Engineering the Production of Major Catechins by Escherichia coli Carrying Metabolite Genes of Camellia sinensis

A mimicked biosynthetic pathway of catechin metabolite genes from C. sinensis, consisting of flavanone 3 hydroxylase (F3H), dihydroflavonol reductase (DFR), and leucoanthocyanidin reductase (LCR), was designed and arranged in two sets of constructs: (a) single promoter in front of F3H and ribosome-b...

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Main Authors: Kabir Mustapha Umar, S. M. Abdulkarim, Son Radu, Azizah Abdul Hamid, Nazamid Saari
Format: Article
Language:English
Published: Wiley 2012-01-01
Series:The Scientific World Journal
Online Access:http://dx.doi.org/10.1100/2012/529031
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author Kabir Mustapha Umar
S. M. Abdulkarim
Son Radu
Azizah Abdul Hamid
Nazamid Saari
author_facet Kabir Mustapha Umar
S. M. Abdulkarim
Son Radu
Azizah Abdul Hamid
Nazamid Saari
author_sort Kabir Mustapha Umar
collection DOAJ
description A mimicked biosynthetic pathway of catechin metabolite genes from C. sinensis, consisting of flavanone 3 hydroxylase (F3H), dihydroflavonol reductase (DFR), and leucoanthocyanidin reductase (LCR), was designed and arranged in two sets of constructs: (a) single promoter in front of F3H and ribosome-binding sequences both in front of DFR and LCR; (b) three different promoters with each in the front of the three genes and ribosome-binding sequences at appropriate positions. Recombinant E. coli BL (DE3) harbouring the constructs were cultivated for 65 h at 26°C in M9 medium consisting of 40 g/L glucose, 1 mM IPTG, and 3 mM eriodictyol. Compounds produced were extracted in ethyl acetate in alkaline conditions after 1 h at room temperature and identified by HPLC. Two of the four major catechins, namely, (−)-epicatechin (0.01 ) and (−)-epicatechin gallate (0.36 mg/L), and two other types ((+)-catechin hydrate (0.13 mg/L) and (−)-catechin gallate (0.04 mg/L)) were successfully produced.
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issn 1537-744X
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publishDate 2012-01-01
publisher Wiley
record_format Article
series The Scientific World Journal
spelling doaj-art-e523415127dd40a28deafba66fdebe6e2025-08-20T02:21:19ZengWileyThe Scientific World Journal1537-744X2012-01-01201210.1100/2012/529031529031Engineering the Production of Major Catechins by Escherichia coli Carrying Metabolite Genes of Camellia sinensisKabir Mustapha Umar0S. M. Abdulkarim1Son Radu2Azizah Abdul Hamid3Nazamid Saari4Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia UPM 43400, Serdang Selangor, MalaysiaDepartment of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia UPM 43400, Serdang Selangor, MalaysiaDepartment of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia UPM 43400, Serdang Selangor, MalaysiaDepartment of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia UPM 43400, Serdang Selangor, MalaysiaDepartment of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia UPM 43400, Serdang Selangor, MalaysiaA mimicked biosynthetic pathway of catechin metabolite genes from C. sinensis, consisting of flavanone 3 hydroxylase (F3H), dihydroflavonol reductase (DFR), and leucoanthocyanidin reductase (LCR), was designed and arranged in two sets of constructs: (a) single promoter in front of F3H and ribosome-binding sequences both in front of DFR and LCR; (b) three different promoters with each in the front of the three genes and ribosome-binding sequences at appropriate positions. Recombinant E. coli BL (DE3) harbouring the constructs were cultivated for 65 h at 26°C in M9 medium consisting of 40 g/L glucose, 1 mM IPTG, and 3 mM eriodictyol. Compounds produced were extracted in ethyl acetate in alkaline conditions after 1 h at room temperature and identified by HPLC. Two of the four major catechins, namely, (−)-epicatechin (0.01 ) and (−)-epicatechin gallate (0.36 mg/L), and two other types ((+)-catechin hydrate (0.13 mg/L) and (−)-catechin gallate (0.04 mg/L)) were successfully produced.http://dx.doi.org/10.1100/2012/529031
spellingShingle Kabir Mustapha Umar
S. M. Abdulkarim
Son Radu
Azizah Abdul Hamid
Nazamid Saari
Engineering the Production of Major Catechins by Escherichia coli Carrying Metabolite Genes of Camellia sinensis
The Scientific World Journal
title Engineering the Production of Major Catechins by Escherichia coli Carrying Metabolite Genes of Camellia sinensis
title_full Engineering the Production of Major Catechins by Escherichia coli Carrying Metabolite Genes of Camellia sinensis
title_fullStr Engineering the Production of Major Catechins by Escherichia coli Carrying Metabolite Genes of Camellia sinensis
title_full_unstemmed Engineering the Production of Major Catechins by Escherichia coli Carrying Metabolite Genes of Camellia sinensis
title_short Engineering the Production of Major Catechins by Escherichia coli Carrying Metabolite Genes of Camellia sinensis
title_sort engineering the production of major catechins by escherichia coli carrying metabolite genes of camellia sinensis
url http://dx.doi.org/10.1100/2012/529031
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