Identification, Recombinant Expression, and Biochemical Analysis of Putative Secondary Product Glucosyltransferases from Citrus paradisi
| Autor: | Starla Kiser, Mark A. Berhow, Zhangfan Lin, Daniel K. Owens, Venkata K. P. S. Mallampalli, Cecilia A. McIntosh, Jennifer Cooke, Tapasree Roy Sarkar, Bruce E. Williams, Lee M. Pike, Anye Wamucho, Shivakumar P. Devaiah, Peri Loftis, Mebrahtu B. Sibhatu, Josephat Asiago, Christy L. Strong, Deborah Hayford |
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| Rok vydání: | 2016 |
| Předmět: |
Limonins
0106 biological sciences 0301 basic medicine Molecular Sequence Data Flavonoid Gene Expression Genes Plant Limonoid 01 natural sciences Pichia Substrate Specificity Pichia pastoris 03 medical and health sciences Glucosyltransferases Phenols Citrus paradisi Coumarins Escherichia coli medicine Primer walking Amino Acid Sequence Phylogeny Flavonoids chemistry.chemical_classification biology food and beverages General Chemistry biology.organism_classification Recombinant Proteins 030104 developmental biology Biochemistry chemistry Seeds biology.protein Glucosyltransferase General Agricultural and Biological Sciences Sequence Alignment 010606 plant biology & botany medicine.drug |
| Zdroj: | Journal of Agricultural and Food Chemistry. 64:1957-1969 |
| ISSN: | 1520-5118 0021-8561 |
| DOI: | 10.1021/acs.jafc.5b05430 |
| Popis: | Flavonoid and limonoid glycosides influence taste properties as well as marketability of Citrus fruit and products, particularly grapefruit. In this work, nine grapefruit putative natural product glucosyltransferases (PGTs) were resolved by either using degenerate primers against the semiconserved PSPG box motif, SMART-RACE RT-PCR, and primer walking to full-length coding regions; screening a directionally cloned young grapefruit leaf EST library; designing primers against sequences from other Citrus species; or identifying PGTs from Citrus contigs in the harvEST database. The PGT proteins associated with the identified full-length coding regions were recombinantly expressed in Escherichia coli and/or Pichia pastoris and then tested for activity with a suite of substrates including flavonoid, simple phenolic, coumarin, and/or limonoid compounds. A number of these compounds were eliminated from the predicted and/or potential substrate pool for the identified PGTs. Enzyme activity was detected in some instances with quercetin and catechol glucosyltransferase activities having been identified. |
| Databáze: | OpenAIRE |
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