Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 56, Issue 15, Pages 6623-6630Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jf800793q
Keywords
cell wall hydrolase; Ellman's reagent; microbial lipase; porcine liver esterase; pectin acetylesterase
Funding
- Engineering and Physical Sciences Research Council [DT/E010415/1] Funding Source: researchfish
- EPSRC [DT/E010415/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council Funding Source: Medline
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Volatile organosulfur compounds (VOSCs) are high impact aroma chemicals characteristic of tropical fruits which are active as both free thiols and the respective thioesters. Using a simple and sensitive colorimetric enzyme assay, a thioesterase activity toward VOSCs has been identified in ripening purple passion fruit (Passiflora edulis Sims). The assay was based on determining the release of free thiols from 2-methyl-3-furanthiol acetate using Ellman's reagent. The major thioesterase in the fruit was found to be a wall-bound protein in the mesocarp. The extracted enzyme activity was purified 150-fold and shown to be associated with a 43 kDa monomeric serine hydrolase which was selectively labeled with a fluorophosphonate suicide probe. MS-MS sequencing identified the thioesterase as a class 13 glycoside hydrolase, most similar to pectin acetylesterase, an enzyme involved in cell wall modifications in the peel of a number of fruit. Our results suggest that cell wall hydrolases in tropical fruit may have additional useful roles in biotransforming VOSCs.
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