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Cold-Adapted Digestive Aspartic Protease of the Clawed Lobsters Homarus americanus and Homarus gammarus: Biochemical Characterization

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Abstract

Aspartic proteinases in the gastric fluid of clawed lobsters Homarus americanus and Homarus gammarus were isolated to homogeneity by single-step pepstatin-A affinity chromatography; such enzymes have been previously identified as cathepsin D-like enzymes based on their deduced amino acid sequence. Here, we describe their biochemical characteristics; the properties of the lobster enzymes were compared with those of its homolog, bovine cathepsin D, and found to be unique in a number of ways. The lobster enzymes demonstrated hydrolytic activity against synthetic and natural substrates at a wider range of pH; they were more temperature-sensitive, showed no changes in the K M value at 4°C, 10°C, and 25°C, and had 20-fold higher k cat /K M values than bovine enzyme. The bovine enzyme was temperature-dependent. We propose that both properties arose from an increase in molecular flexibility required to compensate for the reduction of reaction rates at low habitat temperatures. This is supported by the fast denaturation rates induced by temperature.

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Acknowledgments

We thank Julio Cordoba and Patricia Hernandez for technical guidance. Ira Fogel provided editorial comments. The project was funded by Consejo Nacional de Ciencia y Tecnología (CONACYT grant: 80935). L. R. was a recipient of a CONACYT doctoral fellowship (#200577).

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  1. Centro de Investigaciones Biológicas del Noroeste (CIBNOR), Mar Bermejo 195, Col. Playa Palo de Santa Rita, La Paz, B.C.S., 23096, Mexico

    Liliana Rojo, Fernando García-Carreño & Maria de los Angeles Navarrete del Toro

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  1. Liliana Rojo
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  2. Fernando García-Carreño
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  3. Maria de los Angeles Navarrete del Toro
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Correspondence to Fernando García-Carreño.

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Rojo, L., García-Carreño, F. & de los Angeles Navarrete del Toro, M. Cold-Adapted Digestive Aspartic Protease of the Clawed Lobsters Homarus americanus and Homarus gammarus: Biochemical Characterization. Mar Biotechnol 15, 87–96 (2013). https://doi.org/10.1007/s10126-012-9461-4

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