THAUMATIN

Cas No: 53850-34-3
IUPAC Name: N-[4-(4-cycloheptylpiperazin-1-yl)sulfonylphenyl]acetamide
Molecular Weight: 379.5
Molecular Formula: C19H29N3O3S
EC Number: 258-822-2

Description:
Thaumatin (also known as talin) is a low-calorie sweetener and flavour modifier.
Thaumatin is often used primarily for thaumatin's flavour-modifying properties and not exclusively as a sweetener.
The thaumatins were first found as a mixture of proteins isolated from the katemfe fruit (Thaumatococcus daniellii) (Marantaceae) of West Africa.
Although very sweet, thaumatin's taste is markedly different from sugar's. The sweetness of thaumatin builds very slowly.
Perception lasts a long time, leaving a liquorice-like aftertaste at high concentrations.
Thaumatin is highly water soluble, stable to heating, and stable under acidic conditions.
Thaumatin is a mixture of intensely sweet proteins (thaumatins) extracted with water from the arils of the fruit of the West African perennial plant Thaumatococcus daniellii.
The thaumatins have a normal complement of amino acids, except that histidine is not present.
The molecular weights of the thaumatins are approximately 22,000 and their iso-electric points are in the range of 11.5-12.5.
There are no unusual side-chains, atypical peptide linkages, or end-groups.
Extensive disulfide cross-linking confers to thaumatin thermal stability, resistance to denaturation, and maintenance of the tertiary structure of the polypeptide chain.
The maintenance of tertiary structure is critical to thaumatin's technical function.
Cleavage of just one disulfide bridge results in a loss of sweet taste.
Thaumatin is purified by selective ultrafiltration, but small amounts of organic non-protein impurities remain in the commercial product.
These consist principally of the arabinogalactan and arabinoglucuronoxylan polysaccharides, both of which are normal constituents of plant gums and mucilages.
Thaumatin functions primarily as a flavour enhancer and as a high-intensity sweetener.
The substance was previously evaluated by the Committee at twenty-seventh meeting.
No ADI was allocated at that time, although specifications were prepared.
Thaumatin is an intensely sweet-tasting protein, 100 000 times sweeter than sucrose on a molar basis, found in berries from Thaumatococcus daniellii, a tropical flowering plant known as Katemfe.
Thaumatin is induced by attack by viroids, which are single-stranded unencapsulated RNA molecules that do not code for protein.
Thaumatin consists of about 200 residues and contains 8 disulphide bonds.
Like other PR proteins, thaumatin is predicted to have a mainly beta structure, with a high content of beta-turns and little helix.
Several stress-induced proteins of plants have been found to be related to thaumatins:
-A maize alpha-amylase/trypsin inhibitor
-Two tobacco pathogenesis-related proteins: PR-R major and minor forms, which are induced after infection with viruses
-Salt-induced protein NP24 from tomato
-Osmotin, a salt-induced protein from tobacco 
-Osmotin-like proteins OSML13, OSML15 and OSML81 from potato 
-P21, a leaf protein from soybean
-PWIR2, a leaf protein from wheat 
-Zeamatin, a maize antifungal protein 
The two major sweet-tasting proteins, thaumatin I and II (TI and TIl), were isolated by Van der WeI and his group at Unilever in 1972.
Thaumatin is marketed in the UK by Tate & Lyle pIc as Talln, although the fruit of the plant has been used for centuries by the West Africans as a source of sweetness.
Thaumatin is a natural, low-calorie sweetener isolated from the West African Katemfe fruit (Thaumatococcus daniellii Bennett).
Thaumatin is a mixture of sweet-tasting proteins.
Roughly 2000 times sweeter than table sugar with flavour modifying properties and a tendency to ‘build’ in sweetness leaving a longer-lasting perception of sweetness.
These properties allow it to be used in far smaller amounts than sugar and for it to be used to mask flavours and ‘after-tastes’ in foods and supplemental foods (like protein powders and bars).
In high doses though, it can also leave a liquorice-like aftertaste.
As a sweetener and flavour modifier, thaumatin is highly soluble, and acid- and heat-stable.
Thaumatin has been used as an added sweetener and flavour compound since the 1970s.
Thaumatin is produced from the arils of the fruit.
Thaumatococcus Daniellii grows in the wild abundantly and is available in the rain forests of West African countries like:
Ivory Coast, Ghana, Nigeria, Senegal, Liberia and Central African countries like Cameroon, Gabon etc.
Studies are on to develop it through stem cell technology also. The fruits are available almost 9-10 months in a year and are in abundance and more.
The sweet protein Thaumatin is found in the gel-like aril of the fruit.
About 2-3% Thaumatin can be extracted from the Arils (depending on the factors like maturity of the fruit, area/region from which it is harvested etc).
Cut fruit of Thaumatococcus Daniellii showing white gel-like arils sitting on top of black seeds.
As a food ingredient, thaumatin is considered to be safe for consumption.
In a Swiss chewing gum production plant, thaumatin has been identified as an allergen.
Thaumatin powder led to allergic symptoms in the upper airways in occupationally exposed individuals.
After the factory replaced powdered thaumatin with a liquid form, all of the affected individuals became completely free of symptoms.
Thaumatin interacts with human TAS1R3 receptor to produce a sweet taste. The interacting residues are specific to old world monkeys and apes (including humans); only these animals can perceive it as sweet.
The gel-like aril contains thaumatin.

Chemical Properties of Thaumatin:
Thaumatin is a mixture of sweet proteins occurring in the arils of the fruits of the African plant Thaumatococcus daniellii.
Thaumatins I and II are the main components, but four more thaumatin molecules are known. 
The thaumatins may be extracted with water.
Thaumatin is about 2,000–2,500 times sweeter than sucrose but has a lingering sweetness.
In addition to thaumatin's sweet taste, it has flavor-enhancing properties.
Thaumatin is freely soluble in water and of fairly good stability.
In Europe, thaumatin is approved as E 957 for use as a sweetener.
Thaumatin is also approved in a variety of other countries, but in the United States, GRAS as a flavor enhancer only.
Thaumatin occurs as a pale-brown colored, odorless, hygroscopic powder with an intensely sweet taste.
Thaumatin is odorless but has an intensly sweet taste.
Biological Role of Thaumatin:
Thaumatin production is induced in katemfe in response to an attack upon the plant by viroid pathogens.
Several members of the thaumatin protein family display significant in vitro inhibition of hyphal growth and sporulation by various fungi.
The thaumatin protein is considered a prototype for a pathogen-response protein domain.
This thaumatin domain has been found in species as diverse as rice and Caenorhabditis elegans.
Thaumatins are pathogenesis-related (PR) proteins, which are induced by various agents ranging from ethylene to pathogens themselves, and are structurally diverse and ubiquitous in plants.
They include thaumatin, osmotin, tobacco major and minor PR proteins, alpha-amylase/trypsin inhibitor, and P21 and PWIR2 soybean and wheat leaf proteins.
The proteins are involved in systematically-acquired stress resistance and stress responses in plants, although their precise role is unknown.
Thaumatin is an intensely sweet-tasting protein (on a molar basis about 100,000 times as sweet as sucrose) found in the fruit of the West African plant:
Thaumatococcus daniellii: it is induced by attack by viroids, which are single-stranded unencapsulated RNA molecules that do not code for protein.
The thaumatin protein I consists of a single polypeptide chain of 207 residues.
Like other PR proteins, thaumatin is predicted to have a mainly beta structure, with a high content of beta-turns and little helix.
Tobacco cells exposed to gradually increased salt concentrations develop a greatly increased tolerance to salt, due to the expression of osmotin, a member of the PR protein family.
Wheat plants attacked by barley powdery mildew express a PR protein (PWIR2), which results in resistance against that infection.
The similarity between this PR protein and other PR proteins and the maize alpha-amylase/trypsin inhibitor has suggested that PR proteins may act as some form of inhibitor.
Within West Africa, the katemfe fruit has been locally cultivated and used to flavour foods and beverages for some time.
The fruit's seeds are encased in a membranous sac, or aril, that is the source of thaumatin. In the 1970s, Tate and Lyle began extracting thaumatin from the fruit.
In 1990, researchers at Unilever reported the isolation and sequencing of the two principal proteins found in thaumatin, which they dubbed thaumatin I and thaumatin II.
These researchers were also able to express thaumatin in genetically engineered bacteria.
Thaumatin has been approved as a sweetener in the European Union (E957), Israel, and Japan.
In the United States, it is generally recognized as safe as a flavouring agent (FEMA GRAS 3732) but not as a sweetener.
Since thaumatin crystallizes very quickly and easily in the presence of tartrate ions, thaumatin-tartrate mixtures are frequently used as model systems to study protein crystallization.
The solubility of thaumatin, thaumatin's crystal habit, and mechanism of crystal formation are dependent upon the chirality of precipitant used.
When crystallized with L- tartrate, thaumatin forms bipyramidal crystals and displays a solubility that increases with temperature; with D- and meso-tartrate.
Thaumatin forms stubby and prismatic crystals and displays a solubility that decreases with temperature.
This suggests control of precipitant chirality may be an important factor in protein crystallization in general.
Thaumatin is remarkably stable considering thaumatin's proteinaceous nature.
Thaumatin is successfully applied in canning of pet food where it remains stable at temperatures of 120ºC and in coatings subjected to dry temperatures of 140ºC.
Thaumatin is stable under pasteurisation and UHT conditions.
Further enhancement of its physical and chemical stability can be achieved through reformulation with protective compounds.
Thaumatin's stability under acid conditions, to lower than pH 2, is a useful feature - a result of its molecular structure.
Thaumatin, extracted unmodified from the Katemfe fruit using purely physical extraction methods consists of very closely related Thaumatin proteins.
These have a single polypeptide chain or 207 “normal” amino acid residues linked with 8 disulphide bridges giving a molecular weight of around 22,000.
Thaumatin is completely digested by man and animal, which together with its “normal” amino acid sequence, accounts for its acceptance by regulatory authorities around the world as a safe, natural substance.
Thaumatin has the normal calorific value of a protein (4.1 Kcal/g) but in use is essentially non-calorific due to the very low levels required (parts per million).
Thaumatin is very basic with an iso-electric point of around 11.5 and freely soluble in water.
Thaumatin has a demonstrable effect on various groups of taste receptors, in various species of animal, for which it is used in feeding stuffs.
As well as in man and is assumed to operate with the majority of taste receptors.
This feature is a result of its particular structure.
In the analogy, often used, of flavour molecules being “keys” and the taste receptors being “locks”, thaumatin appears to act as a “master key” in its action with all taste receptors.
Thaumatin’s effect in operating with all the taste receptors is the major reason for its wide range of applicability in situations requiring flavour modifications.
The electrical charge distribution on the thaumatin molecule is believed to be a major factor in the mechanism of interaction with both the taste receptor and the flavour molecules.
This is demonstrated by the specific masking effects on the typically bitter unpleasant taste of metallic ions such as sodium, iron, potassium and its simultaneous enhancement of other free ion, e.g. chloride.
Thaumatin's effect with more complex flavour compounds support the view that thaumatin’s mechanism operates according to the relative nature of the individual molecules present. This is particularly evident in its effect with flavours such as peppermint, ginger, cinnamon and coffee, which it enhances while simultaneously reducing the fiery, peppery or bitter elements associated with them.
Thaumatin’s high and rapid solubility in water and its tendency to increase salivation are additional physical features that increase the effect of flavours and aromas.
The strength of the interaction between the thaumatin molecule and the taste receptors is believed to account not only for its potency effects in prolonging the effects of flavours and sweeteners.

Masking, Synergism, Enhancement of Thaumatin
Thaumatin’s masking effects are an important feature accounting for it’s widespread use in the human and animal food industry and with high intensity sweeteners and in particular with saccharin.
Thaumatin is particularly effective in masking the metallic aftertaste of saccharin, with this combination being used in a wide variety of products from tabletop sweeteners to animal feeds.
In citrus fruit products, thaumatin has been shown to be very effective in masking the bitter elements of natural flavours from the fruit hence its application in products such as juices, yoghurts and deserts.
A good example of thaumatin’s simultaneous action is to be found in applications with vitamin C, often flavoured with citrus and sweetened with high intensity sweeteners.
In this application thaumatin also masks the bitter notes and the sweetener aftertaste, enhances the flavour and also contributes to the sweetening.
At high levels of inclusion but still in the parts per million ranges, thaumatin is effective in masking the taste of some pharmaceutical products in chewables and suspensions.  
A clear synergism is shown when using thaumatin in conjunction with intensity sweeteners and with flavour enhancing compounds MSG and 5’ nucleotides.
With aspartame for example, a commercial application showed sufficient synergy between thaumatin and aspartame that a thaumatin inclusion of 10 parts per million.
The aspartame could be reduced by more than 30% to achieve the original sweetness level.
In addition to the synergistic effects thaumatin improved the sweetness profiles of non nutritive sweeteners giving “body” to the sweeteners and masking the associated aftertaste.
As well as extending the sweetness and flavour profiles.
Thaumatin as the most potent form of sweetness available, and being the only natural high intensity sweetener available, does have some application as a sweetener in its own right.
Due to a liquorice aftertaste being contributed at higher levels, its use as the only source of sweetness is limited to applications where the sweetness requirement is less than the equivalent of 10% of sucrose.
Above this level a taste contribution from thaumatin can become evident, which although desirable in some products, is not generally acceptable.
Applications in conjunction with natural low intensity, non-nutritive sweeteners are of interests in products requiring an “all natural” label.
Thaumatin’s application in products in conjunction with added MSG and nucleotide compounds, or where nucleotides are naturally present, exhibit a high degree of synergy with these compounds.
The additional masking effects of metallic flavours provide a range of applications in low sodium products where NaCl has been replaced in part with KCl.
Thaumatin is effective in enhancing a wide range of flavour compounds, including savoury flavours and, in particular, “aggressive” flavours.
Due to its ability to reduce the aggressive nature of these complex flavour compounds.
Thaumatin provides the opportunity to achieve higher levels of flavour in products without these being too aggressive and for instance, in coffee flavoured products, without generating bitter notes.
In savoury products enhancement is more complex. In addition to enhancement of flavours directly.
Thaumatin synergises with naturally present or added flavour enhancers such as 5’ nucleotides and monosodium glutamate (MSG).
This adds a unique element of what the Japanese industry describes as “Umami” or, approximately translated into English, ”deliciousness”.

Uses of Thaumatic:
Potential low-calorie sweetener.
Thaumatin is a flavor enhancer that is a protein which is approxi- mately 3,000 times as sweet as sucrose. the onset of sweetness may take several seconds and can be affected by heat.
Thaumatin is used in chewing gum.
As a sweetener, thaumatin is used in beverages and desserts, but its applications are limited because of its liquorice taste and delayed sweetness.
In practice, therefore, thaumatin is more commonly used as a partial sweetener, mixed with other more rapidly tasting sweeteners.
Despite its limitations as a sweetener, thaumatin is a powerful flavour enhancer, and magnifies spearmint, cinnamon, wintergreen and peppermint by up to ten times.
This flavour potentiating characteristic can be beneficially used for products such as toothpaste, mouthwash and chewing gum, and for enhancing the masking flavours in medicines.
Thaumatin also boosts the low sweetness of bulk sweeteners added to sugarless gums, without adding calories or cariogenicity.
Thaumatin has been used in Japan since 1979 in a variety of products, where it has been shown to enhance and improve the flavour of coffee and of milk products.
Thaumatin is thus used in coffee-flavoured products, ice-cream, iced milk drinks-on-sticks, and spray-dried milk powders.
Thaumatin also enhances savoury flavours and combinations of thaumatin with nucleotides, spices and/or other flavours may be used to replace monosodium glutamate, an ingredient of current concern with regard to safety.
In addition to its use as a sweetener, thaumatin has become an important protein for basic science research.
We describe below three aspects of thaumatin that are being explored: crystallisation, purity and taste.
In 1994, it was discovered that adding the molecule l-tartrate to a solution of thaumatin causes the protein to crystallise rapidly.
Bipyramidal crystals, shown in the photograph below, form in as little as twenty minutes. 
This is in contrast to most proteins, which takes days, weeks or months to crystallize—if they crystallize at all.
 
Chemical and Physical Properties:
FEMA Number: 3732
Physical State (20 deg.C): Solid
Store Under Inert Gas: Store under inert gas
Condition to Avoid: Hygroscopic
PubChem Substance ID: 87576932
MDL Number: MFCD00132420
InChI Key: IVPOQXWRUMMORK-UHFFFAOYSA-N
XLogP3-AA: 2.4    
Hydrogen Bond Donor Count    : 1    
Hydrogen Bond Acceptor Count: 5    
Rotatable Bond Count: 4    
Exact Mass: 379.19296297    
Monoisotopic Mass: 379.19296297    
Topological Polar Surface Area: 78.1 Ų    
Heavy Atom Count: 26
Formal Charge: 0    
Complexity: 551    
Isotope Atom Count: 0    
Defined Atom Stereocenter Count: 0    
Undefined Atom Stereocenter Count: 0    
Defined Bond Stereocenter Count: 0    
Undefined Bond Stereocenter Count: 0    
Covalently-Bonded Unit Count: 1    
Compound Is Canonicalized: Yes

Synonyms:
Thaumatin B, recombinant
N-[4-(4-cycloheptylpiperazin-1-yl)sulfonylphenyl]acetamide
53850-34-3
Oprea1_565606
Oprea1_669084
CHEBI:175005
STK171150
ZINC54991825
AKOS001637661
MCULE-3623866651
ST45109962
N-[4-(4-cycloheptylpiperazin-1-yl)sulonylphenyl]acetamide
N-[4-(4-cycloheptylpiperazine-1-sulfonyl)phenyl]acetamide
N-{4-[(4-cycloheptylpiperazinyl)sulfonyl]phenyl}acetamide
N-{4-[(4-cycloheptylpiperazin-1-yl)sulfonyl]phenyl}acetamide