Rapid analysis of polyhydroxyalkanoate contents and its monomer compositions by pyrolysis-gas chromatography combined with mass spectrometry (Py-GC/MS)

Tae Uk Khang; Min Jae Kim; Jee In Yoo; Yu Jung Sohn; Sang Goo Jeon; Si Jae Park; Jeong Geol Na

doi:10.1016/j.ijbiomac.2021.01.108

Rapid analysis of polyhydroxyalkanoate contents and its monomer compositions by pyrolysis-gas chromatography combined with mass spectrometry (Py-GC/MS)

Tae Uk Khang, Min Jae Kim, Jee In Yoo, Yu Jung Sohn, Sang Goo Jeon, Si Jae Park, Jeong Geol Na

Department of Chemical Engineering & Materials Science

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

23 Scopus citations

Abstract

Here, we report an analysis method for determining PHA (polyhydroxyalkanoates) contents and their monomer composition in microbial cells based on pyrolysis gas chromatography combined with mass spectrometry (Py-GC/MS). Various kinds of microbial cells accumulating different PHA contents and monomer compositions were prepared through the cultivation of Ralstonia eutropha and recombinant Escherichia coli. Py-GC/MS could analyse these samples in a short time without complicated pretreatment steps. Characteristic peaks such as 2-butenoic acid, 2-pentenoic acid, and hexadecanoic acid regarding PHA compositions and cell components were identified. Considering constituents of cells and ratios of peak areas of dehydrated monomers to hexadecanoic acid, a simple equation for estimation of PHA contents in microbial cells was derived. Also, monomer compositions of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) in R. eutropha could be successfully determined based on peak area of 2-butenoic acid and 2-pentenoic acid of Py-GC/MS, which are the corresponding species of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) in PHBV. Correlation of results between GC-FID and Py-GC/MS could be fitted very well. This method shows similar results for the samples obtained from same experimental conditions, allowing rapid and reliable analysis. Py-GC/MS can be a promising tool to rapidly screen PHA-positive strains based on polymer contents along with monomer compositions.

Original language	English
Pages (from-to)	449-456
Number of pages	8
Journal	International Journal of Biological Macromolecules
Volume	174
DOIs	https://doi.org/10.1016/j.ijbiomac.2021.01.108
State	Published - 31 Mar 2021

Bibliographical note

Funding Information:
This work was supported by “Human Resources Program in Energy Technology” of the Korea Institute of Energy Technology Evaluation and Planning (KETEP), granted financial resource from the Ministry of Trade, Industry and Energy , Republic of Korea. (No. 20174010201150 ) and the National Research Foundation (NRF) grant funded by the Ministry of Science and ICT (MSIT) ( NRF-2020R1A5A1019631 ).

Funding Information:
This work was supported by “Human Resources Program in Energy Technology” of the Korea Institute of Energy Technology Evaluation and Planning (KETEP), granted financial resource from the Ministry of Trade, Industry and Energy, Republic of Korea. (No. 20174010201150) and the National Research Foundation (NRF) grant funded by the Ministry of Science and ICT (MSIT) (NRF-2020R1A5A1019631).

Publisher Copyright:
© 2021 Elsevier B.V.

Keywords

PHB
PHBV
Pyrolysis-GC/MS

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10.1016/j.ijbiomac.2021.01.108

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@article{c12936ae218147bcbcf7188b0305cf5b,

title = "Rapid analysis of polyhydroxyalkanoate contents and its monomer compositions by pyrolysis-gas chromatography combined with mass spectrometry (Py-GC/MS)",

abstract = "Here, we report an analysis method for determining PHA (polyhydroxyalkanoates) contents and their monomer composition in microbial cells based on pyrolysis gas chromatography combined with mass spectrometry (Py-GC/MS). Various kinds of microbial cells accumulating different PHA contents and monomer compositions were prepared through the cultivation of Ralstonia eutropha and recombinant Escherichia coli. Py-GC/MS could analyse these samples in a short time without complicated pretreatment steps. Characteristic peaks such as 2-butenoic acid, 2-pentenoic acid, and hexadecanoic acid regarding PHA compositions and cell components were identified. Considering constituents of cells and ratios of peak areas of dehydrated monomers to hexadecanoic acid, a simple equation for estimation of PHA contents in microbial cells was derived. Also, monomer compositions of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) in R. eutropha could be successfully determined based on peak area of 2-butenoic acid and 2-pentenoic acid of Py-GC/MS, which are the corresponding species of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) in PHBV. Correlation of results between GC-FID and Py-GC/MS could be fitted very well. This method shows similar results for the samples obtained from same experimental conditions, allowing rapid and reliable analysis. Py-GC/MS can be a promising tool to rapidly screen PHA-positive strains based on polymer contents along with monomer compositions.",

keywords = "PHB, PHBV, Pyrolysis-GC/MS",

author = "Khang, {Tae Uk} and Kim, {Min Jae} and Yoo, {Jee In} and Sohn, {Yu Jung} and Jeon, {Sang Goo} and Park, {Si Jae} and Na, {Jeong Geol}",

note = "Funding Information: This work was supported by “Human Resources Program in Energy Technology” of the Korea Institute of Energy Technology Evaluation and Planning (KETEP), granted financial resource from the Ministry of Trade, Industry and Energy , Republic of Korea. (No. 20174010201150 ) and the National Research Foundation (NRF) grant funded by the Ministry of Science and ICT (MSIT) ( NRF-2020R1A5A1019631 ). Funding Information: This work was supported by “Human Resources Program in Energy Technology” of the Korea Institute of Energy Technology Evaluation and Planning (KETEP), granted financial resource from the Ministry of Trade, Industry and Energy, Republic of Korea. (No. 20174010201150) and the National Research Foundation (NRF) grant funded by the Ministry of Science and ICT (MSIT) (NRF-2020R1A5A1019631). Publisher Copyright: {\textcopyright} 2021 Elsevier B.V.",

year = "2021",

month = mar,

day = "31",

doi = "10.1016/j.ijbiomac.2021.01.108",

language = "English",

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T1 - Rapid analysis of polyhydroxyalkanoate contents and its monomer compositions by pyrolysis-gas chromatography combined with mass spectrometry (Py-GC/MS)

AU - Khang, Tae Uk

AU - Kim, Min Jae

AU - Yoo, Jee In

AU - Sohn, Yu Jung

AU - Jeon, Sang Goo

AU - Park, Si Jae

AU - Na, Jeong Geol

N1 - Funding Information: This work was supported by “Human Resources Program in Energy Technology” of the Korea Institute of Energy Technology Evaluation and Planning (KETEP), granted financial resource from the Ministry of Trade, Industry and Energy , Republic of Korea. (No. 20174010201150 ) and the National Research Foundation (NRF) grant funded by the Ministry of Science and ICT (MSIT) ( NRF-2020R1A5A1019631 ). Funding Information: This work was supported by “Human Resources Program in Energy Technology” of the Korea Institute of Energy Technology Evaluation and Planning (KETEP), granted financial resource from the Ministry of Trade, Industry and Energy, Republic of Korea. (No. 20174010201150) and the National Research Foundation (NRF) grant funded by the Ministry of Science and ICT (MSIT) (NRF-2020R1A5A1019631). Publisher Copyright: © 2021 Elsevier B.V.

PY - 2021/3/31

Y1 - 2021/3/31

N2 - Here, we report an analysis method for determining PHA (polyhydroxyalkanoates) contents and their monomer composition in microbial cells based on pyrolysis gas chromatography combined with mass spectrometry (Py-GC/MS). Various kinds of microbial cells accumulating different PHA contents and monomer compositions were prepared through the cultivation of Ralstonia eutropha and recombinant Escherichia coli. Py-GC/MS could analyse these samples in a short time without complicated pretreatment steps. Characteristic peaks such as 2-butenoic acid, 2-pentenoic acid, and hexadecanoic acid regarding PHA compositions and cell components were identified. Considering constituents of cells and ratios of peak areas of dehydrated monomers to hexadecanoic acid, a simple equation for estimation of PHA contents in microbial cells was derived. Also, monomer compositions of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) in R. eutropha could be successfully determined based on peak area of 2-butenoic acid and 2-pentenoic acid of Py-GC/MS, which are the corresponding species of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) in PHBV. Correlation of results between GC-FID and Py-GC/MS could be fitted very well. This method shows similar results for the samples obtained from same experimental conditions, allowing rapid and reliable analysis. Py-GC/MS can be a promising tool to rapidly screen PHA-positive strains based on polymer contents along with monomer compositions.

AB - Here, we report an analysis method for determining PHA (polyhydroxyalkanoates) contents and their monomer composition in microbial cells based on pyrolysis gas chromatography combined with mass spectrometry (Py-GC/MS). Various kinds of microbial cells accumulating different PHA contents and monomer compositions were prepared through the cultivation of Ralstonia eutropha and recombinant Escherichia coli. Py-GC/MS could analyse these samples in a short time without complicated pretreatment steps. Characteristic peaks such as 2-butenoic acid, 2-pentenoic acid, and hexadecanoic acid regarding PHA compositions and cell components were identified. Considering constituents of cells and ratios of peak areas of dehydrated monomers to hexadecanoic acid, a simple equation for estimation of PHA contents in microbial cells was derived. Also, monomer compositions of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) in R. eutropha could be successfully determined based on peak area of 2-butenoic acid and 2-pentenoic acid of Py-GC/MS, which are the corresponding species of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) in PHBV. Correlation of results between GC-FID and Py-GC/MS could be fitted very well. This method shows similar results for the samples obtained from same experimental conditions, allowing rapid and reliable analysis. Py-GC/MS can be a promising tool to rapidly screen PHA-positive strains based on polymer contents along with monomer compositions.

KW - PHB

KW - PHBV

KW - Pyrolysis-GC/MS

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AN - SCOPUS:85100386851

SN - 0141-8130

VL - 174

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EP - 456

JO - International Journal of Biological Macromolecules

JF - International Journal of Biological Macromolecules

ER -

Rapid analysis of polyhydroxyalkanoate contents and its monomer compositions by pyrolysis-gas chromatography combined with mass spectrometry (Py-GC/MS)

Abstract

Bibliographical note

Keywords

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