Separate line and expression vector design Naga in Eukaryotes

Abstract

N-acetyl glucosamine-6-phosphate (GIcNAc6P) deacetylase is an enzyme of amino sugercatabolic pathway that catalyzes the conversion of the GlcNAc6P into glucosamine 6-phosphate (GIcN6P). Depending on the sequence similarities, NagA were divided into 13 families. Theenzyme was found in various species such as Escherichia coli, Bacillus, mouse, and human. Event not similarities in nucleotide lenghs (1.2 - 1.8 bp) and function in own body, but theyshare bioactive property that catalyze convertion of GlcNAc6P into GIcN6P. In this study, the NagA from Escherichia coli have been cloned and the expression vector (pESC-HISINagA) havebeen designed in order to express NagA in Saccharomyces ceriviceae. In general, NagA in vivo,do expression with moderate concentration for the growth and not overexpress if we do notintervene one or more steps in this pathway. Therefore, we can not obtain GlcN6P byfermentation of bacteria in general, but we can produce it by recombinant techniques. The nucleotide sequence of the clone showed the presence of an ORF (nagA gene) lacking introns,with a length of 1149 bp, and coding for a protein of 42 kDa, showing similarity to acetylglucosaminidases.