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标题：Biomanufacturing of Tomato-Derived Nanovesicles
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作者：Ramesh Bokka ; Anna Paulina Ramos ; Immacolata Fiume 等
期刊名称：Foods
电子版ISSN：2304-8158
出版年度：2020
卷号：9
期号：12
页码：1852-1871
DOI：10.3390/foods9121852
出版社：MDPI Publishing
摘要：Micro- and nano-sized vesicles (MVs and NVs, respectively) from edible plant resources are gaining increasing interest as green, sustainable, and biocompatible materials for the development of next-generation delivery vectors. The isolation of vesicles from complex plant matrix is a significant challenge considering the trade-off between yield and purity. Here, we used differential ultracentrifugation (dUC) for the bulk production of MVs and NVs from tomato (Solanum lycopersicum L.) fruit and analyzed their physical and morphological characteristics and biocargo profiles. The protein and phospholipid cargo shared considerable similarities between MVs and NVs. Phosphatidic acid was the most abundant phospholipid identified in NVs and MVs. The bulk vesicle isolates were further purified using sucrose density gradient ultracentrifugation (gUC) or size-exclusion chromatography (SEC). We showed that SEC using gravity column efficiently removed co-purifying matrix components including proteins and small molecular species. dUC/SEC yielded a high yield of purified vesicles in terms of number of particles (2.6 × 1015 particles) and protein quantities (6.9 ± 1.5 mg) per kilogram of tomato. dUC/gUC method separated two vesicle populations on the basis of buoyant density. Proteomics and in silico studies of the SEC-purified MVs and NVs support the presence of different intra- and extracellular vesicles with highly abundant lipoxygenase (LOX), ATPases, and heat shock proteins (HSPs), as well as a set of proteins that overlaps with that previously reported in tomato chromoplast.
关键词：nanovesicles; tomato; size-exclusion chromatography; biomanufacturing; proteomics; phospholipids; Solanum lycopersicum L;; gradient ultracentrifugation; differential ultracentrifugation nanovesicles ; tomato ; size-exclusion chromatography ; biomanufacturing ; proteomics ; phospholipids ; Solanum lycopersicum L; ; gradient ultracentrifugation ; differential ultracentrifugation