Expression of the Lantibiotic Mersacidin in Bacillus amyloliquefaciens FZB42

Abstract

Lantibiotics are small peptide antibiotics that contain the characteristic thioether amino acids lanthionine and methyllanthionine. As ribosomally synthesized peptides, lantibiotics possess biosynthetic gene clusters which contain the structural gene (lanA) as well as the other genes which are involved in lantibiotic modification (lanM, lanB, lanC, lanP), regulation (lanR, lanK), export (lanT(P)) and immunity (lanEFG). The lantibiotic mersacidin is produced by Bacillus sp. HIL Y-85,54728, which is not naturally competent.Methodology/Principal FindingsThe aim of these studies was to test if the production of mersacidin could be transferred to a naturally competent Bacillus strain employing genomic DNA of the producer strain. Bacillus amyloliquefaciens FZB42 was chosen for these experiments because it already harbors the mersacidin immunity genes. After transfer of the biosynthetic part of the gene cluster by competence transformation, production of active mersacidin was obtained from a plasmid in trans. Furthermore, comparison of several DNA sequences and biochemical testing of B. amyloliquefaciens FZB42 and B. sp. HIL Y-85,54728 showed that the producer strain of mersacidin is a member of the species B. amyloliquefaciens.Conclusions/SignificanceThe lantibiotic mersacidin can be produced in B. amyloliquefaciens FZB42, which is closely related to the wild type producer strain of mersacidin. The new mersacidin producer strain enables us to use the full potential of the biosynthetic gene cluster for genetic manipulation and downstream modification approaches.

Class I and Class II Lanthipeptides Produced by Bacillus spp.

“…After MS/ MS analysis, it was predicted that the 2886.3 peptide, referred to as thuricin 4A-4D, is a derivative of thuricin 4A-4 wherein the side chain of Lys7 or Lys10 reacted to an unknown 118 Da substance (Figure 3). The complete thi gene cluster was heterologously expressed in B. thuringiensis BMB171, and both thuricin 4A-4 and thuricin 4A-4D were detected in the supernatant of the transformed strain (designated BMB1661 69 The gene cluster of mersacidin ( Figure 4) contains 10 open reading frames. In addition to the structural gene mrsA, it possesses genes encoding the enzymes required for post-translational modifications of the peptide (mrsD and mrsM), a gene that encodes a bifunctional protein with a transporter and protease domain (mrsT), three immunity genes (mrsF, mrsG, and mrsE), and genes involved in the regulation of the biosynthesis (mrsK2, mrsR2, and mrsR1).…”

“…88 Such medium has often been referred to as mersacidin production medium or 2X BPM. 19,69 Amylolysin. Amylolysin was described by Halimi et al in 2010.…”

BAKTERI ENDOFIT KULIT BATANG TANAMAN CENGKEH (Syzygium aromaticum L.) DAN KEMAMPUANNYA SEBAGAI ANTIBAKTERI

“…Jarak genetik (genetic distance) isolat bakteri endofit kulit batang tanaman cengkeh terhadap bakteri referensi yang diakses dari GenBank. (Herzner et al, 2011).…”

First complete genome sequence of Bacillus glycinifermentans B-27

“…In addition to complete clusters, two partial lantibiotic clusters that might be associated to Mersacidin and Subtilin as well as a partial cluster related to Neotrehalosadiamine were detected. Orthologous genes of the partial gene clusters for Mersacidin and Subtilin were described to be involved in immunity functions of related strains (Herzner et al, 2011;Stein et al, 2005) suggesting that B. glycinifermentans B-27 might also protect itself against these metabolites while not synthesizing them on its own. Genomic features and locations of secondary metabolite clusters were visualized in Figure 3 and summarized in Table 2.…”