A chymotrypsin from the Digestive Tract of California Spiny Lobster, Panulirus interruptus: Purification and Biochemical Characterization

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• 作者：Betsaida Bibo-Verdugo ; Liliana Rojo-Arreola…
• 关键词：Decapod ; Panulirus interruptus ; Serine proteinase ; Chymotrypsin purification ; Biochemical characterization
• 刊名：Marine Biotechnology
• 出版年：2015
• 出版时间：August 2015
• 年：2015
• 卷：17
• 期：4
• 页码：416-427
• 全文大小：1,359 KB
• 参考文献：Balti R, Bougherra F, Bougatef A, Hayet BK, Nedjar-Arroume N, Dhulster P, Guillochon D, Nasri M (2012) Chymotrypsin from the hepatopancreas of cuttlefish (Sepia officinalis) with high activity in the hydrolysis of long chain peptide substrates: purification and biochemical characterisation. Food Chem 130:475-84View Article
  Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248-54PubMed View Article
  Castillo-Ya?ez FJ, Pacheco-Aguilar R, Garcia-Carre?o FL, Navarrete-Del Toro MDLA (2004) Characterization of acidic proteolytic enzymes from Monterey sardine (Sardinops sagax caerulea) viscera. Food Chem 85:343-50View Article
  Castillo-Yá?ez FJ, Pacheco-Aguilar R, García-Carre?o FL, Navarrete-Del Toro MA, Félix López M (2006) Purification and biochemical characterization of chymotrypsin from the viscera of Monterey sardine (Sardinops sagax caeruleus). Food Chem 99:252-59View Article
  Castillo-Ya?ez FJ, Pacheco-Aguilar R, Lugo-Sanchez ME, Garcia-Sanchez G, Quintero Reyes IE (2009) Biochemical characterization of an isoform of chymotrypsin from the viscera of Monterey sardine (Sardinops sagax caerulea), and comparison with bovine chymotrypsin. Food Chem 112:634-39View Article
  Celis-Guerrero LE, García-Carre?o FL, Navarrete Del Toro MA (2004) Characterization of proteases in the digestive system of spiny lobster (Panulirus interruptus). Mar Biotechnol 6:262-69PubMed View Article
  D’Amico S, Claverie P, Collins T, Georlette D, Gratia E, Hoyoux A, Meuwis M-A, Feller G, Gerday C (2002) Molecular basis of cold adaptation. Philos Trans R Soc Lond B Biol Sci 357:917-25PubMed Central PubMed View Article
  DelMar EG, Largman C, Brodrick JW, Geokas MC (1979) A sensitive new substrate for chymotrypsin. Anal Biochem 99:316-20PubMed View Article
  Di Cera E (2009) Serine proteases. IUBMB Life 61:510-15PubMed Central PubMed View Article
  Díaz-Tenorio LM, García-Carre?o FL, Navarrete del Toro MA (2006) Characterization and comparison of digestive proteinases of the Cortez swimming crab, Callinectes bellicosus, and the arched swimming crab, Callinectes arcuatus. Invertebr Biol 125:125-35View Article
  Eberhardt J (1992) Isolation and characterization of 5 serine proteases with trypsin-like, chymotrypsin-like and elastase-like characteristics from the gut of the lugworm Arenicola marina (L.) (Polychaeta). J Comp Physiol B 162:159-67View Article
  El Hadj AN, Hmidet N, Zouari-Fakhfakh N, Ben Khaled H, Nasri M (2010) Alkaline chymotrypsin from striped seabream (Lithognathus mormyrus) viscera: purification and characterization. J Agric Food Chem 58:9787-792View Article
  Fong WP, Chan EY, Lau KK (1998) Isolation of two chymotrypsins from grass carp. Biochem Mol Biol Int 45:409-18PubMed
  García-Carre?o F, Dimes L, Haard N (1993) Substrate-gel electrophoresis for composition of molecular weight of proteinases or protteinaceous proteinase inhibitors. Anal Biochem 214:65-9PubMed View Article
  Geok LP, Razak CNA, Abd Rahman RNZ, Basri M, Salleh AB (2003) Isolation and screening of an extracellular organic solvent-tolerant protease producer. Biochem Eng J 13:73-7View Article
  Gráf L, Szilágyi L, Venekei I (2013) Chymotrypsin. In: Rawlings ND, Salvesen G (eds) Handbook of Proteolytic Enzymes, 3rd edn. Academic Press, San Diego, p 2626-633
  Gupta MN (1992) Enzyme function in organic solvents. Eur J Biochem 203:25-2PubMed View Article
  Hedstrom L (2002) Serine protease mechanism and specificity. Chem Rev 102:4501-523PubMed View Article
  Hernández-Cortés P, Whitaker JR, García-Carre?o F (1997) Purification and characterization of chymotrypsin from Penaeus vannamei. J Food Biochem 21:497-14View Article
  Heu MS, Kim HR, Pyeun JH (1995) Comparison of trypsin and chymotrypsin from the viscera of anchovy, Engraulis japonica. Comp Biochem Physiol B: Biochem Mol Biol 112:557-67View Article
  Klomklao S (2008) Digestive proteinases from marine organisms and their applications. Songklanakarin J Sci Technol 30:37-6
  Kristjansson MM (1991) Purification and characterization of trypsin from the pyloric ceca of rainbow trout (Oncorhynchus mykiss). J Agric Food Chem 39:1738-742View Article
  Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-85PubMed View Article
  Le Chevalier P, Sellos D, Van Wormhoudt A (1995) Purification and partial characterization of chymotrypsin-like proteases from the digestive gland of the scallop Pecten maximus. Comp Biochem Physiol B Biochem Mol Biol 110:777-84View Article
  Li GY, Pukunaga S, Takenouchi K, Nakamura F (2005) Comparative study of the physiological properties of collagen, gelatin and collagen hydrolysate as cosmetic materials. Int J Cosmet Sci 27:101-06PubMed View Article
  Lima CA, Rodrigues PMB, Porto TS, Viana DA, Lima Fihlo JL, Porto ALF, Carn
• 作者单位：Betsaida Bibo-Verdugo (1)
  Liliana Rojo-Arreola (1)
  Maria A. Navarrete-del-Toro (1)
  Fernando García-Carre?o (1)
  
  1. Centro de Investigaciones Biológicas del Noroeste (CIBNOR), Calle IPN 195, La Paz, B.C.S. 23096, Mexico
  
• 刊物类别：Earth and Environmental Science
• 刊物主题：Earth sciences
  Oceanography
  
• 出版者：Springer New York
• ISSN：1436-2236

文摘

A chymotrypsin was purified from the gastric juice of California spiny lobster (Panulirus interrutpus), using preparative electrophoresis and affinity chromatography on agarose-p-aminobenzamidine. The molecular mass was estimated by polyacrylamide gel electrophoresis (SDS-PAGE) under denaturing conditions to be 28?kDa. Chymotrypsin activity was totally inhibited by phenylmethylsulfonyl fluoride (PMSF) and chymostatin. Lobster chymotrypsin had optimal pH?7.0-.0 and temperature of 55?°C. The enzyme is highly stable under a wide range of pH (retaining up to 80?% of activity after 1?h of incubation at pH?3.0, 5.0, and 12.0), showing higher stability at pH?8.0, and was inactivated after 20?min at 55?°C. Lobster chymotrypsin was able to hydrolyze protein substrates at as low as pH?3.0. These results are consistent with the findings of enzyme stability. Activity was assessed after incubation of enzyme with different organic solvents (in the range of 10-0?%); when tested in the presence of acetone, ethanol, propanol, and butanol, lobster chymotrypsin residual activity was >80?%; whereas in the presence of dimethyl sulfoxide (DMSO) and toluene, lobster chymotrypsin residual activity was <80?%. Deduced amino acid sequence, corroborated by mass spectrometry, was determined.