Performing Department
(N/A)
Non Technical Summary
Asian carp have become a highly invasive species since being introduced to the U.S. in the 1970s, threatening local ecosystems, particularly in the Mississippi River Basin and potentially the Great Lakes (billions of dollars from the recreational and fishing industries). Unlike in Asian countries where carp are popular and nutritious table dishes, they are seldom consumed in the U.S., with only a small fraction processed into fillets or surimi products. This project aims to manage the carp population by harvesting them for valuable products, centering on extracting collagen from their skin and converting it into collagen peptides with significant health benefits, presenting opportunities for diverse applications in food, cosmetic, biomedical, and pharmaceutical industries. The objectives of the present project include optimizing collagen extraction and peptide production processes and evaluating the biological activity of collagen peptides including bone and skin protection. By creating profitable products from carp, the initiative seeks to incentivize their removal from U.S. waters, offering economic opportunities to local fishermen and processors, reducing the public concern about the ecology of the U.S. River system, and potentially establishing a sustainable, nationwide Industry centered on carp processing in the long run.
Animal Health Component
0%
Research Effort Categories
Basic
100%
Applied
0%
Developmental
0%
Goals / Objectives
Our long-term goal is to develop niche markets with profitable carp products that have the potential to incentivize the removal of this invasive species from U.S. waters. The specific objectives of this project are 1) to refine and optimize the process for extracting collagen from the skin of carp 2) to optimize the method for preparing low-molecular-weight collagen peptides via enzymatic hydrolysis, and 3) to evaluate the bioaccessibility and bioactivity of the prepared collagen peptides from carp.
Project Methods
Carp (Silver) skin will be used for collagen extraction since it predominates the carp species in the Mississippi River Basin-Kentucky waters. Silver carp skin will be obtained from a Kentucky local fish processor, Two Rivers Fisheries (Wickliffe, KY). Objective #1: To refine and optimize the process for extracting collagen from carp skin. The obtained carp skin will be cut into small pieces, mixed with water, and adjusted to alkaline pH for removing non-collagenous proteins, followed by being sieved and neutralizing the pH. Then, the skin will be dissolved in acetic acid to extract collagen at 4 °C over a certain period. The viscous solution will be salted out by adding NaCl to a final concentration of 0.9 M, with the sediment collected and freeze-dried after refrigerated centrifugation at 10,000 g for 10 min. Specific parameters will be optimized, including the skin size, extraction duration, the strength of acetic acid, solid-to-liquid ratio, and others. The proximate composition of the extracted collagen will be determined. The recovery of collagen and its purity will be measured using sodium dodecyl polyacrylamide sulfate gel electrophoresis (SDS-PAGE) and hydroxyproline assay. Potential fish-borne allergens will be detected. Objective #2: To optimize the preparation of low-molecular-weight collagen peptides via enzymatic hydrolysis. Collagen will be subjected to heat denaturation followed by enzymatic digestion by controlling hydrolysis parameters including initial collagen concentration, time, and enzyme/substrate ratio, followed by the optimization of these parameters based on results of the degree of hydrolysis, hydrolysis yield, peptide purity, SDS-PAGE, and molecular weight distribution. The sequences of collagen peptides will be identified by liquid chromatography with tandem mass spectrometry (LC-MS/MS) and interpreted by de novo sequencing. Objective #3: T to evaluate the bioaccessibility and bioactivity of the prepared collagen peptides from carp. Low-molecular-weight collagen peptides prepared in Objective #2 will be measured using the human Caco-2 transepithelial cell monolayers, an in vitro model mimicking the in vivo transport of nutrients; peptides passing through the Caco-2 cell monolayers will be quantified and further studied on their multiple bioactivities including bone using bone cellsand skin health using human dermal epithelial cells. Data will be expressed as mean values accompanied by standard deviations from at least three to four independent experiments. Data will be subjected to an unpaired t-test or one-way analysis of variance (ANOVA) followed by Duncan's multiple-range tests using IBM SPSS Statistics (version 29, IBM Corporation, NY, USA). The p-value of 0.05 was considered statistically significant.