Biochemical and Molecular Characterization of the Rice Chalcone Isomerase Family
Abstract
:1. Introduction
2. Results
2.1. The CHI Family in Rice
2.2. Analysis of Conserved Residues within CHIs regarding Substrate-Binding and Catalysis
2.3. Cloning and Heterologous Expression of OsCHIs
2.4. CHI Activity and Kinetic Properties of OsCHIs
2.5. Analysis of OsCHI Gene Expression
3. Discussion
3.1. OsCHI3 Is a Bona Fide CHI in the OsCHI Family
3.2. Monocot Bona Fide CHIs Categorized into a Separate Branch within Type I CHIs
3.3. OsCHI3 Participates in UV-Induced Sakuranetin Synthesis in Rice Leaves
3.4. Prospective Role of Non-Catalytic OsCHI Members
4. Materials and Methods
4.1. Plant Growth, UV Treatment, HPLC Analysis, and Materials
4.2. Multiple Sequence Alignment and Phylogenetic Analysis of CHIs
4.3. Cloning of OsCHIs
4.4. Production and Purification of Recombinant OsCHIs
4.5. CHI Activity Assay
4.6. qRT- PCR Analysis of OsCHIs
4.7. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Name | Locus ID | Gene Description in the RGAP Database | ORF (bp) 1 | Protein Size (aa 2) | CHI Type |
---|---|---|---|---|---|
OsCHI1 | Os02g21520 | chalcone isomerase 3, putative | 843 | 280 | III |
OsCHI2 | Os02g53810 | expressed protein | 1284 | 427 | III |
OsCHI3 | Os03g60509 | expressed protein | 702 | 233 | I |
OsCHI4 | Os06g10210 | expressed protein | 1290 | 429 | III |
OsCHI5 | Os07g38390 | chalcone isomerase, putative | 792 | 263 | III |
OsCHI6 | Os11g02440 | chalcone-flavonone isomerase, putative | 651 | 216 | IV |
OsCHI7 | Os12g02370 | chalcone-flavonone isomerase, putative | 627 | 208 | IV |
Substrate | KM (µM) | Vmax (nmol s−1 mg−1) | kcat (s−1) | kcat/KM (M−1 s−1) |
---|---|---|---|---|
Naringenin chalcone | 11.60 ± 0.78 | 2662 ± 207.9 | 69.35 | 5.978 × 106 |
Isoliquiritigenin | 50.95 ± 4.83 | 3.54 × 10−3 ± 1.25 × 10−4 | 9.214 × 10−5 | 1.809 |
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Park, S.-I.; Park, H.-L.; Bhoo, S.-H.; Lee, S.-W.; Cho, M.-H. Biochemical and Molecular Characterization of the Rice Chalcone Isomerase Family. Plants 2021, 10, 2064. https://doi.org/10.3390/plants10102064
Park S-I, Park H-L, Bhoo S-H, Lee S-W, Cho M-H. Biochemical and Molecular Characterization of the Rice Chalcone Isomerase Family. Plants. 2021; 10(10):2064. https://doi.org/10.3390/plants10102064
Chicago/Turabian StylePark, Sang-Il, Hye-Lin Park, Seong-Hee Bhoo, Sang-Won Lee, and Man-Ho Cho. 2021. "Biochemical and Molecular Characterization of the Rice Chalcone Isomerase Family" Plants 10, no. 10: 2064. https://doi.org/10.3390/plants10102064