Molecular and biochemical analysis of phthalate and terephthalate degradation by Rhodococcus sp. strain DK17

Young Choi Ki, Dockyu Kim, Jun Sul Woo, Jong Chan Chae, Gerben J. Zylstra, Min Kim Young, Eungbin Kim

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Alkylbenzene-degrading Rhodococcus sp. strain DK17 is able to utilize phthalate and terephthalate as growth substrates. The genes encoding the transformation of phthalate and terephthalate to protocatechuate are organized as two separate operons, located 6.7 kb away from each other. Interestingly, both the phthalate and terephthalate operons are induced in response to terephthalate while expression of the terephthalate genes is undetectable in phthalate-grown cells. In addition to two known plasmids (380-kb pDK1 and 330-kb pDK2), a third megaplasmid (750-kb pDK3) was newly identified in DK17. The phthalate and terephthalate operons are duplicated and are present on both pDK2 and pDK3. RT-PCR experiments, coupled with sequence analysis, suggest that phthalate and terephthalate degradation in DK17 proceeds through oxygenation at carbons 3 and 4 and at carbons 1 and 2 to form 3,4-dihydro-3,4- dihydroxyphthalate and 1,2-dihydro-1,2-dihydroxyterephthalate, respectively. The 3,4-dihydroxyphthalate pathway was further corroborated through colorometric tests. Apparently, the two dihydrodiol metabolites are subsequently dehydrogenated and decarboxylated to form protocatechuate, which is further degraded by a protocatechuate 3,4-dioxygenase as confirmed by a ring-cleavage enzyme assay.

Original languageEnglish
Pages (from-to)207-213
Number of pages7
JournalFEMS Microbiology Letters
Volume252
Issue number2
DOIs
Publication statusPublished - 2005 Nov 15

Fingerprint

Rhodococcus
Operon
Protocatechuate-3,4-Dioxygenase
Carbon
Enzyme Assays
phthalic acid
terephthalic acid
Sequence Analysis
Plasmids
Gene Expression
Polymerase Chain Reaction
Growth

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology
  • Genetics

Cite this

Ki, Young Choi ; Kim, Dockyu ; Woo, Jun Sul ; Chae, Jong Chan ; Zylstra, Gerben J. ; Young, Min Kim ; Kim, Eungbin. / Molecular and biochemical analysis of phthalate and terephthalate degradation by Rhodococcus sp. strain DK17. In: FEMS Microbiology Letters. 2005 ; Vol. 252, No. 2. pp. 207-213.
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abstract = "Alkylbenzene-degrading Rhodococcus sp. strain DK17 is able to utilize phthalate and terephthalate as growth substrates. The genes encoding the transformation of phthalate and terephthalate to protocatechuate are organized as two separate operons, located 6.7 kb away from each other. Interestingly, both the phthalate and terephthalate operons are induced in response to terephthalate while expression of the terephthalate genes is undetectable in phthalate-grown cells. In addition to two known plasmids (380-kb pDK1 and 330-kb pDK2), a third megaplasmid (750-kb pDK3) was newly identified in DK17. The phthalate and terephthalate operons are duplicated and are present on both pDK2 and pDK3. RT-PCR experiments, coupled with sequence analysis, suggest that phthalate and terephthalate degradation in DK17 proceeds through oxygenation at carbons 3 and 4 and at carbons 1 and 2 to form 3,4-dihydro-3,4- dihydroxyphthalate and 1,2-dihydro-1,2-dihydroxyterephthalate, respectively. The 3,4-dihydroxyphthalate pathway was further corroborated through colorometric tests. Apparently, the two dihydrodiol metabolites are subsequently dehydrogenated and decarboxylated to form protocatechuate, which is further degraded by a protocatechuate 3,4-dioxygenase as confirmed by a ring-cleavage enzyme assay.",
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Molecular and biochemical analysis of phthalate and terephthalate degradation by Rhodococcus sp. strain DK17. / Ki, Young Choi; Kim, Dockyu; Woo, Jun Sul; Chae, Jong Chan; Zylstra, Gerben J.; Young, Min Kim; Kim, Eungbin.

In: FEMS Microbiology Letters, Vol. 252, No. 2, 15.11.2005, p. 207-213.

Research output: Contribution to journalArticle

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AB - Alkylbenzene-degrading Rhodococcus sp. strain DK17 is able to utilize phthalate and terephthalate as growth substrates. The genes encoding the transformation of phthalate and terephthalate to protocatechuate are organized as two separate operons, located 6.7 kb away from each other. Interestingly, both the phthalate and terephthalate operons are induced in response to terephthalate while expression of the terephthalate genes is undetectable in phthalate-grown cells. In addition to two known plasmids (380-kb pDK1 and 330-kb pDK2), a third megaplasmid (750-kb pDK3) was newly identified in DK17. The phthalate and terephthalate operons are duplicated and are present on both pDK2 and pDK3. RT-PCR experiments, coupled with sequence analysis, suggest that phthalate and terephthalate degradation in DK17 proceeds through oxygenation at carbons 3 and 4 and at carbons 1 and 2 to form 3,4-dihydro-3,4- dihydroxyphthalate and 1,2-dihydro-1,2-dihydroxyterephthalate, respectively. The 3,4-dihydroxyphthalate pathway was further corroborated through colorometric tests. Apparently, the two dihydrodiol metabolites are subsequently dehydrogenated and decarboxylated to form protocatechuate, which is further degraded by a protocatechuate 3,4-dioxygenase as confirmed by a ring-cleavage enzyme assay.

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