Expression of thaumatin, a new type of alternative sweetener in rice
DOI:
https://doi.org/10.15835/nbha48311676Keywords:
Agrobacterium tumefaciens; Oryza sativa L; protein expression; taqman PCR; thaumatinAbstract
Sweet proteins are the natural alternative to the artificial sweeteners as well as flavor enhancers. Among other sweet protein, thaumatin protein was isolated from Thaumatococcus daniellii Benth plant fruit. In this study, pinII Ti plasmid vector was constructed with thaumatin gene, where thaumatin was placed under the control of the duel cauliflower mosaic virus 35S promoter into rice (Oryza sativa L. var. japonica cv. ‘Dongjinbyeo’) by Agrobacterium-mediated transformation to generate transgenic plants. Thirteen plant lines were regenerated and the transgenic rice lines were confirmed by different molecular analysis. The genomic PCR result revealed that all of the plant lines were transgenic. The single copy and intergenic plant lines were selected by Taqman PCR analysis and FST analysis, respectively. Expression of thaumatin gene in transgenic rice resulted in the accumulation of thaumatin protein in the leave. Thaumatin protein was also accumulated in leave of T1 generation. Sensory analysis result suggested that the thaumatin protein expressing transgenic lines exerted sweet tasting activity. These results demonstrated that thaumatin was expressed in transgenic rice plants.
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Akter S, Huq MA, Jung YJ, Cho YG, Kang KK (2016). Application of sweet and taste modifying genes for development in plants: current status and prospects. Journal of Plant Biotechnology 43:397-404.
Assadi-Porter FM, Aceti D, Markley JL (2000). Sweetness determinant sites of brazzein, a small, heat-stable, sweettasting protein. Archives of Biochemestry and Biophysics 376(2):259-265.
Baneyx F (1999). Recombinant protein expression in Escherichia coli. Current Opinion in Biotechnology 10:411-421.
Bartoszewski G, Niedziela A, Szwacka M, Niemirowicz-Szczytt K (2003). Modification of tomato taste in transgenic plants carrying a thaumatin gene from Thaumatococcus daniellii Benth. Plant Breeding 122:347-351.
Bradford MM (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248-254.
Colinas J, Schmidler SC, Bohrer G, Iordanov B, Benfey PN (2008). Intergenic and genic sequence lengths have opposite relationships with respect to gene expression. PLoS One 3(11):e3670.
Daniell H, Streatfield SJ, Wycoff K (2001). Medical molecular farming: production of antibodies, biopharmaceuticals and edible vaccines in plants. Trends in Plant Science 6:219-226.
Daniell S, Mellits KH, Faus I, Connerton I (2000). Refolding the sweet-tasting protein thaumatin II from insoluble inclusion bodies synthesised in Escherichia coli. Food Chemistry 71:105-110.
Edens L, Bom L, Ledeboer AM, Maat J, Toonen MY, Visser C, Verrips CT (1984). Synthesis and processing of the plant protein thaumatin in yeast. Cell 37:629-633.
Edens L, Heslinga L, Klok R, Ledeboer AM, Maat J, Toonen MY, … Verrips CT (1982). Cloning of cDNA encoding the sweet-tasting plant protein thaumatin and its expression in Escherichia coli. Gene 18:1-12.
Etheridge K (1994). The sales and marketing of talin. In: Witty M, Higginbotham JD, eds. Thaumatin, Boca Raton: CRC Press 1994:47-59.
Faus I (2000). Recent developments in the characterization and biotechnological production of sweet-tasting proteins. Applied Microbiology and Biotechnology 53:145-151.
Faus I, del Moral C, Adroer N, del Río JL, Patiño C, Sisniega H, … Rubio V (1998). Secretion of the sweet tasting protein thaumatin by recombinant strains of Aspergillus niger var. awamori. Applied Microbiology and Biotechnology 49:393-398.
Faus I, Patiño C, del Río JL, del Moral C, Barroso HS, Bladé J, Rubio V (1997). Expression of a synthetic gene encoding the sweet-tasting protein thaumatin in the filamentous fungus Penicillium roquefortii. Biotechnology Letters 19:1185-1191.
Faus I, Patiño C, Río JL, del Moral C, Barroso HS, Rubio V (1996). Expression of a synthetic gene encoding the sweet-tasting protein thaumatin in Escherichia coli. Biochemical and Biophysical Research Communications 229(1):121-127.
Gibbs BF, Alli I, Mulligan C (1996). Sweet and taste-modifying proteins: a review. Nutrition Research 16:1619-1630.
Hahm YT, Batt CA (1990). Expression and secretion of thaumatin from Aspergillus oryzae. Agricultural and Biological Chemistry 54:2513-2520.
Hannig G, Makrides SC (1998). Strategies for optimizing heterologous protein expression in Escherichia coli. Trends in Biotechnology 16:54-60.
Higginbottom HJD (1985). Talin update. Food Flavourings, Ingredients, Processing & Packaging. 1985, 7(Jan), 39.
Hood EE, Jilka JM (1999). Plant-based production of xenogenic proteins. Current Opinion in Biotechnology 10:382-386.
Hood EE, Woodard SL, Horn ME (2002). Monoclonal antibody manufacturing on transgenic plants- myths and realities. Current Opinion in Biotechnology 13:630-635.
Horn ME, Woodard SL, Howard JA (2004). Plant molecular farming: systems and products. Plant Cell Reports 22:711-720.
Howard JA, Hood E (2005). Bioindustrial and biopharmaceutical products produced in plants. Advances in Agronomy 85:91-124.
Illingworth C, Larson G, Hellekant G (1988). Secretion of the sweet-tasting plant protein thaumatin by Bacillus subtilis. Biotechnology Letters 10:587-592.
Illingworth C, Larson G, Hellekant G (1989). Secretion of the sweet-tasting plant protein thaumatin by Streptomyces lividans. Journal of Industrial Microbiology and Biotechnology 4:37-42.
Kant R (2005). Sweet proteins, potential replacement for artificial low-calorie sweeteners. Nutrition Journal 4:5.
Laemmli UK (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-685.
Lamphear BJ, Barker DK, Brooks CA, Delaney DE, Lane JR, Beifuss K, … Howard JA (2005). Expression of the sweet protein brazzein in maize for production of a new commercial sweetener. Plant Biotechnology Journal 3:103-114.
Lebedev VG, Taran SA, Shmatchenko VV, Dolgov SV (2002). Pear transformation with the gene for super sweet protein thaumatin II. Acta Horticulturae 596:199-202.
Makrides SC (1996). Strategies for achieving high-level expression of genes in Escherichia coli. Microbiological Reviews 60:512-538.
Masuda M, Tamaki S, Kaneko R, Wada R, Fujita Y, Mehta A, Kitabatake N (2004). Cloning, expression, and characterization of recombinant sweet-protein thaumatin II using the methylotrophic yeast Pichia pastoris. Biotechnology and Bioengineering 85:761-769.
Moralejo FJ, Cardoza RE, Gutierrez S, Martin JF (1999). Thaumatin production in Aspergillus awamori by use of expression cassettes with strong fungal promoters and high gene dosage. Applied and Environmental Microbiology 65:1168-1174.
Price JM, Biava CG, Oser BL, Vogin EE, Steinfield J, Ley HL (1970). Bladder tumors in rats fed cyclohexylamine or high doses of a mixture of cyclamate and saccharin. Science 167:1131-1132.
Schestibratov KA, Dolgov SV (2005). Transgenic strawberry plants expressing a thaumatin II gene demonstrate enhanced resistance to Botrytis cinerea. Scientia Horticulturae 106:177-189.
Streatfield SJ, Howard JA (2003). Plant-based vaccines. International Journal for Parasitology 33:479-493.
Streatfield SJ (2007). Approaches to achieve high-level heterologous protein production in plants. Plant Biotechnology Journal 5:2-15.
Sun HJ, Cui ML, Ma B, Ezura H (2006). Functional expression of the taste-modifying protein, miraculin, in transgenic lettuce. FEBS Letter 580:620-626.
Sun HJ, Kataoka H, Yano M, Ezura H (2007). Genetically stable expression of functional miraculin, a new type of alternative sweetener, in transgenic tomato plants. Plant Biotechnology Journal 5:768-777.
Szwacka M, Kryzymowska M, Osuch A, Kowalczyk ME, Malepszy S (2002). Variable properties of transgenic cucumber plants containing the thaumatin II gene from Thaumatococcus daniellii. Acta Physiologiae Plantarum 24:173-185.
Thole V, Alves SC, Worland B, Bevan MW, Vain P (2009). A protocol for efficiently retrieving and characterizing flanking sequence tags (FSTs) in Brachypodium distachyon T-DNA insertional mutants. Nature Protocols 4:650-661.
Van der Wel H, Loeve K (1972). Isolation and characterization of thaumatin I and II, the sweet-tasting proteins from Thaumatococcus daniellii Benth. European Journal of Biochemistry 31:221-225.
Wang X, Jiang D, Yang D (2015). Fast-tracking determination of homozygous transgenic lines and transgene stacking using a reliable quantitative real-time PCR assay. Biotechnology and Applied Biochemistry 175:996-1006.
Weickmann JL, Blair LC, Wilcox GL (1994). High level expression of thaumatin in Saccharomyces cerevisiae. In: Witty M and Higginbotham JD (ed.), Thaumatin. CRC Press, Boca Raton, FL, pp 151-169.
Witty M (1990). Preprothaumatin II is processed to biological activity in Solanum tuberosum. Biotechnology Letter 12:131-136.
Witty M, Higginbotham JD (1994). Thaumatin, CRC Press, Boca Raton, FL.
Witty M, Harvey WJ (1990). Sensory evaluation of transgenic Solanum tuberosum producing r-thaumatin II. New Zealand Journal of Crop and Horticultural Science 18:77-80.
Yoshida K, Shinmyo A (2000). Transgenic expression systems in plants, a natural bioreactor. Journal of Bioscience and Bioengineering 90:353-362.

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