A newly identified enzyme from Japanese common squid Todarodes pacificus has the ability to biosynthesize d-aspartate.

Amino acids exist in two chiral forms, namely L and D. Although l -amino acids are predominant in vivo , certain limited circumstances have reported the usage of d -amino acids. d -aspartate (Asp), among them, plays crucial physiological roles in living organisms and is biosynthesized from L-Asp by the enzyme named aspartate racemase (AspRase). D-Asp is known to accumulate in large amounts in the nervous system of cephalopods. To understand the function of D-Asp in nervous system in more detail, it is necessary to elucidate its metabolic pathway; however, AspRase gene has not been identified in cephalopods as in the case of mammals. In this study, we successfully identified a novel gene encoding AspRase from the optic ganglion of Japanese common squid Todarodes pacificus. Our discovery of the squid AspRase challenges the prevailing assumption that AspRases across different animals share similar structures. Surprisingly, the squid AspRase is a unique enzyme that differs significantly from known AspRases, being structurally and phylogenetically related to aspartate aminotransferase (AST) and possessing both AspRase and AST activities. The optimum pH and temperature for AspRase activity using L-Asp as a substrate are approximately 7.0 and 20 °C, respectively. Moreover, we have found that AspRase activity is enhanced in the presence of 2-oxoacids. These findings have far-reaching implications for the understanding of enzymology and suggest that yet-to-be-identified mammalian AspRases may also be phylogenetically related to AST, rather than conventional AspRases. Furthermore, our results provide valuable insights into the evolution of the D-Asp biosynthetic pathway. [Display omitted] • AspRase is an enzyme with many unknowns involved in the biosynthesis of D-Asp. • We have cloned the AspRase gene from the Japanese common squid for the first time in cephalopods. • Squid AspRase is a unique and novel enzyme that possesses both AspRase and AST activities. • Squid AspRase is phylogenetically related to AST, distinct from AspRases from other animals. • Our findings will help elucidate the D-Asp biosynthesis pathway not only in cephalopods, but also in mammals. [ABSTRACT FROM AUTHOR]