Mapping and characterization of dominant hydrolase enzymes in the digestive tract of Striped marlin (Kajikia audax Philippi, 1887)

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ENDANG ISMAIL
ASEP AWALUDIN PRIHANTO
https://orcid.org/0000-0003-4180-1325
SUKOSO
HARTATI KARTIKANINGSIH

Abstract

Abstract. Ismail E, Prihanto AA, Sukoso, Kartikaningsih H. 2025. Mapping and characterization of dominant hydrolase enzymes in the digestive tract of Striped marlin (Kajikia audax Philippi, 1887). Biodiversitas 26: 1156-1163. The digestive tract of Striped marlin (Kajikia audax Philippi, 1887): a fishery byproduct, contains several hydrolase subclass enzymes, such as protease, lipase, amylase, cellulase, and xylanase, which have practical applications in medicine, agriculture, and various industries, including pharmaceuticals, food, detergents, leather, and cosmetics. This study aimed to identify and characterize the enzymes in the digestive tract of Striped marlin. Enzymes from the hydrolase sub-class in different parts of the digestive tract, such as the stomach, pylorus, pancreas, and intestine, were analyzed separately using qualitative and quantitative methods. Each enzyme was further evaluated to determine its optimal pH and temperature. The results revealed that protease activity was the most dominant among the other enzymes. A clear zone with a diameter of >85 mm was observed on a casein substrate in the stomach and pylorus. The highest protease activity was recorded in the part of the stomach (1,044.71 U/mL): followed by the pylorus (937.21 U/mL): pancreas (492.06) U/mL, and intestine (362.21 U/mL). The optimal pH for enzyme activity was 4 in the stomach, 6 in the pylorus, 7 in the pancreas, and 10 in the intestine. The optimal enzyme activity temperature across all digestive tracts was 40?. This study provides the first comprehensive mapping of hydrolase subclass enzymes in the digestive system of Striped marlin.

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References
Borges S, Odila J, Voss G, Martins R, Rosa A, Couto JA, Almeida A, Pintado M. 2023. Fish by-products: A source of enzymes to generate circular bioactive hydrolysates. Molecules 28 (3): 1155. DOI: 10.3390/molecules28031155.
Chang HY, Sun CL, Yeh SZ, Chang YJ, Su NJ, DiNardo G. 2018. Reproductive biology of female Striped marlin Kajikia audax in the western Pacific Ocean. J Fish Biol 92: 105-130.DOI: 10.1111/jfb.13497.
Cho J, Park JW, Ryu Y, Kim K, Hur S. 2023. Tract of the marbled flounder Pseudopleuronectes yokohamae. Animals 5: 936. DOI: 10.3390/ani13050936.
Chong ASC, Hashim R, Chow-Yang L, Ali AB. 2002. Partial characterization and activities of proteases from the digestive tract of discus fish (Symphysodon aequifasciata). Aquaculture 203: 321-333.DOI: 10.1016/S0044-8486(01)00630-5.
Dahiru MM, State A, Yola P, Abaka AM. 2024. Review: Current perspectives on enzyme applications in medicine, agriculture, and industries. Asian J Trop Biotechnol 21: 10-25. DOI: 10.13057/biotek/c210102.
de la Parra AM, Rosas A, Lazo JP, Viana MT. 2007. Partial characterization of the digestive enzymes of Pacific bluefin tuna Thunnus orientalis under culture conditions. Fish Physiol Biochem 33 (3): 223-231. DOI: 10.1007/s10695-007-9134-9.
Deshavath NN, Mukherjee G, Goud VV, Veeranki VD, Sastri CV. 2020. Pitfalls in the 3, 5-dinitrosalicylic acid (DNS) assay for the reducing sugars: Interference of furfural and 5-hydroxymethylfurfural. Intl J Biol Macromol 156: 180-185. DOI: 10.1016/j.ijbiomac.2020.04.045.
Dhaver P, Pletschke B, Sithole B, Govinden R. 2022. Optimization, purification, and characterization of xylanase production by a newly isolated Trichoderma harzianum strain by a two-step statistical experimental design strategy. Sci Rep 12: 17791. DOI: 10.1038/s41598-022-22723-x.
Ejaz U, Sohail M, Ghanemi A. 2021. Cellulases: From bioactivity to a variety of industrial applications. Biomimetics 6 (3): 44. DOI: 10.3390/biomimetics6030044.
Fahmi Z, Hikmayani Y, Yunanda T, Yudiarso P, Wudianto, Setyadji B. 2019. Indonesia National Report to the Scientific Committee of the Indian Ocean Tuna Commission 2020. https://www.iotc.org/sites/default/ files/documents/2020/11/IOTC-2020-SC23-NR07_-_Indonesia.pdf
García-Meilán I, Herrera-Muñoz JI, Ordóñez-Grande B, Fontanillas R, Gallardo Á. 2023. Growth performance, digestive enzyme activities, and oxidative stress markers in the proximal intestine of European sea bass (Dicentrarchus labrax) fed high starch or lipid diets. Fishes 8 (5): 223. DOI: 10.3390/fishes8050223.
Ghosh A, Mahanta S, Banerjee S, Baishya D. 2020. Exploration of endo-xylanase from novel strain of Bacillus velezensis AG20 isolated from the cave of Meghalaya. BioRxiv 2020: 1-44. DOI: 10.1101/2020.04.06.028878.
Gómez?villegas P, Vigara J, Romero L, Gotor C, Raposo S, Gonçalves B, Léon R. 2021. Biochemical characterization of the amylase activity from the new haloarchaeal strain Haloarcula sp. Hs isolated in the Odiel marshlands. Biology 10: 337. DOI: 10.3390/biology10040337.
Hansen MJ, Krause S, Dhellemmes F, Pacher K, Kurvers RHJM, Domenici P, Krause J. 2022. Mechanisms of prey division in Striped marlin, a marine group hunting predator. Commun Biol 5: 1161. DOI: 10.1038/s42003-022-03951-3.
Hashemzahi A, Makhdoumi A, Asoodeh A, Makhdoumi A. 2020. Culturable Diversity and enzyme production survey of halophilic prokaryotes from a solar saltern on the shore of the Oman Sea. J Genet 6 (1): 1-11. DOI: 10.22080/jgr.2020.17847.1170.
Hatzonikolakis Y, Tsiaras K, Tserpes G, Somarakis S, St. John MA, Peristeraki P, Raitsos DE, Triantafyllou G. 2021. Investigating growth and reproduction of the Mediterranean swordfish Xiphias gladius through a full life cycle bioenergetics model. Mar Ecol Prog Ser 680: 51-77. DOI: 10.3354/meps13861.
Ideia P, Pinto J, Ferreira R, Figueiredo L, Spínola V, Castilho PC. 2020. Fish processing industry residues: A review of valuable products extraction and characterization methods. Waste Biomass Valorization 11 (7): 3223-3246. DOI: 10.1007/s12649-019-00739-1.
Jaswir I, Mahfudh N. 2022. Metodologi Penelitian Riset Bidang Sains Halal MODUL. [Indonesian]
Jenol MA, Chu PH, Ramle IK, Joyce LJW, Lai-Yee P, Ibrahim MF, Alitheen NB, Osman MA, Abd Gani S, Abd-Aziz S. 2024. Feasibility of agricultural biomass in Southeast Asia for enzymes production. Renew Sustain Energy Rev 200: 114601. DOI: 10.1016/j.rser.2024.114601.
Jiao F, Zhang L, Limbu SM, Yin H, Xie Y, Yang Z, Shang Z, Kong L, Rong H. 2023. A comparison of digestive strategies for fishes with different feeding habits: Digestive enzyme activities, intestinal morphology, and gut microbiota. Ecol Evol 13 (9): e10499. DOI: 10.1002/ece3.10499.
Jioe IPJ, Shiesh CC, Lin HL. 2023. Bitterness of papaya milk is related to protein and free amino acid contents, with phenylalanine and tyrosine/tryptophan levels being the most important. HortScience 58 (3): 261-267. DOI: 10.21273/HORTSCI16941-22.
Krishnan S, Antony Pillai T, Chembian Antony Rayappan J, Yagappan T, Rajapandian J. 2024. Diet composition and feeding habits of yellowfin tuna Thunnus albacares (Bonnaterre, 1788) from the Bay of Bengal. Aquatic Living Resources. DOI: 10.1051/alr/2024008.
Krogdahl Å, Sundby A, Holm H. 2015. Characteristics of digestive processes in Atlantic salmon (Salmo salar). Enzyme pH optima, chyme pH, and enzyme activities. Aquaculture 449: 27-36. DOI: 10.1016/j.aquaculture.2015.02.032.
Lal V, Naeem M, Asad M, Tanveer K, Zulfiqar A, Kausar S. 2024. Study of digestive enzymes in marine fish, Terapon jarbua, from Pakistan. Brazilian J Biol 84: e267508. DOI: 10.1590/1519-6984.267508.
Liu KM, Huang LH, Su KY. 2022. Assessment of the impact on 20 pelagic fish species by the Taiwanese small-scale longline fishery in the western north Pacific using ecological risk assessment. Animals 12 (16): 2124. DOI: 10.3390/ani12162124.
Mamoozadeh NR, Graves JE, McDowell JR. 2020. Genome-wide SNPs resolve spatiotemporal patterns of connectivity within Striped marlin (Kajikia audax): A broadly distributed and highly migratory pelagic species. Evol Appl 13 (4): 677-698. DOI: 10.1111/eva.12892.
Miller GL. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31: 426-428. DOI: 10.1021/ac60147a030.
Molitor R, Bollinger A, Kubicki S, Loeschcke A, Jaeger KE, Thies S. 2020. Agar plate-based screening methods for the identification of polyester hydrolysis by Pseudomonas species. Microbial Biotechnol 13 (1): 274-284. DOI: 10.1111/1751-7915.13418.
Nadaf H, Hivrale VK. 2023. Protease isoforms of cotton bollworm Pectinophora gossypiella: A biochemical insight. Biol Forum Intl J 15 (4): 33-39.
Nalinanon S, Benjakul S, Kishimura H. 2010. Biochemical properties of pepsinogen and pepsin from the stomach of albacore tuna (Thunnus alalunga). Food Chem 121: 49-55. DOI: 10.1016/j.foodchem.2009.11.089.
Nero G, Kivirand K, Ben Othman S, Rinken T. 2022. Amperometric method for the determination of cellulase activity and its optimization using response surface method. J Anal Sci Technol 13: 21. DOI: 10.1186/s40543-022-00331-8.
Nolasco-Soria H. 2021. Amylase quantification in aquaculture fish studies: A revision of most used procedures and presentation of a new practical protocol for its assessment. Aquaculture 538: 736536. DOI: 10.1016/j.aquaculture.2021.736536.
Nurhayati T, Abdullah A, Rahmawati S, Kurniawan R. 2024. Characteristics of trypsin isolated from intestines different of fish and correlation toward trypsin activity. Squalen Bull Mar Fish Postharvest Biotechnol 19 (2): 131-143. DOI: 10.15578/squalen.904.
Nurhayati T, Nugraha R, Lihuana DN. 2020. Characterization of ammonium sulphate fraction tripsin isolated from intestine of little tuna. Jurnal Pengolahan Hasil Perikanan Indonesia 23 (2): 372-382. DOI: 10.17844/jphpi.v23i2.32221.
Nurilmala M, Suryamarevita H, Husein Hizbullah H, Jacoeb AM, Ochiai, Y. 2022. Fish skin as a biomaterial for halal collagen and gelatin. Saudi J Biol Sci 29 (2): 1100-1110. DOI: 10.1016/j.sjbs.2021.09.056.
Nurjanah Baharuddin TI, Nurhayati T. 2021. Collagen extraction of yellowfin tuna (Thunnus albacares) skin using pepsin and papain. Jurnal Pengolahan Hasil Perikanan Indonesia 24 (2): 174-187. DOI: 10.17844/jphpi.v24i2.35410.
Pasaribu E, Nurhayati T, Nurilmala M. 2018. Extraction and characterization of pepsin enzyme from tuna (Thunnus albacares) gastric. Jurnal Pengolahan Hasil Perikanan Indonesia 21 (3): 486.DOI: 10.17844/jphpi.v21i3.24727. [Indonesian]
Porto de Souza Vandenberghe L, Karp SG, Binder Pagnoncelli MG, von Linsingen Tavares M, Libardi Junior N, Valladares Diestra K, Viesser JA, Soccol CR. 2020. Classification of Enzymes and Catalytic Properties. Biomass, Biofuels, Biochemicals: Advances in Enzyme Catalysis and Technologies. DOI: 10.1016/B978-0-12-819820-9.00002-8.
Putra WA, Diharmi AR, Karnila R. 2021. Aktivitas ekstrak kasar enzim kolagenase dari organ dalam ikan malong (Congresox talabon) pada pH berbeda. Jurnal Teknologi dan Industri Pertanian Indonesia 13 (1): 27-30. DOI: 10.17969/jtipi.v13i1.18587. [Indonesian]
Santajit S, Kong-Ngoen T, Tunyong W, Pumirat P, Ampawong S, Sookrung N, Indrawattana N. 2022. Occurrence, antimicrobial resistance, virulence, and biofilm formation capacity of Vibrio spp. and Aeromonas spp. isolated from raw seafood marketed in Bangkok, Thailand. Vet World 15 (7): 1887-1895. DOI: 10.14202/vetworld.2022.1887-1895.
Shukla ED Bendre AM Gaikwad S. 2022. Hydrolases: The Most Diverse Class of Enzymes. DOI: 10.5772/intechopen.102350.
Skvortsova E, Golovanova I, Postrash I, Filippov A. 2025. Digestive enzyme activities of African catfish and Nile tilapia reared in RAS. BIO Web Conf 160: 02011. DOI: 10.1051/bioconf/202516002011.
Solovyev MM, Kashinskaya EN, Izvekova GI, Glupov VV. 2015. pH values and activity of digestive enzymes in the gastrointestinal tract of fish in Lake Chany (West Siberia). J Ichthyol 55 (2): 251-258. DOI: 10.1134/S0032945215010208.
Taghizadeh Andevari G, Rezaei M, Tabarsa M, Rustad T. 2019. Extraction, partial purification and characterization of alkaline protease from rainbow trout (Oncorhynchus mykiss) viscera. Aquaculture 500: 458-463. DOI: 10.1016/j.aquaculture.2018.10.052.
Tang SL, Liang XF, He S, Li L, Alam MS, Wu J. 2022. Comparative study of the molecular characterization, evolution, and structure modeling of digestive lipase genes reveal the different evolutionary selection between mammals and fishes. Front Genet 13: 909091. DOI: 10.3389/fgene.2022.909091.
Van Gaelen P, Springael D, Smets I. 2021. Lipid hydrolysis monitoring in wastewater treatment: Proof-of-concept for a high throughput vegetable oil emulsion based assay. Water Practice Technol 16 (2): 605-620. DOI: 10.2166/wpt.2021.022.
Volkoff H, Rønnestad I. 2020. Effects of temperature on feeding and digestive processes in fish. Temperature 7 (4): 307-320. DOI: 10.1080/23328940.2020.1765950.
Wang X, Jiang Y, Liu H, Yuan H, Huang D, Wang T. 2023. Research progress of multi-enzyme complexes based on the design of scaffold protein. Bioresour Bioprocess 10 (1): 72. DOI: 10.1186/s40643-023-00695-8.
Yi Z, Yan J, Ding Z, Xie J. 2023. Purification and characterizations of a novel extracellular protease from Shewanella putrefaciens isolated from bigeye tuna. Food Biosci 52: 102384. DOI: 10.1016/j.fbio.2023.102384.
Young T, Pincin J, Neubauer P, Ortega-García S, Jensen OP. 2018. Investigating diet patterns of highly mobile marine predators using stomach contents, stable isotope, and fatty acid analyses. ICES J Mar Sci 75 (5): 1583-1590. DOI: 10.1093/icesjms/fsy025.
Zhang X, Cao D, Sun X, Sun S, Xu N. 2019. Preparation and identification of antioxidant peptides from protein hydrolysate of marine alga Gracilariopsis lemaneiformis. J Appl Phycol 31 (4): 2585-2596. DOI: 10.1007/s10811-019-1746-9.
Zhang X, Shuai Y, Tao H, Li C, He L. 2021. Novel method for the quantitative analysis of protease activity: The casein plate method and its applications. ACS Omega 6 (5): 3675-3680. DOI: 10.1021/acsomega.0c05192.
Zhou Y, Jiang WD, Zhang JX, Feng L, Wu P, Liu Y, Jiang J, Kuang SY, Tang L, Peng Y, Zhou XQ. 2020. Cinnamaldehyde improves the growth performance and digestion and absorption capacity in grass carp (Ctenopharyngodon idella). Fish Physiol Biochem 46 (4): 1589-1601. DOI: 10.1007/s10695-020-00813-9.
Zou L, Wan Y, Zhang S, Luo J, Li YY, Liu J. 2020. Valorization of food waste to multiple bio-energies based on enzymatic pretreatment: A critical review and blueprint for the future. J Cleaner Prod 277: 124091. DOI: 10.1016/j.jclepro.2020.124091.