A hydrolase from Lactobacillus sakei moonlights as a transaminase.
Enzymatic transamination of amino acids yields α-keto acids and is the initial step for the production of volatile compounds that contribute to the sensory perception of fermented foods such as salami. Lactobacillus sakei is one of the lactic acid bacterial strains commonly used in starter cultures. Although the genome sequence of L. sakei 23K lacks genes encoding typical branched-chain amino acid transaminases, transamination activity and the formation of amino acid-derived volatile metabolites could be demonstrated. A protein purified from L. sakei is held responsible for the transamination activity. By heterologous expression of the corresponding gene in Escherichia coli, we were able to characterize the transamination side activity of an enzyme annotated as a putative acylphosphatase (AcP). A transamination side activity of hen egg white lysozyme (HEWL) was also discovered. Both enzymes showed substrate specificity toward branched-chain and aromatic amino acids. AcP also accepted l-methionine. Activity was optimal at neutral pH for both enzymes, whereas AcP showed a significantly higher temperature optimum (55°C) than that of HEWL (37°C). Kinetic parameters revealed high affinity toward l-leucine for AcP (K(m) = 1.85 mM) and toward l-isoleucine for HEWL (K(m) = 3.79 mM). AcP seems to play a major role in the metabolism of amino acids in L. sakei.
Biotechnology of Natural Products, Technische Universität München, Freising, Germany.
Quirin
Q
Sinz
Simone
S
Freiding
Rudi F
RF
Vogel
Wilfried
W
Schwab
D017766:Acid Anhydride Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007778:Lactobacillus / Q000201:enzymology* / Q000235:genetics; D007930:Leucine / Q000378:metabolism; D009113:Muramidase / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature; D000637:Transaminases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000097024:Acylphosphatase
D017766:Acid Anhydride Hydrolases; D007930:Leucine; D009113:Muramidase; D000637:Transaminases; D000097024:Acylphosphatase; C421864:hen egg lysozyme