Flexible enzymatic activation of artificial polyketide extender units by <i>Streptomyces cinnamonensis</i>
 into the monensin biosynthetic pathway

Möller, David R.; Kushnir, Susanna; Grote, Marius; Ismail-Ali, A.; Koopmans, Kyra R.M.; Calo, F.; Heinrich, Sascha; Diehl, Bernd; Schulz, Frank

doi:10.1111/lam.13039

Cited by 8 publications

(12 citation statements)

References 23 publications

(51 reference statements)

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“…In order to explore the in vivo applicability of MatB_Sc, five malonate derivatives [2‐allyl, 2‐propyl, 2‐propargyl, 2‐butyl and 2‐(3‐chloropropyl)] were supplemented to a fermentation culture of S. cinnamonensis ATCC15413, assuming a co‐expression of MatB_Sc and the monensin PKS, as observed in the case of analogous premonensin fermentations . Five replicates were analyzed independently to allow a computer‐assisted analysis using the MetaboScape software suite for the determination of new metabolites .…”

Section: Resultscontrasting

confidence: 79%

“…We recently reported on the MatB_Sc from S. cinnamonensis . A biocatalytic assay with purified MatB_Sc was performed to further explore its substrate scope.…”

Section: Resultssupporting

confidence: 71%

“…Substrate acceptance was monitored by direct product detection with HRMS/MS 2 . HRMS data for the main H + ‐adduct are shown in the table enclosed, and further adducts, characteristic fragmentation pattern, isotopic pattern and MS 2 data are shown in the Supporting Information (Tables S1 and S2, Figures S1 and S2); 4 – 6 and 11 – 12 were detected in an earlier report on this enzyme . Additional data were obtained for 8 – 16 in Michaelis–Menten kinetics (Figure ).…”

Section: Resultsmentioning

confidence: 67%

“…While direct supplementation of CoA‐activated malonate derivatives is unsuitable due to its limited membrane permeability and synthetic accessibility, N ‐acetylcysteamine esters (SNAC esters), acting as a CoA‐mimic, have been successfully used in feeding experiments . However, significant synthetic effort ahead of the fermentation, toxicity in fermentations and inferior activation compared to CoA‐thioesters have limited their application …”

Section: Introductionmentioning

confidence: 52%

“…We recently introduced a new MatB‐type enzyme from the monensin producer S. cinnamonensis and demonstrated its in vivo activity on a small number of different malonic acid derivatives, outcompeting artificial SNAC activation …”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Exploring the Promiscuous Enzymatic Activation of Unnatural Polyketide Extender Units in Vitro and in Vivo for Monensin Biosynthesis

Grote

Schulz

2019

ChemBioChem

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The incorporation of new‐to‐nature extender units into polyketide synthesis is an important source for diversity yet is restricted by limited availability of suitably activated building blocks in vivo. We here describe a straightforward workflow for the biogenic activation of commercially available new‐to‐nature extender units. Firstly, the substrate scope of a highly flexible malonyl co‐enzyme A synthetase from Streptomyces cinnamonensis was characterized. The results were matched by in vivo experiments in which the said extender units were accepted by both the polyketide synthase and the accessory enzymes of the monensin biosynthetic pathway. The experiments gave rise to a series of predictable monensin derivatives by the exploitation of the innate substrate promiscuity of an acyltransferase and downstream enzyme functions.

show abstract

Section: Resultscontrasting

confidence: 79%

“…We recently reported on the MatB_Sc from S. cinnamonensis . A biocatalytic assay with purified MatB_Sc was performed to further explore its substrate scope.…”

Section: Resultssupporting

confidence: 71%

Section: Resultsmentioning

confidence: 67%

Section: Introductionmentioning

confidence: 52%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Exploring the Promiscuous Enzymatic Activation of Unnatural Polyketide Extender Units in Vitro and in Vivo for Monensin Biosynthesis

Grote

Schulz

2019

ChemBioChem

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Polyether Cyclization Cascade Alterations in Response to Monensin Polyketide Synthase Mutations

et al. 2021

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The targeted manipulation of polyketide synthases has in recent years led to numerous new-to-nature polyketides. For type I polyketide synthases the response of post-polyketide synthases (PKS) processing enzymes onto the most frequently polyketide backbone manipulations is so far insufficiently studied. In particular, complex processes such as the polyether cyclisation in the biosynthesis of ionophores such as monensin pose interesting objects of research. We present here a study of the substrate promiscuity of the polyether cyclisation cascade enzymes in monensin biosynthesis in the conversion of redox derivatives of the nascent polyketide chain. LC-HRMS/MS 2based studies revealed a remarkable flexibility of the post-PKS enzymes. They acted on derivatized polyketide backbones based on the three possible polyketide redox states within two different modules and gave rise to an altered polyether structure. One of these monensin derivatives was isolated and characterized by 2D-NMR spectroscopy, crystallography, and bioactivity studies. Results and DiscussionFor the interrogation of post-PKS processivity, it was first decided to investigate the effect of all three feasible different [a

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Engineering of a Malonyl‐CoA Ligase for Production of Fluorinated Polyketide Extender Units

Paulsel,

Williams

2024

ChemBioChem

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Enzymatic platforms for producing malonyl‐CoA‐based extender units required for polyketide biosynthesis are often based on malonyl‐CoA ligases such as MatB from Rhizobium trifolii and Rhodopseudomonas palustris. However, despite broad interest in the fluorination of polyketides and prior success with engineering MatB homologs, the suitability of MatB for accessing the tertiary substituted fluoromethylmalonyl‐CoA needed to produce flurithromycin and solithromycin has not yet been reported. Herein, we report the structure‐guided engineering of a MatB homolog to optimize the production of fluoromethylmalonyl‐CoA, resulting in a variant with increased conversion and providing a platform to produce a suitable building block mixture for fluorinated macrolide production. Additionally, the mutant demonstrated broad utility for various substituted malonyl‐CoAs. The MatB mutant sets the stage to access fluorinated macrolides by coupling it with altered PKS machinery to install fluorinated malonyl‐CoA into macrolide scaffolds.

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Flexible enzymatic activation of artificial polyketide extender units by Streptomyces cinnamonensis into the monensin biosynthetic pathway

Cited by 8 publications

References 23 publications

Exploring the Promiscuous Enzymatic Activation of Unnatural Polyketide Extender Units in Vitro and in Vivo for Monensin Biosynthesis

Exploring the Promiscuous Enzymatic Activation of Unnatural Polyketide Extender Units in Vitro and in Vivo for Monensin Biosynthesis

Polyether Cyclization Cascade Alterations in Response to Monensin Polyketide Synthase Mutations

Engineering of a Malonyl‐CoA Ligase for Production of Fluorinated Polyketide Extender Units

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