Enrichment of specific monomer in medium-chain-length poly(3-hydroxyalkanoates) by amplification of fadD and fadE genes in recombinant Escherichia coli

Si Jae Park; Jong Pil Park; Sang Yup Lee; Yoshiharu Doi

doi:10.1016/S0141-0229(03)00093-0

Enrichment of specific monomer in medium-chain-length poly(3-hydroxyalkanoates) by amplification of fadD and fadE genes in recombinant Escherichia coli

Si Jae Park, Jong Pil Park, Sang Yup Lee, Yoshiharu Doi

Department of Chemical Engineering & Materials Science

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

21 Scopus citations

Abstract

Recombinant Escherichia coli strains defective in FadA and/or FadB harboring the Pseudomonas sp. 61-3 polyhydroxyalkanoate (PHA) synthase gene (phaC2_Ps) were constructed, and were examined for the production of medium-chain-length (MCL) PHA from sodium decanoate and sodium dodecanoate. All the recombinant E. coli strains accumulated MCL-PHAs mainly composed of C6, C8 and C10 monomers from decanoate and those composed of C8, C10 and C12 monomers from dodecanoate. A new metabolic engineering strategy for enriching specific monomers in MCL-PHA was developed by examining the effect of co-expressing the E. coli fadD_Ec, fadE_Ec and/or fadL_Ec genes along with the PHA synthase gene. Using these engineered E. coli strains, MCL-PHAs enriched in 3-hydroxydecanoate up to 80mol% and 3-hydroxydodecanoate up to 48mol% were produced from sodium decanoate and sodium dodecanoate, respectively. It was found that the amplification of the fadD_Ec, fadE_Ec or fadL_Ec gene had different effect on the monomer composition of MCL-PHA. Among these, the amplification of the fadD_Ec gene had the most significant effect on the alteration of monomer composition of MCL-PHAs.

Original language	English
Pages (from-to)	62-70
Number of pages	9
Journal	Enzyme and Microbial Technology
Volume	33
Issue number	1
DOIs	https://doi.org/10.1016/S0141-0229(03)00093-0
State	Published - 16 Jul 2003

Bibliographical note

Funding Information:
This work was supported by the National Research Laboratory Program (2000-N-NL-01-C-237) of the Korean Ministry of Science and Technology (MOST), Center for Ultramicrochemical Process Systems and by the BK21 project from the Ministry of Education. We thank Dr. Isabelle-S. Hinner (GBF, Germany) for kindly providing plasmid pBBR1MCS. We also thank Dr. G.M. Church (Harvard Medical School, USA) for the kind gift of plasmid pKO3.

Keywords

Acyl-CoA dehydrogenase
Acyl-CoA synthetase
Escherichia coli
Polyhydroxyalkanoates
β-Oxidation pathway

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10.1016/S0141-0229(03)00093-0

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title = "Enrichment of specific monomer in medium-chain-length poly(3-hydroxyalkanoates) by amplification of fadD and fadE genes in recombinant Escherichia coli",

abstract = "Recombinant Escherichia coli strains defective in FadA and/or FadB harboring the Pseudomonas sp. 61-3 polyhydroxyalkanoate (PHA) synthase gene (phaC2Ps) were constructed, and were examined for the production of medium-chain-length (MCL) PHA from sodium decanoate and sodium dodecanoate. All the recombinant E. coli strains accumulated MCL-PHAs mainly composed of C6, C8 and C10 monomers from decanoate and those composed of C8, C10 and C12 monomers from dodecanoate. A new metabolic engineering strategy for enriching specific monomers in MCL-PHA was developed by examining the effect of co-expressing the E. coli fadDEc, fadEEc and/or fadLEc genes along with the PHA synthase gene. Using these engineered E. coli strains, MCL-PHAs enriched in 3-hydroxydecanoate up to 80mol% and 3-hydroxydodecanoate up to 48mol% were produced from sodium decanoate and sodium dodecanoate, respectively. It was found that the amplification of the fadDEc, fadEEc or fadLEc gene had different effect on the monomer composition of MCL-PHA. Among these, the amplification of the fadDEc gene had the most significant effect on the alteration of monomer composition of MCL-PHAs.",

keywords = "Acyl-CoA dehydrogenase, Acyl-CoA synthetase, Escherichia coli, Polyhydroxyalkanoates, β-Oxidation pathway",

author = "Park, {Si Jae} and Park, {Jong Pil} and Lee, {Sang Yup} and Yoshiharu Doi",

note = "Funding Information: This work was supported by the National Research Laboratory Program (2000-N-NL-01-C-237) of the Korean Ministry of Science and Technology (MOST), Center for Ultramicrochemical Process Systems and by the BK21 project from the Ministry of Education. We thank Dr. Isabelle-S. Hinner (GBF, Germany) for kindly providing plasmid pBBR1MCS. We also thank Dr. G.M. Church (Harvard Medical School, USA) for the kind gift of plasmid pKO3.",

year = "2003",

month = jul,

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doi = "10.1016/S0141-0229(03)00093-0",

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T1 - Enrichment of specific monomer in medium-chain-length poly(3-hydroxyalkanoates) by amplification of fadD and fadE genes in recombinant Escherichia coli

AU - Park, Si Jae

AU - Park, Jong Pil

AU - Lee, Sang Yup

AU - Doi, Yoshiharu

N1 - Funding Information: This work was supported by the National Research Laboratory Program (2000-N-NL-01-C-237) of the Korean Ministry of Science and Technology (MOST), Center for Ultramicrochemical Process Systems and by the BK21 project from the Ministry of Education. We thank Dr. Isabelle-S. Hinner (GBF, Germany) for kindly providing plasmid pBBR1MCS. We also thank Dr. G.M. Church (Harvard Medical School, USA) for the kind gift of plasmid pKO3.

PY - 2003/7/16

Y1 - 2003/7/16

N2 - Recombinant Escherichia coli strains defective in FadA and/or FadB harboring the Pseudomonas sp. 61-3 polyhydroxyalkanoate (PHA) synthase gene (phaC2Ps) were constructed, and were examined for the production of medium-chain-length (MCL) PHA from sodium decanoate and sodium dodecanoate. All the recombinant E. coli strains accumulated MCL-PHAs mainly composed of C6, C8 and C10 monomers from decanoate and those composed of C8, C10 and C12 monomers from dodecanoate. A new metabolic engineering strategy for enriching specific monomers in MCL-PHA was developed by examining the effect of co-expressing the E. coli fadDEc, fadEEc and/or fadLEc genes along with the PHA synthase gene. Using these engineered E. coli strains, MCL-PHAs enriched in 3-hydroxydecanoate up to 80mol% and 3-hydroxydodecanoate up to 48mol% were produced from sodium decanoate and sodium dodecanoate, respectively. It was found that the amplification of the fadDEc, fadEEc or fadLEc gene had different effect on the monomer composition of MCL-PHA. Among these, the amplification of the fadDEc gene had the most significant effect on the alteration of monomer composition of MCL-PHAs.

AB - Recombinant Escherichia coli strains defective in FadA and/or FadB harboring the Pseudomonas sp. 61-3 polyhydroxyalkanoate (PHA) synthase gene (phaC2Ps) were constructed, and were examined for the production of medium-chain-length (MCL) PHA from sodium decanoate and sodium dodecanoate. All the recombinant E. coli strains accumulated MCL-PHAs mainly composed of C6, C8 and C10 monomers from decanoate and those composed of C8, C10 and C12 monomers from dodecanoate. A new metabolic engineering strategy for enriching specific monomers in MCL-PHA was developed by examining the effect of co-expressing the E. coli fadDEc, fadEEc and/or fadLEc genes along with the PHA synthase gene. Using these engineered E. coli strains, MCL-PHAs enriched in 3-hydroxydecanoate up to 80mol% and 3-hydroxydodecanoate up to 48mol% were produced from sodium decanoate and sodium dodecanoate, respectively. It was found that the amplification of the fadDEc, fadEEc or fadLEc gene had different effect on the monomer composition of MCL-PHA. Among these, the amplification of the fadDEc gene had the most significant effect on the alteration of monomer composition of MCL-PHAs.

KW - Acyl-CoA dehydrogenase

KW - Acyl-CoA synthetase

KW - Escherichia coli

KW - Polyhydroxyalkanoates

KW - β-Oxidation pathway

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U2 - 10.1016/S0141-0229(03)00093-0

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SN - 0141-0229

VL - 33

SP - 62

EP - 70

JO - Enzyme and Microbial Technology

JF - Enzyme and Microbial Technology

IS - 1

ER -

Enrichment of specific monomer in medium-chain-length poly(3-hydroxyalkanoates) by amplification of fadD and fadE genes in recombinant Escherichia coli

Abstract

Bibliographical note

Keywords

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