agents will meet the current needs of low-fat, low-calorie foods and good texture for frozen and thermally processed foods. In addition, the new manufacturing technique is consistent with the clean production and sustainability of agri-food systems.
Animal Health Component
40%
Research Effort Categories
Basic
50%
Applied
40%
Developmental
10%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
5025010200050%
5015010100050%
Goals / Objectives
The major goal of the proposed project is to provide enzymatic manufacturing solutions based on natural phenomena to create novel fat-like texturizing agents derived from pulse proteins with high amphiphilicity hydrophilic and hydrophobic properties to produce improved texturizes and cross-linked by isopeptide bonds (to result in high tolerance to cold/high temperatures. The new fat texturizing agents can lead to the creation of a wide range of low-fat, reduced-fat, or fat-free processed food products with desired fat texture and healthy values. We will achieve three supporting objectives in the proposed project:Objective 1: Generate amphiphilic polypeptides from pulse proteins via controlled enzymatic hydrolysis with new knowledge on the hydrolysis process - structure - amphiphilicity relationships.Objective 2: Prepare bioinspired hydrogels using amphiphilic pulse polypeptides via site-specific cross-linking by isopeptide bond formation catalyzed using cold-active transglutaminase.Objective 3: Investigate the cold-/heat-tolerance of the newly developed hydrogels as fat texturizing agents, and conduct a case study on its addition as a texturizer in meat analogue products.
Project Methods
In this project, polypeptides with high amphiphilicity will be generated from three pulses, i.e., dry peas, lentils, and chickpeas, using a hydrolytic enzyme after pre-treatment using high-pressure jet processing of the pulse protein isolates. We will investigate and understand the enzymatic hydrolysis process, as well as the polypeptide structure and amphiphilicity relationships of pulse polypeptides via instrumental analyses using high-performance liquid chromatography (HPLC), circular dichroism (CD) spectroscopy, fluorescence spectroscopy. Isopeptide bonds will be subsequently introduced to form cross-links in the polypeptide network via transglutaminase catalysis to allow efficient and stable setting of pulse polypeptide hydrogels. The cross-linking, amphiphilicity, gel strength, and morphological features of hydrogels will be analyzed using liquid chromatography with tandem mass spectrometry (LC-MS/MS), rheometry, scanning electron microscopy (SEM), and other techniques. The newly developed hydrogels will be investigated for their potential to replace fats wholly or partially in frozen and thermally processed foods. Various properties, such as stability and texturizing capacity under low and high-temperature conditions, will be investigated using techniques based on multiple instruments, including a texture analyzer, differential scanning calorimetry (DSC), and Fourier-transform infrared spectroscopy (FTIR). A case study will be performed in which the new texturizing agent will be added to a frozen meat analogue to monitor the texture changes during low-temperature storage and thermal processing.

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

Outputs
Target Audience:The target audience includes graduate students, undergraduate students, and researchers in the food industry. The knowledge generated in this period was delivered to the target audience through classes, short courses, and summer school. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?In this project, one full-time PhD student is being trained and mentored by the PI; two part-time research assistants were trained. In addition, lectures involved the knowledge generated in this project were provided in classes, short courses, and summer school.? How have the results been disseminated to communities of interest?In the first year, the knowledge generated in this project was disseminated to the public through lectures in short courses and summer school.? What do you plan to do during the next reporting period to accomplish the goals?Work on the Objective 2 in the next reporting period.

Impacts
What was accomplished under these goals? We have optimized the extraction of proteins from pulse crops, i.e., chickpeas, peas, and lentils, and the protein isolates presented a high yield and uniform composition. Proteases were applied to partially hydrolyze protein isolates into polypeptides at mild conditions. Functionalities of the produced polypeptides, including solubility, water-holding capacity, oil-binding capacity, foaming, and emulsifying abilities, were studied. The amphiphilicity features of polypeptides were analyzed based on sequences. The amphiphilicity mechanism of selected polypeptides in aqueous-oil phases was explained using bioinformatics. The main goal indicated in Objective 1, i.e., the relationship between enzymatic hydrolysis, protein structure, and amphiphilicity,was elucidated.

Publications

  • Type: Journal Articles Status: Accepted Year Published: 2024 Citation: Zhang, W., Boateng, I. D., Xu, J., & Zhang, Y. (2024). Proteins from Legumes, Cereals, and Pseudo-Cereals: Composition, Modification, Bioactivities, and Applications. Foods, 13(13), 1974.