Identification, Recombinant Expression, and Biochemical Analysis of Putative Secondary Product Glucosyltransferases from Citrus paradisi.

Autor:	Devaiah SP; Department of Biological Sciences, East Tennessee State University , P.O. Box 70703, Johnson City, Tennessee 37614, United States., Owens DK; Natural Products Utilization Research Unit, ARS, U.S. Department of Agriculture , P.O. Box 1848, University, Mississippi 38677, United States., Sibhatu MB; Department of Biological Sciences, East Tennessee State University , P.O. Box 70703, Johnson City, Tennessee 37614, United States., Sarkar TR; Department of Biological Sciences, East Tennessee State University , P.O. Box 70703, Johnson City, Tennessee 37614, United States., Strong CL; Department of Biological Sciences, East Tennessee State University , P.O. Box 70703, Johnson City, Tennessee 37614, United States., Mallampalli VK; Department of Biological Sciences, East Tennessee State University , P.O. Box 70703, Johnson City, Tennessee 37614, United States., Asiago J; Department of Biological Sciences, East Tennessee State University , P.O. Box 70703, Johnson City, Tennessee 37614, United States., Cooke J; Department of Biological Sciences, East Tennessee State University , P.O. Box 70703, Johnson City, Tennessee 37614, United States., Kiser S; Department of Biological Sciences, East Tennessee State University , P.O. Box 70703, Johnson City, Tennessee 37614, United States., Lin Z; Department of Biological Sciences, East Tennessee State University , P.O. Box 70703, Johnson City, Tennessee 37614, United States., Wamucho A; Department of Biological Sciences, East Tennessee State University , P.O. Box 70703, Johnson City, Tennessee 37614, United States., Hayford D; Department of Biological Sciences, East Tennessee State University , P.O. Box 70703, Johnson City, Tennessee 37614, United States., Williams BE; Department of Biological Sciences, East Tennessee State University , P.O. Box 70703, Johnson City, Tennessee 37614, United States., Loftis P; Department of Biological Sciences, East Tennessee State University , P.O. Box 70703, Johnson City, Tennessee 37614, United States., Berhow M; Functional Foods Research Unit, ARS, U.S. Department of Agriculture , Peoria, Illinois 61604, United States., Pike LM; Department of Biological Sciences, East Tennessee State University , P.O. Box 70703, Johnson City, Tennessee 37614, United States., McIntosh CA; Department of Biological Sciences, East Tennessee State University , P.O. Box 70703, Johnson City, Tennessee 37614, United States.; School of Graduate Studies, East Tennessee State University , P.O. Box 70720, Johnson City, Tennessee 37614, United States.
Jazyk:	angličtina
Zdroj:	Journal of agricultural and food chemistry [J Agric Food Chem] 2016 Mar 09; Vol. 64 (9), pp. 1957-69. Date of Electronic Publication: 2016 Feb 29.
DOI:	10.1021/acs.jafc.5b05430
Abstrakt:	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:	MEDLINE
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