Cloning of cDNA encoding the sweet-tasting plant protein thaumatin and its expression in Escherichia coli

Edens, Luppo; Heslinga, L.; Klok, Rob; Am, Ledeboer; Maat, Jaap; My, Toonen; Visser, Chris J.; Ct, Verrips

doi:10.1016/0378-1119(82)90050-6

Cited by 211 publications

(128 citation statements)

References 25 publications

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“…This result is consistent with the mass of thaumatin I (M r ) 22 188 Da) as deduced from the published sequences, assuming that all disulfide bonds are intact. 22 To further characterize the monomer fraction, we examined our samples by quasielastic light scattering (QLS), size-exclusion high performance liquid chromatography, and cation-exchange high performance liquid chromatography (SE-HPLC and CE-HPLC, respectively). Figure 2 shows QLS results for purified Natex and unpurified Sigma thaumatin.…”

Section: Source Of Proteinmentioning

confidence: 99%

“…ESIMS shows that the peak at 27 min has a molecular mass of 22,190 ( 2 Da (Natex) and 22,189 ( 2 Da (Sigma), consistent with that of thaumatin I (22 188 Da); the peak in Sigma thaumatin at 31 min corresponds to a mass of 22 272 ( 2 Da, consistent with that of thaumatin II (22 272 Da). 22 Finally, there is a heterogeneity that can be seen by eye. Sigma thaumatin is contaminated by a pigment that makes the solutions golden-yellow to reddish-brown; as the protein concentration is increased, the color darkens.…”

Section: Source Of Proteinmentioning

confidence: 99%

“…20,21 The two most abundant components are thaumatin I and thaumatin II, which differ by five amino acids. 22 The thaumatins are among the sweetest compounds knownsabout 100 000 times sweeter than sugar on a molar basis 23 sand their physical and chemical properties (in particular, those related to taste) have been studied extensively since thaumatin I and II were first isolated in 1972. 20,[24][25][26] The rise of thaumatin as a model protein began soon after a discovery by Ko et al: the addition of L-tartrate ions to a solution of thaumatin leads to the rapid formation of bipyramidal protein crystals.…”

Section: Introductionmentioning

confidence: 99%

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Effects of Protein Purity and Precipitant Stereochemistry on the Crystallization of Thaumatin

Asherie

Ginsberg

Greenbaum

et al. 2008

Crystal Growth & Design

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ABSTRACT:Thaumatin is frequently used as a model protein in crystallization studies because it rapidly forms crystals in the presence of tartrate ions. The thermodynamic and kinetic properties of thaumatin crystals have been studied for almost 10 years, and the results are contradictory. Here we show that by using a homogeneous preparation of thaumatin and controlling the stereochemistry of the tartrate precipitant, it is possible to achieve consistent results for the protein solubility. To understand the role of protein impurities in the crystallization of thaumatin, we examined two commercial sources of the protein and characterized the heterogeneities therein. To examine the effect of precipitant stereochemistry, we crystallized thaumatin with L, D, and DL (racemic) tartrate ions. We suggest that the inconsistencies among previous results stem in part from the different behavior of thaumatin with the L and D enantiomers: the solubility of thaumatin crystals increases with temperature in L-tartrate, whereas it decreases with temperature in D-tartrate. Our results demonstrate the importance of using pure protein and stereochemically pure precipitants in the crystallization of proteins.

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Section: Source Of Proteinmentioning

confidence: 99%

Section: Source Of Proteinmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effects of Protein Purity and Precipitant Stereochemistry on the Crystallization of Thaumatin

Asherie

Ginsberg

Greenbaum

et al. 2008

Crystal Growth & Design

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“…The nucleotide sequence of thaumatin I and II from cloned cDNA showed that thaumatin was translated as a prepro form with both a 22-amino acid hydrophobic N-terminal extension and an acidic 6-amino acid-long carboxyl terminal extension (Ide et al, 2007b, Edens et al, 1982. The deduced amino acid sequence of thaumatin I is different from that of thaumatin II at five sequence positions (N46K, S63R, K67R, R76Q, and N113D).…”

Section: Introductionmentioning

confidence: 99%

“…Moreover, to elucidate the mechanisms for sweetness of thaumatin, production of homogeneous recombinant thaumatin by microorganism systems would be an optimal approach. In fact, there have been numerous attempts to produce thaumatin by microorganisms (Edens et al, 1982, Daniell et al, 2000, Edens and van der Wel, 1985, Illingworth et al, 1988, Illingworth et al, 1989, Lee et al, 1988, Hahm and Batt, 1990, Faus et al, 1997. However, most of them were insufficient amino acids were obtained from Becton Dickinson.…”

Section: Introductionmentioning

confidence: 99%

High-yield Secretion of the Recombinant Sweet-Tasting Protein Thaumatin I

Masuda

Ide

Ohta

et al. 2010

FSTR

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Thaumatin, an intensely sweet-tasting protein used as a sweetener, was secreted by the methylotrophic yeast Pichia pastoris. Approximately 100 mg L -1 of recombinant thaumatin I was obtained using an expression vector which possesses three copies of the thaumatin gene containing the 22-amino acid presequence. Expression yield was about three-fold higher than when the α-factor secretion signal from Saccharomyces cerevisiae was used. The circular dichroism and tryptophan fluorescence spectra for recombinant thaumatin I were almost the same as those for plant thaumatin. Large amounts of homogeneous recombinant thaumatin allowed for preparation of high-quality crystals in the presence of cryoprotective glycerol used in high-resolution x-ray structural analysis to help further understand the perception of the sweet taste of thaumatin.

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Sweeteners

Lipinski

2000

Ullmann's Encyclopedia of Industrial Chemistry

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The article contains sections titled: 1. Introduction 2. Sensory Properties 2.1. Structural Requirements for Sweetness 2.2. Sweetness Intensity 2.3. Taste Characteristics 2.4. Synergism 3. Uses 3.1. Foods and Beverages 3.2. Table‐Top Sweeteners 3.3. Cosmetics 3.4. Pharmaceuticals 3.5. Feed 3.6. Others 4. General Toxicology and Physiology 4.1. Toxicology 4.2. Metabolism 4.3. Suitability for Diabetics 4.4. Dental Effects 5. Food Legislation 5.1. International Regulations and Assessments 5.2. National Food Legislation 6. Substances Commonly Used as Sweeteners 6.1. Acesulfame K 6.1.1. General Information 6.1.2. Physical and Chemical Properties 6.1.3. Production 6.1.4. Specifications and Analysis 6.1.5. Toxicology and Legislation 6.1.6. Uses 6.2. Aspartame 6.2.1. General Information 6.2.2. Physical and Chemical Properties 6.2.3. Production 6.2.4. Specifications and Analysis 6.2.5. Toxicology and Legislation 6.2.6. Uses 6.3. Cyclamate 6.3.1. General Information 6.3.2. Physical and Chemical Properties 6.3.3. Production 6.3.4. Specifications and Analysis 6.3.5. Toxicology and Legislation 6.3.6. Uses 6.4. Saccharin 6.4.1. General Information 6.4.2. Physical and Chemical Properties 6.4.3. Production 6.4.4. Specifications and Analysis 6.4.5. Toxicology and Legislation 6.4.6. Uses 7. Sweeteners of Lesser Importance 7.1. Glycyrrhizin 7.1.1. General Information and Properties 7.1.2. Production 7.1.3. Toxicology and Legislation 7.1.4. Uses 7.2. Neohesperidin Dihydrochalcone 7.2.1. General Information and Properties 7.2.2. Production 7.2.3. Toxicology and Legislation 7.2.4. Uses 7.3. Stevioside and Rebaudioside 7.3.1. General Information and Properties 7.3.2. Production 7.3.3. Toxicology and Legislation 7.3.4. Uses 7.4. Sucralose 7.4.1. General Information and Properties 7.4.2. Production 7.4.3. Toxicology and Legislation 7.4.4. Uses 7.5. Thaumatin 7.5.1. General Information and Properties 7.5.2. Production 7.5.3. Toxicology and Legislation 7.5.4. Uses 8. New Developments 8.1. Alitame 8.2. Others 9. Substances Formerly Used as Sweeteners 9.1. Dulcin 9.2. Others

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Cloning of cDNA encoding the sweet-tasting plant protein thaumatin and its expression in Escherichia coli

Cited by 211 publications

References 25 publications

Effects of Protein Purity and Precipitant Stereochemistry on the Crystallization of Thaumatin

Effects of Protein Purity and Precipitant Stereochemistry on the Crystallization of Thaumatin

High-yield Secretion of the Recombinant Sweet-Tasting Protein Thaumatin I

Sweeteners

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