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http://dx.doi.org/10.5010/JPB.2018.45.3.257

Stable expression and characterization of brazzein, thaumatin and miraculin genes related to sweet protein in transgenic lettuce  

Jung, Yeo Jin (Department of Horticultural Life Science, Hankyong National University)
Kang, Kwon Kyoo (Department of Horticultural Life Science, Hankyong National University)
Publication Information
Journal of Plant Biotechnology / v.45, no.3, 2018 , pp. 257-265 More about this Journal
Abstract
Sweetener is one of the additives that makes you feel sweet. Artificial sweeteners and sugar are typical examples, and sweetness proteins with sweetness characteristics have been widely studied. These studies elucidated the transformation lettuce cells with Agrobacterium method for stable production of natural sweet proteins, brazzein, thaumatin, and miraculin. In this paper, we report use of a plant expression system for production of sweet proteins. A synthetic gene encoding sweet proteins was placed under the control of constitutive promoters and transferred to lettuce. High and genetically stable expression of sweetener was confirmed in leaves by RT-PCR and Western blot analysis. Sweet proteins expressed in transgenic lettuce had sweetness-inducing activity. Results demonstrate recombinant sweet proteins correctly processed in transgenic lettuce plants, and that this production system could be a viable alternative to production from the native plant.
Keywords
Taste modifying protein; Brazzein; Thaumatin; Miraculin; Transgenic lettuce; Agrobacterium method;
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1 Masuda T, Kitabatake N (2006) Developments in biotechnological production of sweet proteins. J Biosci Bioeng 102:375-389   DOI
2 Masuda T, Ueno Y, Kitabatake N (2001) Sweetness and enzymatic activity of lysozyme. J Agr Food Chem 49:4937-4941   DOI
3 Ming D, Hellekant G (1994) Brazzein, a new high-potency thermostable sweet protein from Pentadiplandra byamarazzeana B. FEBS Lett 355:106-108   DOI
4 Moris JA, Cagan RH (1972) Purification of monellin, the sweet principal for Dioscoreophyllum cumminsii. Biochim Biophys Acta 261:114-122   DOI
5 Rachid A, Belloir C, Chevalier J, Desmetz C, Miller ML, et al. (2009) Optimization of the Production of Recombinant Brazzein Secreted by the Yeast Pichia pastoris. Chem senses 34:A80-A80
6 Ren YL, Zhou YW, Ye YH. 2004. Chemical components of Lactuca and their bioactivites. Acta Pharmaceutica Sinica 39:954-960
7 Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-685   DOI
8 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. J Plant Biotechnol 43:397-404   DOI
9 Assadi-Porter FM, Aceti DJ, Cheng H, Markley JL (2000) Efficient production of recombinant brazzein, a small, heat-stable, sweettasting protein of plant origin. Arch Biochem Biophys 376:252-258   DOI
10 Assadi-Porter FM, Aceti DJ, Markley JL (2000) Sweetness Determinant Sites of Brazzein, a Small, Heat-Stable, Sweet-Tasting Protein. Arch Biochem Biophys 376:259-265.   DOI
11 Assadi-Porter FM, Patry S, Markley JL (2008) Efficient and rapid protein expression and purification of small high disulfide containing sweet protein brazzein in E. coli. Protein Expr Purif 58:263-268   DOI
12 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 4:650-661
13 Shirasuka Y, Nakajima K, Asakura T, Yamashita H, Yamamoto A, Hata S, Nagata S, Abo M, Sorimachi H, Abe K (2004) Neoculin as a new taste-modifying protein occurring in the fruit of Curculigo latifolia. Biosci Biotech Bioch 68:1403-1407   DOI
14 Sun HJ, Cui ML, Ma B, Ezura H (2006) Functional expression of the taste-modifying protein, miraculin, in transgenic lettuce. FEBS Lett 580:620-626   DOI
15 Streatfield SJ (2007) Approaches to achieve high-level heterologous protein production in plants. Plant Biotechnol J 5:2-15   DOI
16 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 Biotechnol J 5:768-777   DOI
17 Streatfield SJ, Howard JA (2003) Plant-based vaccines. Int J Parasitol 33:479-493   DOI
18 Van der Wel H (1972) Isolation and characterization of the sweet principal for Dioscoreophyllum cumminsii (Stapf) Diels. FEBS Lett 21:88-90   DOI
19 Van der Wel H and Loeve K (1972) Isolation and characterization of thaumatin I and II, the sweet-tasting proteins from Thaumatococcus daniellii Benth. Eur J Biochem 31:221-225   DOI
20 Van der Wel H, Larson G, Hladik A, Hladik CM, Hellekant G, Glaser D (1989) Isolation and characterization of pentadin, the sweet principle of Pentadiplandra brazzeana Baillon. Chem Senses 264:6655-6659
21 Yan S, Song H, Pang D, Zou Q, Li L, Yan Q, Fan N, Zhao X, Yu H, Li Z, Wang H, Gao F, Ouyang H, Lai L (2013) Expression of Plant Sweet Protein Brazzein in the Milk of Transgenic Mice. PloS one 8:e76769   DOI
22 Daniell H, Streatfield SJ, Wycoff K (2001) Medical molecular farming: production of antibodies, biopharmaceuticals and edible vaccines in plants. Trends Plant Sci 6:219-226   DOI
23 Berlec A, Jevnikar Z, Majhenic AC, Rogelj I, Strukelj B (2006) Expression of the sweet-tasting plant protein brazzein in Escherichia coli and Lactococcus lactis: a path toward sweet lactic acid bacteria. Appl microbiol biot 73:158-165   DOI
24 Berlec A, Strukelj B (2009) Large increase in brazzein expression achieved by changing the plasmid/strain combination of the NICE system in Lactococcus lactis. Lett Appl Microbiol 48:750-755
25 Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248-254   DOI
26 Faus I (2000) Recent developments in the characterization and biotechnological production of sweet-tasting proteins. Appl Microbiol Biot 53:145-151   DOI
27 Farbman AI, Ogden-Ogle CK, Hellekant G, Simmons SR, Albrecht RM, Van Der Wel H: Labeling of sweet taste binding sites using a colloidal gold-labeled sweet protein, thaumatin. Scanning Microsc. 1987, 1 (1): 351-7
28 Howard JA, Hood E (2005) Bioindustrial and biopharmaceutical products produced in plants. Adv Agron 85:91-124
29 Hwang-Bo J, Kyung Ok Jang, Hayoung Chung, Jong-Hwa Park, Tae Hoon Lee, Jiyoung Kim and In Sik Chung (2016) Antiinflammatory Effect of Lactuca sativa L. Extract in Human Umbilical Vein Endothelial Cells and Improvement of Lipid Levels in Mice Fed a High-fat Diet. Korean J. Food Nutr. 29(6): 998-1007   DOI
30 Yamashita H, Theeraship A, Nakaya T, Nakamura Y, Kurihara Y (1990) Purification and complete amino acid sequence of a new type of sweet protein with taste-modifying activity, curculin. J Biol Chem 265:15770-15775
31 Hood EE, Jilka JM (1999) Plant-based production of xenogenic proteins. Curr Opin Biotech 10:382-386   DOI
32 Jung YJ, Bae SS, Lee GJ, Seo PJ, Cho Y G, Kang KK (2017) A novel method for high-frequency genome editing in rice, using the CRISPR/Cas9 system. J Plant Biotechnol 44:89-96   DOI
33 Yoshida K, Shinmyo A (2000) Transgenic expression systems in plants, a natural bioreactor. J Biosci Bioeng 90:353-362   DOI
34 Zemanek EC, Wasserman BP: Issues and advances in the use of transgenic organisms for the production of thaumatin, the intensely sweet protein from Thaumatococcus danielli. Crit Rev Food Sci Nutr. 1995, 35(5):455-66   DOI
35 Hood EE, Woodard SL, Horn ME (2002) Monoclonal antibody manufacturing on transgenic plants-myths and realities. Curr Opin Biotech 13:630-635   DOI
36 Horn ME, Woodard SL, Howard JA (2004) Plant molecular farming:systems and products. Plant Cell Rep 22:711-720   DOI
37 Jo HJ, Noh JS, Kong KH (2013) Efficient secretory expression of the sweet-tasting protein brazzein in the yeast Kluyveromyces lactis. Protein Expres Purif 90:84-9   DOI
38 Jung YJ, Nogoy FM, Cho Y G, Kang KK (2015) Development of high tryptophan GM rice and its transcriptome analysis. J Plant Biotechnol 42:186-195   DOI
39 Kant R (2005) Sweet proteins, potential replacement for artificial low calorie sweeteners. Nutrition 4:5   DOI
40 Jung YJ, Nou IS, Kang KK (2014) Overexpression of Oshsp16.9 gene encoding small heat shock protein enhances tolerance to abiotic stresses in rice. Plant Breed Biotech 2(4):370-379   DOI
41 Kurihara Y, Beidler LM (1968) Taste-modifying protein from miracle fruit. Science 161:1241-1243   DOI
42 Lamphear BJ, Barker DK, Brooks CA, Delaney DE, Lane JR, Beifuss K, Love R, Thompson K, Mayor J, Clough R, Harkey R, Poage M, Drees C, Horn ME, Streatfield SJ, Nikolov Z, Woodard SL, Hood EE, Jilka JM, Howard JA (2005) Expression of the sweet protein brazzein in maize for production of a new commercial sweetener. Plant Biotechnology Journal 3:103-114
43 Lee JJ, Kong JN, Do HD, Jo DH, Kong KH (2010) Design and efficient soluble expression of a sweet protein, brazzein and minor-form mutant. B Kor Chem Soc 31:3830-3833.   DOI
44 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   DOI
45 Lee YR, Akter S, Lee IH, Jung YJ, Park SY, Cho Y-G, Kang KK, Jung YJ (2018) Stable expression of brazzein protein, a new type of alternative sweetener in transgenic rice. J Plant Biotechnol. 45:63-70   DOI
46 Liu X, Hu Z, Maeda S, Aiuchi T, Nakaya K, Kurihara Y (1993) Purification, complete amino acid sequence and structure characterization of the heat stable sweet protein, mabinlin II. Eur J Biochem 211:281-287   DOI