Iranian Scientific Fisheries Journal

fa مقاله علمی – پژوهشی:‌ بهینه‌‌سازی شرایط هیدرولیز برای تولید پروتئین هیدرولیزشده از ضایعات میگوی موزی (Fenneropenaeus merguiensis) و خصوصیات عملکردی آن Optimization of hydrolysis conditions for protein production from banana shrimp (Fenneropenaeus merguiensis) waste and its functional properties بيوتكنولوژي و فرآوري هاي شيلاتي بيوتكنولوژي و فرآوري هاي شيلاتي پژوهشي Research بازیافت ضایعات حاصل از فراوری موادغذایی یک روش صنعتی مهم برای به‌‌دست ‌آوردن منابع پروتئینی با ارزش افزوده بالاست. هدف از مطالعه حاضر، بهبود یک فرایند آنزیمی مؤثر و سازگار با محیط‌زیست برای تولید پروتئین هیدرولیزشده از ضایعات میگوی موزی بود. بهینه‌سازی شرایط هیدرولیز آنزیمی برای به حداکثر رساندن درجه ‌هیدرولیز و میزان پروتئین هیدرولیزشده از ضایعات سر و پوست میگوی موزی، به طور جداگانه، با استفاده از روش تجزیه‌‌وتحلیل سطح‌ پاسخ  طبق طراحی باکس-بنکن انجام شد. شرایط هیدرولیز شامل: زمان (5، 10، 15 دقیقه)، pH (5/6، 5/7، 5/8)، پلاسمای‌سرد (50، 60، 70 ولت) و نسبت آنزیم به سوبسترا (5/0، 1، 5/1 درصد)، بود. میزان درجه ‌هیدرولیز و پروتئین هیدرولیزشده از ضایعات پوست میگو نسبت به بخش سر آن بیشتر بود. نتایج نشان داد که بیشینه درجه هیدرولیز سر و پوست میگو به‌‌ترتیب با مقدار 47/57 و 11/60 درصد، در تیمار مدت زمان 15 دقیقه، پلاسمای‌سرد 60 ولت، غلظت آنزیم به سوبسترا 5/1 درصد و 5/8 pH= حاصل گردید. بیشینه میزان پروتئین هیدرولیزشده سر و پوست میگو به‌‌ترتیب با مقدار 01/3 و 26/3 درصد حاصل شد. ضریب تعیین R2adj برای درجه هیدرولیز سر، درجه هیدرولیز پوست، پروتئین هیدرولیزشده سر و پروتئین هیدرولیزشده پوست به‌‌ترتیب 9786/0، 9330/0، 9130/0 و 9482/0 بود. تاثیر متغیر مستقل pH بر میزان پروتئین هیدرولیزشده و درجه ‌هیدورلیز سر و پوست میگو موثرتر از سایر شاخص‌‌ها بود. پروتئین هیدرولیزشده از سر و پوست میگو خواص آنتی‌اکسیدانی و ضدمیکروبی خوبی نشان داد. این یافته‌ها بر پتانسیل ضایعات میگوی موزی برای تولید هیدرولیزهای عملکردی و زیست‌فعال با کاربردهای متنوع تأکید دارند. Introduction Shrimp represent the most widely traded product among crustaceans and seafood, primarily due to their appealing flavor and high nutritional value. In 2022, global marine capture fisheries produced 80 million tonnes of aquatic animals, maintaining their position as the principal source of global aquatic animal production, accounting for 43% (FAO, 2024). A considerable amount of waste is produced, contributing to severe environmental problems; this is an issue that needs to be addressed, as these discards have no direct application (Liebana et al., 2025). The dual problem of environmental issues and the less profitable applications for byproducts and waste arising from fish processing industries emphasize the need to achieve valorization from fish processing waste while contributing to a more sustainable fish industry. Protein hydrolysates are produced by breaking peptide bonds in food proteins, generating peptides of varying molecular weights. These hydrolysates are widely used in animal feed, food products, pharmaceuticals, and cosmetics (<a href="https://link.springer.com/article/10.1007/s13399-024-06087-y#auth-Bandela-Dayakar-Aff1" style="color:blue; text-decoration:underline">Dayakar</a> et al., 2025). Enzymatic hydrolysis is particularly advantageous, as it produces hydrolysates with distinct properties depending on the specific peptide bonds cleaved. Recycling of food processing waste is an important industrial method to obtain high-value-added protein sources (Shahidi and Saeid, 2025). During industrial processing, this species generates substantial waste, primarily consisting of cephalothoraxes, which represent 40–60% of the total biomass. The valorization of this underutilized waste is essential to mitigate its environmental impact and recover its rich nutrient content (Pereira et al., 2022). The aim of this study was to improve an efficient and environmentally friendly enzymatic process for protein extraction from banana shrimp waste. Methodology Head and skin sections of banana shrimp (F. merguiensis) were freshly obtained from Ahrar processing centers in Bushehr province. They were then transported to the laboratory on ice in a foam, cleaned, and frozen at -20°C until use. Before preparing the hydrolyzed protein, the samples were transferred to a refrigerator at 4°C. Optimization of enzymatic hydrolysis conditions to maximize the degree of hydrolysis and amount of hydrolyzed protein from banana shrimp head and skin waste, separately, was carried out using response surface methodology (RSM) according to Box-Behnken design. The hydrolysis conditions were determined as follows: time (5, 10, 15 min), pH (6.5, 7.5, 8.5), cold plasma (50, 60, 70 V) and enzyme/substrate ratio (0.5, 1, 1.5%). Three experimental replicates of each case were performed, and mean values have been expressed for the experimental responses. The experimental studies were optimized to minimize unexpected variability effects in the observed responses. The 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) radicals scavenging, activity was assayed. The ability of fractions to reduce iron was measured. Inhibition zone of bacterial strains was measured according to disk diffusion test. The optimization tests were analyzed using experimental design modules BBD from the RSM. The same software was used to analyze data obtained through RSM to determine the optimized condition. Results The results showed that the maximum degree of hydrolysis of shrimp head and skin was 57.47 and 60.11%, respectively, in the treatment time of 15 min, cold plasma of 60 volts, enzyme concentration to substrate of 1.5% and pH = 8.5. The degree of hydrolysis and amount of hydrolyzed protein from shrimp skin waste were higher than from shrimp head part. The maximum amount of hydrolyzed protein of shrimp head and skin was 3.01 and 3.26%, respectively. The coefficient of determination Radj2 for the degree of hydrolysis of head, degree of hydrolysis of skin, hydrolyzed protein of head and hydrolyzed protein of skin were 0.9786, 0.9330, 0.9130 and 0.9482, respectively. The effect of the independent variable pH on the amount of hydrolyzed protein and degree of hydrolysis of shrimp head and skin was more effective than other factors. Both hydrolyzed proteins from shrimp head and skin showed good antioxidant and antimicrobial functional properties. From the examination of Table 3, it is clear that the linear effect of pH, enzyme to substrate, cold plasma, time and the interaction effect of pH-enzyme to substrate, pH-cold plasma, enzyme to substrate-cold plasma, cold plasma-time and also the quadratic effect of pH-enzyme had a significant effect (p<0.05) on the degree of hydrolysis of protein obtained from shrimp heads, and only the interaction effect of pH-time, enzyme to substrate-time and the quadratic effect of cold plasma had no significant effect (p>0.05). The study of the effect of variables on the degree of hydrolysis of banana shrimp skin showed that the analysis of variance of the linear effects of pH, enzyme to substrate, cold plasma, time and the interaction effects of pH-enzyme to substrate, cold plasma-time and the quadratic effect of pH on the degree of hydrolysis of protein obtained from shrimp skin were significant (p<0.05). However, the interaction effects of pH-cold plasma, pH-time, enzyme-to-substrate-cold plasma, enzyme-to-substrate-time, and the quadratic effects of enzyme-to-substrate, cold plasma, and time on the degree of protein hydrolysis from shrimp skin were not significant (p>0.05). <div aria-label="Page Break" class="cke_pagebreak" contenteditable="false" data-cke-display-name="pagebreak" data-cke-pagebreak="1" style="page-break-after:always" title="Page Break"></div>Discussion and conclusion BBD design was used to determine the degree of hydrolysis using papain enzyme to produce hydrolyzed protein. Hydrolyzed protein production experiments were conducted to evaluate the effects of variables at 4 levels. The selection of independent variables including time, pH and enzyme to substrate ratio was because these variables significantly affect the peptide composition of hydrolyzed protein and their optimization is essential to achieve the desired degree of hydrolysis. BBD was chosen for the optimization of independent variables for two main reasons. First, to obtain extensive information while conducting a limited number of experiments and then to analyze the interactive effects between the independent variables and the response variable (Dayakar et al., 2024). On the other hand, BBD requires that all the values of the independent factors align with the three levels (-1, 0 and +1) and the distances between them are equal (Dayakar et al., 2025), a requirement that was met in our study. A three-level BBD was applied in the response surface methodology study by creating an equation to determine the effective conditions for the highest degree of hydrolysis. The results showed that this model can estimate the percentage of hydrolysis degree of proteins obtained from wastes in different combinations of experimental parameters affecting hydrolysis. The results showed that the highest degree of hydrolysis was achieved by the effect of pH-time at 54.91% for the head part and 55.01% for the skin part. In a study, it was shown that the optimal conditions for the highest degree of hydrolysis were achieved at 73.91% within 1 hour of hydrolysis time and 1% enzyme concentration. The findings of this study showed that temperature had a significant effect on the degree of hydrolysis, but the amount of enzyme did not have a significant effect P (Korkmaz and Tokur, 2022). Conflict of interest The authors declare no conflict of interest. Acknowledgment The authors extend their gratitude to the Ahvaz Branch, Islamic Azad University, for providing the necessary facilities and support. هیدرولیز, پاپائین, پروتئین هیدرولیزشده, ضایعات, میگوی موزی Hydrolysis, Papain, Protein hydrolysate, Shrimp waste, Fenneropenaeus merguiensis 27 49 http://isfj.ir/browse.php?a_code=A-10-206-16&slc_lang=fa&sid=1 K. Ghorbani Birgani کبری قربانی بیرگانی kobraghorbanibirgani@gmail.com 100319475328460042126 100319475328460042126 No Department of Food Science and Technology, Ahv. C., Islamic Azad University, Ahvaz, Iran گروه علوم و صنایع غذایی، واحد اهواز، دانشگاه آزاد اسلامی، اهواز، ایران L. Roomiani لاله رومیانی Laleh.roomiani@iau.ac.ir 100319475328460042127 100319475328460042127 Yes Department of Fisheries, Ahv. C., Islamic Azad University, Ahvaz, Iran گروه شیلات، واحد اهواز، دانشگاه آزاد اسلامی، اهواز، ایران