Structural Analysis of Class I Lanthipeptides from <i>Pedobacter lusitanus</i> NL19 Reveals an Unusual Ring Pattern

Bothwell, Ian R.; Caetano, Tânia; Sarksian, Raymond; Mendo, Sónia; Donk, Wilfred A. van der

doi:10.1021/acschembio.1c00106

Cited by 33 publications

(62 citation statements)

References 87 publications

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“… 9 More recent additional examples of the ll -(Me)Lan stereochemistry were identified in class I lanthipeptides where the conserved Dhb-Dhx-Xxx-Xxx-Cys motif was absent suggesting that anti- addition to the Re -face is more common than previously thought. 7 , 8 …”

Section: Discussionmentioning

confidence: 99%

Unexpected Methyllanthionine Stereochemistry in the Morphogenetic Lanthipeptide SapT

et al. 2022

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Lanthipeptides are polycyclic peptides characterized by the presence of lanthionine (Lan) and/or methyllanthionine (MeLan). They are members of the ribosomally synthesized and post-translationally modified peptides (RiPPs). The stereochemical configuration of (Me)Lan cross-links is important for the bioactivity of lanthipeptides. To date, MeLan residues in characterized lanthipeptides have either the 2 S ,3 S or 2 R ,3 R stereochemistry. Herein, we reconstituted in Escherichia coli the biosynthetic pathway toward SapT, a class I lanthipeptide that exhibits morphogenetic activity. Through the synthesis of standards, the heterologously produced peptide was shown to possess three MeLan residues with the 2 S ,3 R stereochemistry ( d - allo - l -MeLan), the first time such stereochemistry has been observed in a lanthipeptide. Bioinformatic analysis of the biosynthetic enzymes suggests this stereochemistry may also be present in other lanthipeptides. Analysis of another gene cluster in Streptomyces coelicolor that is widespread in actinobacteria confirmed another example of d - allo - l -MeLan and verified the bioinformatic prediction. We propose a mechanism for the origin of the unexpected stereochemistry and provide support using site-directed mutagenesis.

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Section: Discussionmentioning

confidence: 99%

Unexpected Methyllanthionine Stereochemistry in the Morphogenetic Lanthipeptide SapT

et al. 2022

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“…And no clear conserved motif was found among leader peptides of similar peptides except for Lys rich region (2nd-11th amino acids). In the previous report (Bothwell et al, 2021), Lys rich region in the leader peptide was reported in the biosynthesis of a class I lanthipeptide PedA15.1. We proposed that this Lys rich region may be the modification enzyme recognition motif of this group of class I lanthipeptides.…”

Section: Discussionmentioning

confidence: 90%

“…Glutamyl residue is provided from glutamyl tRNA (tRNA-Glu). In the case of heterologous production of class I lanthipeptide, tRNA-Glu and glutamyl tRNA synthase (GluRS) are required from the same bacterium which have the dehydratase (LanB) because lanthionine dehydratase (LanB) is indicated to recognize specific tRNA-Glu of same bacterial origin (Bothwell et al, 2021;Hudson et al, 2015;Ozaki et al, 2016). In this report, we F I G U R E 3 Proposed biosynthetic pathway for thalassomonasin A, underlined sequence is Lys rich region possibly recognized by modification enzymes observed production of class I lanthipeptides thalassomonasins A and B without introducing tRNA-Glu and GluRS of T. actiniarum into the co-expression system.…”

Section: Discussionmentioning

confidence: 99%

“…And the BGCs of pinensin‐like lanthipeptides were indicated to exist in the genomes of Bacteroidetes (Caetano et al, 2020). The class I lanthipeptide PedA15.1 was produced by heterologous expression using BGC of Pedobacter lusitanus which belongs to Bacteroides (Bothwell et al, 2021). To the best of our knowledge, there are just two reports about lanthipeptides derived from Gram‐negative bacterial origins (Bothwell et al, 2021; Caetano et al, 2020), although there are many BGCs of lanthipeptides found in the genome data of Gram‐negative bacteria (Walker et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

“…The class I lanthipeptide PedA15.1 was produced by heterologous expression using BGC of Pedobacter lusitanus which belongs to Bacteroides (Bothwell et al, 2021). To the best of our knowledge, there are just two reports about lanthipeptides derived from Gram‐negative bacterial origins (Bothwell et al, 2021; Caetano et al, 2020), although there are many BGCs of lanthipeptides found in the genome data of Gram‐negative bacteria (Walker et al, 2020). On the basis of these backgrounds, we performed heterologous expression of a cryptic gene cluster of a marine proteobacterium Thalassomonas actiniarum (Hosoya et al, 2009) using E. coli as the host.…”

Section: Introductionmentioning

confidence: 99%

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Heterologous expression of a cryptic gene cluster from a marine proteobacterium Thalassomonas actiniarum affords new lanthipeptides thalassomonasins A and B

Thetsana

Ijichi

Kaweewan

et al. 2022

Journal of Applied Microbiology

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Aims The aim of this study was to utilize a cryptic biosynthetic gene cluster (BGC) of a marine proteobacterium Thalassomonas actiniarum for production of new lanthipeptides by heterologous expression system. Methods and Results Based on genome mining, a new BGC of class I lanthipeptide was found in the genome sequence of a marine proteobacterium T. actiniarum. Molecular cloning was performed to construct an expression vector derived from commercially available plasmid pET‐41a(+). Heterologous production of new lanthipeptides named thalassomonasins A and B was performed using the host Escherichia coli BL21(DE3) harbouring the expression vector. The structure of thalassomonasin A was determined by the interpretation of NMR and MS data. As a result, thalassomonasin A was determined to be a lanthipeptide with three units of lanthionine. The bridging pattern of the lanthionine rings in thalassomonasin A was determined by interpretation of NOESY data. The structure of thalassomonasin B was proposed by MS/MS experiment. Conclusions We succeeded in heterologous production of new class I lanthipeptides using a BGC of a marine proteobacterium T. actiniarum. Significance and Impact of the Study To the best of our knowledge, this is the first report of heterologous production of lanthipeptides derived from proteobacterial origin. There are many cryptic biosynthetic gene clusters (BCGs) of this class of lanthipeptides in proteobacterial genomes. This study may lead to the production of new lanthipeptides by utilizing the BCGs.

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Elucidation of the Bridging Pattern of the Lantibiotic Pseudomycoicidin

et al. 2022

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Lantibiotics are post-translationally modified antibiotic peptides with lanthionine thioether bridges that represent potential alternatives to conventional antibiotics. The lantibiotic pseudomycoicidin is produced by Bacillus pseudomycoides DSM 12442 and is effective against many Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus. While prior work demonstrated that pseudomycoicidin possesses one disulfide bridge and four thioether bridges, the ring topology has so far remained unclear. Here, we analyzed several pseudomycoicidin analogues that are affected in ring formation via MALDI-TOF-MS and tandem mass spectrometry with regard to their dehydration and fragmentation patterns, respectively. As a result, we propose a bridging pattern involving Thr8 and Cys13, Thr10 and Cys16, Ser18 and Cys21, and Ser20 and Cys26, thus, forming two double ring systems. Additionally, we localized the disulfide bridge to connect Cys3 and Cys7 and, therefore, fully elucidated the bridging pattern of pseudomycoicidin.

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Structural Analysis of Class I Lanthipeptides from Pedobacter lusitanus NL19 Reveals an Unusual Ring Pattern

Cited by 33 publications

References 87 publications

Unexpected Methyllanthionine Stereochemistry in the Morphogenetic Lanthipeptide SapT

Unexpected Methyllanthionine Stereochemistry in the Morphogenetic Lanthipeptide SapT

Heterologous expression of a cryptic gene cluster from a marine proteobacterium Thalassomonas actiniarum affords new lanthipeptides thalassomonasins A and B

Elucidation of the Bridging Pattern of the Lantibiotic Pseudomycoicidin

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