Isolation and properties of AMP deaminase from jumbo squid (Dosidicus gigas) mantle muscle from the Gulf of California, Mexico

E Marquez-Rios; R Pacheco-Aguilar; F J Castillo-Yañez; C G Figueroa-Soto; J M Ezquerra-Brauer; T Gollas-Galvan

doi:10.1016/j.foodchem.2008.01.059

Isolation and properties of AMP deaminase from jumbo squid (Dosidicus gigas) mantle muscle from the Gulf of California, Mexico

Food Chem. 2008 Sep 1;110(1):69-75. doi: 10.1016/j.foodchem.2008.01.059. Epub 2008 Feb 6.

Authors

E Marquez-Rios¹, R Pacheco-Aguilar², F J Castillo-Yañez³, C G Figueroa-Soto¹, J M Ezquerra-Brauer³, T Gollas-Galvan¹

Affiliations

¹ Centro de Investigación en Alimentación y Desarrollo, A.C. P.O. Box 1735, Hermosillo, Sonora 83000, Mexico.
² Centro de Investigación en Alimentación y Desarrollo, A.C. P.O. Box 1735, Hermosillo, Sonora 83000, Mexico. Electronic address: rpacheco@cascabel.ciad.mx.
³ Departamento de Ciencias Químico Biológicas, Universidad de Sonora, Encinas y Rosales s/n, Hermosillo, Sonora 83000, Mexico.

PMID: 26050167
DOI: 10.1016/j.foodchem.2008.01.059

Abstract

Adenosine monophosphate (AMP) deaminase was purified from jumbo squid mantle muscle by chromatography in cellulose phosphate, Q-Fast and 5'-AMP sepharose. Specific activity of 2.5U/mg protein, 4.5% recovery and 133.68 purification fold were obtained at the end of the experiment. SDS-PAGE showed a single band with 87kDa molecular mass, native PAGE proved a band of 178kDa, whereas gel filtration detected a 180kDa protein, suggesting the homodimeric nature of this enzyme, in which subunits are not linked by covalent forces. Isoelectric focusing of this enzyme showed a pI of 5.76, which agrees with pI values of AMP deaminase from other invertebrate organisms. AMP deaminase presented a kinetic sigmoidal plot with Vmax of 1.16μM/min/mg, Km of 13mM, Kcat of 3.48μM.s(-1) and a Kcat/Km of 267 (mol/L)(-1).s(-1). The apparent relative low catalytic activity of jumbo squid muscle AMP deaminase in the absence of positive effectors is similar to that reported for homologous enzymes in other invertebrate organisms.

Keywords: AMP deaminase; Jumbo squid mantle; Purification.