Re-engineering of genetic circuit for 2-deoxystreptamine (2-DOS) biosynthesis in Escherichia coli BL21 (DE3)

Amit Kumar Chaudhary; Je Won Park; Yeo Joon Yoon; Byung Gee Kim; Jae Kyung Sohng

doi:10.1007/s10529-012-1077-2

Re-engineering of genetic circuit for 2-deoxystreptamine (2-DOS) biosynthesis in Escherichia coli BL21 (DE3)

Amit Kumar Chaudhary, Je Won Park, Yeo Joon Yoon, Byung Gee Kim, Jae Kyung Sohng

Research output: Contribution to journal › Article › peer-review

15 Citations (Scopus)

Abstract

Various approaches for monocistronic constructions of genetic circuits have been designed for metabolite production but there has been no attempt to apply such methodology for aminoglycosides biosynthesis. Here, a simple and commercially available bio-part, despite the current trend focusing on the standardized BioBricks bio-parts available in the registry, is used. A 181-bp nucleotide fragment was designed for the efficient construction of an expression vector for monocistronic assembly of genes. Furthermore, a single vector with multi-monocistronic assembled genes for 2-deoxystreptamine (2-DOS) synthesis was constructed for production in engineered Escherichia coli. The working efficiency of model vector was concluded by reporter assay whereas the expressions of biosynthesis genes were confirmed by RT-PCR and SDS-PAGE. Production of 2-DOS was confirmed by TLC, LC-ELSD, and ESI-MS/MS.

Original language	English
Pages (from-to)	285-293
Number of pages	9
Journal	Biotechnology letters
Volume	35
Issue number	2
DOIs	https://doi.org/10.1007/s10529-012-1077-2
Publication status	Published - 2013 Feb
Externally published	Yes

Bibliographical note

Funding Information:
Acknowledgments This study was supported by the Intelligent Synthetic Biology Center of Global Frontier Project funded by the Ministry of Education, Science and Technology (2011-0031960) and by the grant from the Next-Generation BioGreen 21 Program (PJ008013), Rural Development Administration, and Republic of Korea.

Keywords

Multi-monocistronic
Polycistronic
Pseudo-operon
Re-engineering of genetic circuit
Synthetic biology

ASJC Scopus subject areas

Biotechnology
Bioengineering
Applied Microbiology and Biotechnology

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10.1007/s10529-012-1077-2

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title = "Re-engineering of genetic circuit for 2-deoxystreptamine (2-DOS) biosynthesis in Escherichia coli BL21 (DE3)",

abstract = "Various approaches for monocistronic constructions of genetic circuits have been designed for metabolite production but there has been no attempt to apply such methodology for aminoglycosides biosynthesis. Here, a simple and commercially available bio-part, despite the current trend focusing on the standardized BioBricks bio-parts available in the registry, is used. A 181-bp nucleotide fragment was designed for the efficient construction of an expression vector for monocistronic assembly of genes. Furthermore, a single vector with multi-monocistronic assembled genes for 2-deoxystreptamine (2-DOS) synthesis was constructed for production in engineered Escherichia coli. The working efficiency of model vector was concluded by reporter assay whereas the expressions of biosynthesis genes were confirmed by RT-PCR and SDS-PAGE. Production of 2-DOS was confirmed by TLC, LC-ELSD, and ESI-MS/MS.",

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note = "Funding Information: Acknowledgments This study was supported by the Intelligent Synthetic Biology Center of Global Frontier Project funded by the Ministry of Education, Science and Technology (2011-0031960) and by the grant from the Next-Generation BioGreen 21 Program (PJ008013), Rural Development Administration, and Republic of Korea.",

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doi = "10.1007/s10529-012-1077-2",

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AU - Kim, Byung Gee

AU - Sohng, Jae Kyung

N1 - Funding Information: Acknowledgments This study was supported by the Intelligent Synthetic Biology Center of Global Frontier Project funded by the Ministry of Education, Science and Technology (2011-0031960) and by the grant from the Next-Generation BioGreen 21 Program (PJ008013), Rural Development Administration, and Republic of Korea.

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Re-engineering of genetic circuit for 2-deoxystreptamine (2-DOS) biosynthesis in Escherichia coli BL21 (DE3)

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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