In vivo 15N-enrichment of metabolites in suspension cultured cells and its application to metabolomics

Kazuo Harada, Eiichiro Fukusaki, Takeshi Bamba, Fumihiko Sato, Akio Kobayashi

研究成果: ジャーナルへの寄稿記事

29 引用 (Scopus)

抄録

The incorporation of stable isotopes in suspension cultured cells is very simple and useful as a preliminary experimental method in the experimental scene of plant metabolomics to elucidate the metabolic profiles of mutants and transformants. Stable isotope methods would afford a dynamic explanation of turnover speed that would concern the metabolic flux. Utilization of suspension cultured cells allows genes to be easily induced or suppressed, culture conditions to be controlled, and samples to be easily prepared. Stable isotope tracing allows an index of metabolic flux to be obtained. Here we present an experiment feeding 15N-labeled inorganic salts to Arabidopsis (cell line T87) and Coptis cultured cells. Results of a comparison of 15N labeling ratios of amino acids derived from T87 cells cultured under light with those cultured in the dark corresponded to transcriptional expressions revealed by microarray experiments published previously, demonstrating the validity of this procedure. Furthermore, 15N labeling ratios of Coptis cultured cells revealed arginine and lysine metabolism inhibition, which should result in inhibition of polyamine biosynthesis and cell division. This very simple experiment allowed us to uncover metabolic dynamic features of the plant cell. Therefore this method is very useful for forming working hypotheses and experimental design.

元の言語英語
ページ(範囲)1003-1011
ページ数9
ジャーナルBiotechnology Progress
22
発行部数4
DOI
出版物ステータス出版済み - 7 1 2006
外部発表Yes

Fingerprint

Metabolomics
metabolomics
cultured cells
Cultured Cells
Suspensions
metabolites
Coptis
Isotopes
stable isotopes
inorganic salts
Metabolome
Polyamines
Plant Cells
polyamines
Arabidopsis
Cell Division
Lysine
arginine
Arginine
cell division

All Science Journal Classification (ASJC) codes

  • Biotechnology

これを引用

In vivo 15N-enrichment of metabolites in suspension cultured cells and its application to metabolomics. / Harada, Kazuo; Fukusaki, Eiichiro; Bamba, Takeshi; Sato, Fumihiko; Kobayashi, Akio.

:: Biotechnology Progress, 巻 22, 番号 4, 01.07.2006, p. 1003-1011.

研究成果: ジャーナルへの寄稿記事

Harada, Kazuo ; Fukusaki, Eiichiro ; Bamba, Takeshi ; Sato, Fumihiko ; Kobayashi, Akio. / In vivo 15N-enrichment of metabolites in suspension cultured cells and its application to metabolomics. :: Biotechnology Progress. 2006 ; 巻 22, 番号 4. pp. 1003-1011.
@article{102c7796290d4c15bebcbf66b382ffae,
title = "In vivo 15N-enrichment of metabolites in suspension cultured cells and its application to metabolomics",
abstract = "The incorporation of stable isotopes in suspension cultured cells is very simple and useful as a preliminary experimental method in the experimental scene of plant metabolomics to elucidate the metabolic profiles of mutants and transformants. Stable isotope methods would afford a dynamic explanation of turnover speed that would concern the metabolic flux. Utilization of suspension cultured cells allows genes to be easily induced or suppressed, culture conditions to be controlled, and samples to be easily prepared. Stable isotope tracing allows an index of metabolic flux to be obtained. Here we present an experiment feeding 15N-labeled inorganic salts to Arabidopsis (cell line T87) and Coptis cultured cells. Results of a comparison of 15N labeling ratios of amino acids derived from T87 cells cultured under light with those cultured in the dark corresponded to transcriptional expressions revealed by microarray experiments published previously, demonstrating the validity of this procedure. Furthermore, 15N labeling ratios of Coptis cultured cells revealed arginine and lysine metabolism inhibition, which should result in inhibition of polyamine biosynthesis and cell division. This very simple experiment allowed us to uncover metabolic dynamic features of the plant cell. Therefore this method is very useful for forming working hypotheses and experimental design.",
author = "Kazuo Harada and Eiichiro Fukusaki and Takeshi Bamba and Fumihiko Sato and Akio Kobayashi",
year = "2006",
month = "7",
day = "1",
doi = "10.1021/bp060139z",
language = "English",
volume = "22",
pages = "1003--1011",
journal = "Biotechnology Progress",
issn = "8756-7938",
publisher = "John Wiley and Sons Ltd",
number = "4",

}

TY - JOUR

T1 - In vivo 15N-enrichment of metabolites in suspension cultured cells and its application to metabolomics

AU - Harada, Kazuo

AU - Fukusaki, Eiichiro

AU - Bamba, Takeshi

AU - Sato, Fumihiko

AU - Kobayashi, Akio

PY - 2006/7/1

Y1 - 2006/7/1

N2 - The incorporation of stable isotopes in suspension cultured cells is very simple and useful as a preliminary experimental method in the experimental scene of plant metabolomics to elucidate the metabolic profiles of mutants and transformants. Stable isotope methods would afford a dynamic explanation of turnover speed that would concern the metabolic flux. Utilization of suspension cultured cells allows genes to be easily induced or suppressed, culture conditions to be controlled, and samples to be easily prepared. Stable isotope tracing allows an index of metabolic flux to be obtained. Here we present an experiment feeding 15N-labeled inorganic salts to Arabidopsis (cell line T87) and Coptis cultured cells. Results of a comparison of 15N labeling ratios of amino acids derived from T87 cells cultured under light with those cultured in the dark corresponded to transcriptional expressions revealed by microarray experiments published previously, demonstrating the validity of this procedure. Furthermore, 15N labeling ratios of Coptis cultured cells revealed arginine and lysine metabolism inhibition, which should result in inhibition of polyamine biosynthesis and cell division. This very simple experiment allowed us to uncover metabolic dynamic features of the plant cell. Therefore this method is very useful for forming working hypotheses and experimental design.

AB - The incorporation of stable isotopes in suspension cultured cells is very simple and useful as a preliminary experimental method in the experimental scene of plant metabolomics to elucidate the metabolic profiles of mutants and transformants. Stable isotope methods would afford a dynamic explanation of turnover speed that would concern the metabolic flux. Utilization of suspension cultured cells allows genes to be easily induced or suppressed, culture conditions to be controlled, and samples to be easily prepared. Stable isotope tracing allows an index of metabolic flux to be obtained. Here we present an experiment feeding 15N-labeled inorganic salts to Arabidopsis (cell line T87) and Coptis cultured cells. Results of a comparison of 15N labeling ratios of amino acids derived from T87 cells cultured under light with those cultured in the dark corresponded to transcriptional expressions revealed by microarray experiments published previously, demonstrating the validity of this procedure. Furthermore, 15N labeling ratios of Coptis cultured cells revealed arginine and lysine metabolism inhibition, which should result in inhibition of polyamine biosynthesis and cell division. This very simple experiment allowed us to uncover metabolic dynamic features of the plant cell. Therefore this method is very useful for forming working hypotheses and experimental design.

UR - http://www.scopus.com/inward/record.url?scp=33747154732&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33747154732&partnerID=8YFLogxK

U2 - 10.1021/bp060139z

DO - 10.1021/bp060139z

M3 - Article

C2 - 16889377

AN - SCOPUS:33747154732

VL - 22

SP - 1003

EP - 1011

JO - Biotechnology Progress

JF - Biotechnology Progress

SN - 8756-7938

IS - 4

ER -