title: One-pot synthesis of class II lanthipeptide bovicin HJ50 via an engineered lanthipeptide synthetase.
abstract: Lanthipeptides are a large class of bacteria-produced, ribosomally-synthesized and post-translationally modified peptides. They are recognized as peptide antibiotics because most of them exhibit potent antimicrobial activities against Gram-positive bacteria especially those that are phylogenetically related to producers. Maturation of class II lanthipeptide like bovicin HJ50 undergoes precursor modification by LanM and a subsequent leader peptide cleavage by LanT. Herein, via co-expression of precursor gene bovA, modification gene bovM and transporter gene bovT in Escherichia coli C43 (DE3), bioactive bovicin HJ50 was successfully produced and secreted. To further achieve in vitro one-pot synthesis of bovicin HJ50, an engineered bovicin HJ50 synthetase BovT150M was obtained by fusing the peptidase domain of BovT (BovT150) to the N-terminus of BovM. BovT150M exhibited dual functions of precursor modification and leader peptide cleavage to release mature bovicin HJ50. Under the guidance of BovA leader peptide, BovT150M exhibited substrate tolerance to modify non-native substrates including suicin and lacticin 481. This work exemplifies the feasibility of enzyme chimera of peptidase domain (LanT150) and modification enzyme (LanM) as a one-pot lanthipeptide synthetase.
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, P. R. China.|University of Chinese Academy of Sciences, Beijing, 100039, P. R. China.
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, P. R. China.|University of Chinese Academy of Sciences, Beijing, 100039, P. R. China.
authors.forename: Xiaoxuan
authors.initials: X
authors.lastname: Ge
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, P. R. China.|University of Chinese Academy of Sciences, Beijing, 100039, P. R. China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, P. R. China.
authors.forename: Kunling
authors.initials: K
authors.lastname: Teng
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, P. R. China.|University of Chinese Academy of Sciences, Beijing, 100039, P. R. China.
authors.forename: Jin
authors.initials: J
authors.lastname: Zhong
mesh_terms: D000595:Amino Acid Sequence; D001430:Bacteriocins / Q000138:chemical synthesis* / Q000737:chemistry / Q000235:genetics; D002384:Catalysis; D060326:Chemistry Techniques, Synthetic; D004252:DNA Mutational Analysis; D004789:Enzyme Activation; D008958:Models, Molecular; D009154:Mutation; D010453:Peptide Synthases / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation
chemical_list: D001430:Bacteriocins; C000591653:bovicin HJ50, Streptococcus bovis; D010453:Peptide Synthases