L-amino acid oxidase as biocatalyst: a dream too far?
L-amino acid oxidase (LAAO) is a flavoenzyme containing non-covalently bound flavin adenine dinucleotide, which catalyzes the stereospecific oxidative deamination of l-amino acids to α-keto acids and also produces ammonia and hydrogen peroxide via an imino acid intermediate. LAAOs purified from snake venoms are the best-studied members of this family of enzymes, although a number of LAAOs from bacterial and fungal sources have been also reported. From a biochemical point of view, LAAOs from different sources are distinguished by molecular mass, substrate specificity, post-translational modifications and regulation. In analogy to the well-known biotechnological applications of d-amino acid oxidase, important results are expected from the availability of suitable LAAOs; however, these expectations have not been fulfilled yet because none of the "true" LAAOs has successfully been expressed as a recombinant protein in prokaryotic hosts, such as Escherichia coli. In enzyme biotechnology, recombinant production of a protein is mandatory both for the production of large amounts of the catalyst and to improve its biochemical properties by protein engineering. As an alternative, flavoenzymes active on specific l-amino acids have been identified, e.g., l-aspartate oxidase, l-lysine oxidase, l-phenylalanine oxidase, etc. According to presently available information, amino acid oxidases with "narrow" or "strict" substrate specificity represent as good candidates to obtain an enzyme more suitable for biotechnological applications by enlarging their substrate specificity by means of protein engineering.
Loredano
L
Pollegioni
Paolo
P
Motta
Gianluca
G
Molla
D000596:Amino Acids / Q000378:metabolism*; D000641:Ammonia / Q000378:metabolism; D001419:Bacteria / Q000201:enzymology; D001709:Biotechnology / Q000379:methods; D003001:Cloning, Molecular; D005658:Fungi / Q000201:enzymology; D015870:Gene Expression; D006861:Hydrogen Peroxide / Q000378:metabolism; D007651:Keto Acids / Q000378:metabolism*; D050830:L-Amino Acid Oxidase / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D015202:Protein Engineering; D012910:Snake Venoms / Q000201:enzymology; D013379:Substrate Specificity
D000596:Amino Acids; D007651:Keto Acids; D012910:Snake Venoms; D000641:Ammonia; D006861:Hydrogen Peroxide; D050830:L-Amino Acid Oxidase