Purification and characterization of recombinant supersweet protein thaumatin II from tomato fruit.

Autor: Firsov A; Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the RAS, Moscow Region, Pushchino, 142290, Russia. Electronic address: aleksey_firsov@mail.ru., Shaloiko L; Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the RAS, Moscow Region, Pushchino, 142290, Russia., Kozlov O; Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the RAS, Moscow Region, Pushchino, 142290, Russia., Vinokurov L; Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the RAS, Moscow Region, Pushchino, 142290, Russia., Vainstein A; The Hebrew University of Jerusalem, Robert H. Smith Faculty of Agriculture, Food and Environment, POB 12, Rehovot, 76100, Israel., Dolgov S; Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the RAS, Moscow Region, Pushchino, 142290, Russia.
Jazyk: angličtina
Zdroj: Protein expression and purification [Protein Expr Purif] 2016 Jul; Vol. 123, pp. 1-5. Date of Electronic Publication: 2016 Mar 08.
DOI: 10.1016/j.pep.2016.03.002
Abstrakt: Thaumatin, a supersweet protein from the African plant katemfe (Thaumatococcus daniellii Benth.), is a promising zero-calorie sweetener for use in the food and pharmaceutical industries. Due to limited natural sources of thaumatin, its production using transgenic plants is an advantageous alternative. We report a simple protocol for purification of recombinant thaumatin II from transgenic tomato. Thaumatin was extracted from ripe tomato fruit in a low-salt buffer and purified on an SP-Sephacryl column. Recombinant thaumatin yield averaged 50 mg/kg fresh fruit. MALDI-MS analysis showed correct processing of thaumatin in tomato plants. The recombinant thaumatin was indistinguishable from the native protein in a taste test. The purified tomato-derived thaumatin had an intrinsic sweetness with a threshold value in taste tests of around 50 nM. These results demonstrate the potential of an expression system based on transgenic tomato plants for production of recombinant thaumatin for the food and pharmaceutical industries.
(Copyright © 2016 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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