Molecular cloning and in silico analysis of chalcone isomerase from Polygonum minus
Background: Chalcone isomerase (CHI; EC 5.5.1.6) is one of the key enzymes in the flavonoid biosynthetic pathway that is responsible for the intramolecular cyclization of chalcones into specific 2S-flavanones. Methods and results: In this study, the open reading frame (ORF) of CHI was successfully i...
| Main Authors: | , , , , , , |
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| Format: | Article |
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Springer
2023
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| Online Access: | http://psasir.upm.edu.my/id/eprint/108679/ |
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| author | Shah, Fatin Lyana Azman Baharum, Syarul Nataqain Goh, Hoe-Han Leow, Thean Chor Ramzi, Ahmad Bazli Oslan, Siti Nurbaya Sabri, Suriana |
| author_facet | Shah, Fatin Lyana Azman Baharum, Syarul Nataqain Goh, Hoe-Han Leow, Thean Chor Ramzi, Ahmad Bazli Oslan, Siti Nurbaya Sabri, Suriana |
| author_sort | Shah, Fatin Lyana Azman |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Background: Chalcone isomerase (CHI; EC 5.5.1.6) is one of the key enzymes in the flavonoid biosynthetic pathway that is responsible for the intramolecular cyclization of chalcones into specific 2S-flavanones. Methods and results: In this study, the open reading frame (ORF) of CHI was successfully isolated from the cDNA of Polygonum minus at 711-bp long, encoding for 236 amino acid residues, with a predicted molecular weight of 25.4 kDa. Multiple sequence alignment and phylogenetic analysis revealed that the conserved residues (Thr50, Tyr108, Asn115, and Ser192) in the cleft of CHI enzyme group active site are present in PmCHI protein sequence and classified as type I. PmCHI comprises more hydrophobic residues without a signal peptide and transmembrane helices. The three-dimensional (3D) structure of PmCHI predicted through homology modeling was validated by Ramachandran plot and Verify3D, with values within the acceptable range of a good model. PmCHI was cloned into pET-28b(+) plasmid, expressed in Escherichia coli BL21(DE3) at 16 °C and partially purified. Conclusion: These findings contribute to a deeper understanding of the PmCHI protein and its potential for further characterization of its functional properties in the flavonoid biosynthetic pathway. |
| first_indexed | 2025-11-15T14:00:47Z |
| format | Article |
| id | upm-108679 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-15T14:00:47Z |
| publishDate | 2023 |
| publisher | Springer |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-1086792024-09-26T08:24:43Z http://psasir.upm.edu.my/id/eprint/108679/ Molecular cloning and in silico analysis of chalcone isomerase from Polygonum minus Shah, Fatin Lyana Azman Baharum, Syarul Nataqain Goh, Hoe-Han Leow, Thean Chor Ramzi, Ahmad Bazli Oslan, Siti Nurbaya Sabri, Suriana Background: Chalcone isomerase (CHI; EC 5.5.1.6) is one of the key enzymes in the flavonoid biosynthetic pathway that is responsible for the intramolecular cyclization of chalcones into specific 2S-flavanones. Methods and results: In this study, the open reading frame (ORF) of CHI was successfully isolated from the cDNA of Polygonum minus at 711-bp long, encoding for 236 amino acid residues, with a predicted molecular weight of 25.4 kDa. Multiple sequence alignment and phylogenetic analysis revealed that the conserved residues (Thr50, Tyr108, Asn115, and Ser192) in the cleft of CHI enzyme group active site are present in PmCHI protein sequence and classified as type I. PmCHI comprises more hydrophobic residues without a signal peptide and transmembrane helices. The three-dimensional (3D) structure of PmCHI predicted through homology modeling was validated by Ramachandran plot and Verify3D, with values within the acceptable range of a good model. PmCHI was cloned into pET-28b(+) plasmid, expressed in Escherichia coli BL21(DE3) at 16 °C and partially purified. Conclusion: These findings contribute to a deeper understanding of the PmCHI protein and its potential for further characterization of its functional properties in the flavonoid biosynthetic pathway. Springer 2023-05-06 Article PeerReviewed Shah, Fatin Lyana Azman and Baharum, Syarul Nataqain and Goh, Hoe-Han and Leow, Thean Chor and Ramzi, Ahmad Bazli and Oslan, Siti Nurbaya and Sabri, Suriana (2023) Molecular cloning and in silico analysis of chalcone isomerase from Polygonum minus. Molecular Biology Reports, 50 (6). pp. 5283-5294. ISSN 0301-4851; ESSN: 1573-4978 https://link.springer.com/article/10.1007/s11033-023-08417-1?error=cookies_not_supported&code=2a4a92d0-4112-4a0c-8972-a4e431a89ead 10.1007/s11033-023-08417-1 |
| spellingShingle | Shah, Fatin Lyana Azman Baharum, Syarul Nataqain Goh, Hoe-Han Leow, Thean Chor Ramzi, Ahmad Bazli Oslan, Siti Nurbaya Sabri, Suriana Molecular cloning and in silico analysis of chalcone isomerase from Polygonum minus |
| title | Molecular cloning and in silico analysis of chalcone isomerase from Polygonum minus |
| title_full | Molecular cloning and in silico analysis of chalcone isomerase from Polygonum minus |
| title_fullStr | Molecular cloning and in silico analysis of chalcone isomerase from Polygonum minus |
| title_full_unstemmed | Molecular cloning and in silico analysis of chalcone isomerase from Polygonum minus |
| title_short | Molecular cloning and in silico analysis of chalcone isomerase from Polygonum minus |
| title_sort | molecular cloning and in silico analysis of chalcone isomerase from polygonum minus |
| url | http://psasir.upm.edu.my/id/eprint/108679/ http://psasir.upm.edu.my/id/eprint/108679/ http://psasir.upm.edu.my/id/eprint/108679/ |