TDP-L-megosamine Biosynthesis Pathway Elucidation and Megalomicin a Production in Escherichia Coli

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 2010 Apr 27

PMID 20418422

Citations 10

Authors

Mariana Useglio

Salvador Peiru

Eduardo Rodriguez

Guillermo R Labadie

John R Carney

Hugo Gramajo

Affiliations

Soon will be listed here.

Abstract

In vivo reconstitution of the TDP-l-megosamine pathway from the megalomicin gene cluster of Micromonospora megalomicea was accomplished by the heterologous expression of its biosynthetic genes in Escherichia coli. Mass spectrometric analysis of the TDP-sugar intermediates produced from operons containing different sets of genes showed that the production of TDP-l-megosamine from TDP-4-keto-6-deoxy-d-glucose requires only five biosynthetic steps, catalyzed by MegBVI, MegDII, MegDIII, MegDIV, and MegDV. Bioconversion studies demonstrated that the sugar transferase MegDI, along with the helper protein MegDVI, catalyzes the transfer of l-megosamine to either erythromycin C or erythromycin D, suggesting two possible routes for the production of megalomicin A. Analysis in vivo of the hydroxylation step by MegK indicated that erythromycin C is the intermediate of megalomicin A biosynthesis.

Citing Articles

Antibacterial activity and biosynthetic potential of Streptomyces sp. PBR19, isolated from forest rhizosphere soil of Assam.

Mazumdar R, Thakur D Braz J Microbiol. 2024; 55(4):3335-3352.

PMID: 38985434 PMC: 11711432. DOI: 10.1007/s42770-024-01454-3.

Nematicidal glycosylated resorcylic acid lactones from the fungus PC-170 and their key biosynthetic genes.

Li Z, Luo N, Zhang W, Khan R, Ling J, Zhao J Front Microbiol. 2024; 15:1385255.

PMID: 38638906 PMC: 11024724. DOI: 10.3389/fmicb.2024.1385255.

OSMAC Method to Assess Impact of Culture Parameters on Metabolomic Diversity and Biological Activity of Marine-Derived Actinobacteria.

Le Loarer A, Dufosse L, Bignon J, Frederich M, Ledoux A, Fouillaud M Mar Drugs. 2024; 22(1).

PMID: 38248648 PMC: 10817652. DOI: 10.3390/md22010023.

New Insights on Biological Activities, Chemical Compositions, and Classifications of Marine Actinomycetes Antifouling Agents.

Morgan R, Al Ali A, Alshahrani M, Aboshanab K Microorganisms. 2023; 11(10).

PMID: 37894102 PMC: 10609280. DOI: 10.3390/microorganisms11102444.

Prioritization of Microorganisms Isolated from the Indian Ocean Sponge Based on Metabolomic Diversity and Biological Activity for the Discovery of Natural Products.

Le Loarer A, Marcellin-Gros R, Dufosse L, Bignon J, Frederich M, Ledoux A Microorganisms. 2023; 11(3).

PMID: 36985270 PMC: 10057949. DOI: 10.3390/microorganisms11030697.

References

Alarcon B, Gonzalez M, Carrasco L . Megalomycin C, a macrolide antibiotic that blocks protein glycosylation and shows antiviral activity. FEBS Lett. 1988; 231(1):207-11. DOI: 10.1016/0014-5793(88)80732-4. View

Chen H, Thomas M, Hubbard B, Losey H, WALSH C, Burkart M . Deoxysugars in glycopeptide antibiotics: enzymatic synthesis of TDP-L-epivancosamine in chloroeremomycin biosynthesis. Proc Natl Acad Sci U S A. 2000; 97(22):11942-7. PMC: 17274. DOI: 10.1073/pnas.210395097. View

Hwang B, Lee H, Yang Y, Joo H, Kim B . Characterization and investigation of substrate specificity of the sugar aminotransferase WecE from E. coli K12. Chem Biol. 2004; 11(7):915-25. DOI: 10.1016/j.chembiol.2004.04.015. View

Lu W, Leimkuhler C, Gatto Jr G, Kruger R, Oberthur M, Kahne D . AknT is an activating protein for the glycosyltransferase AknS in L-aminodeoxysugar transfer to the aglycone of aclacinomycin A. Chem Biol. 2005; 12(5):527-34. DOI: 10.1016/j.chembiol.2005.02.016. View

Bonay P, Fresno M, Alarcon B, Alcina A . Antiparasitic effects of the intra-Golgi transport inhibitor megalomicin. Antimicrob Agents Chemother. 1998; 42(10):2668-73. PMC: 105916. DOI: 10.1128/AAC.42.10.2668. View

Hong J, Park S, Parajuli N, Park S, Koh H, Jung W . Functional analysis of desVIII homologues involved in glycosylation of macrolide antibiotics by interspecies complementation. Gene. 2006; 386(1-2):123-30. DOI: 10.1016/j.gene.2006.08.021. View

Weinstein M, Wagman G, Marquez J, Testa R, Oden E, Waitz J . Megalomicin, a new macrolide antibiotic complex produced by Micromonospora. J Antibiot (Tokyo). 1969; 22(6):253-8. DOI: 10.7164/antibiotics.22.253. View

Williams G, Gantt R, Thorson J . The impact of enzyme engineering upon natural product glycodiversification. Curr Opin Chem Biol. 2008; 12(5):556-64. PMC: 4552347. DOI: 10.1016/j.cbpa.2008.07.013. View

Tamura K, Dudley J, Nei M, Kumar S . MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol. 2007; 24(8):1596-9. DOI: 10.1093/molbev/msm092. View

10.

Borisova S, Zhao L, Melancon Iii C, Kao C, Liu H . Characterization of the glycosyltransferase activity of desVII: analysis of and implications for the biosynthesis of macrolide antibiotics. J Am Chem Soc. 2004; 126(21):6534-5. DOI: 10.1021/ja049967j. View

11.

Peiru S, Menzella H, Rodriguez E, Carney J, Gramajo H . Production of the potent antibacterial polyketide erythromycin C in Escherichia coli. Appl Environ Microbiol. 2005; 71(5):2539-47. PMC: 1087553. DOI: 10.1128/AEM.71.5.2539-2547.2005. View

12.

Yuan Y, Chung H, Leimkuhler C, Walsh C, Kahne D, Walker S . In vitro reconstitution of EryCIII activity for the preparation of unnatural macrolides. J Am Chem Soc. 2005; 127(41):14128-9. PMC: 2525786. DOI: 10.1021/ja053704n. View

13.

San Jose E, Munoz-Fernandez M, Alarcon B . Megalomicin inhibits HIV-1 replication and interferes with gp160 processing. Virology. 1998; 239(2):303-14. DOI: 10.1006/viro.1997.8872. View

14.

Chen H, Yamase H, Murakami K, Chang C, Zhao L, Zhao Z . Expression, purification, and characterization of two N,N-dimethyltransferases, tylM1 and desVI, involved in the biosynthesis of mycaminose and desosamine. Biochemistry. 2002; 41(29):9165-83. DOI: 10.1021/bi020245j. View

15.

Jaret R, Mallams A, Vernay H . The megalomicins. V. Mass spectral studies. J Chem Soc Perkin 1. 1973; 13:1389-400. DOI: 10.1039/p19730001389. View

16.

Jaret R, Mallams A, Reimann H . The megalomicins. IV. The structures of megalomicins A, B, C1, and C2. J Chem Soc Perkin 1. 1973; 13:1374-88. DOI: 10.1039/p19730001374. View

17.

Peiru S, Rodriguez E, Tran C, Carney J, Gramajo H . Characterization of the heterodimeric MegBIIa:MegBIIb aldo-keto reductase involved in the biosynthesis of L-mycarose from Micromonospora megalomicea. Biochemistry. 2007; 46(27):8100-9. DOI: 10.1021/bi700396n. View

18.

Wang Y, Xu Y, Perepelov A, Qi Y, Knirel Y, Wang L . Biochemical characterization of dTDP-D-Qui4N and dTDP-D-Qui4NAc biosynthetic pathways in Shigella dysenteriae type 7 and Escherichia coli O7. J Bacteriol. 2007; 189(23):8626-35. PMC: 2168959. DOI: 10.1128/JB.00777-07. View

19.

Bonay P, Fresno M, Alarcon B . Megalomicin disrupts lysosomal functions. J Cell Sci. 1997; 110 ( Pt 16):1839-49. DOI: 10.1242/jcs.110.16.1839. View

20.

Peiru S, Rodriguez E, Menzella H, Carney J, Gramajo H . Metabolically engineered Escherichia coli for efficient production of glycosylated natural products. Microb Biotechnol. 2011; 1(6):476-86. PMC: 3815289. DOI: 10.1111/j.1751-7915.2008.00046.x. View