WO2004039174A2 - Procede - Google Patents

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Publication number
WO2004039174A2
WO2004039174A2 PCT/IB2003/005278 IB0305278W WO2004039174A2 WO 2004039174 A2 WO2004039174 A2 WO 2004039174A2 IB 0305278 W IB0305278 W IB 0305278W WO 2004039174 A2 WO2004039174 A2 WO 2004039174A2
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WO
WIPO (PCT)
Prior art keywords
foodstuff
acrylamide
enzyme
formation
process according
Prior art date
Application number
PCT/IB2003/005278
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English (en)
Other versions
WO2004039174A3 (fr
Inventor
Jørn Borch SØE
Charlotte Horsmans Poulsen
Dana L Boll
Original Assignee
Danisco A/S
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from GB0225236A external-priority patent/GB0225236D0/en
Application filed by Danisco A/S filed Critical Danisco A/S
Priority to AU2003276613A priority Critical patent/AU2003276613A1/en
Publication of WO2004039174A2 publication Critical patent/WO2004039174A2/fr
Publication of WO2004039174A3 publication Critical patent/WO2004039174A3/fr
Priority to US11/048,230 priority patent/US8163317B2/en
Priority to US13/433,470 priority patent/US8956670B2/en
Priority to US14/619,149 priority patent/US20150223499A1/en

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
    • C12Y101/03—Oxidoreductases acting on the CH-OH group of donors (1.1) with a oxygen as acceptor (1.1.3)
    • C12Y101/03004—Glucose oxidase (1.1.3.4)
    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
    • A21D8/00—Methods for preparing or baking dough
    • A21D8/02—Methods for preparing dough; Treating dough prior to baking
    • A21D8/04—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes
    • A21D8/042—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes with enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C19/00—Cheese; Cheese preparations; Making thereof
    • A23C19/06—Treating cheese curd after whey separation; Products obtained thereby
    • A23C19/063—Addition of, or treatment with, enzymes or cell-free extracts of microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
    • A23L19/10—Products from fruits or vegetables; Preparation or treatment thereof of tuberous or like starch containing root crops
    • A23L19/12—Products from fruits or vegetables; Preparation or treatment thereof of tuberous or like starch containing root crops of potatoes
    • A23L19/18—Roasted or fried products, e.g. snacks or chips
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
    • A23L5/25—Removal of unwanted matter, e.g. deodorisation or detoxification using enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
    • C12Y101/03—Oxidoreductases acting on the CH-OH group of donors (1.1) with a oxygen as acceptor (1.1.3)
    • C12Y101/03005—Hexose oxidase (1.1.3.5)

Definitions

  • the present invention relates to the control of the formation of acrylamide in a foodstuff.
  • Acrylamide and polyacrylamide are used in industry for the production of plastics. It has been supposed that the main exposure for acrylamide in the general population has been through drinking water and tobacco smoking. Exposure via drinking water is small and the EU has determined maximum levels of 0.1 microgram per litre water.
  • Acrylamide is water soluble and is quickly absorbed in the digestive tract. Excretion via the urine is fast and half of acrylamide is cleared from the body in a few hours.
  • acrylamide The toxicological effects of acrylamide are well known. It causes DNA damage and at high doses neurological and reproductive effects have been observed. Glycidamide, a metabolite of acrylamide, binds to DNA and can cause genetic damage. Prolonged exposure has induced tumours in rats, but cancer in man has not been convincingly shown.
  • IARC International Agency for Research on Cancer
  • acrylamide a "probably carcinogenic to humans" (Group 2A).
  • Acrylamide has been shown to induce gene mutations in cultured animal cells and also in animals treated in vivo. Thus it is assumed that exposure also to very low doses of acrylamide increases the risk for mutation and cancer.
  • tumours were most evident in specific organs, e.g. mammary gland, uterus, adrenal gland, scrotal mesothelium. In mice there was an increase of lung and skin tumours. These cancer studies have been used for the assessment of the risk of cancer in humans due to acrylamide exposure.
  • the levels of acrylamide vary considerably between single foodstuffs within food groups, but potato crisps and French fries generally contained high levels compared to many other food groups.
  • the average content in potato crisps is approximately 1000 microgram/kg and in French fries approximately 500 microgram/kg.
  • Other food groups which may contain low as well as high levels of acrylamide are crisp bread, breakfast cereals, fried potato products, biscuits, cookies and snacks, e.g. popcorn.
  • Foodstuffs which are not fried, deep fried or oven-baked during production or preparation are not considered to contain any appreciable levels of acrylamide. No levels could be detected in any of the raw foodstuffs or foods cooked by boiling investigated so far (potato, rice, pasta, flour and bacon).
  • the background level has been estimated to account for a daily intake corresponding to approximately 100 microgram per day.
  • the present invention alleviates the problems of the prior art.
  • the present invention provides a process for the prevention and/or reduction of acrylamide formation and/or acrylamide precursor formation in a foodstuff containing (i) a protein, a peptide or an amino acid and (ii) a reducing sugar, the process comprising contacting the foodstuff with an enzyme capable of oxidising a reducing group of the sugar.
  • the present invention provides use of an enzyme for the prevention and/or reduction of acrylamide formation and/or acrylamide precursor formation in a foodstuff containing (i) a protein, a peptide or an amino acid and (ii) a reducing sugar, wherein the enzyme is capable of oxidising a reducing group of the sugar.
  • Acrylamide formation and/or acrylamide precursor formation in cooked foodstuffs in particular starch foodstuffs and foodstuffs containing a protein/amino acid/peptide and reducing sugar is described in Appendices 1 and 2, for example by the Amadori reaction, and is known in the art.
  • a sugar such as glucose, galactose and/or maltose may react with an amino acid such as asparagine, glutamic acid, lysine, or arginine.
  • Any primary amine capable of nucleophilic attack on the carbonyl group of a reducing sugar may be involved This reaction may be an important step in the formation of acrylamide.
  • the present invention prevents and/or reduces the problematic condensation reactions between amino acids, in particular the amino group thereof, and reducing sugars which result in acrylamide or acrylamide precursor formation. These reactions may comprise the Amadori reaction, Heynes rearrangements, or reaction cascades resulting from the Maillard reaction.
  • the present invention may prevent and/or reduce the reaction which directly results in acrylamide formation. It may also prevent and/or reduce reaction(s) which provide materials which further react to provide acrylamide, namely acrylamide precursors.
  • Acrylamide precursors are often provided by degradation of carbohydrates. A typical acrylamide precursor is 2-propenal.
  • the problems of the formation of acrylamide and/or acrylamide precursor formation in foodstuffs containing a protein and a reducing sugar such as baked food products, in particular formation caused either completely or in part by the Amadori reaction, can be controlled by contacting the foodstuff with an enzyme capable of oxidising the reducing group of the sugar.
  • This is a novel approach in which reducing sugar is oxidised to avoid acrylamide formation and/or acrylamide precursor formation by bringing the foodstuff into contact with an enzyme which is capable of performing the necessary oxidation and thereby eliminating the reducing sugar from the foodstuff by conversion.
  • prevention and/or reduction of acrylamide formation it is meant that the amount of acrylamide produced is reduced and/or the period of time required for formation of a given amount of acrylamide is increased.
  • the process prevents and/or reduces Amadori reaction in a foodstuff.
  • the present invention provides a process for the prevention and/or reduction of Amadori reaction in a foodstuff containing (i) a protein, a peptide or an amino acid and (ii) a reducing sugar, the process comprising contacting the foodstuff with an enzyme capable of oxidising a reducing group of the sugar.
  • the present invention provides use of an enzyme for the prevention and/or reduction of Amadori reaction in a foodstuff containing (i) a protein, a peptide or an amino acid and (ii) a reducing sugar, wherein the enzyme is capable of oxidising a reducing group of the sugar.
  • prevention and/or reduction of Amadori reaction it is meant that the extent of a Amadori reaction is reduced and/or the period of time required for completion of a Amadori reaction is increased.
  • the enzyme is capable of oxidising the reducing group of a monosaccharide and the reducing group of a disaccharide.
  • the enzyme is hexose oxidase (EC1.1.3.5) or glucose oxidase (EC1.1.3.4). In a highly preferred aspect the enzyme is hexose oxidase.
  • the HOX is obtained or prepared in accordance with WO 96/40935.
  • the HOX is DairyHOXTM available from Danisco A/S, Denmark.
  • the enzyme may oxidise matlodextrins and/or celludextrins.
  • the enzyme is a carbohydrate oxidase which may oxidise matlodextrins and/or celludextrins.
  • the carbohydrate oxidase is obtained or prepared in accordance with WO 99/31990.
  • Hexose oxidase is a carbohydrate oxidase originally obtained from the red alga Chondrus crispus. As discussed in WO 96/39851 HOX catalyses the reaction between oxygen and carbohydrates such as glucose, galactose, lactose and maltose. Compared with other oxidative enzymes such as glucose oxidase, hexose oxidase not only catalyse the oxidation of monosaccharides but also disaccharides are oxidised. (Biochemica et Biophysica Acta 309 (1973), 11 -22).
  • gluconolactone In an aqueous environment the gluconolactone is subsequently hydrolysed to form gluconic acid.
  • HOX oxidises the carbohydrate at the reducing end at carbon 1 and thus eliminates the possible involvement of the carbohydrate in acrylamide formation and/or acrylamide precursor formation by Amadori rearrangement or later reaction with a ketoseamine or aldoseamine to a diketoseamine or a diaminosugar respectively.
  • the enzyme is capable of oxidising the sugar of the foodstuff at the 1 position.
  • This aspect is advantageous because it ensures that the reducing sugar is oxidised such that the reducing part of the sugar is no longer available to undergo a condensation reaction with an amino acid such the Amadori reaction.
  • the reducing sugar is selected from lactose, galactose, glucose, xylose, mannose, cellobiose and maltose.
  • the reducing sugar is lactose or galactose.
  • the reducing sugar is galactose.
  • the foodstuff is selected from bakery goods including bread and cakes, pasta, rice, fish, sausages, meat including beef and pork, biscuits, cookies, crisp bread, cereals, pizza, beverages including coffee, and products based on potatoes, maize and flour, including potato flour and potato starch products.
  • the foodstuff is a beverage.
  • the foodstuff is a starch containing foodstuff.
  • the foodstuff is a cereal or part of a cereal.
  • the foodstuff is selected from a dairy foodstuff; milk based or milk containing foodstuff, such as gratin; an egg based foodstuff; an egg containing foodstuff; bakery foodstuffs including toasts, bread, cakes; and shallow or deep fried foodstuff such as spring rolls.
  • the foodstuff is a dairy foodstuff it may be cheese, such as mozzarella cheese.
  • the foodstuff is a potato or a part of a potato.
  • Typical potato products in which the present invention may be applied are French fries, potato chips (crisps), coated French fries and coated potato chips, for example French fries or potato chips coated with corn starch, and potato flour and potato starch products.
  • the enzyme may be contacted with foodstuff during its preparation or it may be contacted with the foodstuff after the foodstuff has been prepared yet before the food stuff is subjected to conditions which may result in the undesirable acrylamide formation and/or acrylamide precursor formation.
  • the enzyme will be incorporated in the foodstuff.
  • the enzyme will be present on the surface of the foodstuff. When present on the surface acrylamide formation and/or acrylamide precursor formation is still prevented as it is the surface of a material exposed to drying and atmospheric oxygen which undergoes the predominant acrylamide formation and/or acrylamide precursor formation.
  • the enzyme When contacted with foodstuff during its preparation the enzyme may be contacted at any suitable stage during its production.
  • the foodstuff is a dairy product it may be contacted with the milk during acidification of the milk and precipitation of the milk curd.
  • the enzyme (such as HOX) is not active during the anaerobic conditions created during the acidification and milk protein precipitation, but will be active in the dairy product such as cheese when aerobic conditions are created.
  • the enzyme oxidise the reducing sugar and reduce the tendency to acrylamide formation and/or acrylamide precursor formation.
  • the enzyme for application of the enzyme to the surface of the foodstuff, one may apply the enzyme in any suitable manner.
  • the enzyme is provided in a solution or dispersion and sprayed on the foodstuff.
  • the solution/dispersion may comprise the enzyme in an amount of 1-50 units enzyme/ml, such as 1-50 units Hexose Oxidase/ml.
  • the enzyme may also be added in dry or powder form. When in wet or dry form the enzyme may be combined with other components for contact with the foodstuff. For example when the enzyme is in dry form it may be combined with an anticaking agent.
  • Typical amounts of enzyme which may be contacted with the foodstuff are from 0.05 to 50 U/g (units of enzyme per gram of foodstuff), from 0.05 to 10 U/g, from 0.05 to 5 U/g, from 0.05 to 3 U/g, from 0.05 to 2 U/g, from 0.1 to 2 U/g, from 0.1 to 1.5 U/g, and from 0.5 to 1.5 U/g,
  • the use/process of the present invention further comprises use of a catalase or contacting a catalase with a foodstuff to remove oxygen and thereby prevent and/or reduce acrylamide formation and/or acrylamide precursor formation (such as 2-propenal formation).
  • the foodstuff contains an amino acid.
  • the amino acid is asparagine. It has been identified that asparagine is particularly important in the formation of acrylamide in foodstuffs.
  • the enzyme prevents and/or inhibits Amadori reactions and subsequent reactions with asparagine resulting in the formation of acrylamide.
  • the foodstuff contains a protein. In some aspects the foodstuff contains a peptide.
  • Acrylamide formation and/or acrylamide precursor formation in a foodstuff may take place during the heating thereof or may take place during storage of the foodstuff.
  • acrylamide formation and/or acrylamide precursor formation can happen upon storage of any kind of seeds without heating.
  • the enzyme of the present invention such as HOX, may still be useful however in removing a second mole of aldose or ketose sugar which may react with the already formed Amadori product to yield the diketoseamine or diaminosugar.
  • system of the present invention may prevent loss of the nutritionally important Lysine in foods.
  • reducing sugars may play an important role in the initiation of Amadori and Maillard reactions at certain moisture levels of the foodstuff (8-12%), but that lipid auto-oxidation, which is also known to initiate Amadori reactions, becomes increasingly common at low moisture levels (6%) (McDonald 1999). Lipid oxidation may actually be the primary cause for the initiation of Amadori or Maillard reactions when reducing sugars are absent.
  • the present enzyme such as HOX, may serve the dual purpose of removing both reducing sugars and oxygen and thereby preventing lipid oxidation as well as sugar hydrolysis at all moisture levels.
  • the samples in accordance with the present invention have a lower content of acrylamide than the control samples.
  • 75g shortening (mp. 35°C) and 100 g flour are heated in a pot during mixing.
  • 350 ml skim milk (preheated to 90°C) is added during continued mixing.
  • Salt and pepper is added.
  • 4 eggs are divided into yolk and egg white.
  • the egg yolks are added individually.
  • the egg white is whipped to a foam with 10 gram baking powder and mixed carefully into the dough.
  • the dough is placed in 2 aluminium trays. One of the trays is sprayed with a solution of hexose oxidase 7.5 Units/ml and kept at room temperature for 30 minutes.
  • the gratin is then baked in a air circulating oven at 175°C for 20 minutes. After baking the gratin is evaluated.
  • fried potato as French fries (pommes frites) and potato chips (crisps) has increased significantly during the past two decades.
  • One of the important parameters in the production of fried potatoes is level of reducing sugar. The level should remain low, because high level of reducing sugar contribute to higher levels of acrylamide.
  • potatoes are often sprayed with a herbicide called chlorpropham, which prevents the potato from sprouting. Sprouting induces amylases in the potato which in turn form reducing sugars.
  • Organic grown potatoes are used in order to ensure that no herbicides has been used.
  • the potatoes are peeled and sliced into 2 mm thick slices using a food processor.
  • Half of the slices are immersed in a water solution of HOX containing 100 Units/ml for 3 minutes.
  • the other half of the potato slices are immersed in water for 3 minutes.
  • the slices are then stored in a closed container for over night (16 hours) and then fried in vegetable oil for 2 minutes at 180 °C.
  • the samples in accordance with the present invention have a lower content of acrylamide than the control samples.
  • 1 glucose oxidase (GOX) unit corresponds to the amount of enzyme which under the specified conditions results in the conversion of 1 ⁇ mole glucose per minute, with resultant generation of 1 ⁇ mole of hydrogen peroxide (H 2 O 2 )
  • 1 hexose oxidase (HOX) unit corresponds to the amount of enzyme which under the specified conditions results in the conversion of 1 ⁇ mole of glucose per minute, with resultant generation of 1 ⁇ mole of hydrogen peroxide (H 2 O 2 )
  • the commonly used horse radish peroxidase dye substrate ABTS was incorporated into an assay, measuring the production of H 2 O 2 produced by HOX or GOX respectively.
  • ABTS serves as a chromogenic substrate for peroxidase.
  • Peroxidase in combination with H 2 O 2 facilitates the electron transport from the chromogenic dye, which is oxidised to an intensely green/blue compound.
  • An assay mixture contained 266 ⁇ l ⁇ -D-glucose (Sigma P-5504, 0.055 M in 0.1 M sodium phosphate buffer, pH 6.3), 11.6 ⁇ l 2,2'-Azino-bis(3-ethylbenzothiozoline-6-Sulfonic acid)(ABTS)(Sigma A-9941 , 5 mg/ml aqueous solution), 11.6 ⁇ l peroxidase (POD)(Sigma P-6782, 0.1 mg/ml in 0.1 M sodium phosphate buffer, pH 6.3) and 10 ⁇ l enzyme (HOX or GOX) aqueous solution.
  • the incubation was started by the addition of glucose at 25° C.
  • the absorbance was monitored at 405 nm in an ELISA reader.
  • a standard curve, based on varying concentrations of H 2 O 2 was used for calculation of enzyme activity according to the definition above.
  • Reaction (1) is catalysed by enzyme (HOX or GOX)
  • Reaction (2) is catalysed by enzyme (POD)
  • Example 6 Use of Hexose Oxidase and Glucose Oxidase to Reduce the Amount of Acrylamide Developed by Frying Potato Chips. Frying
  • Italian potatoes of the sort Nicola were peeled and sliced into pieces of approximately (3 mm x 30 mm x 40 mm) . Portions of approx. 30 g of sliced potatoes were treated with 40 mL of one of the incubation solutions as described below. During treatment it was made sure that all potatoes were covered with solution and the incubating beakers were stirred at RT for 4 hours in total.
  • Portions of app 50 g were treated with 100 mL of incubation solution and incubated for 15 min, while stirring at RT. During treatment it was made sure that all potatoes were covered with solution.
  • the potato slices where air dried for approx. 30 min and baked in a pre-heated oven for 30 min at 175° C.
  • the baking plate was divided into 9 segments of equal size. Potatoes treated as in (1)-(3) (see below), were divided into 9 equal fractions and 1 fraction from each was placed in each segment to a total of 3 fractions per segment. This was done to minimize the chance of faulty results as a consequence of uneven heating in the oven. Subsequently the potatoes were spread on tissue paper and allowed to cool for approx. 30 min. They were stored dark in closed containers at -20° C.
  • Oasis MAX (6cc, 150 mg, Part No. 186000370)
  • Oasis MCX (6cc, 150mg, Part No.
  • the HPLC system consisted of a quaternary pump (G1311A), autosampler (G1313A), column compartment (G1316A) all from Agilent Technologies (Waldbronn, Germany).
  • the flow rate was 0.20 ml/min.
  • Calibration standards (acrylamide) were prepared with the following concentrations: 500, 150, 50, 15, 5 ng/ml in water.
  • concentration of internal standard (acrylamide-1 ,2,3- 13 C 3 ) was maintained at 40 ng/ml.
  • the sample to be analysed was coarsely ground with a knife.
  • An aliquot (1 g) was homogenized (Ultra-Turrax T25) with 15 ml of internal standard, (ISTD, 1000 ng acrylamide 1 ,2,3- 13 C 3 /15 ml H 2 O) in a 100 ml beaker.
  • Oasis MAX cartridge and an Oasis MCX cartridge were each conditioned with 5 ml methanol followed by 2*5 ml water. After conditioning, they were combined in series with Oasis MAX on top.
  • acrylamide has been reported to be present in plant material (potatoes, carrots, radish, lettuce,
  • acrolein is a very probable precursor of acrylamide.
  • Simple, fundamental chemical transformations (such as reaction with ammonia liberated from amino acids) can then convert acrolein (or a derivative from it) into acrylamide.
  • the production of acrylamide through the reaction of acrolein with ammonia has been demonstrated in model systems (22).
  • glycerol When oil is heated at temperatures above the smoke point, glycerol is degraded to acrolein, the unpleasant acrid black and irritating smoke (26-29).
  • the formation of acrolein is known to increase with the increase in unsaturation in the oil and to lead to a lowering of the smoke point.
  • the smoke point is higher for oils with higher content of saturated fatty acids and lower content of polyunsaturated acids.
  • the smoke points for some of the main oils and fats are as follows: palm 240° C, peanut 220° C, olive: 210° C, lard and copra 180° C, sunflower and soybean 170° C, corn 160° C, margarine 150° C, and butter 110° C.
  • acrolein can also be produced as a result of oxidation of polyunsaturated fatty acids and their degradation products (31-34).
  • a number of aldehydic products including malondialdehyde, C3-C10 straight chain aldehydes, and ⁇ , ⁇ -unsaturated aldehydes, such as 4-hydroxynonenal and acrolein) are known to form as secondary oxidation products of lipids (35).
  • Acrolein was also found to form in vivo by the metal- catalyzed oxidation of polyunsaturated fatty acids including arachidonic acid (36).
  • acrolein Several sources for the formation of acrolein are known. It may arise from degradation of amino acids and proteins (37, 38), from degradation of carbohydrates (39), and from the Maillard reaction between amino acids or proteins and carbohydrates (40, 41). Many possible routes for the formation of this three-carbon aldehyde - taking the starting point from many different sugars or amino acids - may be proposed. Its formation from methionine by the Strecker degradation in the frame of the Maillard reaction is one example. Alanine, with its tree-carbon skeleton, has also been suggested as a possible source. However, fission reactions of longer carbon chains are common and well-known, so at present there is no basis to give priority to any specific reaction routes.
  • acrolein can be converted into acrylamide by a series of fundamental reactions.
  • both acrolein and acrylamide are reactive, because of their double bonds and the amino group of acrylamide. They can readily react further with other reactive groups present in the food matrix or formed during the heating process.
  • acrylamide can react with small reactive molecules, such as urea (CO(NH2) 2 ) and formaldehyde (HCHO), or with glyoxal ((CHO)2), aldehydes (RCHO), amines (R2NH), thiols (RSH) etc.
  • the products shown in the following scheme can even react further in the same mode of reaction.
  • reactive functional groups may also be found in macromolecules, such as proteins, for instance.
  • macromolecules such as proteins, for instance.
  • the presence or absence of reactive groups (or its concentration) in the food matrix may thus be one explanation of differences in final acrylamide content in different food systems.
  • the resulting acrylamide level may be due to a balance between formation and further reactions.
  • the low acrylamide levels in heated meat products could, for instance, depend on adduct formation between acrylamide (or acrolein) and proteins.
  • Acrolein may be formed from the glycerol part of triglycerides or through oxidation of fatty acids. This means that factors favouring lipid hydrolysis as well as factors favouring lipid oxidation would promote acrolein formation. Temperature is an important factor for both these reactions. Regarding hydrolysis, pH may also be of importance and high as well as low pH may be supposed to favour acrolein formation. Regarding oxidation, lipid composition is of key importance; the higher the degree of unsaturation, the lower the stability. Protection against oxygen and light will limit the oxidation and prooxidants , such as metals, should be avoided. The protective effect of antioxidants should also be taken into account.
  • the Maillard reaction has been proposed as a route for acrolein formation. Also the direct formation of acrylamide through amino acid transformations has been proposed. These amino acid transformations also involve reactions common in the Maillard reaction system.
  • the Maillard reaction is one of the most important chemical reactions in food processing, with influence on several aspects of food quality. Flavour, colour and nutritional value may be affected and certain reaction products have been noticed to be antioxidative, antimicrobial, genotoxic etc.
  • the practical applications of Maillard chemistry in food processing are, therefore, a matter of balance between favourable and unfavourable effects, and the aim of the food manufacturer is to find an optimum in this balance. This may be accomplished by influencing the main variables affecting the MR (42).
  • the Maillard reaction takes place in 3 major stages and is dependent upon factors, such as concentrations of reactants and reactant type, pH, time, temperature, and water activity. Free radicals and antioxidants are also involved (43).
  • the early stage involves the condensation of a free amino group (from free amino acids and/or proteins) with a reducing sugar to form Amadori or Heyns rearrangement products.
  • the advanced stage means degradation of the Amadori or Heyns rearrangement products via different alternative routes involving deoxyosones, fission or Strecker degradation.
  • a complex series of reactions including dehydration, elimination, cyclization, fission and fragmentation result in a pool of flavour intermediates and flavour compounds.
  • key intermediates and flavour chemicals can be identified.
  • Strecker degradation in which amino acids react with dicarbonyls (formed by the Maillard reaction) to generate a wealth of reactive intermediates.
  • Typical Strecker degradation products are aldehydes, e.g. formaldehyde, acetaldehyde, and possibly propenaldehyde (acrolein).
  • Strecker degradation results in degradation of amino acids to aldehydes, ammonia and carbon dioxide (44) and takes place in foods at higher concentrations of free amino acids and under more drastic reactions, e.g. at higher temperatures or under pressure (45).
  • Fig 1 Pathways of formation of key flavour intermediates and products in the Maillard reaction (43).
  • stage 3 of the MR is characterized by the formation of brown nitrogenous polymers and co-polymers. While the development of colour is an important feature of the reaction, relatively little is known about the chemical nature of the compounds responsible. Colour compounds can be grouped into two general classes - low molecular weight colour compounds, which comprise two to four linked rings, and the melanoidins, which have much higher molecular weights.
  • Factors that are particularly important for the MR are the starting reactants, e.g. type of sugar and amino acid (protein), time, temperature and water activity. Presence of metal salts (pro-oxidants), and inhibitors, like antioxidants and sulphite, might all have an impact. Starting reactants - reducing sugar and amino acids/proteins
  • MR requires reducing sugars, i.e. sugars containing keto- or aldehydes (free carbonyl groups).
  • sugars containing keto- or aldehydes free carbonyl groups.
  • Aldoses are more reactive than ketoses both in aqueous solution model systems and at storage (low water content) - Among isomeric sugars, stereochemistry is important. Thus ribose is more reactive than xylose monitored as lysine losses.
  • All monosacharides are reducing sugars. (Sugar alcohols do not participate in MR.) Among the disaccharides all sugars except sucrose are reducing sugars. In oligosaccharides and starch only the end-terminal monosaccharide is a reducing sugar. Starch and sugars, such as sucrose, lactose, maltose etc can easily hydrolyse upon heating above 100°C at slightly acidic pH, resulting in the formation of monosaccharides (reducing sugars). Thus, thermal processing often result in a continuous supply of reducing sugar formed from complex carbohydrates.
  • the temperature dependence of chemical reactions is often expressed as the activation energy, Ea, in the Arrhenius equation.
  • Ea activation energy
  • Activation energy data for the MR have been reported within a wide range, 10-160 kJ/mole, depending on, among other things, water activity and pH and what effect of the reaction has been measured.
  • the temperature dependence of the MR is also influenced by the participating reactants. The temperature effect is also affected by the other variables and different aspects of the MR thus differ in temperature dependence (42).
  • Water has both an inhibitory and an accelerating impact on the MR. Water acts partly as a reactant and partly as a solvent and transporting medium of reactants (reactant mobility). In the initial steps of the MR, 3 moles of water are formed per mol carbohydrate. Thus the reaction occurs less readily in foods with a high aw value. Water might depress the initial glucosylamine reaction, but enhance the deamination step later in the reaction.
  • the MR itself has a strong influence on pH. Therefore, aqueous model systems based on reflux boiling of sugars and amino acids need to be buffered since the pH quickly drops from 7 to 5. Low pH values ( ⁇ 7) favour the formation of furfurals (from Amadori rearrangement products), while the routes for reductones and fission products are preferred at a high pH.
  • Measures to inhibit the Maillard reaction in cases where it is undesirable involve lowering of the pH value, maintenance of lowest possible temperatures and avoidance of critical water contents (moistures below 30%, during processing and storage), use of non- reducing sugars, and addition of sulphite (45).
  • the use of the inhibitor, sulphur dioxide constitutes an important way of controlling the Maillard reaction. It may combine with early intermediates. However, sulphite only delays colour formation and it is interesting to note that the colour formed in sulphite-treated systems is less red and more yellow than in untreated systems.
  • the most common route for formation of flavours via the MR comprises the interaction of a-dicarbonyl compounds (intermediate products in the MR, stage 2) with amino acids through the Strecker degradation reactions.
  • Alkyl pyrazines and Strecker aldehydes belong to commonly found flavour compounds from MR.
  • low levels of pyrazines are formed during the processing of potato flakes when the temperature is less than 130°C, but increases tenfold when the temperature is increased to 160°C, and decreases at 190°C, probably due to evaporation or binding to macromolecules.
  • the aroma profile varies with the temperature and the time of heating. At any given temperature-time combination, a unique aroma, which is not likely to be produced at any other combination of heating conditions, is produced. Temperature also affects the development of aroma during extrusion cooking
  • the coloured products of the Maillard reaction are of two types: the high molecular weight macromolecule materials commonly referred to as the melanoidines, and the low molecular weight coloured compounds, containing two or three heterocyclic rings (48).
  • the concentration of reducing sugar has the greatest impact on colour development.
  • lysine gives the largest contribution to colour formation and cysteine has the least effect on colour formation.
  • Loss in protein quality is often associated with the MR, especially in cereal products and milk powder produced by heat-treatment.
  • the essential amino acid having an extra free amino group e.g. lysine
  • the essential amino acid also is the nutritionally limiting amino acid
  • the influence of MR on the protein quality is substantial. This is not a problem in cooking meat and fish, since these food items are very rich in protein.
  • Loss of protein quality in terms of nutritional value is a more serious problems for heat-treatment and dehydration of especially cereals, milk and their mixtures (breakfast cereals, gruels, bread, biscuits), since carbohydrates dominates over proteins in these food items and the proteins levels are also generally low.
  • gravies prepared from dried meat- juice collected from pan-residues or oven-roasting could be rich in heterocyclic amines.
  • Pro-oxidants, water activity in the optimal range for the MR, and high temperatures (200- 400°C) enhance their yield.
  • the average daily exposure for heterocyclic amines is around 0.5 ⁇ g/day and person, with a range between 0-20 ⁇ g.
  • Antioxidants, excess of carbohydrates, cooking temperatures below 200°C and moisture contents above 30% reduce the occurrence of heterocyclic amines.
  • heterocyclic amines rarely occur in plant foods even during well-done cooking (52).
  • the nonenzymic browning or Maillard reaction is ( ⁇
  • the following steps involve the ⁇ -elimination of the hydroxyl initial products of condensation are fluorescent and continuation of group on carbon No. 3.
  • the enolamine is in the free base form the reaction results in the formation of the brown melanoidins. and converts to the Schiff base (5).
  • the Schiff base may undergo These polymers are of non-distinct composition and contain varying hydrolysis and form the enolform (7) of 3-deoxyosulose (8). In levels of nitrogen.
  • the composition varies with the nature of the another step the Schiff base (5) may lose a proton and the hydroxyl reaction partners, pH, temperature and other conditions.
  • the amino acid is converted into an aldehyde with one less carbon atom (Sch ⁇ nberg and Moubacher 1952).
  • Some of the compounds of browning flavor have been described by Hodge et al. (1972).
  • Corny, - nutty, bready and crackery-aroma compounds consist of planar unsaturated heterocyclic compounds with one or two nitrogen atoms in the ring.
  • Other important members of this group are partially saturated N-heterocyclics with alkyl or acetyl group substituents.
  • Compounds that contribute to pungent, burnt aromas are listed in Table 3.7. These are mostly vicinal polycarbonyl compounds and ⁇ ,
  • the Strecker degradation aldehydes contribute to the aroma of bread, peanuts, cocoa and other roasted foods.
  • Temperature also affects the composition of the The nature of the sugars in a nonenzymic browning reaction deterpigment formed. At higher temperatures, the carbon content of the mines their reactivity. Reactivity is related to their conformational pigment increases and more pigment is formed per mole of carbon stability or to the amount of open-chain structure present in soludioxide released. Color intensity of the pigment increases with intion. Pentoses are more reactive than hexoses, and hexoses more creasing temperature. The effect of temperature on the reaction rate than reducing disaccharides. Nonreducing disaccharides only react of D-glucose with DL-leucine is demonstrated in Fig. 3.14. a ⁇ feThydr ⁇ lysis has taken place. The order of reactivity of some of
  • Methods of preventing browning could consist of measures inand the amide groups of asparagine or glutamine, with the reacting tended to slow reaction rates, such as control of moisture, temperaunits present either in the same peptide chain or in neighboring ones ture or pH or removal of an active intermediate. Generally, it is (Pig. 3.16). easier to use an inhibitor.
  • One of the most effective inhibitors of Some amino acids may be oxidized by reacting with free radicals browning is sulfur dioxide. The action of sulfur dioxide is unique formed by lipid oxidation. Methionine can react with a lipid perox- and no other suitable inhibitor has been found. It is known that ide to yield methionine sulfoxide. Cysteine can be decomposed by a sulfite can combine with the carbonyl group of an aldose to give an lipid free radical according to the following scheme: addition compound:

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Abstract

L'invention porte sur un procédé de prévention et/ou réduction de la formation d'acrylamide et/ou de son précurseur dans un aliment contenant: (i) une protéine; un peptide et/ou un acide aminé, et (ii) un sucre réducteur. Ledit procédé consiste à mettre en contact l'aliment avec une enzyme susceptible d'oxyder le groupe réducteur du sucre.
PCT/IB2003/005278 2000-11-17 2003-10-24 Procede WO2004039174A2 (fr)

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AU2003276613A AU2003276613A1 (en) 2002-10-30 2003-10-24 A method of preventing acrylamide formation in a foodstuff
US11/048,230 US8163317B2 (en) 2000-11-17 2005-02-01 Method
US13/433,470 US8956670B2 (en) 2000-11-17 2012-03-29 Method for the control of the formation of acrylamide in a foodstuff
US14/619,149 US20150223499A1 (en) 2000-11-17 2015-02-11 Method for the control of the formation of acrylamide in a foodstuff

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Cited By (6)

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WO2005004629A1 (fr) * 2003-06-25 2005-01-20 The Procter & Gamble Company Procede de reduction de l'acrylamide dans les aliments consistant a reduire le niveau des sucres reducteurs, aliments presentant des niveaux reduits d'acrylamide, et article de commerce
US7267834B2 (en) 2003-02-21 2007-09-11 Frito-Lay North America, Inc. Method for reducing acrylamide formation in thermally processed foods
US7393550B2 (en) 2003-02-21 2008-07-01 Frito-Lay North America, Inv. Method for reducing acrylamide formation in thermally processed foods
US7811618B2 (en) 2002-09-19 2010-10-12 Frito-Lay North America, Inc. Method for reducing asparagine in food products
ES2376117A1 (es) * 2009-07-28 2012-03-09 Leng-D'or, S.A. Procedimiento para reducir la formación de acrilamida en alimentos obtenidos a partir de pellets vegetales.
WO2018169055A1 (fr) 2017-03-16 2018-09-20 学校法人北里研究所 Nouveau composé pochoniolide et son utilisation

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US8486684B2 (en) 2007-08-13 2013-07-16 Frito-Lay North America, Inc. Method for increasing asparaginase activity in a solution
US8284248B2 (en) 2009-08-25 2012-10-09 Frito-Lay North America, Inc. Method for real time detection of defects in a food product
US8158175B2 (en) 2008-08-28 2012-04-17 Frito-Lay North America, Inc. Method for real time measurement of acrylamide in a food product
US9095145B2 (en) 2008-09-05 2015-08-04 Frito-Lay North America, Inc. Method and system for the direct injection of asparaginase into a food process
US9215886B2 (en) 2008-12-05 2015-12-22 Frito-Lay North America, Inc. Method for making a low-acrylamide content snack with desired organoleptical properties

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7811618B2 (en) 2002-09-19 2010-10-12 Frito-Lay North America, Inc. Method for reducing asparagine in food products
US7267834B2 (en) 2003-02-21 2007-09-11 Frito-Lay North America, Inc. Method for reducing acrylamide formation in thermally processed foods
US7393550B2 (en) 2003-02-21 2008-07-01 Frito-Lay North America, Inv. Method for reducing acrylamide formation in thermally processed foods
US8114463B2 (en) 2003-02-21 2012-02-14 Frito-Lay North America, Inc. Method for reducing acrylamide formation in thermally processed foods
WO2005004629A1 (fr) * 2003-06-25 2005-01-20 The Procter & Gamble Company Procede de reduction de l'acrylamide dans les aliments consistant a reduire le niveau des sucres reducteurs, aliments presentant des niveaux reduits d'acrylamide, et article de commerce
KR100848519B1 (ko) 2003-06-25 2008-07-25 더 프록터 앤드 갬블 캄파니 환원당의 수준을 감소시키는 단계를 포함하는 식품에서의아크릴아미드의 감소 방법, 아크릴아미드의 수준이 감소된식품 및 시판 물품
ES2376117A1 (es) * 2009-07-28 2012-03-09 Leng-D'or, S.A. Procedimiento para reducir la formación de acrilamida en alimentos obtenidos a partir de pellets vegetales.
WO2018169055A1 (fr) 2017-03-16 2018-09-20 学校法人北里研究所 Nouveau composé pochoniolide et son utilisation

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