Large scale production of cyclohexanone monooxygenase from Escherichia coli TOP10 pQR239
The cyclohexanone monooxygenase (CHMO) from Acinetobacter calcoaceticus NCIMB 9871 has been cloned into Escherichia coli in an L-arabinose inducible vector. The recombinant E. coli containing the L-arabinose inducible CHMO was grown at 1.5 litres under controlled conditions to determine the paramete...
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| Veröffentlicht in: | Enzyme and microbial technology 2001-02, Vol.28 (2), p.265-274 |
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| creator | O’Sullivan, Lisa M Patel, Sejal Ward, John M Woodley, John M Doig, Steven D |
| description | The cyclohexanone monooxygenase (CHMO) from
Acinetobacter calcoaceticus NCIMB 9871 has been cloned into
Escherichia coli in an L-arabinose inducible vector. The recombinant
E. coli containing the L-arabinose inducible CHMO was grown at 1.5 litres under controlled conditions to determine the parameters for growth and induction. It was found that induction with 0.1% (w/v) L-arabinose at late logarithmic phase of growth and growth for a further 2.5 to 3 h gave the optimal CHMO titre (∼3500 U.l
−1, 630 U · g dry cell weight
−1). High dissolved oxygen concentrations were shown to be deleterious to the CHMO titre. This influenced the strategy for growth and induction, and was optimal when the oxygen uptake rate was maximized but the dissolved oxygen concentration was zero. Finally, a 300 litre scale fermentation was carried out giving a total CHMO titre of >8 × 10
5 U. |
| doi_str_mv | 10.1016/S0141-0229(00)00320-3 |
| format | Article |
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Acinetobacter calcoaceticus NCIMB 9871 has been cloned into
Escherichia coli in an L-arabinose inducible vector. The recombinant
E. coli containing the L-arabinose inducible CHMO was grown at 1.5 litres under controlled conditions to determine the parameters for growth and induction. It was found that induction with 0.1% (w/v) L-arabinose at late logarithmic phase of growth and growth for a further 2.5 to 3 h gave the optimal CHMO titre (∼3500 U.l
−1, 630 U · g dry cell weight
−1). High dissolved oxygen concentrations were shown to be deleterious to the CHMO titre. This influenced the strategy for growth and induction, and was optimal when the oxygen uptake rate was maximized but the dissolved oxygen concentration was zero. Finally, a 300 litre scale fermentation was carried out giving a total CHMO titre of >8 × 10
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Acinetobacter calcoaceticus NCIMB 9871 has been cloned into
Escherichia coli in an L-arabinose inducible vector. The recombinant
E. coli containing the L-arabinose inducible CHMO was grown at 1.5 litres under controlled conditions to determine the parameters for growth and induction. It was found that induction with 0.1% (w/v) L-arabinose at late logarithmic phase of growth and growth for a further 2.5 to 3 h gave the optimal CHMO titre (∼3500 U.l
−1, 630 U · g dry cell weight
−1). High dissolved oxygen concentrations were shown to be deleterious to the CHMO titre. This influenced the strategy for growth and induction, and was optimal when the oxygen uptake rate was maximized but the dissolved oxygen concentration was zero. Finally, a 300 litre scale fermentation was carried out giving a total CHMO titre of >8 × 10
5 U.</description><subject>Acinetobacter calcoaceticus</subject><subject>arabinose</subject><subject>Biocatalyst</subject><subject>Cyclohexanone monooxygenase</subject><subject>E.coli</subject><subject>Escherichia coli</subject><subject>L-arabinose</subject><issn>0141-0229</issn><issn>1879-0909</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><recordid>eNqFkE1vEzEQhi0EoqHwE0A-oXJYmPHXrk8IVYVWilQ-isTN8nonjdHuOrUT1Pz7bpqovdHTaEbPzDt6GHuL8BEBzadfgAorEMKeAHwAkAIq-YzNsKltBRbsczZ7QI7Yq1L-AkwDBS_ZESIa0wgxY3_mPl8TL8H3xFc5dZuwjmnkacHDNvRpSbd-TCPxIY0p3W6vafSF-CKngZ-VsKQcwzJ6HlIf-dXldwS--vFTSPuavVj4vtCbQz1mv7-eXZ2eV_PLbxenX-ZVUMqsKy1qrJUhbVBJ00IbAiGGxlgtWhE6Zc3U1AR1ozwKrUKrhdekfWNr6YU8Zif7u9PzNxsqazfEEqjv_UhpUxw22kpVi7qZ0Pf_R6cMhXJ3U-_BkFMpmRZulePg89YhuJ19d2_f7dQ6AHdv38lp790hYNMO1D1uHXRPwOc9QJORf5GyKyHSGKiLmcLadSk-EXEHRBWR0g</recordid><startdate>20010201</startdate><enddate>20010201</enddate><creator>O’Sullivan, Lisa M</creator><creator>Patel, Sejal</creator><creator>Ward, John M</creator><creator>Woodley, John M</creator><creator>Doig, Steven D</creator><general>Elsevier Inc</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20010201</creationdate><title>Large scale production of cyclohexanone monooxygenase from Escherichia coli TOP10 pQR239</title><author>O’Sullivan, Lisa M ; Patel, Sejal ; Ward, John M ; Woodley, John M ; Doig, Steven D</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c446t-5271746e561436b0bcce11c86952b2cd496c867e0784a1254cb52a5e5a8973a23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Acinetobacter calcoaceticus</topic><topic>arabinose</topic><topic>Biocatalyst</topic><topic>Cyclohexanone monooxygenase</topic><topic>E.coli</topic><topic>Escherichia coli</topic><topic>L-arabinose</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>O’Sullivan, Lisa M</creatorcontrib><creatorcontrib>Patel, Sejal</creatorcontrib><creatorcontrib>Ward, John M</creatorcontrib><creatorcontrib>Woodley, John M</creatorcontrib><creatorcontrib>Doig, Steven D</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Enzyme and microbial technology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>O’Sullivan, Lisa M</au><au>Patel, Sejal</au><au>Ward, John M</au><au>Woodley, John M</au><au>Doig, Steven D</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Large scale production of cyclohexanone monooxygenase from Escherichia coli TOP10 pQR239</atitle><jtitle>Enzyme and microbial technology</jtitle><addtitle>Enzyme Microb Technol</addtitle><date>2001-02-01</date><risdate>2001</risdate><volume>28</volume><issue>2</issue><spage>265</spage><epage>274</epage><pages>265-274</pages><issn>0141-0229</issn><eissn>1879-0909</eissn><abstract>The cyclohexanone monooxygenase (CHMO) from
Acinetobacter calcoaceticus NCIMB 9871 has been cloned into
Escherichia coli in an L-arabinose inducible vector. The recombinant
E. coli containing the L-arabinose inducible CHMO was grown at 1.5 litres under controlled conditions to determine the parameters for growth and induction. It was found that induction with 0.1% (w/v) L-arabinose at late logarithmic phase of growth and growth for a further 2.5 to 3 h gave the optimal CHMO titre (∼3500 U.l
−1, 630 U · g dry cell weight
−1). High dissolved oxygen concentrations were shown to be deleterious to the CHMO titre. This influenced the strategy for growth and induction, and was optimal when the oxygen uptake rate was maximized but the dissolved oxygen concentration was zero. Finally, a 300 litre scale fermentation was carried out giving a total CHMO titre of >8 × 10
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| fulltext | fulltext |
| identifier | ISSN: 0141-0229 |
| ispartof | Enzyme and microbial technology, 2001-02, Vol.28 (2), p.265-274 |
| issn | 0141-0229 1879-0909 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_1859347278 |
| source | ScienceDirect Journals (5 years ago - present) |
| subjects | Acinetobacter calcoaceticus arabinose Biocatalyst Cyclohexanone monooxygenase E.coli Escherichia coli L-arabinose |
| title | Large scale production of cyclohexanone monooxygenase from Escherichia coli TOP10 pQR239 |
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