Functional Characterization of a Regiospecific Sugar-<i>O</i>-Methyltransferase from<i>Nocardia</i>

Poudel, Purna Bahadur; Pandey, Ramesh Prasad; Dhakal, Dipesh; Kim, Taesu; Nguyen, Trang Thi Huyen; Jung, Hye Jin; Shin, Hee Jeong; Timalsina, Binod; Sohng, Jae Kyung

doi:10.1128/aem.00754-22

Cited by 3 publications

(3 citation statements)

References 33 publications

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“…CS682 were taken to prepare E . coli S2 [ 30 ]. E. coli S2 was generated by transforming pET32a ± 7665(Am) CRISPRi-S1(Cm r ) and pCDFDuet-metK-thnM1(Sm) into an E. coli strain harboring the rhamnose cassette piBR181-tgs.dh.ep.kr.pgm2.glf.glk (Km) [ 31 ].…”

Section: Methodsmentioning

confidence: 99%

Microbial Biosynthesis of Chrysazin Derivatives in Recombinant Escherichia coli and Their Biological Activities

Poudel

Dhakal²,

Magar³

et al. 2022

Molecules

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Anthraquinone and its derivatives show remarkable biological properties such as anticancer, antibacterial, antifungal, and antiviral activities. Hence, anthraquinones derivatives have been of prime interest in drug development. This study developed a recombinant Escherichia coli strain to modify chrysazin to chrysazin-8-O-α-l-rhamnoside (CR) and chrysazin-8-O-α-l-2′-O-methylrhamnoside (CRM) using rhamnosyl transferase and sugar-O-methyltransferase. Biosynthesized CR and CRM were structurally characterized using HPLC, high-resolution mass spectrometry, and various nuclear magnetic resonance analyses. Antimicrobial effects of chrysazin, CR, and CRM against 18 superbugs, including 14 Gram-positive and 4 Gram-negative pathogens, were investigated. CR and CRM exhibited antimicrobial activities against nine pathogens, including methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA) in a disk diffusion assay at a concentration of 40 µg per disk. There were MIC and MBC values of 7.81–31.25 µg/mL for CR and CRM against methicillin-sensitive S. aureus CCARM 0205 (MSSA) for which the parent chrysazin is more than >1000 µg/mL. Furthermore, the anti-proliferative properties of chrysazin, CR, and CRM were assayed using AGS, Huh7, HL60, and HaCaT cell lines. CR and CRM showed higher antibacterial and anticancer properties than chrysazin.

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Section: Methodsmentioning

confidence: 99%

Microbial Biosynthesis of Chrysazin Derivatives in Recombinant Escherichia coli and Their Biological Activities

Poudel

Dhakal²,

Magar³

et al. 2022

Molecules

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“…CS682 genome [ 21 ]. Although its biological function is currently unknown, ThnA has been implicated in the biosynthesis of 1-( α -L-(2- O -methyl)-6-deoxymannopyranosyloxy)-3,6,8-trimethoxynaphthalene (IBR-3) [ 22 ]. Nocardia sp.…”

Section: Introductionmentioning

confidence: 99%

“…Different enzymes like methyltransferases and glycosyltransferase are involved in the post-modification of 1,3,6,8-tetrahydroxynaphthalene (THN) during the formation of IBR-3 ( Fig. 2 ) [ 15 , 21 , 22 ]. Due to the crucial role played by ThnA in forming the core THN scaffold, we carried out a sequence alignment of ThnA with established type III PKSs.…”

Section: Introductionmentioning

confidence: 99%

Identification of 1,3,6,8-Tetrahydroxynaphthalene Synthase (ThnA) from Nocardia sp. CS682

Poudel

Magar

Bridget

et al. 2023

J. Microbiol. Biotechnol.

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Type III polyketide synthase (PKS) found in bacteria is known as 1,3,6,8-tetrahydroxynaphthalene synthase (THNS). Microbial type III PKSs synthesize various compounds that possess crucial biological functions and significant pharmaceutical activities. Based on our sequence analysis, we have identified a putative type III polyketide synthase from Nocardia sp. CS682 was named as ThnA. The role of ThnA, in Nocardia sp. CS682 during the biosynthesis of 1,3,6,8 tetrahydroxynaphthalene (THN), which is the key intermediate of 1-( α -L-(2- O -methyl)-6-deoxymannopyranosyloxy)-3,6,8-trimethoxynaphthalene (IBR-3) was characterized. ThnA utilized five molecules of malonyl-CoA as a starter substrate to generate the polyketide 1,3,6,8-tetrahydroxynaphthalene, which could spontaneously be oxidized to the red flaviolin compound 2,5,7-trihydroxy-1,4-naphthoquinone. The amino acid sequence alignment of ThnA revealed similarities with a previously identified type III PKS and identified Cys 138 , Phe 188 , His 270 , and Asn 303 as four highly conserved active site amino acid residues, as found in other known polyketide synthases. In this study, we report the heterologous expression of the type III polyketide synthase thnA in S. lividan TK24 and the identification of THN production in a mutant strain. We also compared the transcription level of thnA in S. lividan TK24 and S. lividan pIBR25- thnA and found that thnA was only transcribed in the mutant.

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Functional characterization of a naphthalene-O-methyltransferase from Nocardia sp. CS682

Poudel,

Dhakal,

Lee

et al. 2024

Enzyme and Microbial Technology

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Functional Characterization of a Regiospecific Sugar-O-Methyltransferase fromNocardia

Cited by 3 publications

References 33 publications

Microbial Biosynthesis of Chrysazin Derivatives in Recombinant Escherichia coli and Their Biological Activities

Microbial Biosynthesis of Chrysazin Derivatives in Recombinant Escherichia coli and Their Biological Activities

Identification of 1,3,6,8-Tetrahydroxynaphthalene Synthase (ThnA) from Nocardia sp. CS682

Functional characterization of a naphthalene-O-methyltransferase from Nocardia sp. CS682

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