Identification of extracellular N-acylhomoserine lactone acylase from a Streptomyces sp. and its application to quorum quenching.
N-acylhomoserine lactones (AHLs) play an important role in regulating virulence factors in pathogenic bacteria. Recently, the enzymatic inactivation of AHLs, which can be used as antibacterial targets, has been identified in several soil bacteria. In this study, strain M664, identified as a Streptomyces sp., was found to secrete an AHL-degrading enzyme into a culture medium. The ahlM gene for AHL degradation from Streptomyces sp. strain M664 was cloned, expressed heterologously in Streptomyces lividans, and purified. The enzyme was found to be a heterodimeric protein with subunits of approximately 60 kDa and 23 kDa. A comparison of AhlM with known AHL-acylases, Ralstonia strain XJ12B AiiD and Pseudomonas aeruginosa PAO1 PvdQ, revealed 35% and 32% identities in the deduced amino acid sequences, respectively. However, AhlM was most similar to the cyclic lipopeptide acylase from Streptomyces sp. strain FERM BP-5809, exhibiting 93% identity. A mass spectrometry analysis demonstrated that AhlM hydrolyzed the amide bond of AHL, releasing homoserine lactone. AhlM exhibited a higher deacylation activity toward AHLs with long acyl chains rather than short acyl chains. Interestingly, AhlM was also found to be capable of degrading penicillin G by deacylation, showing that AhlM has a broad substrate specificity. The addition of AhlM to the growth medium reduced the accumulation of AHLs and decreased the production of virulence factors, including elastase, total protease, and LasA, in P. aeruginosa. Accordingly, these results suggest that AHL-acylase, AhlM could be effectively applied to the control of AHL-mediated pathogenicity.
Park
Sun-Yang
SY
R&D Center, INBIONET Corporation, Daejeon 305-390, Korea.
Kang
Hye-Ok
HO
Jang
Hak-Sun
HS
Lee
Jung-Kee
JK
Koo
Bon-Tag
BT
Yum
Do-Young
DY
D015107:4-Butyrolactone / Q000031:analogs & derivatives* / Q000378:metabolism*; D000581:Amidohydrolases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D011550:Pseudomonas aeruginosa / Q000187:drug effects; D015398:Signal Transduction; D013302:Streptomyces / Q000201:enzymology*; D013379:Substrate Specificity; D037521:Virulence Factors / Q000096:biosynthesis
D037521:Virulence Factors; C088386:homoserine lactone; D000581:Amidohydrolases; C019665:amidase; D015107:4-Butyrolactone