Coordinated Action of NRPS, Baeyer-Villiger Monooxygenase, and Methyltransferase Ensures the Economical Biosynthesis of Bohemamines in Streptomyces sp. CB02009

2020 ◽  
Author(s):  
Ling Liu ◽  
Sainan Li ◽  
Runze Sun ◽  
Xiangjing Qin ◽  
Jianhua Ju ◽  
...  
Keyword(s):  
Methyl Groups ◽  
Ribosome Engineering ◽  
Methyl Group ◽  
Proteinogenic Amino Acid ◽  
Streptomyces Sp ◽  
Coordinated Action ◽  
Peptide Synthetase ◽  
Identification And Characterization ◽  
Shed Light

<p> Bohemamines (BHMs) are bacterial alkaloids containing a pyrrolizidine core with two unprecedented methyl groups. Herein we report the activation of BHMs biosynthesis in <i>Streptomyces </i>sp. CB02009 using a ribosome engineering approach. Identification and characterization of the <i>bhm</i> gene cluster reveals a coordinated action of nonribosomal peptide synthetase BhmJ, Baeyer-villiger monooxygenase BhmK and methyltransferase BhmG for BHMs biosynthesis. BhmG is responsible for the C-methylation on C-7, while the C-9 methyl group is from a non-proteinogenic amino acid (2<i>S</i>,5<i>S</i>)-5-methylproline, required for BHMs production in three model <i>Streptomyces </i>hosts. Our study shed light on the intricate interaction of BhmJ/BhmK/BhmG for the economical biosynthesis of BHMs in their native producer, and also unraveled that BhmJ and BhmK are competent biocatalysts in <i>S</i><i>treptomyce </i><i>albus</i>.</p>

Coordinated Action of NRPS, Baeyer-Villiger Monooxygenase, and Methyltransferase Ensures the Economical Biosynthesis of Bohemamines in Streptomyces sp. CB02009

2020 ◽  
Author(s):  
Ling Liu ◽  
Sainan Li ◽  
Runze Sun ◽  
Xiangjing Qin ◽  
Jianhua Ju ◽  
...  
Keyword(s):  
Methyl Groups ◽  
Ribosome Engineering ◽  
Methyl Group ◽  
Proteinogenic Amino Acid ◽  
Streptomyces Sp ◽  
Coordinated Action ◽  
Peptide Synthetase ◽  
Identification And Characterization ◽  
Shed Light

<p> Bohemamines (BHMs) are bacterial alkaloids containing a pyrrolizidine core with two unprecedented methyl groups. Herein we report the activation of BHMs biosynthesis in <i>Streptomyces </i>sp. CB02009 using a ribosome engineering approach. Identification and characterization of the <i>bhm</i> gene cluster reveals a coordinated action of nonribosomal peptide synthetase BhmJ, Baeyer-villiger monooxygenase BhmK and methyltransferase BhmG for BHMs biosynthesis. BhmG is responsible for the C-methylation on C-7, while the C-9 methyl group is from a non-proteinogenic amino acid (2<i>S</i>,5<i>S</i>)-5-methylproline, required for BHMs production in three model <i>Streptomyces </i>hosts. Our study shed light on the intricate interaction of BhmJ/BhmK/BhmG for the economical biosynthesis of BHMs in their native producer, and also unraveled that BhmJ and BhmK are competent biocatalysts in <i>S</i><i>treptomyce </i><i>albus</i>.</p>

Identification and characterization of a novel spermidine/spermine acetyltransferase encoded by gene ste26 from Streptomyces sp. 139

Biochimie ◽  
2011 ◽  
Vol 93 (9) ◽  
pp. 1401-1407 ◽  
Author(s):  
Liping Bai ◽  
Ming Chang ◽  
Junjie Shan ◽  
Rong Jiang ◽  
Yang Zhang ◽  
...  
Keyword(s):  
Streptomyces Sp ◽  
Identification And Characterization

scholarly journals Activation and Identification of a Griseusin Cluster in Streptomyces sp. CA-256286 by Employing Transcriptional Regulators and Multi-Omics Methods

Molecules ◽  
2021 ◽  
Vol 26 (21) ◽  
pp. 6580
Author(s):  
Charlotte Beck ◽  
Tetiana Gren ◽  
Francisco Javier Ortiz-López ◽  
Tue Sparholt Jørgensen ◽  
Daniel Carretero-Molina ◽  
...  
Keyword(s):  
Biosynthetic Pathway ◽  
Genome Mining ◽  
Gene Clusters ◽  
Transcriptional Regulators ◽  
Biosynthetic Gene ◽  
Biosynthetic Gene Clusters ◽  
Heterologous Host ◽  
Streptomyces Sp ◽  
Identification And Characterization

Streptomyces are well-known producers of a range of different secondary metabolites, including antibiotics and other bioactive compounds. Recently, it has been demonstrated that “silent” biosynthetic gene clusters (BGCs) can be activated by heterologously expressing transcriptional regulators from other BGCs. Here, we have activated a silent BGC in Streptomyces sp. CA-256286 by overexpression of a set of SARP family transcriptional regulators. The structure of the produced compound was elucidated by NMR and found to be an N-acetyl cysteine adduct of the pyranonaphtoquinone polyketide 3′-O-α-d-forosaminyl-(+)-griseusin A. Employing a combination of multi-omics and metabolic engineering techniques, we identified the responsible BGC. These methods include genome mining, proteomics and transcriptomics analyses, in combination with CRISPR induced gene inactivations and expression of the BGC in a heterologous host strain. This work demonstrates an easy-to-implement workflow of how silent BGCs can be activated, followed by the identification and characterization of the produced compound, the responsible BGC, and hints of its biosynthetic pathway.

scholarly journals Identification and characterization of functional homologs of nitrogenase cofactor biosynthesis protein NifB from methanogens

2015 ◽  
Vol 112 (48) ◽  
pp. 14829-14833 ◽  
Author(s):  
Aaron W. Fay ◽  
Jared A. Wiig ◽  
Chi Chung Lee ◽  
Yilin Hu
Keyword(s):  
Azotobacter Vinelandii ◽  
Methanosarcina Acetivorans ◽  
Radical Sam ◽  
Methyl Group ◽  
C Terminus ◽  
Cofactor Biosynthesis ◽  
Monomeric Proteins ◽  
Cluster Biosynthesis ◽  
Identification And Characterization

Nitrogenase biosynthesis protein NifB catalyzes the radical S-adenosyl-L-methionine (SAM)-dependent insertion of carbide into the M cluster, the cofactor of the molybdenum nitrogenase from Azotobacter vinelandii. Here, we report the identification and characterization of two naturally “truncated” homologs of NifB from Methanosarcina acetivorans (NifBMa) and Methanobacterium thermoautotrophicum (NifBMt), which contain a SAM-binding domain at the N terminus but lack a domain toward the C terminus that shares homology with NifX, an accessory protein in M cluster biosynthesis. NifBMa and NifBMt are monomeric proteins containing a SAM-binding [Fe4S4] cluster (designated the SAM cluster) and a [Fe4S4]-like cluster pair (designated the K cluster) that can be processed into an [Fe8S9] precursor to the M cluster (designated the L cluster). Further, the K clusters in NifBMa and NifBMt can be converted to L clusters upon addition of SAM, which corresponds to their ability to heterologously donate L clusters to the biosynthetic machinery of A. vinelandii for further maturation into the M clusters. Perhaps even more excitingly, NifBMa and NifBMt can catalyze the removal of methyl group from SAM and the abstraction of hydrogen from this methyl group by 5′-deoxyadenosyl radical that initiates the radical-based incorporation of methyl-derived carbide into the M cluster. The successful identification of NifBMa and NifBMt as functional homologs of NifB not only enabled classification of a new subset of radical SAM methyltransferases that specialize in complex metallocluster assembly, but also provided a new tool for further characterization of the distinctive, NifB-catalyzed methyl transfer and conversion to an iron-bound carbide.

scholarly journals The role of the central melanocortin system in the regulation of food intake and energy homeostasis: lessons from mouse models

2006 ◽  
Vol 361 (1471) ◽  
pp. 1265-1274 ◽  
Author(s):  
Kate L.J Ellacott ◽  
Roger D Cone
Keyword(s):  
Energy Homeostasis ◽  
Neural Circuitry ◽  
Neural Systems ◽  
Melanocortin System ◽  
Genes Encoding ◽  
Obesity Genes ◽  
Identification And Characterization ◽  
Shed Light

A little more than a decade ago, the molecular basis of the lipostat was largely unknown. At that time, many laboratories were at work attempting to clone the genes encoding the obesity , diabetes , fatty , tubby and agouti loci, with the hope that identification of these obesity genes would help shed light on the process of energy homeostasis, appetite and energy expenditure. Characterization of obesity and diabetes elucidated the nature of the adipostatic hormone leptin and its receptor, respectively, while cloning of the agouti gene eventually led to the identification and characterization of one of the key neural systems upon which leptin acts to regulate intake and expenditure. In this review, we describe the neural circuitry known as the central melanocortin system and discuss the current understanding of its role in feeding and other processes involved in energy homeostasis.

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