Artificial Caprolactam-Specific Riboswitch as an Intracellular Metabolite Sensor

Sungyeon Jang, Sungho Jang, Dae Kyun Im, Taek Jin Kang, Min-Kyu Oh, Gyoo Yeol Jung

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Caprolactam is a monomer used for the synthesis of nylon-6, and a recombinant microbial strain for biobased production of nylon-6 was recently developed. An intracellular biosensor for caprolactam can facilitate high-throughput metabolic engineering of recombinant microbial strains. Because of the mixed production of caprolactam and valerolactam in the recombinant strain, a caprolactam biosensor should be highly specific for caprolactam. However, a highly specific caprolactam sensor has not been reported. Here, we developed an artificial riboswitch that specifically responds to caprolactam. This riboswitch was prepared using a coupled in vitro-in vivo selection strategy with a heterogeneous pool of RNA aptamers obtained from in vitro selection to construct a riboswitch library used in in vivo selection. The caprolactam riboswitch successfully discriminated caprolactam from valerolactam. Moreover, the riboswitch was activated by 3.36-fold in the presence of 50 mM caprolactam. This riboswitch enabled caprolactam-dependent control of cell growth, which will be useful for improving caprolactam production and is a valuable tool for metabolic engineering.

Original languageEnglish
Pages (from-to)1276-1283
Number of pages8
JournalACS Synthetic Biology
Volume8
Issue number6
DOIs
Publication statusPublished - 2019 Jun 21

Fingerprint

Caprolactam
Riboswitch
Metabolites
Metabolic engineering
Biosensors
Sensors
Cell growth
RNA
Monomers
Throughput
Metabolic Engineering
Biosensing Techniques
Nucleotide Aptamers

Keywords

  • caprolactam
  • high-throughput screening
  • metabolite biosensor
  • riboswitch
  • SELEX

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

Cite this

Artificial Caprolactam-Specific Riboswitch as an Intracellular Metabolite Sensor. / Jang, Sungyeon; Jang, Sungho; Im, Dae Kyun; Kang, Taek Jin; Oh, Min-Kyu; Jung, Gyoo Yeol.

In: ACS Synthetic Biology, Vol. 8, No. 6, 21.06.2019, p. 1276-1283.

Research output: Contribution to journalArticle

Jang, Sungyeon ; Jang, Sungho ; Im, Dae Kyun ; Kang, Taek Jin ; Oh, Min-Kyu ; Jung, Gyoo Yeol. / Artificial Caprolactam-Specific Riboswitch as an Intracellular Metabolite Sensor. In: ACS Synthetic Biology. 2019 ; Vol. 8, No. 6. pp. 1276-1283.
@article{d8007b876fd142e1b8cf4bc5c5a815a2,
title = "Artificial Caprolactam-Specific Riboswitch as an Intracellular Metabolite Sensor",
abstract = "Caprolactam is a monomer used for the synthesis of nylon-6, and a recombinant microbial strain for biobased production of nylon-6 was recently developed. An intracellular biosensor for caprolactam can facilitate high-throughput metabolic engineering of recombinant microbial strains. Because of the mixed production of caprolactam and valerolactam in the recombinant strain, a caprolactam biosensor should be highly specific for caprolactam. However, a highly specific caprolactam sensor has not been reported. Here, we developed an artificial riboswitch that specifically responds to caprolactam. This riboswitch was prepared using a coupled in vitro-in vivo selection strategy with a heterogeneous pool of RNA aptamers obtained from in vitro selection to construct a riboswitch library used in in vivo selection. The caprolactam riboswitch successfully discriminated caprolactam from valerolactam. Moreover, the riboswitch was activated by 3.36-fold in the presence of 50 mM caprolactam. This riboswitch enabled caprolactam-dependent control of cell growth, which will be useful for improving caprolactam production and is a valuable tool for metabolic engineering.",
keywords = "caprolactam, high-throughput screening, metabolite biosensor, riboswitch, SELEX",
author = "Sungyeon Jang and Sungho Jang and Im, {Dae Kyun} and Kang, {Taek Jin} and Min-Kyu Oh and Jung, {Gyoo Yeol}",
year = "2019",
month = "6",
day = "21",
doi = "10.1021/acssynbio.8b00452",
language = "English",
volume = "8",
pages = "1276--1283",
journal = "ACS Synthetic Biology",
issn = "2161-5063",
publisher = "American Chemical Society",
number = "6",

}

TY - JOUR

T1 - Artificial Caprolactam-Specific Riboswitch as an Intracellular Metabolite Sensor

AU - Jang, Sungyeon

AU - Jang, Sungho

AU - Im, Dae Kyun

AU - Kang, Taek Jin

AU - Oh, Min-Kyu

AU - Jung, Gyoo Yeol

PY - 2019/6/21

Y1 - 2019/6/21

N2 - Caprolactam is a monomer used for the synthesis of nylon-6, and a recombinant microbial strain for biobased production of nylon-6 was recently developed. An intracellular biosensor for caprolactam can facilitate high-throughput metabolic engineering of recombinant microbial strains. Because of the mixed production of caprolactam and valerolactam in the recombinant strain, a caprolactam biosensor should be highly specific for caprolactam. However, a highly specific caprolactam sensor has not been reported. Here, we developed an artificial riboswitch that specifically responds to caprolactam. This riboswitch was prepared using a coupled in vitro-in vivo selection strategy with a heterogeneous pool of RNA aptamers obtained from in vitro selection to construct a riboswitch library used in in vivo selection. The caprolactam riboswitch successfully discriminated caprolactam from valerolactam. Moreover, the riboswitch was activated by 3.36-fold in the presence of 50 mM caprolactam. This riboswitch enabled caprolactam-dependent control of cell growth, which will be useful for improving caprolactam production and is a valuable tool for metabolic engineering.

AB - Caprolactam is a monomer used for the synthesis of nylon-6, and a recombinant microbial strain for biobased production of nylon-6 was recently developed. An intracellular biosensor for caprolactam can facilitate high-throughput metabolic engineering of recombinant microbial strains. Because of the mixed production of caprolactam and valerolactam in the recombinant strain, a caprolactam biosensor should be highly specific for caprolactam. However, a highly specific caprolactam sensor has not been reported. Here, we developed an artificial riboswitch that specifically responds to caprolactam. This riboswitch was prepared using a coupled in vitro-in vivo selection strategy with a heterogeneous pool of RNA aptamers obtained from in vitro selection to construct a riboswitch library used in in vivo selection. The caprolactam riboswitch successfully discriminated caprolactam from valerolactam. Moreover, the riboswitch was activated by 3.36-fold in the presence of 50 mM caprolactam. This riboswitch enabled caprolactam-dependent control of cell growth, which will be useful for improving caprolactam production and is a valuable tool for metabolic engineering.

KW - caprolactam

KW - high-throughput screening

KW - metabolite biosensor

KW - riboswitch

KW - SELEX

UR - http://www.scopus.com/inward/record.url?scp=85067652557&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85067652557&partnerID=8YFLogxK

U2 - 10.1021/acssynbio.8b00452

DO - 10.1021/acssynbio.8b00452

M3 - Article

C2 - 31074964

AN - SCOPUS:85067652557

VL - 8

SP - 1276

EP - 1283

JO - ACS Synthetic Biology

JF - ACS Synthetic Biology

SN - 2161-5063

IS - 6

ER -