Enantioselective microbial reduction of 6-oxo-8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5]decane-7,9-dione: Cloning and expression of reductases
The enantioselective microbial reduction of 6-oxo-8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5]decane-7,9-dione ( 1) to either of the corresponding ( S)- and ( R)-6-hydroxy-8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5]decane-7,9-diones ( 2 and 3, respectively) is descri...
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| Veröffentlicht in: | Enzyme and microbial technology 2006-11, Vol.39 (7), p.1441-1450 |
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| creator | Goldberg, Steven L. Nanduri, Venkata B. Chu, Linda Johnston, Robert M. Patel, Ramesh N. |
| description | The enantioselective microbial reduction of 6-oxo-8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5]decane-7,9-dione (
1) to either of the corresponding (
S)- and (
R)-6-hydroxy-8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5]decane-7,9-diones (
2 and
3, respectively) is described. The NADP
+-dependent (
R)-reductase (RHBR) which catalyzes the reduction of 6-ketobuspirone (
1) to (
R)-6-hydroxybuspirone (
3) was purified to homogeneity from cell extracts of
Hansenula polymorpha SC 13845. The subunit molecular weight of the enzyme is 35,000
kDa based on sodium dodecyl sulfate gel electrophoresis and the molecular weight of the enzyme is 37,000
kDa as estimated by gel filtration chromatography. (
R)-reductase from
H. polymorpha was cloned and expressed in
Escherichia coli. To regenerate the cofactor NADPH required for reduction we have cloned and expressed the glucose-6-phosphate dehydrogenase gene from
Saccharomyces cerevisiae in
E. coli. The NAD
+-dependent (
S)-reductase (SHBR) which catalyzes the reduction of 6-ketobuspirone (
1) to (
S)-6-hydroxybuspirone (
2) was purified to homogeneity from cell extracts of
Pseudomonas putida SC 16269. The subunit molecular weight of the enzyme is 25,000
kDa based on sodium dodecyl sulfate gel electrophoresis. The (
S)-reductase from
P. putida was cloned and expressed in
E. coli. To regenerate the cofactor NADH required for reduction we have cloned and expressed the formate dehydrogenase gene from
Pichia pastoris in
E. coli. Recombinant
E. coli expressing (
S)-reductase and (
R)-reductase catalyzed the reduction of
1 to (
S)-6-hyroxybuspirone (
2) and (
R)-6-hyroxybuspirone (
3), respectively, in >98% yield and >99.9% e.e. |
| doi_str_mv | 10.1016/j.enzmictec.2006.03.033 |
| format | Article |
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1) to either of the corresponding (
S)- and (
R)-6-hydroxy-8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5]decane-7,9-diones (
2 and
3, respectively) is described. The NADP
+-dependent (
R)-reductase (RHBR) which catalyzes the reduction of 6-ketobuspirone (
1) to (
R)-6-hydroxybuspirone (
3) was purified to homogeneity from cell extracts of
Hansenula polymorpha SC 13845. The subunit molecular weight of the enzyme is 35,000
kDa based on sodium dodecyl sulfate gel electrophoresis and the molecular weight of the enzyme is 37,000
kDa as estimated by gel filtration chromatography. (
R)-reductase from
H. polymorpha was cloned and expressed in
Escherichia coli. To regenerate the cofactor NADPH required for reduction we have cloned and expressed the glucose-6-phosphate dehydrogenase gene from
Saccharomyces cerevisiae in
E. coli. The NAD
+-dependent (
S)-reductase (SHBR) which catalyzes the reduction of 6-ketobuspirone (
1) to (
S)-6-hydroxybuspirone (
2) was purified to homogeneity from cell extracts of
Pseudomonas putida SC 16269. The subunit molecular weight of the enzyme is 25,000
kDa based on sodium dodecyl sulfate gel electrophoresis. The (
S)-reductase from
P. putida was cloned and expressed in
E. coli. To regenerate the cofactor NADH required for reduction we have cloned and expressed the formate dehydrogenase gene from
Pichia pastoris in
E. coli. Recombinant
E. coli expressing (
S)-reductase and (
R)-reductase catalyzed the reduction of
1 to (
S)-6-hyroxybuspirone (
2) and (
R)-6-hyroxybuspirone (
3), respectively, in >98% yield and >99.9% e.e.</description><identifier>ISSN: 0141-0229</identifier><identifier>EISSN: 1879-0909</identifier><identifier>DOI: 10.1016/j.enzmictec.2006.03.033</identifier><identifier>CODEN: EMTED2</identifier><language>eng</language><publisher>Amsterdam: Elsevier Inc</publisher><subject>( R)-Reductase ; ( S)-Reductase ; Biocatalysis ; Biological and medical sciences ; Biotechnology ; Cloning ; Enantioselective ; Escherichia coli ; Expression ; Fundamental and applied biological sciences. Psychology ; Hansenula polymorpha ; Pichia pastoris ; Pseudomonas putida ; Reduction ; Saccharomyces cerevisiae</subject><ispartof>Enzyme and microbial technology, 2006-11, Vol.39 (7), p.1441-1450</ispartof><rights>2006 Elsevier Inc.</rights><rights>2007 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c376t-7b60dce39111bf004f06b3b44cc860fe1f9b7c8570d9ebcf82ec63b4d08442163</citedby><cites>FETCH-LOGICAL-c376t-7b60dce39111bf004f06b3b44cc860fe1f9b7c8570d9ebcf82ec63b4d08442163</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0141022906001748$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=18154816$$DView record in Pascal Francis$$Hfree_for_read</backlink></links><search><creatorcontrib>Goldberg, Steven L.</creatorcontrib><creatorcontrib>Nanduri, Venkata B.</creatorcontrib><creatorcontrib>Chu, Linda</creatorcontrib><creatorcontrib>Johnston, Robert M.</creatorcontrib><creatorcontrib>Patel, Ramesh N.</creatorcontrib><title>Enantioselective microbial reduction of 6-oxo-8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5]decane-7,9-dione: Cloning and expression of reductases</title><title>Enzyme and microbial technology</title><description>The enantioselective microbial reduction of 6-oxo-8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5]decane-7,9-dione (
1) to either of the corresponding (
S)- and (
R)-6-hydroxy-8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5]decane-7,9-diones (
2 and
3, respectively) is described. The NADP
+-dependent (
R)-reductase (RHBR) which catalyzes the reduction of 6-ketobuspirone (
1) to (
R)-6-hydroxybuspirone (
3) was purified to homogeneity from cell extracts of
Hansenula polymorpha SC 13845. The subunit molecular weight of the enzyme is 35,000
kDa based on sodium dodecyl sulfate gel electrophoresis and the molecular weight of the enzyme is 37,000
kDa as estimated by gel filtration chromatography. (
R)-reductase from
H. polymorpha was cloned and expressed in
Escherichia coli. To regenerate the cofactor NADPH required for reduction we have cloned and expressed the glucose-6-phosphate dehydrogenase gene from
Saccharomyces cerevisiae in
E. coli. The NAD
+-dependent (
S)-reductase (SHBR) which catalyzes the reduction of 6-ketobuspirone (
1) to (
S)-6-hydroxybuspirone (
2) was purified to homogeneity from cell extracts of
Pseudomonas putida SC 16269. The subunit molecular weight of the enzyme is 25,000
kDa based on sodium dodecyl sulfate gel electrophoresis. The (
S)-reductase from
P. putida was cloned and expressed in
E. coli. To regenerate the cofactor NADH required for reduction we have cloned and expressed the formate dehydrogenase gene from
Pichia pastoris in
E. coli. Recombinant
E. coli expressing (
S)-reductase and (
R)-reductase catalyzed the reduction of
1 to (
S)-6-hyroxybuspirone (
2) and (
R)-6-hyroxybuspirone (
3), respectively, in >98% yield and >99.9% e.e.</description><subject>( R)-Reductase</subject><subject>( S)-Reductase</subject><subject>Biocatalysis</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cloning</subject><subject>Enantioselective</subject><subject>Escherichia coli</subject><subject>Expression</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Hansenula polymorpha</subject><subject>Pichia pastoris</subject><subject>Pseudomonas putida</subject><subject>Reduction</subject><subject>Saccharomyces cerevisiae</subject><issn>0141-0229</issn><issn>1879-0909</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><recordid>eNqFUVFrFDEQXkTBs_U3uC-KQrOdbHLZjW_lqFUo-FKfSgnZZCI59pI12Su9-zX-VHPeoY-FIWEm33zfZL6qekehoUDF5brBsN94M6NpWgDRACvBXlQL2neSgAT5sloA5ZRA28rX1Zuc1wClwGFR_b4OOsw-ZhzRzP4R60KV4uD1WCe021KLoY6uFiQ-RdKTe36Ijy2ZdslvvPVhN34ilEx-wqT3h_Rh2M7lLGC913nyKd7zZvlg0eiApLuQxBZS_Fyvxhh8-FnrYGt8mhLmfFI7SuuM-bx65fSY8e3pPqt-fLm-W30lt99vvq2ubolhnZhJNwiwBpmklA4OgDsQAxs4N6YX4JA6OXSmX3ZgJQ7G9S0aUd4t9Jy3VLCz6sORd0rx1xbzrDY-GxzHMnPcZkVlz5adpAXYHYFlTTkndGoqi9BppyiogyNqrf45og6OKGAlWOl8f5LQ2ejRJR2Mz__be7rk_d9Rro44LP999JhUNh6DQetTMUnZ6J_V-gMY2Kfh</recordid><startdate>20061103</startdate><enddate>20061103</enddate><creator>Goldberg, Steven L.</creator><creator>Nanduri, Venkata B.</creator><creator>Chu, Linda</creator><creator>Johnston, Robert M.</creator><creator>Patel, Ramesh N.</creator><general>Elsevier Inc</general><general>Elsevier Science</general><scope>IQODW</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QO</scope><scope>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope></search><sort><creationdate>20061103</creationdate><title>Enantioselective microbial reduction of 6-oxo-8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5]decane-7,9-dione: Cloning and expression of reductases</title><author>Goldberg, Steven L. ; Nanduri, Venkata B. ; Chu, Linda ; Johnston, Robert M. ; Patel, Ramesh N.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c376t-7b60dce39111bf004f06b3b44cc860fe1f9b7c8570d9ebcf82ec63b4d08442163</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>( R)-Reductase</topic><topic>( S)-Reductase</topic><topic>Biocatalysis</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Cloning</topic><topic>Enantioselective</topic><topic>Escherichia coli</topic><topic>Expression</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Hansenula polymorpha</topic><topic>Pichia pastoris</topic><topic>Pseudomonas putida</topic><topic>Reduction</topic><topic>Saccharomyces cerevisiae</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Goldberg, Steven L.</creatorcontrib><creatorcontrib>Nanduri, Venkata B.</creatorcontrib><creatorcontrib>Chu, Linda</creatorcontrib><creatorcontrib>Johnston, Robert M.</creatorcontrib><creatorcontrib>Patel, Ramesh N.</creatorcontrib><collection>Pascal-Francis</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</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>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Enzyme and microbial technology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Goldberg, Steven L.</au><au>Nanduri, Venkata B.</au><au>Chu, Linda</au><au>Johnston, Robert M.</au><au>Patel, Ramesh N.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Enantioselective microbial reduction of 6-oxo-8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5]decane-7,9-dione: Cloning and expression of reductases</atitle><jtitle>Enzyme and microbial technology</jtitle><date>2006-11-03</date><risdate>2006</risdate><volume>39</volume><issue>7</issue><spage>1441</spage><epage>1450</epage><pages>1441-1450</pages><issn>0141-0229</issn><eissn>1879-0909</eissn><coden>EMTED2</coden><abstract>The enantioselective microbial reduction of 6-oxo-8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5]decane-7,9-dione (
1) to either of the corresponding (
S)- and (
R)-6-hydroxy-8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5]decane-7,9-diones (
2 and
3, respectively) is described. The NADP
+-dependent (
R)-reductase (RHBR) which catalyzes the reduction of 6-ketobuspirone (
1) to (
R)-6-hydroxybuspirone (
3) was purified to homogeneity from cell extracts of
Hansenula polymorpha SC 13845. The subunit molecular weight of the enzyme is 35,000
kDa based on sodium dodecyl sulfate gel electrophoresis and the molecular weight of the enzyme is 37,000
kDa as estimated by gel filtration chromatography. (
R)-reductase from
H. polymorpha was cloned and expressed in
Escherichia coli. To regenerate the cofactor NADPH required for reduction we have cloned and expressed the glucose-6-phosphate dehydrogenase gene from
Saccharomyces cerevisiae in
E. coli. The NAD
+-dependent (
S)-reductase (SHBR) which catalyzes the reduction of 6-ketobuspirone (
1) to (
S)-6-hydroxybuspirone (
2) was purified to homogeneity from cell extracts of
Pseudomonas putida SC 16269. The subunit molecular weight of the enzyme is 25,000
kDa based on sodium dodecyl sulfate gel electrophoresis. The (
S)-reductase from
P. putida was cloned and expressed in
E. coli. To regenerate the cofactor NADH required for reduction we have cloned and expressed the formate dehydrogenase gene from
Pichia pastoris in
E. coli. Recombinant
E. coli expressing (
S)-reductase and (
R)-reductase catalyzed the reduction of
1 to (
S)-6-hyroxybuspirone (
2) and (
R)-6-hyroxybuspirone (
3), respectively, in >98% yield and >99.9% e.e.</abstract><cop>Amsterdam</cop><pub>Elsevier Inc</pub><doi>10.1016/j.enzmictec.2006.03.033</doi><tpages>10</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0141-0229 |
| ispartof | Enzyme and microbial technology, 2006-11, Vol.39 (7), p.1441-1450 |
| issn | 0141-0229 1879-0909 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_19835791 |
| source | Elsevier ScienceDirect Journals |
| subjects | ( R)-Reductase ( S)-Reductase Biocatalysis Biological and medical sciences Biotechnology Cloning Enantioselective Escherichia coli Expression Fundamental and applied biological sciences. Psychology Hansenula polymorpha Pichia pastoris Pseudomonas putida Reduction Saccharomyces cerevisiae |
| title | Enantioselective microbial reduction of 6-oxo-8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5]decane-7,9-dione: Cloning and expression of reductases |
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