Effect of Different Biosynthetic Precursors on the Production of Nargenicin A1 from Metabolically Engineered Nocardia sp. CS682

Koju, Dinesh; Maharjan, Sushila; Dhakal, Dipesh; Yoo, Jin Cheol; Sohng, Jae Kyung

doi:10.4014/jmb.1202.02027

Cited by 13 publications

(10 citation statements)

References 28 publications

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“…Previously, the enhancement of nargenicin A 1 was successfully obtained to 3.8-fold with metabolic engineering approach [29]. Further, by using methyl oleate, sodium propionate, and sodium acetate as sources for enhancing the pool of short fatty acids, the level of nargenicin A 1 was enhanced to the level of 6.99-fold [23]. In this study, the effect of further cheap sources for enhancing the level of short chain fatty acids and specific precursor for unique pyrrole moiety of nargenicin A 1 was evaluated.…”

Section: Discussionmentioning

confidence: 96%

“…CS682 without feeding. Previously, there has been ample reports of enhancement of different secondary metabolites by heterologous expression of metK1-sp and ACCase in different actinomycetes with media optimization and exogenous supplementation [23,[31][32][33]. Thus, directing the precursor flux by metabolic engineering and supplementing the sources, which in turn contribute for an enhanced precursor titer can be a rational strategy for enhancing the molecules of importance.…”

Section: Discussionmentioning

confidence: 97%

“…The primary metabolite precursors such as acetate are directed to the pool of malonyl-CoA, and propionate contributes for methylmalonyl-CoA. Thus, there has been considerable interest in enhancing the yield of polyketides by increasing the respective precursors because intracellular availability of biosynthetic precursors may play an influential role in productivity [23]. Previously, the enhancement of nargenicin A 1 was successfully obtained to 3.8-fold with metabolic engineering approach [29].…”

Section: Discussionmentioning

confidence: 98%

“…The strains were seeded in brain heart infusion (BHI) broth at 37°C for 5 days. Nargenicin A 1 aglycon has been shown to be derived from common precursors as acetate and propionate [23], and the pyrrole moiety is presumed to be derived from L-proline [24,25]. In the present study, the influence of precursor substrates including ethanol, glucose, and glycerol as source of acetate, propanol as source of propionate, and L-proline contributing for pyrrole moiety, was evaluated on nargenicin A 1 biosynthesis.…”

Section: Bacterial Strains and Growth Conditionsmentioning

confidence: 99%

“…ACC18 (Nocardia sp. CS682 harboring acetyl-CoA carboxylase complex, accA2 and accBE from Streptomyces coelicolor) [23] were used for the study. The strains were seeded in brain heart infusion (BHI) broth at 37°C for 5 days.…”

Section: Bacterial Strains and Growth Conditionsmentioning

confidence: 99%

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Enhanced Production of Nargenicin A1 and Generation of Novel Glycosylated Derivatives

Dhakal

Pandey

et al. 2015

Appl Biochem Biotechnol

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Nargenicin A1, an antibacterial polyketide macrolide produced by Nocardia sp. CS682, was enhanced by increasing the pool of precursors using different sources. Furthermore, by using engineered strain Nocardia sp. ACC18 and supplementation of glucose and glycerol, enhancement was ~7.1 fold in comparison to Nocardia sp. CS682 without supplementation of any precursors. The overproduced compound was validated by mass spectrometry and nuclear magnetic resonance analyses. The novel glycosylated derivatives of purified nargenicin A1 were generated by efficient one-pot reaction systems in which the syntheses of uridine diphosphate (UDP)-α-D-glucose and UDP-α-D-2-deoxyglucose were modified and combined with glycosyltransferase (GT) from Bacillus licheniformis. Nargenicin A1 11-O-β- D-glucopyranoside, nargenicin A1 18-O-β-D-glucopyranoside, nargenicin A111 18-O-β-D- diglucopyranoside, and nargenicin 11-O-β-D-2-deoxyglucopyranoside were generated. Nargenicin A1 11-O-β-D-glucopyranoside was structurally elucidated by ultra-high performance liquid chromatography-photodiode array (UPLC-PDA) conjugated with high-resolution quantitative time-of-flight-electrospray ionization mass spectroscopy (HR-QTOF ESI-MS/MS), supported by one- and two-dimensional nuclear magnetic resonance studies, whereas other nargenicin A1 glycosides were characterized by UPLC-PDA and HR-QTOF ESI-MS/MS analyses. The overall conversion studies indicated that the one-pot synthesis system is a highly efficient strategy for production of glycosylated derivatives of compounds like macrolides as well. Furthermore, assessment of solubility indicated that there was enhanced solubility in the case of glycoside, although a substantial increase in activity was not observed.

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Section: Discussionmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 98%

Section: Bacterial Strains and Growth Conditionsmentioning

confidence: 99%

Section: Bacterial Strains and Growth Conditionsmentioning

confidence: 99%

See 3 more Smart Citations

Enhanced Production of Nargenicin A1 and Generation of Novel Glycosylated Derivatives

Dhakal

Pandey

et al. 2015

Appl Biochem Biotechnol

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Genetic Manipulation ofNocardiaSpecies

et al. 2016

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Nocardia spp. are aerobic, Gram-positive, catalase positive, and non-motile actinomycetes. They are associated with human infections. However, some species produce important natural products, degrade toxic chemicals, and are involved in biotransformation of valuable products. The lack of robust genetic tools has hindered detailed studies and advanced research. This unit describes the major genetic engineering approaches using Nocardia sp. CS682 as a prototype. These methods will certainly help in understanding the basis of their pathogenicity as well as biosynthetic and biotransforming abilities. It can be expected that knowledge of the biochemistry behind their pathogenicity will be crucial in developing effective treatment strategies. These genetic tools can be utilized to develop rational metabolic engineering approaches for crafting host strains with higher production or biotransformation ability.

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Enhancement of toyocamycin production through increasing supply of precursor GTP in Streptomyces diastatochromogenes 1628

Zhang

et al. 2022

J Basic Microbiol

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The nucleoside antibiotic toyocamycin (TM), which is produced by Streptomyces diastatochromogenes 1628, exhibits potent activity against a broad range of phytopathogenic fungi. TM was synthesized through a multi-step reaction, using guanosine triphosphate (GTP) as precursor. Based on a comparison of proteomics data from S. diastatochromogenes 1628 and rifamycin-resistant mutant 1628-T15 with high yield of TM, we determined that the differentially expressed protein X0NBV6 called ribose-phosphate pyrophosphokinase (RHP), which is a rate-limiting enzyme involved in the de novo biosynthesis of GTP, exhibits a higher expression level in mutant 1628-T15. In this study, to elucidate the relationships between RHP, GTP, and TM production, the gene rhp sd encoding RHP was cloned and overexpressed in S. diastatochromogenes strain 1628. The recombinant strain S. diastatochromogenes 1628-RHP exhibited better performance at the transcriptional level of the rhp sd gene, as well as RHP enzymatic activity, intracellular GTP concentration, and TM production, compared to S. diastatochromogenes 1628. Finally, the yield of TM produced by S. diastatochromogenes 1628-RHP (340.2 mg/L) was 133.3% higher than that produced by S. diastatochromogenes1628. Moreover, the transcriptional level of toy genes involved in TM biosynthesis was enhanced due to the overexpression of the rhp sd gene.

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Effect of Different Biosynthetic Precursors on the Production of Nargenicin A1 from Metabolically Engineered Nocardia sp. CS682

Cited by 13 publications

References 28 publications

Enhanced Production of Nargenicin A1 and Generation of Novel Glycosylated Derivatives

Enhanced Production of Nargenicin A1 and Generation of Novel Glycosylated Derivatives

Genetic Manipulation ofNocardiaSpecies

Enhancement of toyocamycin production through increasing supply of precursor GTP in Streptomyces diastatochromogenes 1628

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