title: The midgut trypsins of shrimp (Penaeus monodon). High efficiency toward native protein substrates including collagens.
abstract: Two shrimp trypsins have been purified from the midguts of Penaeid shrimps by various chromatographies and HPLC. The molecular masses of them are 27 and 29 kDa, respectively. They show the typical specificity of trypsin for substrates and inhibitors, and their N-terminal amino-acid sequences are homologous to those of other trypsins. The shrimp enzymes are very acidic (pI less than or equal to 2.4), and show distinctively low Km for the synthetic amide substrates. They also hydrolyse various native proteins more efficiently than bovine trypsin in vitro. However, the anionic shrimp trypsins do not have special preference for basic protein substrates over the acidic one. Collagenolytic activity of the midgut extract was mainly due to serine proteases. The collagenolytic activity of the purified shrimp trypsin was confirmed by assays with either soluble or insoluble native type I collagens. In comparison with the other trypsins from the Crustacean decapods, the shrimp enzymes have four pairs of disulfide bonds, intermediary between the crayfish trypsin (three pairs) and the crab trypsin (five pairs), and are immunochemically different from them.
authors.affiliation: Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan, Republic of China.
authors.forename: P J
authors.initials: PJ
authors.lastname: Lu
authors.forename: H C
authors.initials: HC
authors.lastname: Liu
authors.forename: I H
authors.initials: IH
authors.lastname: Tsai
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003386:Brachyura / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid; D003094:Collagen / Q000378:metabolism; D004064:Digestive System / Q000737:chemistry / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D007700:Kinetics; D003012:Microbial Collagenase / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D033561:Penaeidae / Q000378:metabolism*; D012967:Sodium Dodecyl Sulfate; D013379:Substrate Specificity; D014357:Trypsin / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D014361:Trypsin Inhibitors / Q000378:metabolism
chemical_list: D014361:Trypsin Inhibitors; D012967:Sodium Dodecyl Sulfate; D003094:Collagen; D014357:Trypsin; D003012:Microbial Collagenase