Imine reduction by an Ornithine cyclodeaminase/μ-crystallin homolog purified from Candida parapsilosis ATCC 7330
We report a stereospecific imine reductase from Candida parapsilosis ATCC 7330 (CpIM1), a versatile biocatalyst and a rich source of highly stereospecific oxidoreductases. The recombinant gene was overexpressed in Escherichia coli and the protein CpIM1 was purified to homogeneity. This protein belon...
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| Format: | Article |
| Language: | English |
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Elsevier
2021-09-01
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| Series: | Biotechnology Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215017X21000801 |
| _version_ | 1819136390704660480 |
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| author | V.N.M. Uma Mahesh Anju Chadha |
| author_facet | V.N.M. Uma Mahesh Anju Chadha |
| author_sort | V.N.M. Uma Mahesh |
| collection | DOAJ |
| description | We report a stereospecific imine reductase from Candida parapsilosis ATCC 7330 (CpIM1), a versatile biocatalyst and a rich source of highly stereospecific oxidoreductases. The recombinant gene was overexpressed in Escherichia coli and the protein CpIM1 was purified to homogeneity. This protein belongs to the Ornithine cyclodeaminase/ μ-crystallin (OCD-Mu) family of proteins which has only a few characterized members. CpIM1 catalyzed the alkylamination of α-keto acids/esters producing exclusively (S)-N-alkyl amino acids/esters e.g. N-methyl-l-alanine with > 90% conversion and > 99% enantiomeric excess (ee). The enzyme showed the highest activity for the alkylamination of pyruvate and methylamine leading to N-methyl-l-alanine with an apparent KM of 15.04 ± 2.8 mM and Vmax of 13.75 ± 1.07 μmol/min/mg. CpIM1 also catalyzed (i) the reduction of imines e.g. 2-methyl-1-pyrroline to (S)-2-methylpyrrolidine with ∼30% conversion and 75% ee and (ii) the dehydrogenation of cyclic amino acids e.g. l-Proline (as monitered by reduction of cofactor NADP+ spectrophotometrically). |
| first_indexed | 2024-12-22T10:34:13Z |
| format | Article |
| id | doaj.art-84c8da01141440e09f2d75e0857c06c4 |
| institution | Directory Open Access Journal |
| issn | 2215-017X |
| language | English |
| last_indexed | 2024-12-22T10:34:13Z |
| publishDate | 2021-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biotechnology Reports |
| spelling | doaj.art-84c8da01141440e09f2d75e0857c06c42022-12-21T18:29:15ZengElsevierBiotechnology Reports2215-017X2021-09-0131e00664Imine reduction by an Ornithine cyclodeaminase/μ-crystallin homolog purified from Candida parapsilosis ATCC 7330V.N.M. Uma Mahesh0Anju Chadha1Laboratory of Bioorganic Chemistry, Department of Biotechnology, Indian Institute of Technology Madras, Chennai 600 036, IndiaLaboratory of Bioorganic Chemistry, Department of Biotechnology, Indian Institute of Technology Madras, Chennai 600 036, India; National Center for Catalysis Research, Indian Institute of Technology Madras, Chennai 600 036, India; Corresponding author.We report a stereospecific imine reductase from Candida parapsilosis ATCC 7330 (CpIM1), a versatile biocatalyst and a rich source of highly stereospecific oxidoreductases. The recombinant gene was overexpressed in Escherichia coli and the protein CpIM1 was purified to homogeneity. This protein belongs to the Ornithine cyclodeaminase/ μ-crystallin (OCD-Mu) family of proteins which has only a few characterized members. CpIM1 catalyzed the alkylamination of α-keto acids/esters producing exclusively (S)-N-alkyl amino acids/esters e.g. N-methyl-l-alanine with > 90% conversion and > 99% enantiomeric excess (ee). The enzyme showed the highest activity for the alkylamination of pyruvate and methylamine leading to N-methyl-l-alanine with an apparent KM of 15.04 ± 2.8 mM and Vmax of 13.75 ± 1.07 μmol/min/mg. CpIM1 also catalyzed (i) the reduction of imines e.g. 2-methyl-1-pyrroline to (S)-2-methylpyrrolidine with ∼30% conversion and 75% ee and (ii) the dehydrogenation of cyclic amino acids e.g. l-Proline (as monitered by reduction of cofactor NADP+ spectrophotometrically).http://www.sciencedirect.com/science/article/pii/S2215017X21000801Imine reductaseμ-crystallin/Ornithine cyclodeaminase familyCandida parapsilosisAlkylaminationN-alkyl amino acid/estersChiral amines |
| spellingShingle | V.N.M. Uma Mahesh Anju Chadha Imine reduction by an Ornithine cyclodeaminase/μ-crystallin homolog purified from Candida parapsilosis ATCC 7330 Biotechnology Reports Imine reductase μ-crystallin/Ornithine cyclodeaminase family Candida parapsilosis Alkylamination N-alkyl amino acid/esters Chiral amines |
| title | Imine reduction by an Ornithine cyclodeaminase/μ-crystallin homolog purified from Candida parapsilosis ATCC 7330 |
| title_full | Imine reduction by an Ornithine cyclodeaminase/μ-crystallin homolog purified from Candida parapsilosis ATCC 7330 |
| title_fullStr | Imine reduction by an Ornithine cyclodeaminase/μ-crystallin homolog purified from Candida parapsilosis ATCC 7330 |
| title_full_unstemmed | Imine reduction by an Ornithine cyclodeaminase/μ-crystallin homolog purified from Candida parapsilosis ATCC 7330 |
| title_short | Imine reduction by an Ornithine cyclodeaminase/μ-crystallin homolog purified from Candida parapsilosis ATCC 7330 |
| title_sort | imine reduction by an ornithine cyclodeaminase μ crystallin homolog purified from candida parapsilosis atcc 7330 |
| topic | Imine reductase μ-crystallin/Ornithine cyclodeaminase family Candida parapsilosis Alkylamination N-alkyl amino acid/esters Chiral amines |
| url | http://www.sciencedirect.com/science/article/pii/S2215017X21000801 |
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