Show legend Show as text

Structure type: monomer
Trivial name: lincomycin, lincomycin A
Compound class: glycoside

• Article ID: 5564
  Janata J, Kadlcik S, Koberska M, Ulanova D, Kamenik Z, Novak P, Kopecky J, Novotna J, Radojevic B, Plhackova K, Gazak R, Najmanova L "Lincosamide synthetase - a unique condensation system combining elements of nonribosomal peptide synthetase and mycothiol metabolism" - PLoS One 10(3) (2015) ID e0118850
  

  In the biosynthesis of lincosamide antibiotics lincomycin and celesticetin, the amino acid and amino sugar units are linked by an amide bond. The respective condensing enzyme lincosamide synthetase (LS) is expected to be an unusual system combining nonribosomal peptide synthetase (NRPS) components with so far unknown amino sugar related activities. The biosynthetic gene cluster of celesticetin was sequenced and compared to the lincomycin one revealing putative LS coding ORFs shared in both clusters. Based on a bioassay and production profiles of S. lincolnensis strains with individually deleted putative LS coding genes, the proteins LmbC, D, E, F and V were assigned to LS function. Moreover, the newly recognized N-terminal domain of LmbN (LmbN-CP) was also assigned to LS as a NRPS carrier protein (CP). Surprisingly, the homologous CP coding sequence in celesticetin cluster is part of ccbZ gene adjacent to ccbN, the counterpart of lmbN, suggesting the gene rearrangement, evident also from still active internal translation start in lmbN, and indicating the direction of lincosamide biosynthesis evolution. The in vitro test with LmbN-CP, LmbC and the newly identified S. lincolnensis phosphopantetheinyl transferase Slp, confirmed the cooperation of the previously characterized NRPS A-domain LmbC with a holo-LmbN-CP in activation of a 4-propyl-L-proline precursor of lincomycin. This result completed the functional characterization of LS subunits resembling NRPS initiation module. Two of the four remaining putative LS subunits, LmbE/CcbE and LmbV/CcbV, exhibit low but significant homology to enzymes from the metabolism of mycothiol, the NRPS-independent system processing the amino sugar and amino acid units. The functions of particular LS subunits as well as cooperation of both NRPS-based and NRPS-independent LS blocks are discussed. The described condensing enzyme represents a unique hybrid system with overall composition quite dissimilar to any other known enzyme system

  biosynthesis, antibiotics, lincosamide, Streptomyces lincolnensis

  NCBI PubMed ID: 25741696
  Publication DOI: 10.1371/journal.pone.0118850
  Journal NLM ID: 101285081
  Publisher: San Francisco, CA: Public Library of Science
  Correspondence: janata@biomed.cas.cz
  Institutions: Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech, Oceanography Section, Science Research Center, Kochi University, IMT-MEXT, Oko-cho, Japan
  Methods: DNA techniques, biological assays, extraction, LC-MS, cell growth, antibacterial assay
  
   
  
  Streptomyces lincolnensis ATCC 25466
  CSDB ID 47330 (all data & tools)
• Article ID: 5597
  Wang M, Zhao Q, Liu W "The versatile low-molecular-weight thiols: Beyond cell protection" - BioEssays: News and Reviews in Molecular, Cellular and Developmental Biology 37(12) (2015) 1262-1267
  

  Low-molecular-weight (LMW) thiols are extensively involved in the maintenance of cellular redox potentials and the protection of cells from a variety of reactive chemical and electrophilic species. However, we recently found that the metabolic coupling of two LMW thiols - mycothiol (MSH) and ergothioneine (EGT) - programs the biosynthesis of the anti-infective agent lincomycin A. Remarkably, such a constructive role of the thiols in the biosynthesis of natural products has so far received relatively little attention. We speculate that the unusual thiol EGT might function as a chiral thiolation carrier (for modification) and a novel activator (for glycosylation) of sugar. Additionally, we examine recent evidence for LMW thiols (MSH and others) as sulfur donors of sulfur-containing natural products. Clearly, the LMW thiols have more diverse activities beyond cell protection, and more attention should be paid to the correlation of their functions with thiol-dependent enzymes

  ergothioneine, low-molecular-weight thiols, mycothiol, non-Leloir pathway, sulfur incorporation, thiolation carrier

  NCBI PubMed ID: 26515639
  Publication DOI: 10.1002/bies.201500067
  Journal NLM ID: 8510851
  Publisher: Hoboken, N.J.: Wiley
  Correspondence: wliu@mail.sioc.ac.cn
  Institutions: State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China, Huzhou Center of Bio-Synthetic Innovation, Huzhou, China
  
   
  
  Streptomyces lincolnensis var. lincolnensis
  CSDB ID 47721 (all data & tools)
• Article ID: 5606
  Zhao Q, Wang M, Xu D, Zhang Q, Liu W "Metabolic coupling of two small-molecule thiols programs the biosynthesis of lincomycin A" - Nature 518(7537) (2015) 115-119
  

  Low-molecular-mass thiols in organisms are well known for their redox-relevant role in protection against various endogenous and exogenous stresses. In eukaryotes and Gram-negative bacteria, the primary thiol is glutathione (GSH), a cysteinyl-containing tripeptide. In contrast, mycothiol (MSH), a cysteinyl pseudo-disaccharide, is dominant in Gram-positive actinobacteria, including antibiotic-producing actinomycetes and pathogenic mycobacteria. MSH is equivalent to GSH, either as a cofactor or as a substrate, in numerous biochemical processes, most of which have not been characterized, largely due to the dearth of information concerning MSH-dependent proteins. Actinomycetes are able to produce another thiol, ergothioneine (EGT), a histidine betaine derivative that is widely assimilated by plants and animals for variable physiological activities. The involvement of EGT in enzymatic reactions, however, lacks any precedent. Here we report that the unprecedented coupling of two bacterial thiols, MSH and EGT, has a constructive role in the biosynthesis of lincomycin A, a sulfur-containing lincosamide (C8 sugar) antibiotic that has been widely used for half a century to treat Gram-positive bacterial infections. EGT acts as a carrier to template the molecular assembly, and MSH is the sulfur donor for lincomycin maturation after thiol exchange. These thiols function through two unusual S-glycosylations that program lincosamide transfer, activation and modification, providing the first paradigm for EGT-associated biochemical processes and for the poorly understood MSH-dependent biotransformations, a newly described model that is potentially common in the incorporation of sulfur, an element essential for life and ubiquitous in living systems

  biosynthesis, lincomycin A, S-glycoside, mycothiols

  NCBI PubMed ID: 25607359
  Publication DOI: 10.1038/nature14137
  Journal NLM ID: 0410462
  Publisher: Basingstoke: Nature Publishing Group
  Correspondence: wliu@mail.sioc.ac.cn
  Institutions: State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China, Huzhou Center of Bio-Synthetic Innovation, Huzhou, China
  Methods: 13C NMR, 1H NMR, PCR, DNA techniques, ESI-MS, chemical synthesis, HPLC, CC, HR-ESI-MS, enzymatic assay, evaporation, centrifugation, HILIC
  
   
  
  Streptomyces lincolnensis, Streptomyces lincolnensis (ΔlmbE mutant), Streptomyces lincolnensis (ΔmshAlin)
  CSDB ID 47749 (all data & tools)

Show legend Show as text

Structure type: monomer
Trivial name: celesticetin
Compound class: glycoside

• Article ID: 5564
  Janata J, Kadlcik S, Koberska M, Ulanova D, Kamenik Z, Novak P, Kopecky J, Novotna J, Radojevic B, Plhackova K, Gazak R, Najmanova L "Lincosamide synthetase - a unique condensation system combining elements of nonribosomal peptide synthetase and mycothiol metabolism" - PLoS One 10(3) (2015) ID e0118850
  

  In the biosynthesis of lincosamide antibiotics lincomycin and celesticetin, the amino acid and amino sugar units are linked by an amide bond. The respective condensing enzyme lincosamide synthetase (LS) is expected to be an unusual system combining nonribosomal peptide synthetase (NRPS) components with so far unknown amino sugar related activities. The biosynthetic gene cluster of celesticetin was sequenced and compared to the lincomycin one revealing putative LS coding ORFs shared in both clusters. Based on a bioassay and production profiles of S. lincolnensis strains with individually deleted putative LS coding genes, the proteins LmbC, D, E, F and V were assigned to LS function. Moreover, the newly recognized N-terminal domain of LmbN (LmbN-CP) was also assigned to LS as a NRPS carrier protein (CP). Surprisingly, the homologous CP coding sequence in celesticetin cluster is part of ccbZ gene adjacent to ccbN, the counterpart of lmbN, suggesting the gene rearrangement, evident also from still active internal translation start in lmbN, and indicating the direction of lincosamide biosynthesis evolution. The in vitro test with LmbN-CP, LmbC and the newly identified S. lincolnensis phosphopantetheinyl transferase Slp, confirmed the cooperation of the previously characterized NRPS A-domain LmbC with a holo-LmbN-CP in activation of a 4-propyl-L-proline precursor of lincomycin. This result completed the functional characterization of LS subunits resembling NRPS initiation module. Two of the four remaining putative LS subunits, LmbE/CcbE and LmbV/CcbV, exhibit low but significant homology to enzymes from the metabolism of mycothiol, the NRPS-independent system processing the amino sugar and amino acid units. The functions of particular LS subunits as well as cooperation of both NRPS-based and NRPS-independent LS blocks are discussed. The described condensing enzyme represents a unique hybrid system with overall composition quite dissimilar to any other known enzyme system

  biosynthesis, antibiotics, lincosamide, Streptomyces lincolnensis

  NCBI PubMed ID: 25741696
  Publication DOI: 10.1371/journal.pone.0118850
  Journal NLM ID: 101285081
  Publisher: San Francisco, CA: Public Library of Science
  Correspondence: janata@biomed.cas.cz
  Institutions: Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech, Oceanography Section, Science Research Center, Kochi University, IMT-MEXT, Oko-cho, Japan
  Methods: DNA techniques, biological assays, extraction, LC-MS, cell growth, antibacterial assay
  
   
  
  Streptomyces caelestis ATCC 15084
  CSDB ID 47331 (all data & tools)
• Article ID: 5597
  Wang M, Zhao Q, Liu W "The versatile low-molecular-weight thiols: Beyond cell protection" - BioEssays: News and Reviews in Molecular, Cellular and Developmental Biology 37(12) (2015) 1262-1267
  

  Low-molecular-weight (LMW) thiols are extensively involved in the maintenance of cellular redox potentials and the protection of cells from a variety of reactive chemical and electrophilic species. However, we recently found that the metabolic coupling of two LMW thiols - mycothiol (MSH) and ergothioneine (EGT) - programs the biosynthesis of the anti-infective agent lincomycin A. Remarkably, such a constructive role of the thiols in the biosynthesis of natural products has so far received relatively little attention. We speculate that the unusual thiol EGT might function as a chiral thiolation carrier (for modification) and a novel activator (for glycosylation) of sugar. Additionally, we examine recent evidence for LMW thiols (MSH and others) as sulfur donors of sulfur-containing natural products. Clearly, the LMW thiols have more diverse activities beyond cell protection, and more attention should be paid to the correlation of their functions with thiol-dependent enzymes

  ergothioneine, low-molecular-weight thiols, mycothiol, non-Leloir pathway, sulfur incorporation, thiolation carrier

  NCBI PubMed ID: 26515639
  Publication DOI: 10.1002/bies.201500067
  Journal NLM ID: 8510851
  Publisher: Hoboken, N.J.: Wiley
  Correspondence: wliu@mail.sioc.ac.cn
  Institutions: State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China, Huzhou Center of Bio-Synthetic Innovation, Huzhou, China
  
   
  
  Streptomyces caelestis
  CSDB ID 47722 (all data & tools)

Show legend Show as text

Structure type: monomer
Trivial name: celesticetin
Compound class: glycoside

• Article ID: 5606
  Zhao Q, Wang M, Xu D, Zhang Q, Liu W "Metabolic coupling of two small-molecule thiols programs the biosynthesis of lincomycin A" - Nature 518(7537) (2015) 115-119
  

  Low-molecular-mass thiols in organisms are well known for their redox-relevant role in protection against various endogenous and exogenous stresses. In eukaryotes and Gram-negative bacteria, the primary thiol is glutathione (GSH), a cysteinyl-containing tripeptide. In contrast, mycothiol (MSH), a cysteinyl pseudo-disaccharide, is dominant in Gram-positive actinobacteria, including antibiotic-producing actinomycetes and pathogenic mycobacteria. MSH is equivalent to GSH, either as a cofactor or as a substrate, in numerous biochemical processes, most of which have not been characterized, largely due to the dearth of information concerning MSH-dependent proteins. Actinomycetes are able to produce another thiol, ergothioneine (EGT), a histidine betaine derivative that is widely assimilated by plants and animals for variable physiological activities. The involvement of EGT in enzymatic reactions, however, lacks any precedent. Here we report that the unprecedented coupling of two bacterial thiols, MSH and EGT, has a constructive role in the biosynthesis of lincomycin A, a sulfur-containing lincosamide (C8 sugar) antibiotic that has been widely used for half a century to treat Gram-positive bacterial infections. EGT acts as a carrier to template the molecular assembly, and MSH is the sulfur donor for lincomycin maturation after thiol exchange. These thiols function through two unusual S-glycosylations that program lincosamide transfer, activation and modification, providing the first paradigm for EGT-associated biochemical processes and for the poorly understood MSH-dependent biotransformations, a newly described model that is potentially common in the incorporation of sulfur, an element essential for life and ubiquitous in living systems

  biosynthesis, lincomycin A, S-glycoside, mycothiols

  NCBI PubMed ID: 25607359
  Publication DOI: 10.1038/nature14137
  Journal NLM ID: 0410462
  Publisher: Basingstoke: Nature Publishing Group
  Correspondence: wliu@mail.sioc.ac.cn
  Institutions: State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China, Huzhou Center of Bio-Synthetic Innovation, Huzhou, China
  Methods: 13C NMR, 1H NMR, PCR, DNA techniques, ESI-MS, chemical synthesis, HPLC, CC, HR-ESI-MS, enzymatic assay, evaporation, centrifugation, HILIC
  
   
  
  Streptomyces caelestis
  CSDB ID 47750 (all data & tools)

Show legend Show as text

Structure type: oligomer
Contained glycoepitopes: IEDB_141807,IEDB_150900,IEDB_151531

• Article ID: 5606
  Zhao Q, Wang M, Xu D, Zhang Q, Liu W "Metabolic coupling of two small-molecule thiols programs the biosynthesis of lincomycin A" - Nature 518(7537) (2015) 115-119
  

  Low-molecular-mass thiols in organisms are well known for their redox-relevant role in protection against various endogenous and exogenous stresses. In eukaryotes and Gram-negative bacteria, the primary thiol is glutathione (GSH), a cysteinyl-containing tripeptide. In contrast, mycothiol (MSH), a cysteinyl pseudo-disaccharide, is dominant in Gram-positive actinobacteria, including antibiotic-producing actinomycetes and pathogenic mycobacteria. MSH is equivalent to GSH, either as a cofactor or as a substrate, in numerous biochemical processes, most of which have not been characterized, largely due to the dearth of information concerning MSH-dependent proteins. Actinomycetes are able to produce another thiol, ergothioneine (EGT), a histidine betaine derivative that is widely assimilated by plants and animals for variable physiological activities. The involvement of EGT in enzymatic reactions, however, lacks any precedent. Here we report that the unprecedented coupling of two bacterial thiols, MSH and EGT, has a constructive role in the biosynthesis of lincomycin A, a sulfur-containing lincosamide (C8 sugar) antibiotic that has been widely used for half a century to treat Gram-positive bacterial infections. EGT acts as a carrier to template the molecular assembly, and MSH is the sulfur donor for lincomycin maturation after thiol exchange. These thiols function through two unusual S-glycosylations that program lincosamide transfer, activation and modification, providing the first paradigm for EGT-associated biochemical processes and for the poorly understood MSH-dependent biotransformations, a newly described model that is potentially common in the incorporation of sulfur, an element essential for life and ubiquitous in living systems

  biosynthesis, lincomycin A, S-glycoside, mycothiols

  NCBI PubMed ID: 25607359
  Publication DOI: 10.1038/nature14137
  Journal NLM ID: 0410462
  Publisher: Basingstoke: Nature Publishing Group
  Correspondence: wliu@mail.sioc.ac.cn
  Institutions: State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China, Huzhou Center of Bio-Synthetic Innovation, Huzhou, China
  Methods: 13C NMR, 1H NMR, PCR, DNA techniques, ESI-MS, chemical synthesis, HPLC, CC, HR-ESI-MS, enzymatic assay, evaporation, centrifugation, HILIC
  
   
  
  Streptomyces lincolnensis (ΔlmbE mutant), Streptomyces lincolnensis (ΔlmbE-E3457)
  CSDB ID 47751 (all data & tools)

Show legend Show as text

Structure type: monomer
Contained glycoepitopes: IEDB_150900

• Article ID: 5606
  Zhao Q, Wang M, Xu D, Zhang Q, Liu W "Metabolic coupling of two small-molecule thiols programs the biosynthesis of lincomycin A" - Nature 518(7537) (2015) 115-119
  

  Low-molecular-mass thiols in organisms are well known for their redox-relevant role in protection against various endogenous and exogenous stresses. In eukaryotes and Gram-negative bacteria, the primary thiol is glutathione (GSH), a cysteinyl-containing tripeptide. In contrast, mycothiol (MSH), a cysteinyl pseudo-disaccharide, is dominant in Gram-positive actinobacteria, including antibiotic-producing actinomycetes and pathogenic mycobacteria. MSH is equivalent to GSH, either as a cofactor or as a substrate, in numerous biochemical processes, most of which have not been characterized, largely due to the dearth of information concerning MSH-dependent proteins. Actinomycetes are able to produce another thiol, ergothioneine (EGT), a histidine betaine derivative that is widely assimilated by plants and animals for variable physiological activities. The involvement of EGT in enzymatic reactions, however, lacks any precedent. Here we report that the unprecedented coupling of two bacterial thiols, MSH and EGT, has a constructive role in the biosynthesis of lincomycin A, a sulfur-containing lincosamide (C8 sugar) antibiotic that has been widely used for half a century to treat Gram-positive bacterial infections. EGT acts as a carrier to template the molecular assembly, and MSH is the sulfur donor for lincomycin maturation after thiol exchange. These thiols function through two unusual S-glycosylations that program lincosamide transfer, activation and modification, providing the first paradigm for EGT-associated biochemical processes and for the poorly understood MSH-dependent biotransformations, a newly described model that is potentially common in the incorporation of sulfur, an element essential for life and ubiquitous in living systems

  biosynthesis, lincomycin A, S-glycoside, mycothiols

  NCBI PubMed ID: 25607359
  Publication DOI: 10.1038/nature14137
  Journal NLM ID: 0410462
  Publisher: Basingstoke: Nature Publishing Group
  Correspondence: wliu@mail.sioc.ac.cn
  Institutions: State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China, Huzhou Center of Bio-Synthetic Innovation, Huzhou, China
  Methods: 13C NMR, 1H NMR, PCR, DNA techniques, ESI-MS, chemical synthesis, HPLC, CC, HR-ESI-MS, enzymatic assay, evaporation, centrifugation, HILIC
  
   
  
  Streptomyces lincolnensis (ΔmshAlin mutant)
  CSDB ID 47752 (all data & tools)

Show legend Show as text

Structure type: monomer

• Article ID: 5606
  Zhao Q, Wang M, Xu D, Zhang Q, Liu W "Metabolic coupling of two small-molecule thiols programs the biosynthesis of lincomycin A" - Nature 518(7537) (2015) 115-119
  

  Low-molecular-mass thiols in organisms are well known for their redox-relevant role in protection against various endogenous and exogenous stresses. In eukaryotes and Gram-negative bacteria, the primary thiol is glutathione (GSH), a cysteinyl-containing tripeptide. In contrast, mycothiol (MSH), a cysteinyl pseudo-disaccharide, is dominant in Gram-positive actinobacteria, including antibiotic-producing actinomycetes and pathogenic mycobacteria. MSH is equivalent to GSH, either as a cofactor or as a substrate, in numerous biochemical processes, most of which have not been characterized, largely due to the dearth of information concerning MSH-dependent proteins. Actinomycetes are able to produce another thiol, ergothioneine (EGT), a histidine betaine derivative that is widely assimilated by plants and animals for variable physiological activities. The involvement of EGT in enzymatic reactions, however, lacks any precedent. Here we report that the unprecedented coupling of two bacterial thiols, MSH and EGT, has a constructive role in the biosynthesis of lincomycin A, a sulfur-containing lincosamide (C8 sugar) antibiotic that has been widely used for half a century to treat Gram-positive bacterial infections. EGT acts as a carrier to template the molecular assembly, and MSH is the sulfur donor for lincomycin maturation after thiol exchange. These thiols function through two unusual S-glycosylations that program lincosamide transfer, activation and modification, providing the first paradigm for EGT-associated biochemical processes and for the poorly understood MSH-dependent biotransformations, a newly described model that is potentially common in the incorporation of sulfur, an element essential for life and ubiquitous in living systems

  biosynthesis, lincomycin A, S-glycoside, mycothiols

  NCBI PubMed ID: 25607359
  Publication DOI: 10.1038/nature14137
  Journal NLM ID: 0410462
  Publisher: Basingstoke: Nature Publishing Group
  Correspondence: wliu@mail.sioc.ac.cn
  Institutions: State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China, Huzhou Center of Bio-Synthetic Innovation, Huzhou, China
  Methods: 13C NMR, 1H NMR, PCR, DNA techniques, ESI-MS, chemical synthesis, HPLC, CC, HR-ESI-MS, enzymatic assay, evaporation, centrifugation, HILIC
  
   
  
  Streptomyces lincolnensis (ΔlmbV mutant), Streptomyces lincolnensis (ΔmshAlin mutant)
  CSDB ID 47753 (all data & tools)