Biochemical characterization of recombinant Penaeus vannamei trypsinogen

• Published in: Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 2019-12, Vol.238, p.110337-110337, Article 110337
• Main Authors: Guerrero-Olazarán, Martha, Castillo-Galván, Mauricio, Gallegos-López, Juan Antonio, Fuentes-Garibay, José Antonio, Viader-Salvadó, José María
• Format: Article
• Language: English
• Subjects: Penaeus vannamei; Serine proteases; Trypsinogen autoactivation; Zymogen autoactivation
• ISSN: 1096-4959; 1879-1107
• DOI: 10.1016/j.cbpb.2019.110337

Trypsinogens are the inactive precursors of trypsins (EC 3.4.21.4), which are digestive serine proteases. Despite knowing the properties of trypsins from Pacific white shrimp, Penaeus vannamei, the biochemical properties of shrimp trypsinogens including activation mechanisms and kinetics are unknown, due to difficulties isolating them from natural sources. In the present work, we describe the purification and biochemical characterization of four trypsinogen-like isoforms from recombinant P. vannamei trypsinogen, with a special emphasis on understanding its activation kinetics. The major trypsinogen-like isoform had an apparent molecular mass of 29 kDa. The other three forms of recombinant trypsinogen were: an N-glycosylated form of 32 kDa, a possibly O-glycosylated form of 41 kDa, and a likely double-chain form with a subunit of 23 kDa. The autoactivation profile of three-recombinant trypsinogen-like isoforms showed increased trypsin activity at a rate that was higher than that of bovine trypsinogen. This confirms the hypothesis proposed in the literature of a rapid trypsinogen autoactivation in the absence of aspartates in the activation peptide as it is for P. vannamei trypsinogen. [Display omitted] •Four trypsinogen isoforms from recombinant shrimp trypsinogen are characterized.•The major trypsinogen-like isoform has an apparent molecular mass of 29 kDa.•One of the trypsinogen isoforms is a double-stranded form with a subunit of 23 kDa.•Autoactivation kinetics profiles of P. vannamei trypsinogen isoforms are described.•Rapid autoactivation hypothesis in the absence of D in the pro region is confirmed.