Pepsinogen and pepsin from the stomach of smooth hound (Mustelus mustelus): Purification, characterization and amino acid terminal sequences

Bougatef, Ali; Balti, Rafik; Zaied, Saïda Ben; Souissi, Nabil; Nasri, Monçef

doi:10.1016/j.foodchem.2007.08.077

Cited by 55 publications

(68 citation statements)

References 38 publications

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“…The ATPS method gave much higher SA, PF and RY, compared to those of the ASF. There is no literature on the comparison of two methods, but the ASF using similar saturations for PG purification gave a SA of ~ 0.88-3.0 U, PF of ~ 1.1-3.7 and RY of ~ 64-75% [31,32,34,37]. The values of SA, PF and RY obtained in the present study were in these ranges.…”

Section: Comparing Atps and Asf Methodssupporting

confidence: 58%

“…Zhou et al [31] homogenized stomach samples using a homogenizer with phosphate buffer, centrifuged the homogenate to collect the crude enzymes from sea bream, and reported decreases in the A E , C P and RY of 35%, 55% and 36%, and increases in the SA and PF of 42% and 40% during extraction steps, respectively. Bougatef et al [34] homogenized stomach samples of smooth hound (Mustelus mustelus), using a homogenizer with Tris buffer, centrifuged the homogenate to collect the crude enzymes, and reported A E , C P and RY decreases of 29%, 81% and 40%, and SA and PF increases of 272% and 271% during extraction steps, respectively.…”

Section: Extraction Of Crude Pepsinogenmentioning

confidence: 99%

“…Pepsin, as well as its zymogen, pepsinogen (PG), has been widely purified from several fish species, including arctic fish capelin (Mallotus villosus) [27], rainbow trout (Salmo gairdneri) [28], Atlantic cod (Gadus morhua) [29], bolti fish (Tilapia nilotica) [7], Antarctic rock cod (Trematomus bernacchii) [30], sea bream (Sparuslatus houttuyn) [31], African coelacanth (Latimeria chalumnae) [32], Mandarin fish (Siniperca chuatsi) [33], smooth hound (Mustelus mustelus) [34], orange-spotted grouper (Epinephelus coioides) [35], albacore tuna (Thunnus alalunga) [36], and European eel (Anguilla anguilla) [37]. During the purification of proteases, several conventional techniques such as Ammonium Sulfate Fractionation (ASF), Gel Filtration Chromatography (GFC) and ion exchange chromatography (IEC), are frequently performed.…”

Section: Introductionmentioning

confidence: 99%

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Partition of Pepsinogen from the Stomach of Red Perch (Sebastes marinus) by Aqueous Two Phase Systems: Effects of PEG Molecular Weight and Concentration

Zhao¹,

Budge²,

Ghaly³

et al. 2013

Enz Eng

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Fish processing waste can be used to produce commercially valuable by-products, such as pepsinogen, which has application in food, manufacturing industries, collagen extraction, gelatin extraction, and in regulating digestibility. An important acidic protease, pepsin, is synthesized and secreted in the gastric membrane in an inactive state called pepsinogen (PG). In the present study, the purification of pepsinogen from the stomach of red perch, using aqueous two phase systems (ATPS) formed by polyethylene glycol (PEG) and salt at 4°C, was optimized.The effects of PEG molecular weight (PEG 1000, 1500, 3000 and 4000) and concentration (16, 18, 20, 22 and 24%) on the partitioning of PG were studied, and parameters including total volume (TV), volume ratio (V R ), total enzyme activity (A E ), protein content (C p ), specific enzyme activity (SA), partition coefficient (K p ), purification fold (PF), and recovery yield (RY) were evaluated. PEG molecular weight and PEG concentration also had significant effects on each parameter. TV and V R decreased with increased salt concentration, since salt formed hydrogen bonds with water molecules and formed a more compact and ordered water structure. PG partitioned predominantly in the PEG-rich top phase due to its negative charge. A E , C P , SA, PF and RY increased with increased salt concentration and then decreased, while K P had an opposite pattern. The PEG 3000 (20%), PEG 1000 (24%), PEG 4000 (16%) and PEG 1000 (18%) concentrations gave the highest TV, V R , C P and K P , respectively. PEG 1500 with 18% concentration gave the highest A E, SA, PF and RY (86.2%) . As PEG 1500 at 18% concentration gave the highest RY (86.2%). It was selected as the optimum PEG molecular weight and PEG concentration. (NH 4 ) 2 SO 4 at 15%, which gave the highest RY (71.7%), was selected as the optimum salt type and salt concentration. 15% (NH 4 ) 2 SO 4 18% PEG 1500 was the optimal ATPS combination, and presented a better partition. The values of SA and PF and RY obtained with ATPS method were much higher (2 fold in case of SA and PF, and 1.2 fold in case of RY), than those obtained with the Ammonium Sulphate Fractionation (ASF) method.

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Section: Comparing Atps and Asf Methodssupporting

confidence: 58%

Section: Extraction Of Crude Pepsinogenmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Partition of Pepsinogen from the Stomach of Red Perch (Sebastes marinus) by Aqueous Two Phase Systems: Effects of PEG Molecular Weight and Concentration

Zhao¹,

Budge²,

Ghaly³

et al. 2013

Enz Eng

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show abstract

“…The main inhibitors and activators of pepsin are shown in table 2. The enzymatic activity of pepsin is almost completely inhibited by pepstatin A (aspartic proteinase competitive inhibitor that binds strongly to residues of the active site of the enzyme) and partially inhibited by phenyl methyl sulfonyl fluoride (PMSF, serinoproteinase inhibitor), L-3-carboxytrans-2, 3-epoxy-propionyl-L-leucyl-4-guanidino-butylamide (E-64), cysteine, molybdate and EDTA (Ethylenediamine tetra acetic acid, specific for metalloproteases) (DIAZ-LOPEZ et al, 1998;CASTILLO-YAÑEZ et al, 2004;TANJI et al, 2007;BOUGATEF et al, 2008;KLOMKLAO et al, 2007;NALINANON et al, 2008;NALINANON et al, 2010;WU et al, 2009;CAO et al, 2011;ZHAO et al, 2011). In contrast, the activity increases in the presence of divalent cations such as CaCl 2 , MgCl 2 and CoCl 2 (El-BELTAGY et al, 2004;KLOMKLAO et al, 2007).…”

Section: Factors Affecting the Activity (Ph Temperature Activators mentioning

confidence: 99%

“…Temperature and proper pH are essential during the extraction process. The temperature of extraction should be between 0-4°C to prevent protein denaturation and autolysis enzymes (DIAZ-LOPEZ et al, 1998;BOUGATEF et al, 2008;NALINANON et al, 2008;NALINANON et al, 2010;ZHAO et al, 2011). …”

Section: Factors Affecting the Activity (Ph Temperature Activators mentioning

confidence: 99%

Fish pepsin: basic characteristics, extraction, determination and biotechnological applications

2015

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Enzyme technology has been employed in various segments of the industry, food production and consumer goods. Fish viscera are potential sources of proteins with enzymatic properties appreciable in this market. Among them, pepsin (EC 3.4.23.x) deserves special attention. The active enzyme has at least five isoforms which are precursor groups of pepsins type: A, B, and F, gastricsin and chymosin. Their zymogen (pepsinogen) is activated by autocatalysis at acidic pH in stomach. Hemoglobin is their specific substrate. Features like optimal pH and temperature vary according to species and even within the same species may be found different values. The molecular weight also varies according to species and is generally between 25 and 35 kDa. The protocols for extraction and determination of pepsin fish follow a line of standardization, varying concentrations of analysis. These data are basic steps to detect the presence of the enzyme targeting a future purification process for its effective biotechnological applications. KEYWORDS:Digestive; Enzyme; Fish; Pepsin; Biotechnology. PEPSINA DE PEIXES: CARACTERÍSTICAS BÁSICAS, EXTRAÇÃO, DETERMINAÇÃO E APLICAÇÕES BIOTECNOLÓGICAS RESUMOTecnologia enzimática vem sendo empregada em diversos segmentos da indústria, da produção de alimentos e bens de consumo. Vísceras de peixes são potenciais fontes de proteínas com propriedades enzimáticas apreciáveis neste mercado. Entre eles, a pepsina (EC 3.4.23.x) merece atenção especial. A enzima ativa tem, pelo menos, cinco isoformas, sendo elas precursoras do tipo pepsinas: A, B e F, gastricsina e quimosina. O zimogênio (pepsinogênio) é ativado por autocatálise em pH ácido do estômago que vem do ácido clorídrico. A hemoglobina é seu substrato específico. Características como temperatura e pH ótimos variam de acordo com as espécies e até mesmo dentro da mesma espécie podem ser encontrados valores diferentes. O peso molecular varia também de acordo com a espécie e situa-se geralmente, entre 25 e 35 kDa. Os protocolos de extração e de determinação de pepsina de peixe seguem uma linha de normalização, com várias concentrações de análise. Estes dados são passos básicos para detectar a presença da enzima destinada a um futuro processo de purificação para aplicações biotecnológicas eficazes.

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Seafood Enzymes: Biochemical Properties and Their Impact on Quality

Klomklao

Benjakul

Simpson

2012

Food Biochemistry and Food Processing

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Pepsinogen and pepsin from the stomach of smooth hound (Mustelus mustelus): Purification, characterization and amino acid terminal sequences

Cited by 55 publications

References 38 publications

Partition of Pepsinogen from the Stomach of Red Perch (Sebastes marinus) by Aqueous Two Phase Systems: Effects of PEG Molecular Weight and Concentration

Partition of Pepsinogen from the Stomach of Red Perch (Sebastes marinus) by Aqueous Two Phase Systems: Effects of PEG Molecular Weight and Concentration

Fish pepsin: basic characteristics, extraction, determination and biotechnological applications

Seafood Enzymes: Biochemical Properties and Their Impact on Quality

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