Substrate specificity, physicochemical and kinetic properties of a trypsin from the giant Amazonian fish pirarucu (Arapaima gigas)

Freitas-Júnior, A. C. V.; Costa, Helane Maria Silva; Marcuschi, Marina; Icimoto, Marcelo Yudi; Machado, Marcelo Ferreira Marcondes; Machado, Maurício F.M.; Ferreira, Juliana C.; Oliveira, Vitor; Buarque, Diego S.; Bezerra, Ranilson S.

doi:10.1016/j.bcab.2021.102073

Cited by 6 publications

(7 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A gradual decrease in the activity of both trypsins was recorded at 55 • C, whereas enzymatic activity sharply decreased at 60 • C. After heating the ICEs at 70 • C, the relative activities for both trypsins were only about 0.9% and 1.6% of their initial activity for sevruga and beluga, respectively. These results were in accordance with those of sardinelle, common kilka, mrigal carp, and pirarucu, which were exhibited to be stable up to 50 • C [40,48,49,57]. The trypsins from beluga and sevruga showed to be more stable at high temperatures in comparison with those reported for the Monterey sardine, chinook salmon, bluefish, Tunisian barbell, and common dolphinfish that the enzymatic activity was rapidly lost at temperatures above 40 • C [54,55,58,68,69].…”

Section: Optimum Temperature and Thermostabilitysupporting

confidence: 87%

“…In general, trypsins have molecular weights in the range of 20-30 kDa [47]. In particular, different molecular weights for trypsins have been reported in various fish species such as 21.7 kDa for mrigal carp [48], 23.2 kDa for common kilka [40], 23.5 kDa for pirarucu [49], 24 kDa for small red scorpion fish [50], 21 and 24 kDa for liver of albacore tuna [51], 24 kDa for catfish [52], 24.4 kDa for gulf corvina [53], 25 kDa for Monterey sardine [54], 26 kDa for common dolphinfish [55], 27 kDa for zebra blenny [56], 28.8 kDa for sardinelle [57], 29 kDa for Atlantic bonito [58], 38.5 kDa for tambaqui [59], and 42 kDa for skipjack tuna [60]. However, several reasons such as different habitat and climate, autolytic degradation, and genetic variation among fish species may explain why trypsins from various sources have different molecular weights [60,61].…”

Section: Optimum Temperature and Thermostabilitymentioning

confidence: 99%

See 1 more Smart Citation

Physicochemical and Biochemical Properties of Trypsin-like Enzyme from Two Sturgeon Species

Zamani

Khajavi

Kenari

et al. 2023

Animals

View full text Add to dashboard Cite

This work aimed to determine the physicochemical and biochemical properties of trypsin from beluga Huso huso and sevruga Acipenser stellatus, two highly valuable sturgeon species. According to the results obtained from the methods of casein-zymogram and inhibitory activity staining, the molecular weight of trypsin for sevruga and beluga was 27.5 and 29.5 kDa, respectively. Optimum pH and temperature values for both trypsins were recorded at 8.5 and 55 °C by BAPNA (a specific substrate), respectively. The stability of both trypsins was well-preserved at pH values from 6.0 to 11.0 and temperatures up to 50 °C. TLCK and SBTI, two specific trypsin inhibitors, showed a significant inhibitory effect on the enzymatic activity of both trypsins (p < 0.05). The enzyme activity was significantly increased in the presence of Ca+2 and surfactants and decreased by oxidizing agents, Cu+2, Zn+2, and Co+2 (p < 0.05). However, univalent ions Na+ and K+ did not show any significant effect on the activity of both trypsins (p > 0.05). The results of our study show that the properties of trypsin from beluga and sevruga are in agreement with data reported in bony fish and can contribute to the clear understanding of trypsin activity in these primitive species.

show abstract

Section: Optimum Temperature and Thermostabilitysupporting

confidence: 87%

Section: Optimum Temperature and Thermostabilitymentioning

confidence: 99%

Physicochemical and Biochemical Properties of Trypsin-like Enzyme from Two Sturgeon Species

Zamani

Khajavi

Kenari

et al. 2023

Animals

View full text Add to dashboard Cite

show abstract

“…In general, trypsins have MWs in the range of 20-42 kDa [27]. In particular, different MWs for trypsins have been documented in numerous fish species, including 21.7 kDa for trypsin from the Cirrhinus mrigala digestive tract [12], 23.2 kDa for trypsin from the Clupeonella cultriventris caspia pyloric caeca [13], 23.5 kDa for trypsin from the Arapaima gigas pyloric caeca [14], 24 kDa from trypsin from the Scorpaena notata intestine [15], 21 and 24 kDa for trypsin from the Thunnus alalunga liver [16], etc. Differences in the MWs of trypsin from various fish species might be due to varying factors, including genetic differences, the organs used, autolytic degradation, a different climate and habitat, etc.…”

Section: Sodium Dodecyl Sulphate-polyacrylamide Gel Electrophoresis (...mentioning

confidence: 99%

“…Trypsin is a very important enzyme, with several commercial uses in food companies. Trypsin has been isolated, purified, and characterized from various species of fish, including the mrigal carp (Cirrhinus mrigala) [12], common kilka (Clupeonella cultriventris caspia) [13], pirarucu (Arapaima gigas) [14], small red scorpion fish (Scorpaena notata) [15], albacore tuna (Thunnus alalunga) [16], catfish (Luphiosilurus alexandri) [17], gulf corvina (Cynoscion parvipinnis) [18], Monterey sardine (Sardinops sagax caerulea) [19], common dolphinfish (Coryphaena hippurus) [20], skipjack tuna (Katsuwonus pelamis) [21], zebra blenny (Salaria basilisca) [22], sardinelle (Sardinella aurita) [23], Atlantic bonito (Sarda sarda) [24], tambaqui (Colossoma macropomum) [25], yellowfin tuna (Thunnus albacores) [26], beluga (Huso huso), and sevruga (Acipenser stellatus) [27]. Trypsin was used to hydrolyze gelatin from the skin of giant catfish, in which antioxidative peptides were generated [28].…”

Section: Introductionmentioning

confidence: 99%

Trypsin from Pyloric Caeca of Asian Seabass: Purification, Characterization, and Its Use in the Hydrolysis of Acid-Soluble Collagen

Patil

Baloch

Nile

et al. 2023

Foods

View full text Add to dashboard Cite

The study aimed to purify trypsin from the pyloric caeca of Asian seabass (Lates calcarifer), and investigate its proteolytic capability toward acid-soluble collagen (ASC) in comparison with commercial porcine trypsin (CPT). Trypsin was purified from pyloric caeca, a leftover from the evisceration process, via ammonium sulphate (40–60% saturation) precipitation, and a soybean trypsin inhibitor (SBTI)–Sepharose 4B column. A 18.5-fold purification and a yield of 15.2% were obtained. SDS-PAGE analysis confirmed a single band of trypsin with a molecular weight of 23.5 kDa. Purified trypsin also showed the single band in native-PAGE. The optimal pH and temperature of trypsin for BAPNA (the specific substrate for amidase) hydrolysis were 8.5 and 60 °C, respectively. The trypsin was stable within the pH range of 7.0–9.5 and temperature range of 25–55 °C. Protease inhibition study confirmed that the purified enzyme was trypsin. The purified trypsin had a Michaelis–Menten constant (Km) and catalytic constant (kcat) of 0.078 mM and 5.4 s−1, respectively, when BAPNA was used. For the hydrolysis of TAME (the specific substrate for esterase), the Km and Kcat were 0.09 mM and 4.8 s−1, respectively. Partially purified seabass trypsin (PPST) had a slightly lower hydrolysis capacity toward ASC than CPT, as evidenced by the lower degree of hydrolysis and protein degradation when the former was used. Both the α-chain and β-chain became more degraded as the hydrolysis time increased. Based on MALDI-TOP, peptides with MW of 2992-2970 Da were dominant in the hydrolysates. Therefore, seabass trypsin could be used in the production of hydrolyzed collagen. It could have economic importance to the market, by replacing some commercial proteases, which have religious constraints.

show abstract

“…Serine proteases are present in the intestines, pyloric caeca, and the pancreatic tissues of fish and shrimp and have optimal activity at alkaline pH (de Freitas‐Júnior et al., 2021). These enzymes (i.e., trypsin and chymotrypsin) belong to endopeptidases, and have a serine residue, an imidazole, and an aspartic carboxyl group in their catalytic site.…”

Section: Endogenous Proteases: Sources Enzyme Types and Molecular Cha...mentioning

confidence: 99%

Protein hydrolysates derived from aquaculture and marine byproducts through autolytic hydrolysis

Nikoo

Benjakul

Gavlighi

2022

Comp Rev Food Sci Food Safe

View full text Add to dashboard Cite

Autolysis technology has shown potential for protein hydrolysates production from marine and aquaculture byproducts. Viscera are a source of cheap proteolytic enzymes for producing protein hydrolysates from the whole fish or processing byproducts of the most valuable commercial species by applying autolysis technology. The use of autolysis allows economical production of protein hydrolysate and provides an opportunity to valorize downstream fish and shellfish processing byproducts at a lower cost. As a result, production and application of marine byproduct autolysates is increasing in the global protein hydrolysates market. Nevertheless, several restrictions occur with autolysis, including lipid and protein oxidation mediated by the heterogeneous composition of byproducts. The generally poor storage and handling of byproducts may increase the formation of undesirable metabolites during autolysis, which can be harmful. The formation of nitrogenous compounds (i.e., biogenic amines), loss of freshness, and process of autolysis in the byproducts could increase the rate of quality and safety loss and lead to more significant concern about the use of autolysates for human food applications. The current review focuses on the autolysis process, which is applied for the hydrolysis of aquaculture and marine discards to obtain peptides as functional or nutritive ingredients. It further addresses the latest findings on the mechanisms and factors contributing the deterioration of byproducts and possible ways to control oxidation and other food quality and safety issues in raw materials and protein hydrolysates.

show abstract

Substrate specificity, physicochemical and kinetic properties of a trypsin from the giant Amazonian fish pirarucu (Arapaima gigas)

Cited by 6 publications

References 34 publications

Physicochemical and Biochemical Properties of Trypsin-like Enzyme from Two Sturgeon Species

Physicochemical and Biochemical Properties of Trypsin-like Enzyme from Two Sturgeon Species

Trypsin from Pyloric Caeca of Asian Seabass: Purification, Characterization, and Its Use in the Hydrolysis of Acid-Soluble Collagen

Protein hydrolysates derived from aquaculture and marine byproducts through autolytic hydrolysis

Contact Info

Product

Resources

About