Cloning and molecular characterization of the betaine aldehyde dehydrogenase involved in the biosynthesis of glycine betaine in white shrimp ( Litopenaeus vannamei )

• DOI: 10.1016/j.cbi.2017.02.006
• Publication Date: 2017-02-15T16:01:27Z
• Authors: María F. Delgado-...; Jesús A. Rosas-Ro...; Gloria Yepiz-Plas...; Ciria G. Figueroa...; Elisa M. Valenzue...
• Keywords: Models, Molecular; 0303 health sciences; Binding Sites; Amino Acid Motifs; Molecular Sequence Data; Betaine-Aldehyde Dehydrogenase; Recombinant Proteins; Protein Structure, Tertiary; Betaine; 03 medical and health sciences; Penaeidae; Biocatalysis; Animals; Humans; Amino Acid Sequence; Cloning, Molecular; Sequence Alignment; Phylogeny

The enzyme betaine aldehyde dehydrogenase (BADH) catalyzes the irreversible oxidation of betaine aldehyde to glycine betaine (GB), a very efficient osmolyte accumulated during osmotic stress. In this study, we determined the nucleotide sequence of the cDNA for the BADH from the white shrimp Litopenaeus vannamei (LvBADH). The cDNA was 1882 bp long, with a complete open reading frame of 1524 bp, encoding 507 amino acids with a predicted molecular mass of 54.15 kDa and a pI of 5.4. The predicted LvBADH amino acid sequence shares a high degree of identity with marine invertebrate BADHs. Catalytic residues (C-298, E-264 and N-167) and the decapeptide VTLELGGKSP involved in nucleotide binding and highly conserved in BADHs were identified in the amino acid sequence. Phylogenetic analyses classified LvBADH in a clade that includes ALDH9 sequences from marine invertebrates. Molecular modeling of LvBADH revealed that the protein has amino acid residues and sequence motifs essential for the function of the ALDH9 family of enzymes. LvBADH modeling showed three potential monovalent cation binding sites, one site is located in an intra-subunit cavity; other in an inter-subunit cavity and a third in a central-cavity of the protein. The results show that LvBADH shares a high degree of identity with BADH sequences from marine invertebrates and enzymes that belong to the ALDH9 family. Our findings suggest that the LvBADH has molecular mechanisms of regulation similar to those of other BADHs belonging to the ALDH9 family, and that BADH might be playing a role in the osmoregulation capacity of L. vannamei.