TY - JOUR
T1 - Purification and characterization of recombinant supersweet protein thaumatin II from tomato fruit
AU - Firsov, Aleksey
AU - Shaloiko, Lyubov
AU - Kozlov, Oleg
AU - Vinokurov, Leonid
AU - Vainstein, Alexander
AU - Dolgov, Sergey
N1 - Publisher Copyright:
© 2016 Elsevier Inc. All rights reserved.
PY - 2016/7/1
Y1 - 2016/7/1
N2 - 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.
AB - 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.
KW - Recombinant thaumatin II purification
KW - Supersweet proteins
KW - Thaumatin II
KW - Transgenic tomato
UR - http://www.scopus.com/inward/record.url?scp=84960941029&partnerID=8YFLogxK
U2 - 10.1016/j.pep.2016.03.002
DO - 10.1016/j.pep.2016.03.002
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C2 - 26965414
AN - SCOPUS:84960941029
SN - 1046-5928
VL - 123
SP - 1
EP - 5
JO - Protein Expression and Purification
JF - Protein Expression and Purification
ER -