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bottromycin A2

naturalproductmech:mibig-d717cc8774 MINTED SEEDED

Grounding review: No 3-star ChEBI entry shares this structure, so the record keeps a content-hashed identifier.
What these status tags mean

Curation status describes a workflow state. SEEDED means the record entered through the source-seeding workflow; it does not imply that no later curation occurred. PROPOSED marks an addition awaiting review; REVIEWED records curator sign-off on identity, structure, filing class and producer claims; DEPRECATED retains a superseded or out-of-scope record. These are workflow decisions, not independently verified human review. Agent-assisted contributions may appear in the record history.

Grounding describes the identifier mapping: EXACT uses an ontology identity asserted by the source; MINTED keeps a local identifier when no defensible ontology identity is available; REVIEW_NEEDED flags an exact structure whose ontology identity needs adjudication. The grounding note retains any unresolved alternatives. These tags do not establish the strength of producer or activity evidence.

Structure

Standard InChIKeyKSIZLOPUXFSFNR-XZNJZDOZSA-N
SMILES[H][C@@]12[C@H](C)CCN1C(=O)CN\C(=N/[C@H](C(=O)N[C@@H]([C@@H](C)C1=CC=CC=C1)C(=O)N[C@H](CC(=O)OC)C1=NC=CS1)C(C)(C)C)[C@@H](NC(=O)[C@@H](NC2=O)C(C)C)C(C)(C)C
Stereochemistryfully defined
Cross-referencesnpatlas:NPA04746, pubchem:3081434

Filing pathway

Amino acids and peptides — computed by NPClassifier npclassifier.gnps2.org@2026-09-07, which is why it is pinned rather than recomputed. The gene cluster's asserted class is ribosomal, from MIBiG.

Producer organisms 2

A claim that this taxon makes the compound. The basis says what the evidence addressed — a knockout and a database assertion are not the same claim.

TaxonBasisClusterEvidence
Streptomyces bottropensis DSM40262
NCBITaxon:42235		 BGC_CHARACTERIZED mibig:BGC0000468 PMID:19115340
Streptomyces sp. BC16019
NCBITaxon:1109705		 SOURCE_ASSERTION mibig:BGC0000469 PMID:23021914

Occurrences 0

None cited.

Biosynthetic gene clusters 2

AccessionHostLocus evidenceGenome
mibig:BGC0000468 Streptomyces bottropensis DSM40262
NCBITaxon:42235 		CLUSTER_DEMONSTRATED genbank:HE983629.1
mibig:BGC0000469 Streptomyces sp. BC16019
NCBITaxon:1109705 		CLUSTER_DEMONSTRATED genbank:JX235926.1

A locus claim and a taxon claim are different questions. Heterologous expression settles the first and leaves the second where the isolation report left it.

Biosynthesis curated

Written by a curator with claim-level evidence on each step; not derived from a source. This is what the rest of the record exists to support.

#EnzymeReactionSubstrate → productEvidence
1 botOMT
UniProtKB:K4MJZ1		 ungrounded → ungrounded DOI:10.1016/j.chembiol.2012.08.013
2 botRMT1
UniProtKB:K4MHV5		 ungrounded → ungrounded DOI:10.1016/j.chembiol.2012.08.013
3 botRMT2
UniProtKB:K4MJZ5		 ungrounded → ungrounded DOI:10.1016/j.chembiol.2012.08.013
4 botRMT3
UniProtKB:K4MHG1		 ungrounded → ungrounded DOI:10.1016/j.chembiol.2012.08.013
5 BotCYP
UniProtKB:K4MJU2		 ungrounded → ungrounded DOI:10.1021/jacs.0c10361
6 BotP
UniProtKB:K4MHW2		 ungrounded → ungrounded DOI:10.1002/cbic.201600406
7 BotH
UniProtKB:K4MHV9		 ungrounded → ungrounded DOI:10.1038/s41589-020-0569-y

Entry 1: BC16019 botOMT/AFV25479.1 is curated as an exact aspartate methyl ester formation contributor to bottromycin A2 maturation. The exact free substrate/product pair for this RiPP tailoring reaction is not represented in the record.

Entry 2: BC16019 botRMT1/AFV25480.1 is curated as an exact radical-SAM C-methylation contributor to bottromycin A2 maturation. The exact free substrate/product pair for this RiPP tailoring reaction is not represented in the record.

Entry 3: BC16019 botRMT2/AFV25484.1 is curated as an exact radical-SAM C-methylation contributor to bottromycin A2 maturation. The exact free substrate/product pair for this RiPP tailoring reaction is not represented in the record.

Entry 4: BC16019 botRMT3/AFV25488.1 is curated as an exact radical-SAM C-methylation contributor to bottromycin A2 maturation. The exact free substrate/product pair for this RiPP tailoring reaction is not represented in the record.

Entry 5: BC16019 BotCYP/AFV25487.2 is curated as the exact cytochrome P450 that oxidatively decarboxylates the terminal thiazoline of a bottromycin precursor. The exact free substrate/product pair for this RiPP tailoring reaction is not represented in the record.

Entry 6: BC16019 BotP/AFV25490.1 is curated as the exact peptidase that removes the N-terminal methionine from a bottromycin precursor peptide, exposing an N-terminal glycine. The exact modified precursor substrate and product are not represented in the record.

Entry 7: BC16019 BotH/AFV25485.1 epimerizes L-Asp7 to D-Asp7 in a post-follower-cleavage bottromycin precursor; that exact substrate/product pair is not represented.

Mechanism & Biosynthesis Overview

PRODUCERS & OCCURRENCE 2 producers (1 causal) 0 cited occurrences BIOSYNTHETIC LOCUS 2 gene clusters 7 curated pathway steps NATURAL PRODUCT bottromycin A2 naturalproductmech:mibig-d717cc8774 Amino acids and peptides TARGETS & BIOACTIVITY 0 molecular targets 0 bioactivities · 8 graphs

Causal graph — biosynthesis 8 nodes, 7 edges

Partial Streptomyces sp. BC16019 Bot RiPP graph for the precursor peptide, the three radical-SAM C-methyltransferases, and BotOMT that contribute methylations to bottromycin A2 maturation.

SubjectPredicateObjectNotesEvidence
BotA precursor peptide is precursor of bottromycin A2 BC16019 botA encodes the 44 aa bottromycin precursor peptide; the graph leaves downstream proteolysis, thiazole formation, amidine-ring formation, and oxidation unmodeled until exact protein-to-step evidence is curated.
DOI:10.1016/j.chembiol.2012.08.013
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
https://www.ncbi.nlm.nih.gov/nuccore/JX235926.1
https://www.ncbi.nlm.nih.gov/protein/AFV25481.1
https://www.uniprot.org/uniprotkb/K4MV53
  • reference
    DOI:10.1016/j.chembiol.2012.08.013
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    identified the biosynthetic gene cluster
    notes
    Huo et al. identified the Streptomyces sp. BC16019 bottromycin cluster, established heterologous bottromycin production from the cloned locus, and genetically implicated radical-SAM C-methylation plus aspartate methyl-ester formation in bottromycin biosynthesis.
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
  • reference
    https://www.ncbi.nlm.nih.gov/nuccore/JX235926.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
    snippet
    bottromycin biosynthetic gene cluster
  • reference
    https://www.ncbi.nlm.nih.gov/protein/AFV25481.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
    snippet
    BotA
    notes
    NCBI Protein AFV25481.1 is the 44 aa Streptomyces sp. BC16019 botA precursor-peptide coding sequence from the JX235926.1 bottromycin biosynthetic gene cluster.
  • reference
    https://www.uniprot.org/uniprotkb/K4MV53
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
    notes
    UniProtKB:K4MV53 is the 44 aa Streptomyces sp. BC16019 botA entry cross-referenced to EMBL JX235926; an exact GenPept accession search for AFV25481.1 resolved to this UniProtKB entry on 2026-09-27.
BotRMT1 radical-SAM methyltransferase contributes to radical-SAM C-methylation botRMT1 is one of the three BC16019 radical-SAM methyltransferases required for unactivated-carbon methylations during bottromycin maturation.
DOI:10.1016/j.chembiol.2012.08.013
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
https://www.ncbi.nlm.nih.gov/nuccore/JX235926.1
https://www.ncbi.nlm.nih.gov/protein/AFV25480.1
https://www.uniprot.org/uniprotkb/K4MHV5
  • reference
    DOI:10.1016/j.chembiol.2012.08.013
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    three radical SAM methyltransferase-encoding genes
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
  • reference
    https://www.ncbi.nlm.nih.gov/nuccore/JX235926.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
    snippet
    bottromycin biosynthetic gene cluster
  • reference
    https://www.ncbi.nlm.nih.gov/protein/AFV25480.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
    snippet
    botRMT1
  • reference
    https://www.uniprot.org/uniprotkb/K4MHV5
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
BotRMT2 radical-SAM methyltransferase contributes to radical-SAM C-methylation botRMT2 is one of the three BC16019 radical-SAM methyltransferases required for unactivated-carbon methylations during bottromycin maturation.
DOI:10.1016/j.chembiol.2012.08.013
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
https://www.ncbi.nlm.nih.gov/nuccore/JX235926.1
https://www.ncbi.nlm.nih.gov/protein/AFV25484.1
https://www.uniprot.org/uniprotkb/K4MJZ5
  • reference
    DOI:10.1016/j.chembiol.2012.08.013
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    three radical SAM methyltransferase-encoding genes
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
  • reference
    https://www.ncbi.nlm.nih.gov/nuccore/JX235926.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
    snippet
    bottromycin biosynthetic gene cluster
  • reference
    https://www.ncbi.nlm.nih.gov/protein/AFV25484.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
    snippet
    botRMT2
  • reference
    https://www.uniprot.org/uniprotkb/K4MJZ5
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
BotRMT3 radical-SAM methyltransferase contributes to radical-SAM C-methylation botRMT3 is one of the three BC16019 radical-SAM methyltransferases required for unactivated-carbon methylations during bottromycin maturation.
DOI:10.1016/j.chembiol.2012.08.013
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
https://www.ncbi.nlm.nih.gov/nuccore/JX235926.1
https://www.ncbi.nlm.nih.gov/protein/AFV25488.1
https://www.uniprot.org/uniprotkb/K4MHG1
  • reference
    DOI:10.1016/j.chembiol.2012.08.013
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    three radical SAM methyltransferase-encoding genes
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
  • reference
    https://www.ncbi.nlm.nih.gov/nuccore/JX235926.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
    snippet
    bottromycin biosynthetic gene cluster
  • reference
    https://www.ncbi.nlm.nih.gov/protein/AFV25488.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
    snippet
    botRMT3
  • reference
    https://www.uniprot.org/uniprotkb/K4MHG1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
BotOMT methyltransferase contributes to aspartate methyl-ester formation BC16019 BotOMT is assigned to the methyltransferase gene involved in aspartate methyl-ester formation.
DOI:10.1016/j.chembiol.2012.08.013
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
https://www.ncbi.nlm.nih.gov/nuccore/JX235926.1
https://www.ncbi.nlm.nih.gov/protein/AFV25479.1
https://www.uniprot.org/uniprotkb/K4MJZ1
  • reference
    DOI:10.1016/j.chembiol.2012.08.013
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    gene involved in aspartate methyl-ester formation
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
  • reference
    https://www.ncbi.nlm.nih.gov/nuccore/JX235926.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
    snippet
    bottromycin biosynthetic gene cluster
  • reference
    https://www.ncbi.nlm.nih.gov/protein/AFV25479.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
    snippet
    botOMT
  • reference
    https://www.uniprot.org/uniprotkb/K4MJZ1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
radical-SAM C-methylation contributes to bottromycin A2
DOI:10.1016/j.chembiol.2012.08.013
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
  • reference
    DOI:10.1016/j.chembiol.2012.08.013
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    identified the biosynthetic gene cluster
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27
aspartate methyl-ester formation contributes to bottromycin A2
DOI:10.1016/j.chembiol.2012.08.013
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
  • reference
    DOI:10.1016/j.chembiol.2012.08.013
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    identified the biosynthetic gene cluster
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-09-27

Every edge carries its own citation; an uncited edge is refused by the corpus tests.

Causal graph — biosynthesis 4 nodes, 3 edges

Partial Streptomyces sp. BC16019 Bot RiPP graph for BotCYP-catalyzed oxidative decarboxylation of the terminal thiazoline in a bottromycin precursor.

SubjectPredicateObjectNotesEvidence
terminal-thiazoline bottromycin precursor is substrate for terminal thiazoline oxidative decarboxylation MITE represents the BotCYP substrate as a bottromycin intermediate whose terminal thiazoline is consumed by this reaction; that precursor remains ungrounded until it has a stable chemical identifier.
DOI:10.1021/jacs.0c10361
https://mite.bioinformatics.nl/repository/MITE0000044
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
https://www.ncbi.nlm.nih.gov/protein/AFV25487.2
https://www.uniprot.org/uniprotkb/K4MJU2
  • reference
    DOI:10.1021/jacs.0c10361
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    converts a C-terminal thiazoline to a thiazole
  • reference
    https://mite.bioinformatics.nl/repository/MITE0000044
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    P450-catalyzed oxidative decarboxylation of terminal thiazoline
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    MITE0000044: Decarboxylation, Oxidation
  • reference
    https://www.ncbi.nlm.nih.gov/protein/AFV25487.2
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    BotCYP [Streptomyces sp. BC16019]
  • reference
    https://www.uniprot.org/uniprotkb/K4MJU2
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    SubName: Full=BotCYP
BotCYP cytochrome P450 catalyzes terminal thiazoline oxidative decarboxylation The BotCYP protein is grounded to the BC16019 GenPept AFV25487.2 sequence, its UniProtKB:K4MJU2 cross-reference, and the 100% identical BotCYP feature in MIBiG BGC0000469.6.
DOI:10.1021/jacs.0c10361
https://mite.bioinformatics.nl/repository/MITE0000044
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
https://www.ncbi.nlm.nih.gov/protein/AFV25487.2
https://www.uniprot.org/uniprotkb/K4MJU2
  • reference
    DOI:10.1021/jacs.0c10361
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    converts a C-terminal thiazoline to a thiazole
  • reference
    https://mite.bioinformatics.nl/repository/MITE0000044
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    P450-catalyzed oxidative decarboxylation of terminal thiazoline
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    MITE0000044: Decarboxylation, Oxidation
  • reference
    https://www.ncbi.nlm.nih.gov/protein/AFV25487.2
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    BotCYP [Streptomyces sp. BC16019]
  • reference
    https://www.uniprot.org/uniprotkb/K4MJU2
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    SubName: Full=BotCYP
terminal thiazoline oxidative decarboxylation contributes to bottromycin A2 This edge records the BotCYP oxidative decarboxylation as a BC16019 bottromycin A2 tailoring reaction; methyl-ester formation and radical-SAM C-methylations are modeled in a separate partial graph.
DOI:10.1021/jacs.0c10361
https://mite.bioinformatics.nl/repository/MITE0000044
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
https://www.ncbi.nlm.nih.gov/protein/AFV25487.2
https://www.uniprot.org/uniprotkb/K4MJU2
  • reference
    DOI:10.1021/jacs.0c10361
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    converts a C-terminal thiazoline to a thiazole
  • reference
    https://mite.bioinformatics.nl/repository/MITE0000044
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    P450-catalyzed oxidative decarboxylation of terminal thiazoline
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    MITE0000044: Decarboxylation, Oxidation
  • reference
    https://www.ncbi.nlm.nih.gov/protein/AFV25487.2
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    BotCYP [Streptomyces sp. BC16019]
  • reference
    https://www.uniprot.org/uniprotkb/K4MJU2
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    SubName: Full=BotCYP

Every edge carries its own citation; an uncited edge is refused by the corpus tests.

Causal graph — biosynthesis 5 nodes, 4 edges

Partial Streptomyces sp. BC16019 Bot RiPP graph for BotP-catalyzed N-terminal methionine cleavage in a bottromycin precursor.

SubjectPredicateObjectNotesEvidence
N-terminal Met bottromycin precursor is substrate for N-terminal methionine cleavage MITE represents the BotP substrate as a short N-terminal Met-Gly-Pro-Xn bottromycin precursor; the complete BotP peptide substrate remains ungrounded until it has a stable chemical identifier.
DOI:10.1002/cbic.201600406
https://mite.bioinformatics.nl/repository/MITE0000048
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
https://www.ncbi.nlm.nih.gov/protein/AFV25490.1
https://www.uniprot.org/uniprotkb/K4MHW2
  • reference
    DOI:10.1002/cbic.201600406
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    responsible for the removal of the N-terminal methionine
  • reference
    https://mite.bioinformatics.nl/repository/MITE0000048
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    Cleavage of N-terminal Met from a NH2-Met-Gly-Pro-Xn chain, exposing a N-terminal Gly
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    MITE0000048: Hydrolysis
  • reference
    https://www.ncbi.nlm.nih.gov/protein/AFV25490.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    BotP [Streptomyces sp. BC16019]
  • reference
    https://www.uniprot.org/uniprotkb/K4MHW2
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
BotP peptidase catalyzes N-terminal methionine cleavage The BotP protein is grounded to the BC16019 GenPept AFV25490.1 sequence, its UniProtKB:K4MHW2 cross-reference, and the 100% identical BotP feature in MIBiG BGC0000469.6.
DOI:10.1002/cbic.201600406
https://mite.bioinformatics.nl/repository/MITE0000048
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
https://www.ncbi.nlm.nih.gov/protein/AFV25490.1
https://www.uniprot.org/uniprotkb/K4MHW2
  • reference
    DOI:10.1002/cbic.201600406
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    responsible for the removal of the N-terminal methionine
  • reference
    https://mite.bioinformatics.nl/repository/MITE0000048
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    Cleavage of N-terminal Met from a NH2-Met-Gly-Pro-Xn chain, exposing a N-terminal Gly
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    MITE0000048: Hydrolysis
  • reference
    https://www.ncbi.nlm.nih.gov/protein/AFV25490.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    BotP [Streptomyces sp. BC16019]
  • reference
    https://www.uniprot.org/uniprotkb/K4MHW2
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
N-terminal methionine cleavage produces N-terminal Gly bottromycin precursor The in vitro reaction removes the N-terminal methionine and exposes the N-terminal glycine of the bottromycin precursor.
DOI:10.1002/cbic.201600406
https://mite.bioinformatics.nl/repository/MITE0000048
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
https://www.ncbi.nlm.nih.gov/protein/AFV25490.1
https://www.uniprot.org/uniprotkb/K4MHW2
  • reference
    DOI:10.1002/cbic.201600406
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    responsible for the removal of the N-terminal methionine
  • reference
    https://mite.bioinformatics.nl/repository/MITE0000048
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    Cleavage of N-terminal Met from a NH2-Met-Gly-Pro-Xn chain, exposing a N-terminal Gly
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    MITE0000048: Hydrolysis
  • reference
    https://www.ncbi.nlm.nih.gov/protein/AFV25490.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    BotP [Streptomyces sp. BC16019]
  • reference
    https://www.uniprot.org/uniprotkb/K4MHW2
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
N-terminal methionine cleavage contributes to bottromycin A2 This edge records BotP-catalyzed precursor processing as one bottromycin maturation reaction upstream of the final Streptomyces sp. BC16019 bottromycin A2 scaffold.
DOI:10.1002/cbic.201600406
https://mite.bioinformatics.nl/repository/MITE0000048
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
https://www.ncbi.nlm.nih.gov/protein/AFV25490.1
https://www.uniprot.org/uniprotkb/K4MHW2
  • reference
    DOI:10.1002/cbic.201600406
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    responsible for the removal of the N-terminal methionine
  • reference
    https://mite.bioinformatics.nl/repository/MITE0000048
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    Cleavage of N-terminal Met from a NH2-Met-Gly-Pro-Xn chain, exposing a N-terminal Gly
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    MITE0000048: Hydrolysis
  • reference
    https://www.ncbi.nlm.nih.gov/protein/AFV25490.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    BotP [Streptomyces sp. BC16019]
  • reference
    https://www.uniprot.org/uniprotkb/K4MHW2
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03

Every edge carries its own citation; an uncited edge is refused by the corpus tests.

Causal graph — biosynthesis 5 nodes, 4 edges

Partial Streptomyces sp. BC16019 Bot RiPP graph for BotH Asp7 epimerization in a bottromycin precursor.

SubjectPredicateObjectNotesEvidence
L-Asp7 bottromycin precursor is substrate for Asp7 epimerization MITE represents the BotH substrate as the post-follower-cleavage L-Asp7 bottromycin precursor; that intermediate remains ungrounded.
DOI:10.1038/s41589-020-0569-y
https://mite.bioinformatics.nl/repository/MITE0000051
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
https://www.ncbi.nlm.nih.gov/protein/AFV25485.1
https://www.uniprot.org/uniprotkb/K4MHV9
https://www.rcsb.org/structure/6T6X
  • reference
    DOI:10.1038/s41589-020-0569-y
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    epimerization of L-Asp to D-Asp
  • reference
    https://mite.bioinformatics.nl/repository/MITE0000051
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    epimerization of L-Asp7 to D-Asp7 of modified bottromycin precursor peptide
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    MITE0000051: Epimerization
  • reference
    https://www.ncbi.nlm.nih.gov/protein/AFV25485.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    BotH [Streptomyces sp. BC16019]
  • reference
    https://www.uniprot.org/uniprotkb/K4MHV9
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
  • reference
    https://www.rcsb.org/structure/6T6X
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    Structure of the Bottromycin epimerase BotH in complex with substrate
BotH epimerase catalyzes Asp7 epimerization BotH is grounded to BC16019 GenPept AFV25485.1, UniProtKB:K4MHV9, MIBiG BGC0000469.6, and BotH PDB structures.
DOI:10.1038/s41589-020-0569-y
https://mite.bioinformatics.nl/repository/MITE0000051
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
https://www.ncbi.nlm.nih.gov/protein/AFV25485.1
https://www.uniprot.org/uniprotkb/K4MHV9
https://www.rcsb.org/structure/6T6X
  • reference
    DOI:10.1038/s41589-020-0569-y
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    epimerization of L-Asp to D-Asp
  • reference
    https://mite.bioinformatics.nl/repository/MITE0000051
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    epimerization of L-Asp7 to D-Asp7 of modified bottromycin precursor peptide
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    MITE0000051: Epimerization
  • reference
    https://www.ncbi.nlm.nih.gov/protein/AFV25485.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    BotH [Streptomyces sp. BC16019]
  • reference
    https://www.uniprot.org/uniprotkb/K4MHV9
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
  • reference
    https://www.rcsb.org/structure/6T6X
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    Structure of the Bottromycin epimerase BotH in complex with substrate
Asp7 epimerization produces D-Asp7 bottromycin precursor BotH epimerizes Asp7 in the bottromycin precursor from L to D.
DOI:10.1038/s41589-020-0569-y
https://mite.bioinformatics.nl/repository/MITE0000051
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
https://www.ncbi.nlm.nih.gov/protein/AFV25485.1
https://www.uniprot.org/uniprotkb/K4MHV9
https://www.rcsb.org/structure/6T6X
  • reference
    DOI:10.1038/s41589-020-0569-y
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    epimerization of L-Asp to D-Asp
  • reference
    https://mite.bioinformatics.nl/repository/MITE0000051
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    epimerization of L-Asp7 to D-Asp7 of modified bottromycin precursor peptide
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    MITE0000051: Epimerization
  • reference
    https://www.ncbi.nlm.nih.gov/protein/AFV25485.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    BotH [Streptomyces sp. BC16019]
  • reference
    https://www.uniprot.org/uniprotkb/K4MHV9
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
  • reference
    https://www.rcsb.org/structure/6T6X
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    Structure of the Bottromycin epimerase BotH in complex with substrate
Asp7 epimerization contributes to bottromycin A2 BotH-catalyzed Asp7 epimerization is one bottromycin A2 maturation reaction in the BC16019 pathway.
DOI:10.1038/s41589-020-0569-y
https://mite.bioinformatics.nl/repository/MITE0000051
https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
https://www.ncbi.nlm.nih.gov/protein/AFV25485.1
https://www.uniprot.org/uniprotkb/K4MHV9
https://www.rcsb.org/structure/6T6X
  • reference
    DOI:10.1038/s41589-020-0569-y
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    epimerization of L-Asp to D-Asp
  • reference
    https://mite.bioinformatics.nl/repository/MITE0000051
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    epimerization of L-Asp7 to D-Asp7 of modified bottromycin precursor peptide
  • reference
    https://mibig.secondarymetabolites.org/repository/BGC0000469.6/index.html
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    MITE0000051: Epimerization
  • reference
    https://www.ncbi.nlm.nih.gov/protein/AFV25485.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    BotH [Streptomyces sp. BC16019]
  • reference
    https://www.uniprot.org/uniprotkb/K4MHV9
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
  • reference
    https://www.rcsb.org/structure/6T6X
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    snippet
    Structure of the Bottromycin epimerase BotH in complex with substrate

Every edge carries its own citation; an uncited edge is refused by the corpus tests.

Causal graph — biosynthesis 14 nodes, 16 edges

In-vitro processing of the BC16019 BotA bottromycin A2 precursor by S. ipomoeae IpoC and S. purpureus PurCD/PurAH. These are the assayed homologs, not BC16019 BotC/BotCD/BotAH. The recombinant substrates lack native C-methylations; products are intermediates, not mature bottromycin A2. No native production by the enzyme-source species is asserted.

SubjectPredicateObjectNotesEvidence
BotA(P): N-terminal Met removed is substrate for In-vitro thiazoline formation BotP-processed wild-type precursor; ATP/Mg2+ supplied. Figure 1 and SI Figures S4-S6.
DOI:10.1021/jacs.7b09898
  • reference
    DOI:10.1021/jacs.7b09898
    evidence type
    PRIMARY_EXPERIMENT
S. ipomoeae IpoC catalyzes In-vitro thiazoline formation
DOI:10.1021/jacs.7b09898
DOI:10.1021/jacs.7b09898.s001
https://www.ncbi.nlm.nih.gov/protein/WP_048819674.1
  • reference
    DOI:10.1021/jacs.7b09898
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    IpoC installs a cysteine-derived thiazoline
  • reference
    DOI:10.1021/jacs.7b09898.s001
    evidence type
    PRIMARY_EXPERIMENT
    notes
    SI p. S38 identifies codon-optimized WP_048819674.1 as IpoC; pHisSUMOTEV expression in E. coli Lemo21(DE3), followed by tag cleavage (p. S40).
  • reference
    https://www.ncbi.nlm.nih.gov/protein/WP_048819674.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    notes
    387-aa S. ipomoeae sequence. No exact UniProt match resolved in accession, name, or organism/YcaO searches on 2026-10-03.
In-vitro thiazoline formation produces BotA(PC): thiazoline, follower retained
DOI:10.1021/jacs.7b09898
  • reference
    DOI:10.1021/jacs.7b09898
    evidence type
    PRIMARY_EXPERIMENT
BotA(PC): thiazoline, follower retained is substrate for In-vitro macroamidine formation
DOI:10.1021/jacs.7b09898
  • reference
    DOI:10.1021/jacs.7b09898
    evidence type
    PRIMARY_EXPERIMENT
S. purpureus PurCD catalyzes In-vitro macroamidine formation ATP/Mg2+-dependent reaction without PurAH; not an obligate two-protein complex.
DOI:10.1021/jacs.7b09898
DOI:10.1021/jacs.7b09898.s001
https://www.ncbi.nlm.nih.gov/protein/WP_019887079.1
  • reference
    DOI:10.1021/jacs.7b09898
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    PurCD alone is sufficient to create the macroamidine structure
  • reference
    DOI:10.1021/jacs.7b09898.s001
    evidence type
    PRIMARY_EXPERIMENT
    notes
    SI p. S38 identifies codon-optimized WP_019887079 as PurCD; pHisTEV expression in E. coli Lemo21(DE3), followed by tag cleavage (p. S40).
  • reference
    https://www.ncbi.nlm.nih.gov/protein/WP_019887079.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    notes
    Version 1 is a 445-aa S. purpureus sequence. No exact UniProt match resolved in the bounded searches; the unrelated Francisella purCD result was rejected.
In-vitro macroamidine formation produces BotA(PCCD): thiazoline and macroamidine
DOI:10.1021/jacs.7b09898
  • reference
    DOI:10.1021/jacs.7b09898
    evidence type
    PRIMARY_EXPERIMENT
BotA(PCCD): thiazoline and macroamidine is substrate for Follower-retaining macroamidine reopening
DOI:10.1021/jacs.8b12231
  • reference
    DOI:10.1021/jacs.8b12231
    evidence type
    PRIMARY_EXPERIMENT
S. purpureus PurCD catalyzes Follower-retaining macroamidine reopening Purified follower-retaining BotA(PCCD), PurCD and ATP/MgCl2; Figure S4.
DOI:10.1021/jacs.8b12231
  • reference
    DOI:10.1021/jacs.8b12231
    evidence type
    PRIMARY_EXPERIMENT
Follower-retaining macroamidine reopening produces BotA(PC): thiazoline, follower retained
DOI:10.1021/jacs.8b12231
  • reference
    DOI:10.1021/jacs.8b12231
    evidence type
    PRIMARY_EXPERIMENT
BotA(PCCD): thiazoline and macroamidine is substrate for Thiazoline-adjacent follower cleavage Preferred substrate, not exclusive: BotA(PC) also undergoes slower cleavage in Figure 1a.
DOI:10.1021/jacs.8b12231
  • reference
    DOI:10.1021/jacs.8b12231
    evidence type
    PRIMARY_EXPERIMENT
S. purpureus PurAH catalyzes Thiazoline-adjacent follower cleavage Cleavage follows the thiazoline, not a thiazole; Figure 1 and SI Figures S1-S2.
DOI:10.1021/jacs.8b12231
DOI:10.1021/jacs.8b12231.s001
https://www.ncbi.nlm.nih.gov/protein/WP_019887083.1
https://www.uniprot.org/uniprotkb/A0A5S8WF49
https://www.rcsb.org/structure/6I5S
  • reference
    DOI:10.1021/jacs.8b12231
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    removes the follower peptide
  • reference
    DOI:10.1021/jacs.8b12231.s001
    evidence type
    PRIMARY_EXPERIMENT
    notes
    SI p. 3 specifies full-length, codon-optimized WP_019887083.1 in pET-28b, expressed in Rosetta(DE3) and purified by His affinity and size exclusion.
  • reference
    https://www.ncbi.nlm.nih.gov/protein/WP_019887083.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    notes
    474-aa S. purpureus sequence; exact match to UniProt A0A5S8WF49 and the canonical PDB 6I5S entity-1 sequence.
  • reference
    https://www.uniprot.org/uniprotkb/A0A5S8WF49
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    notes
    PDB-derived S. purpureus bottromycin amidohydrolase entry, not BC16019 BotAH/K4MV58.
  • reference
    https://www.rcsb.org/structure/6I5S
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    notes
    Chain A maps all 474 canonical residues to A0A5S8WF49; carboxylated Lys183 is represented as KCX. This is not a substrate-bound structure.
Thiazoline-adjacent follower cleavage produces Core 2: macroamidine, follower removed
DOI:10.1021/jacs.8b12231
  • reference
    DOI:10.1021/jacs.8b12231
    evidence type
    PRIMARY_EXPERIMENT
Thiazoline-adjacent follower cleavage releases Released BotA follower peptide
DOI:10.1021/jacs.8b12231
  • reference
    DOI:10.1021/jacs.8b12231
    evidence type
    PRIMARY_EXPERIMENT
Core 2: macroamidine, follower removed is tested in PurCD test on follower-free core 2
DOI:10.1021/jacs.8b12231
DOI:10.1021/jacs.8b12231.s001
  • reference
    DOI:10.1021/jacs.8b12231
    evidence type
    PRIMARY_EXPERIMENT
  • reference
    DOI:10.1021/jacs.8b12231.s001
    evidence type
    PRIMARY_EXPERIMENT
    notes
    SI pp. 4-5 and Figure S4: 50 uM core 2, 10 uM PurCD and 10 mM ATP/MgCl2, pH 8.5, 16 h at room temperature; main text instead says 37 C.
S. purpureus PurCD is supplied in PurCD test on follower-free core 2
DOI:10.1021/jacs.8b12231
  • reference
    DOI:10.1021/jacs.8b12231
    evidence type
    PRIMARY_EXPERIMENT
PurCD test on follower-free core 2 has no reported detectable formation of Core 1: no macroamidine, follower removed Condition-specific lack of macroamidine reopening after follower removal; not proof of universal chemical irreversibility.
DOI:10.1021/jacs.8b12231
  • reference
    DOI:10.1021/jacs.8b12231
    evidence type
    PRIMARY_EXPERIMENT

Every edge carries its own citation; an uncited edge is refused by the corpus tests.

Causal graph — biosynthesis 9 nodes, 8 edges

Parallel in-vitro processing of unmethylated BmbC by MBP-BmbD or MBP-BmbE without partners; no native order, joint product or mature bottromycin A2 is asserted. SI Table S1 accessions identify S. bottropensis DSM 40262 proteins on HE983629.1, not BC16019 homologs.

SubjectPredicateObjectNotesEvidence
S. bottropensis BmbC (44 aa) provides sequence for BmbC: initiator Met removed (43 aa) TEV-released recombinant substrate lacks initiator Met; native removal enzyme not asserted.
DOI:10.1021/jacs.7b09899.s001
https://www.ncbi.nlm.nih.gov/protein/CCM09443.1
  • reference
    DOI:10.1021/jacs.7b09899.s001
    evidence type
    PRIMARY_EXPERIMENT
  • reference
    https://www.ncbi.nlm.nih.gov/protein/CCM09443.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    notes
    44-aa precursor; SI Table S4 wild-type assay peptide exactly matches residues 2-44.
BmbC: initiator Met removed (43 aa) is substrate for In-vitro thiazoline formation Figure 2; SI pp. S4-S5: 50 uM precursor, 10 uM MBP enzyme, 5 mM ATP, 20 mM MgCl2, pH 8.5, 25 C, 5 h.
DOI:10.1021/jacs.7b09899
  • reference
    DOI:10.1021/jacs.7b09899
    evidence type
    PRIMARY_EXPERIMENT
MBP-BmbD catalyzes In-vitro thiazoline formation SI Table S3 codon-optimized BmbD translates exactly to CCM09444.1 and K0P4N8 (410 aa). MBP fusion expressed in E. coli; tag retained.
DOI:10.1021/jacs.7b09899
DOI:10.1021/jacs.7b09899.s001
https://www.uniprot.org/uniprotkb/K0P4N8
  • reference
    DOI:10.1021/jacs.7b09899
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    thiazoline formation to BmbD
  • reference
    DOI:10.1021/jacs.7b09899.s001
    evidence type
    PRIMARY_EXPERIMENT
  • reference
    https://www.uniprot.org/uniprotkb/K0P4N8
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
In-vitro thiazoline formation produces BmbC thiazoline intermediate Labelling/MS/MS locate Cys8 cyclodehydration; supporting NMR used trypsin-treated BmbC-N15K.
DOI:10.1021/jacs.7b09899
  • reference
    DOI:10.1021/jacs.7b09899
    evidence type
    PRIMARY_EXPERIMENT
BmbC: initiator Met removed (43 aa) is substrate for In-vitro macrolactamidine formation Same standard conditions; no prior BmbD step.
DOI:10.1021/jacs.7b09899
  • reference
    DOI:10.1021/jacs.7b09899
    evidence type
    PRIMARY_EXPERIMENT
MBP-BmbE catalyzes In-vitro macrolactamidine formation SI Table S3 BmbE translates exactly to CCM09445.1 and K0NZY4 (463 aa). MBP fusion retained; ATP-dependent Gly1-Val4 macrocyclization.
DOI:10.1021/jacs.7b09899
DOI:10.1021/jacs.7b09899.s001
https://www.uniprot.org/uniprotkb/K0NZY4
  • reference
    DOI:10.1021/jacs.7b09899
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    macrolactamidine formation to BmbE
  • reference
    DOI:10.1021/jacs.7b09899.s001
    evidence type
    PRIMARY_EXPERIMENT
  • reference
    https://www.uniprot.org/uniprotkb/K0NZY4
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
In-vitro macrolactamidine formation produces BmbC macrolactamidine intermediate Figures S6-S8 distinguish this isobaric -18 Da product from the thiazoline product; NMR used trypsin-treated BmbC-N15K.
DOI:10.1021/jacs.7b09899
  • reference
    DOI:10.1021/jacs.7b09899
    evidence type
    PRIMARY_EXPERIMENT
S. bottropensis BmbC (44 aa) has a sequence-identical UniProtKB example BmbC / BotA (S. bottropensis UniProtKB example) UniProtKB:K0P8E3 (Streptomyces bottropensis botA / QBA35_11730) shares 100% amino-acid sequence identity (44 aa, UPI0001B7FB89) with Streptomyces bottropensis BmbC (CCM09443.1).
https://www.uniprot.org/uniprotkb/K0P8E3
https://www.uniprot.org/uniparc/UPI0001B7FB89
https://www.ncbi.nlm.nih.gov/protein/CCM09443.1
  • reference
    https://www.uniprot.org/uniprotkb/K0P8E3
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-17
    notes
    Active Streptomyces bottropensis UniProtKB entry (K0P8E3) for the 44-aa bottromycin precursor peptide.
  • reference
    https://www.uniprot.org/uniparc/UPI0001B7FB89
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-17
    notes
    UniParc UPI0001B7FB89 links EMBL CCM09443.1 (bmbC) and active Streptomyces bottropensis UniProtKB:K0P8E3 under a single 100%-identical 44-aa sequence.
  • reference
    https://www.ncbi.nlm.nih.gov/protein/CCM09443.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-17
    notes
    EMBL/GenPept CCM09443.1 is the 44-aa Streptomyces bottropensis BmbC precursor peptide.

Every edge carries its own citation; an uncited edge is refused by the corpus tests.

Causal graph — biosynthesis 7 nodes, 6 edges

Fluorescence-polarization binding to engineered BmbC-C8S/S43C with a C-terminal His-tag extension and 6-(iodoacetamido)fluorescein label (SI Figure S12). All binding proteins remain MBP-tagged. BmbB/F nodes represent excised RRE domains, not assayed full-length methyltransferases; no methylation or native-substrate affinity is inferred. SI p. S7: 25 nM tracer, serial protein dilution, 50 mM HEPES pH 7.5, 300 mM NaCl, 2.5% glycerol, 0.5 mM TCEP, 30 min at 25 C, three independent titrations.

SubjectPredicateObjectNotesEvidence
S. bottropensis BmbC (44 aa) provides sequence for BmbC-C8S/S43C-His fluorescent tracer Engineered construct, not a biological conversion: native Met absent; C8S/S43C use 43-aa assay numbering (C9S/S44C in CCM09443.1). His-tag extension and fluorophore are absent from the canonical accession.
DOI:10.1021/jacs.7b09899.s001
  • reference
    DOI:10.1021/jacs.7b09899.s001
    evidence type
    PRIMARY_EXPERIMENT
MBP-BmbB RRE (492-593) binds BmbC-C8S/S43C-His fluorescent tracer Figure S13C: Kd 876 +/- 67 nM by fluorescence polarization. SI Table S3 domain sequence exactly matches residues 492-593 of the 630-aa canonical protein.
DOI:10.1021/jacs.7b09899.s001
https://www.ncbi.nlm.nih.gov/protein/CCM09442.1
https://rest.uniprot.org/uniparc/UPI00027EA686.json
  • reference
    DOI:10.1021/jacs.7b09899.s001
    evidence type
    PRIMARY_EXPERIMENT
  • reference
    https://www.ncbi.nlm.nih.gov/protein/CCM09442.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    notes
    Canonical BmbB RRE parent.
  • reference
    https://rest.uniprot.org/uniparc/UPI00027EA686.json
    evidence type
    DATABASE_ASSERTION
    notes
    UniParc UPI00027EA686 (630 aa) links active EMBL CCM09442.1 (Streptomyces bottropensis, taxon 42239, gene bmbB) and inactive UniProtKB/TrEMBL K0P1P7 (bmbB), with zero active UniProtKB cross-references.
    retrieved on
    2026-10-30
MBP-BmbF RRE (526-670) binds BmbC-C8S/S43C-His fluorescent tracer Figure S13D: Kd 597 +/- 25 nM by fluorescence polarization. SI Table S3 domain sequence exactly matches residues 526-670 of the 670-aa canonical protein.
DOI:10.1021/jacs.7b09899.s001
https://www.ncbi.nlm.nih.gov/protein/CCM09446.1
https://rest.uniprot.org/uniparc/UPI00027EA689.json
  • reference
    DOI:10.1021/jacs.7b09899.s001
    evidence type
    PRIMARY_EXPERIMENT
  • reference
    https://www.ncbi.nlm.nih.gov/protein/CCM09446.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    notes
    Canonical BmbF RRE parent.
  • reference
    https://rest.uniprot.org/uniparc/UPI00027EA689.json
    evidence type
    DATABASE_ASSERTION
    notes
    UniParc UPI00027EA689 (670 aa) links active EMBL CCM09446.1 (Streptomyces bottropensis, taxon 42239, gene bmbF) and inactive UniProtKB/TrEMBL K0P4U1 (bmbF), M3FCL1 and A0A7K3EJA3, with zero active UniProtKB cross-references.
    retrieved on
    2026-10-30
MBP-BmbD shows weak binding to BmbC-C8S/S43C-His fluorescent tracer Main text estimates Kd >50 uM; Figure S13A labels >=50 uM. This is a lower bound, not an exact 50 uM affinity.
DOI:10.1021/jacs.7b09899
DOI:10.1021/jacs.7b09899.s001
  • reference
    DOI:10.1021/jacs.7b09899
    evidence type
    PRIMARY_EXPERIMENT
  • reference
    DOI:10.1021/jacs.7b09899.s001
    evidence type
    PRIMARY_EXPERIMENT
MBP-BmbE shows weak binding to BmbC-C8S/S43C-His fluorescent tracer Main text estimates Kd >50 uM; Figure S13B labels >=50 uM. This is a lower bound, not a no-binding claim.
DOI:10.1021/jacs.7b09899
DOI:10.1021/jacs.7b09899.s001
  • reference
    DOI:10.1021/jacs.7b09899
    evidence type
    PRIMARY_EXPERIMENT
  • reference
    DOI:10.1021/jacs.7b09899.s001
    evidence type
    PRIMARY_EXPERIMENT
S. bottropensis BmbC (44 aa) has a sequence-identical UniProtKB example BmbC / BotA (S. bottropensis UniProtKB example) UniProtKB:K0P8E3 (Streptomyces bottropensis botA / QBA35_11730) shares 100% amino-acid sequence identity (44 aa, UPI0001B7FB89) with Streptomyces bottropensis BmbC (CCM09443.1).
https://www.uniprot.org/uniprotkb/K0P8E3
https://www.uniprot.org/uniparc/UPI0001B7FB89
https://www.ncbi.nlm.nih.gov/protein/CCM09443.1
  • reference
    https://www.uniprot.org/uniprotkb/K0P8E3
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-17
    notes
    Active Streptomyces bottropensis UniProtKB entry (K0P8E3) for the 44-aa bottromycin precursor peptide.
  • reference
    https://www.uniprot.org/uniparc/UPI0001B7FB89
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-17
    notes
    UniParc UPI0001B7FB89 links EMBL CCM09443.1 (bmbC) and active Streptomyces bottropensis UniProtKB:K0P8E3 under a single 100%-identical 44-aa sequence.
  • reference
    https://www.ncbi.nlm.nih.gov/protein/CCM09443.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-17
    notes
    EMBL/GenPept CCM09443.1 is the 44-aa Streptomyces bottropensis BmbC precursor peptide.

Every edge carries its own citation; an uncited edge is refused by the corpus tests.

Causal graph — biosynthesis 4 nodes, 4 edges

BmbG or BmbH individually supplemented MBP-BmbD/K0P4N8 or MBP-BmbE/K0NZY4 reactions on Met-deleted BmbC. SI Figure S9: 16-h endpoint MALDI-TOF-MS, no enhanced cyclodehydration. This does not exclude native functions or establish inactivity, inhibition, or a joint G+H test. Figure S3 documents MBP-fused S. bottropensis proteins; G/H are not BC16019 BotH/BotAH.

SubjectPredicateObjectNotesEvidence
MBP-BmbG was reported not to enhance BmbD 16-h cyclodehydration assay BmbG supplied alone as supplement; comparison is endpoint conversion to the -18 Da species.
DOI:10.1021/jacs.7b09899.s001
https://www.ncbi.nlm.nih.gov/protein/CCM09447.1
https://rest.uniprot.org/uniparc/UPI00027EA68A.json
  • reference
    DOI:10.1021/jacs.7b09899.s001
    evidence type
    PRIMARY_EXPERIMENT
  • reference
    https://www.ncbi.nlm.nih.gov/protein/CCM09447.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    notes
    333-aa BmbG; identity from SI Table S1, purified protein in Figure S3.
  • reference
    https://rest.uniprot.org/uniparc/UPI00027EA68A.json
    evidence type
    DATABASE_ASSERTION
    notes
    UniParc UPI00027EA68A (333 aa) links active EMBL CCM09447.1 (Streptomyces bottropensis, taxon 42239, gene bmbG) and inactive UniProtKB/TrEMBL K0P1P9 (bmbG), with zero active UniProtKB cross-references.
    retrieved on
    2026-10-30
MBP-BmbH was reported not to enhance BmbD 16-h cyclodehydration assay BmbH supplied alone as supplement; no claim of statistical equivalence or universal lack of effect.
DOI:10.1021/jacs.7b09899.s001
https://www.ncbi.nlm.nih.gov/protein/CCM09448.1
https://rest.uniprot.org/uniparc/UPI00027EA68B.json
  • reference
    DOI:10.1021/jacs.7b09899.s001
    evidence type
    PRIMARY_EXPERIMENT
  • reference
    https://www.ncbi.nlm.nih.gov/protein/CCM09448.1
    evidence type
    DATABASE_ASSERTION
    retrieved on
    2026-10-03
    notes
    415-aa BmbH; identity from SI Table S1, purified protein in Figure S3.
  • reference
    https://rest.uniprot.org/uniparc/UPI00027EA68B.json
    evidence type
    DATABASE_ASSERTION
    notes
    UniParc UPI00027EA68B (415 aa) links active EMBL CCM09448.1 (Streptomyces bottropensis, taxon 42239, gene bmbH) and inactive UniProtKB/TrEMBL K0P8F1 (bmbH), with zero active UniProtKB cross-references.
    retrieved on
    2026-10-30
MBP-BmbG was reported not to enhance BmbE 16-h cyclodehydration assay BmbG individually added to the BmbE reaction.
DOI:10.1021/jacs.7b09899
DOI:10.1021/jacs.7b09899.s001
  • reference
    DOI:10.1021/jacs.7b09899
    evidence type
    PRIMARY_EXPERIMENT
    snippet
    Neither addition of BmbH or BmbG resulted in more product formation
  • reference
    DOI:10.1021/jacs.7b09899.s001
    evidence type
    PRIMARY_EXPERIMENT
MBP-BmbH was reported not to enhance BmbE 16-h cyclodehydration assay BmbH individually added to the BmbE reaction.
DOI:10.1021/jacs.7b09899.s001
  • reference
    DOI:10.1021/jacs.7b09899.s001
    evidence type
    PRIMARY_EXPERIMENT

Every edge carries its own citation; an uncited edge is refused by the corpus tests.

Explore across X-Mech

AntibioticMech (bottromycin A2) ↗ MediaIngredientMech (naturalproductmech:mibig-d717cc8774) ↗ TaxonMech: Streptomyces bottropensis DSM40262 (NCBITaxon:42235) ↗ TaxonMech: Streptomyces sp. BC16019 (NCBITaxon:1109705) ↗ PathwayMech (mibig:BGC0000468) ↗ PathwayMech (mibig:BGC0000469) ↗

Provenance

Source conceptSourceVersion
BGC0000468MIBIG5
BGC0000469MIBIG6

This page is generated from data/natural_products/amino_acids_and_peptides/bottromycin-a2-2.yaml, which is generated from the committed inventories. Neither is edited by hand.