Premium
Probing the acyl carrier protein–Enzyme interactions within terminal alkyne biosynthetic machinery
Author(s) -
Su Michael,
Zhu Xuejun,
Zhang Wenjun
Publication year - 2018
Publication title -
aiche journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.958
H-Index - 167
eISSN - 1547-5905
pISSN - 0001-1541
DOI - 10.1002/aic.16355
Subject(s) - alkyne , chemistry , biosynthesis , dna ligase , biochemistry , stereochemistry , chemical biology , enzyme , combinatorial chemistry , catalysis
The alkyne functionality has attracted much interest due to its diverse chemical and biological applications. We recently elucidated an acyl carrier protein (ACP)‐dependent alkyne biosynthetic pathway, however, little is known about ACP interactions with the alkyne biosynthetic enzymes, an acyl‐ACP ligase (JamA) and a membrane‐bound bifunctional desaturase/acetylenase (JamB). Here, we showed that JamB has a more stringent interaction with ACP than JamA. In addition, site‐directed mutagenesis of a non‐cognate ACP significantly improved its compatibility with JamB, suggesting a possible electrostatic interaction at the ACP‐JamB interface. Finally, error‐prone PCR and screening of a second non‐cognate ACP identified hot spots on the ACP that are important for interacting with JamB and yielded mutants which were better recognized by JamB. Our data thus not only provide insights into the ACP interactions in alkyne biosynthesis, but it also potentially aids in future combinatorial biosynthesis of alkyne‐tagged metabolites for chemical and biological applications. © 2018 American Institute of Chemical Engineers AIChE J , 64: 4255–4262, 2018