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Identification of two secondary acyltransferases of lipid A in Pseudomonas putida KT 2442
Author(s) -
Zhu L.,
Li Y.,
Wang J.,
Wang X.
Publication year - 2017
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.13499
Subject(s) - pseudomonas putida , acyltransferases , acyltransferase , biochemistry , escherichia coli , biology , chemistry , electrospray ionization , biosynthesis , enzyme , gene , mass spectrometry , chromatography
Aims Identify the secondary acyltransferases of lipid A in Pseudomonas putida . Methods and Results Two homologues of Escherichia coli acyltransferase LpxL, encoded by PP _0063 and PP _1735 , exist in P. putida . Pseudomonas putida mutant strains KWZ 001 and KWZ 002 were constructed by deleting the genes PP _0063 and PP _1735 respectively. Lipid A species were isolated from P. putida KT 2442, KWZ 001 and KWZ 002, respectively, and analysed by using electrospray ionization/mass spectrometry (ESI/MS). The results suggest that both PP _0063 and PP _1735 encode secondary acyltransferase of lipid A in P. putida . To further study the site‐specificity of these two acyltransferases, PP _0063 , PP _1735 and Escherichia coli lpxL were overexpressed in KWZ 001 and KWZ 002 respectively. Lipid A species isolated from these recombinant strains were analysed by using ESI/MS, and the results suggest that the acyltransferase encoded by PP _0063 catalyses the addition of the 2‐ OH ‐C 12:0 chain at the 2‐position and the acyltransferase encoded by PP _1735 catalyses the addition of the C 12:0 chain to the 2′‐position of lipid A in P. putida . Conclusion The two acyltransferases encoded by PP _0063 and PP _1735 , respectively, are responsible for the site‐specific additions of the two secondary acyl chains at the 2‐ and 2′‐positions of lipid A in P. putida . Significance and Impact of the Study Understanding the lipid A structure variation in P. putida might provide new clues for the survival of P. putida under various stress conditions.