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: Umar, Kabir Mustapha, Mohammed, Abdulkarim Sabo, Radu, Son, Abdul Hamid, Azizah, Saari, Nazamid
Format: Article
Language:English
Published: Hindawi Publishing Corporation 2012
Online Access:http://psasir.upm.edu.my/id/eprint/24140/
http://psasir.upm.edu.my/id/eprint/24140/1/24140.pdf
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author Umar, Kabir Mustapha
Mohammed, Abdulkarim Sabo
Radu, Son
Abdul Hamid, Azizah
Saari, Nazamid
author_facet Umar, Kabir Mustapha
Mohammed, Abdulkarim Sabo
Radu, Son
Abdul Hamid, Azizah
Saari, Nazamid
author_sort Umar, Kabir Mustapha
building UPM Institutional Repository
collection Online Access
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 65h at 26°C in M9 medium consisting of 40g/L glucose, 1mM IPTG, and 3mM eriodictyol. Compounds produced were extracted in ethyl acetate in alkaline conditions after 1h at room temperature and identified by HPLC. Two of the four major catechins, namely, (-)-epicatechin (0.01) and (-)-epicatechin gallate (0.36mg/L), and two other types ((+)-catechin hydrate (0.13mg/L) and (-)-catechin gallate (0.04mg/L)) were successfully produced.
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spelling upm-241402018-09-28T07:46:33Z http://psasir.upm.edu.my/id/eprint/24140/ Engineering the production of major catechins by Escherichia coli carrying metabolite genes of Camellia sinensis Umar, Kabir Mustapha Mohammed, Abdulkarim Sabo Radu, Son Abdul Hamid, Azizah Saari, Nazamid 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 65h at 26°C in M9 medium consisting of 40g/L glucose, 1mM IPTG, and 3mM eriodictyol. Compounds produced were extracted in ethyl acetate in alkaline conditions after 1h at room temperature and identified by HPLC. Two of the four major catechins, namely, (-)-epicatechin (0.01) and (-)-epicatechin gallate (0.36mg/L), and two other types ((+)-catechin hydrate (0.13mg/L) and (-)-catechin gallate (0.04mg/L)) were successfully produced. Hindawi Publishing Corporation 2012 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/24140/1/24140.pdf Umar, Kabir Mustapha and Mohammed, Abdulkarim Sabo and Radu, Son and Abdul Hamid, Azizah and Saari, Nazamid (2012) Engineering the production of major catechins by Escherichia coli carrying metabolite genes of Camellia sinensis. The Scientific World Journal, 2012. art. no. 529031. pp. 1-7. ISSN 2356-6140; ESSN: 1537-744X https://www.hindawi.com/journals/tswj/2012/529031/abs/ 10.1100/2012/529031
spellingShingle Umar, Kabir Mustapha
Mohammed, Abdulkarim Sabo
Radu, Son
Abdul Hamid, Azizah
Saari, Nazamid
Engineering the production of major catechins by Escherichia coli carrying metabolite genes of Camellia sinensis
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://psasir.upm.edu.my/id/eprint/24140/
http://psasir.upm.edu.my/id/eprint/24140/
http://psasir.upm.edu.my/id/eprint/24140/
http://psasir.upm.edu.my/id/eprint/24140/1/24140.pdf