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INVESTIGATOR: Osburn, W.; Miller, RH, K..; Wu, GU, .; Ramanathan, RA, .; Chitlapilly Dass, SA, .; Kerth, CH, R.

PERFORMING INSTITUTION:
TEXAS A&M UNIVERSITY
750 AGRONOMY RD STE 2701
COLLEGE STATION, TEXAS 77843-0001

AN AMINO ACID ALTERNATIVE CURING SYSTEM

NON-TECHNICAL SUMMARY: The primary, long-term term goal of the PDs is to determine the feasibility of utilizing the endothelial nitric oxide synthase system (NOS) in post rigor skeletal muscle as an innovative method to generate nitric oxide and residual nitrite to cure meat and poultry products. We hypothesize that the addition of the amino acid L-arginine activates the NOS system in post rigor skeletal muscle to generate nitric oxide (NO) for cured meat color development and residual nitrite for product shelf life and safety as an antimicrobial.The outcomes of this project are: 1) Develop a better understanding of the factors that impact the ability of the NOS system to generate nitric oxide and residual nitrite in the manufacture of cured meat products and 2) Develop processing/operating procedures/parameters for meat processors to effectively use the NOS system to cure meat and poultry products in a consistent and predictable manner and 3) Verify that this system does not compromise food quality or safety.This project will improve our knowledge and understanding of the chemical, physical, and biological properties of meat products cured via this novel amino acid based alternative curing method by eliminating the need for direct addition of sodium nitrite. These efforts will allow for better understanding of the interconnected biochemical mechanisms contributing to the functionality of the NOS system in post rigor skeletal muscle. It is expected that this innovative alternative curing system will improve the safety, quality, shelf-life, convenience, nutrient profile and sensory attributes of cured meat and poultry products.

OBJECTIVES: Long-term Goals. The primary, long-term term goal of the PDs is to determine the feasibility of utilizing the semi-essential amino acid L-arginine as the substrate to activate the endothelial nitric oxide synthase (eNOS) system in post rigor skeletal muscle as an innovative novel method to generate nitric oxide and sodium nitrite to cure meat and poultry products. We hypothesize that the addition of the amino acid L-arginine as a substrate can activate the eNOS system in post rigor skeletal muscle to generate nitric oxide (NO) to form nitrosohemachromagen for cured meat color development and residual nitrite as an antimicrobial/antioxidant for product shelf life and safety.The outcomes forthis project are: 1) Develop a better understanding of the factors that impact the ability of the NOS system to generate NO and residual nitrite in the manufacture of cured meat products; 2) Develop processing/operating procedures/parameters for meat processors to effectively use the NOS system to cure meat and poultry products in a consistent and predictable manner and 3) Verify that the amino acid alternative curing system (AAACS) does not compromise food quality or safety.This knowledge will form the basis for a novel amino acid based alternative curing method that eliminates the need for direct or indirect addition of sodium nitrite in cured meat products. Importantly, these efforts will allow for better understanding of the interconnected biochemical mechanisms contributing to the functionality of the NOS system in post rigor skeletal muscle as well as providing additional insight into evaluating the efficacy of the proposed alternative system to eliminate the need for sodium nitrite addition to cure meat and poultry products.Objectives:Year 1: Elucidate the factors that eNOS activity to generate nitric oxide and residual nitrite in in post rigor skeletal muscleYear 2: Determine if the AAACS is comparable to conventionally cured (sodium nitrite) methods in the manufacture of cured meat products.Year 3: Determine if AAACS can control pathogenic growth compared to conventionally cured meat products during thermal processing and chilling.

APPROACH: Year 1: Elucidate the factors that impact eNOS activity to generate nitric oxide and residual nitrite in post rigor skeletal muscleEfforts include investigating the mechanism of eNOS in post rigor meat to determine the factors that affect the functionality/enzyme activity of eNOS in post rigor muscle (production of residual nitrite). Questions to be answered include: How well does eNOS utilize L-arginine as a substrate under standard processing conditions? Is L-arginine the only amino acid (L-citrulline) that activates eNOS to generate residual nitrite in post rigor skeletal muscle? How does concentration/temperature/time affect the ability of the eNOS system to generate residual nitrite compared to meat samples containing sodium nitrite? Research activities include: 1) Formulation and manufacture of sodium nitrite, L-arginine and L-citrulline beef, pork and poultry samples; 2) Nitrate and nitrite analysis 3) Amino acid analysis 4) NOS activity.Expected ResultsExperimental results will determine if L-arginine or L-citrulline result in the generation of nitric oxide and residual nitrite comparable to sodium nitrite treated samples. These results will provide important info for increasing knowledge on how amino acid type, concentration and hold times impact NOS activity and how the AAACS might be used in commercial cured meat manufacturing.Year 2: Determine if the AAACS is comparable to conventionally cured (sodium nitrite) methods in the manufacture of cured meat products.Efforts include the manufacture of a selected cured meat product using conventional and AAACS methods and evaluating the products for the following attributes: color development and stability, shelf life extension (lipid oxidation and microbial stability) and cured meat flavor. This investigation will provide data to determine processing parameters to optimize product performance. Research activities include: 1) Formulation and manufacture of conventionally cured (sodium nitrite) and amino acid (L-arginine and/or L-citrulline) meat product 2) Nitrate and nitrite analyses 3) Color determinations/analyses and 4) shelf life and sensory attributes.Expected ResultsExperimental results will determine if L-arginine treated ham samples or comparable to a conventionally cured ham product with respect to color, flavor and shelf life properties. These results will provide important information for increasing knowledge on how the AAACS might be used in commercial cured meat product.Year 3: Determine if AAACS can control pathogenic growth compared to conventionally cured meat products during thermal processing and chilling.Efforts include the manufacture of a selected cured meat product using conventional and AAACS methods and evaluating the products for the following attributes: growth of Clostridium species and other/or other organisms in the microbiome during extended storage times. This data is vital to determine if the AAACS can cure meat products without compromising food safety (i.e., pathogen growth. Research activities include: 1) Formulation and manufacture of conventionally cured (sodium nitrite) and amino acid (L-arginine and/or L-citrulline) meat product 2) Nitrate and nitrite analyses and 3) Microbial analysisExpected ResultsExperimental results will determine if L-arginine treated ham samples are comparable to a conventionally cured ham product with respect to discouraging pathogen colonization and will help determine community succession patterns. These results will provide important information for increasing knowledge on how the AAACS might be used to safely cure commercial meat products.

PROGRESS: 2023/01 TO 2023/12
Target Audience:The target audience for this reporting period include Animal Science and Food Science undergraduate and graduate students and professionals in the discipline of meat science as well as professionals in the meat industry - specifically in the area of valued added/further processed meat and poultry products and the general public during this period. Efforts to deliver science based knowledge from this project forward will be through, on campus and in-plant training workshops, international seminars and workshops, scientific conferences, consulting, undergraduate and graduate level courses and through publication of peer reviewed manuscripts in scientific journals. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project provided training for 1 PhD and 4 MSc students in product manufacturing, nitricoxide synthase activity and nitrate/nitriteanalyses. How have the results been disseminated to communities of interest?Results of this project have been communicatedto: Texas Association of Meat Processors (Annual Convention), The American Meat Science Association (Annual Convention), Tyson Foods (invitation), Texas Beef Cattle Producers (Annual Beef Cattle Shortcourse), and Latin American Meat Processors (Tenth Latin American Meat Technology Course). What do you plan to do during the next reporting period to accomplish the goals?The next reporting period is focused on Year 3 objectives to determine?if the amino acid alternative curing system can control pathogenic growth (surrogate Clostridium species) compared to conventionally cured meat products during thermal processing and chilling. Manuscripts from year 1 and 2 objectives will be submitted for reviewand publication.

IMPACT: 2023/01 TO 2023/12
What was accomplished under these goals? Year 2 objective was thefocus of this reporting period.This study investigated which amino acid and/or concentration most effectively cures restructured hams via endothelial nitric oxide synthase system (eNOS). Boneless pork semimembranosus muscle were manufactured with 20% brine and the addition of L-arginine (Arg), L-citrulline (Cit) or combination of the two amino acids (Arg/Cit) at varying concentrations of 1000, 3000 or 5000ppm and a nitrite (NaNO2) control. Hams were analyzed on day 1, 7, 28 and 56 of refrigerated (40°C) storage for residual nitrate (RNO3) nitrite (RNO2) and nitroslyhemochromagen (NOheme), with a trained sensory panel and objective textural measurements analyzed on day 28 storage. For RNO3 a significant interaction (p=0.0001) occurred between amino acid and concentration. From the data it can be concluded 1000ppm is a sufficient concentration for eNOS to generate NO3. There were no significant interactions for RNO2, no amino acid or concentration produced significantly greater RNO2 values in the restructured hams however all amino acids produced ~2/3 of the amount of RNO2 NaNO2 produced. There was a significant interaction (p=0.01) between amino acid and concentration in NOheme values. There was also a significant interaction (p=0.001) between amino acid and storage day in NOheme values, however, overall, no singular amino acid or concentration was more effective at producing higher NOheme values. Regarding sensory, there was a significant difference between amino acid for Cured Ham ID (p=0.004) and Ham Flavor Aftertaste (p=0.006) with Arg having the closest value to NaNO2 for both attributes. There was a significant difference between amino acid regarding Soured Aromatic (p=0.03) and Chemical/Medicinal/Metallic (p=0.001) with Arg having the lowest and closest values NaNO2 and Cit having the highest values for both attributes. There was a significant interaction (p=0.0001) between amino acid and concentration on Hardness with Arg values increasing as concentration increased and Arg having closest values to NaNO2. There was a significant difference (p=0.0009) between amino acids regarding chewiness with Arg having the least and closest value to NaNO2. This data suggests that Arg most effectively cures restructured hams by activation of the eNOS system and that a concentration of 1000ppm is sufficient for generation of eNOS in a restructured ham product.

PUBLICATIONS (not previously reported): 2023/01 TO 2023/12
No publications reported this period.