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ENZYMES & LYVEN PRODUCTS PRESENTATION. presentation enzymes 2007 2 ENZYMES = PROTEINS Biochemicals Molecules composed of different amino-acid and other.

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Presentation on theme: "ENZYMES & LYVEN PRODUCTS PRESENTATION. presentation enzymes 2007 2 ENZYMES = PROTEINS Biochemicals Molecules composed of different amino-acid and other."— Presentation transcript:

1 ENZYMES & LYVEN PRODUCTS PRESENTATION

2 presentation enzymes ENZYMES = PROTEINS Biochemicals Molecules composed of different amino-acid and other radicals Produce from a microorganism or cell Thermolabile Proteins able to work in or out the production cell. Active site is the operational part of the enzyme

3 presentation enzymes ENZYMES=PROTEINS Active site

4 presentation enzymes Side Activity Primary activity Side activity Other proteins Chromatography of a xylanase extraction

5 presentation enzymes Micro-organisms Two kinds of micro-organisms can produce enzyme: Fungal or Bacterial way Different side activities Fungal/YeastBacterial pHAcid-NeutralNeutral-Alkaline TemperatureLow Temperature High Temperature

6 presentation enzymes Enzymes: Enzymatic Catalysis Agent Catalysis:Increase in the velocity of a chemical reaction or process produced by the presence of a substance that is not consumed in the net chemical reaction or process S+E E+P S=Substrate P=Product E=Enzyme

7 presentation enzymes Enzymatic Action Scheme G G G G G G G G E E G G E Maltose Glucose Enzyme GG GG

8 presentation enzymes Enzymes: Specific Catalysts Each enzyme catalyses one and only one reaction Nowaday, near enzymes were discovered but only 25 to 30 enzymes are industrialy produced

9 presentation enzymes Enzymatic Action Oxidoreductases (EC 1): oxidation–reduction reactions, which entail the transfer of electrons from a substrate to another substrat Transferases (EC 2): move a chemical group from one compound to another compound Hydrolases (EC 3): Hydrolysis of a chemical bond Lyases (EC 4): formation of a double bond Isomerases (EC 5): catalyzes substrate to an isomeric form Ligases (EC 6): catalyzes a reaction that joins two substrates

10 presentation enzymes Lock and Key Model: One and only one molecule could match with one enzyme Substrate Active Site Enzymes changes shape slightly as substrate binds Products Substrate entering active site of enzyme Enzyme/Substrate complex Products leaving active site of enzyme

11 presentation enzymes EFFECTOR INHIBITOR S I E I S E E: Enzymes I: Inhibitor S: Substrate e.g: Cu, Hg, Fe inhibit amylase

12 presentation enzymes EFFECTOR ACTIVATOR A E S S E: Enzymes A: Activator S: Substrate e.g: Calcium activates amylase A+E

13 presentation enzymes Enzymatic catalysis principle Any reaction catalysed by an enzyme can occur naturally but with a more important energy cost. Source wikipedia A, B: Substrate E: Enzyme AB: Product

14 presentation enzymes Factors acting on the enzymatic activity Substrate concentration Water: Enzymes work only in aqueous phase pH: There is always an optimum pH Temperature: There is an optimum and over 70°C, main enzymes are distorted Duration

15 presentation enzymes pH influences e.g: Amylase activity

16 presentation enzymes Temperature influences e.g: Amylase activity

17 presentation enzymes Parameters Variation p.H T°C Time Dose Optimum p.H T°C Time Dose DOSE=COST

18 presentation enzymes Enzymes deactivation By thermic treatment (e.g: Baking…) By settling/filtration (e.g: Fruit Juice…) By increasing pH with Sodium carbonate or decreasing pH with acid (e.g: Textile…)

19 presentation enzymes Enzyme=Processing Aids Normally enzymes are not present in final products, they were destroyed or degraded during the process (Specifications from FAO, WHO, JECFA, FCC and French law 19/10/2006)

20 presentation enzymes Enzymes in Food-Industry FRUIT JUICE BAKING BREWING WINE FEED

21 presentation enzymes ENZYMES ACTION ON FRUIT

22 presentation enzymes Fruit Structure Vegetable raw material is composed of polysaccharides more or less complex. The main are: –Cellulose – Hémi-cellulose –Lignine –Pectins

23 presentation enzymes The Vegetal Cell Wall of a Fruit Source: Dean MADDEN, 2000

24 presentation enzymes Chemical Constitution of Vegetal Cell Wall Source: Bayou & Cortes CYTOPLASMCYTOPLASM Lignine Middle Lamella Secondary Cell Wall Primary Cell Wall Cellulose Hémicellulose PECTINS OTHER CELLOTHER CELL Vegetal Cell

25 presentation enzymes PECTINS Fruit Juice cloudiness at the end of the pressing step is caused by pectins

26 presentation enzymes Pectin A carbohydrate consisting of one primary chain of galacturonic acid and branched out with several sugar molecules including rhamnose, galactose, arabinose Galacturonic acid Méthyl Galacturonic acid A detail of pectin Source: Jussieu University Galacturonic acid Methyl ac. Gal. Rhamnose Other oses Calcium ions

27 presentation enzymes Pectinases Three basical enzymatic activities are present in commercial preparation : »PG: PolyGalacturonase »PL: PectineLyase »PME: Pectine Methyl Esterase

28 presentation enzymes How work Pectinases: PL: breaks links between esterified galacturoni acid and residues Non esterified galacturonic acid PE: add a methyl group Galacturonic acid residue PG: creates galacturonic acid residue from non esterified galacturonic acid Non-reducing end of a polygalacturonic acid chain Source: Dean MADDEN, 2000

29 presentation enzymes FRUIT JUICE WINE

30 presentation enzymes ENZYMES IN WINE

31 presentation enzymes Grape Berry: Structure Berry Stem (Pedicel) Brush Skin AnthocyanesFlavones Flesh Seed Vascular Bundle Red GrapeWhite Grape Glucose, Fructose Tannins, Oil

32 presentation enzymes Red Wine Making Process CRUSHER Maceration + Alcoholic Fermentation Maceration + Alcoholic Fermentation PRESSING « Vin de Goutte » MALOLACTIC FERMENTATION MALOLACTIC FERMENTATION Blending « Vin de Presse » SKIN DESTEMMER Stem Grapes PECLYVE V Extraction PECLYVE V CLARIFICATION

33 presentation enzymes Red Wine Making Process – flash détente CRUSHER PRESSING FERMENTATION SKIN DESTEMMER Stem Grape Berries PECLYVE V EXTRACTION PECLYVE V CLARIFICATION CLARIFICATION

34 presentation enzymes PECLYVE V EXTRACTION in Red Wine Allows to extract faster the color from the red grape berry Avoid to extract too much tannins Give less astringent wine

35 presentation enzymes Action of Peclyve V extr Tannins Extraction % Days Color extraction with Peclyve V extraction Color extraction without Peclyve V extraction

36 presentation enzymes DECANTATION STEP WITH PECLYVE V CLARIFICATION CLOUDY JUICE CLEAR JUICE MUST ENZYMATIC ACTION PECLYVE V Clarification protein pectin

37 presentation enzymes White Wine Making Process CRUSHER MACERATION PRESSING ALCOHOLIC FERMENTATION DESTEMMER Stem Grape Berries PECLYVE V EXTRACTION PECLYVE VP LYVAROME A5 CLARIFICATION PECLYVE V Clarification

38 presentation enzymes PECLYVE V EXTR in White Wine Allows to extract faster the flavour from the white grape berry Avoid to extract too much tannins Give less astringent wine

39 presentation enzymes Flavour in Wine Flavour in the wine comes from terpenols which are derived from terpen. Sometimes, terpens must be transformed in terpenols by enzymes help. G G T T T T TL G: Glucose T: Terpen TL: Terpenol YeastEnzymes

40 presentation enzymes LYVAROME A5 At the last quarter of the fermentation, adding LYVAROME A5 will transform Terpen in Terpenol, giving more flavour to the wine OPP GÉRANYL DIPHOSPHATE NERYLE DIPHOSPHATE LINALYL DIPHOSPHATE LYVAROMEA5LYVAROMEA5 CH 2 OH OH CH 2 OH GERANIOL NEROL LINALOL

41 presentation enzymes

42 presentation enzymes Action of Peclyve V extraction Tannins Extraction % Days Flavour extraction with Peclyve V extr Flavour extraction without Peclyve V extr

43 presentation enzymes PECLYVE VP Sometimes, Pectinases enzymes with Cinnamoyl-Esterase (CIE) could lead to release vinyl-phenols. Vinyl-phenol could give a bad profil aroma to the white wine Peclyve VP with a very low level of CIE activity avoid this problem

44 presentation enzymes Optimum pH

45 presentation enzymes Thermostability

46 presentation enzymes Washing Efficiency

47 presentation enzymes ENZYMES FOR FRUIT JUICE

48 presentation enzymes Pectinases in Fruit Juice Process The commercial preparations of pectinase are adapted to the process and to the types of fruit Different preparations exist for pulp treatment and for Juice All pectinases from LYVEN are from Aspergillus niger and are GMO free

49 presentation enzymes FRUIT JUICE PROCESS MACERATION STEP PRESSING DECANTATION STEP CLEAR JUICE PACKING CONCENTRATION STEP CONCENTRATION STEP MASHING PECLYVE PR PECLYVE CP PECLYVE UF+ AMYLYVE TC SUPER PECLYVE CP PECLYVE UF+ AMYLYVE TC SUPER

50 presentation enzymes PECLYVE PR PECLYVE PR is well balanced in PG, PL & PE to increase the yield during pressing Pulp Treatment Fresh Apple Frozen Apple Without enzyme 80-82%70-74% With PECLYVE PR 90-92%85-90%

51 presentation enzymes AMYLYVE TC SUPER When the fruit is harvested early in the season, some residual starch could remain in the fruit causing cloudiness at the pasteurisation and filtration step. AMYLYVE TC Super allows to degrade this residual starch.

52 presentation enzymes DECANTATION STEP WITH PECLYVE CP CLOUDY JUICE CLEAR JUICE MUD ENZYMATIC ACTION PECLYVE CP protein pectin

53 presentation enzymes Pectin Test To check presence of pectin, a test with alcohol could be make.

54 presentation enzymes OTHER FRUITS FRUITPECTIN(%)REMARK APPLE PEAR ARABAN LEMON GRAPEFRUIT/ ORANGE CLOUDY JUICE STRAWBERRY CRANBERRY ACID

55 presentation enzymes PEAR Due to araban, a haze could appear. PEARLYVE is specially formulated to avoid that.

56 presentation enzymes CITRUS To reduce viscosity, improving the concentration, PECLYVE CITRUS JUICE is the best product. To clarify the juice (e.g lemon) PECLYVE CITRUS CLAR is highly recommended

57 presentation enzymes CITRUS To increase yield and extract the residual juice of the pulp, PECLYVE CITRUS WASH is the ideal product. For extraction of essential oil: PECLYVE CITRUS OIL To remove peel from citrus without damage for the general aspect of the fruit: PECLYVE CITRUS PEEL is perfectly adapted.

58 presentation enzymes FRUIT FIRMING Thanks to its high content in Pectin Methyl Esterase, PECLYVE PME PLUS allows an improvement of fruit consistency (e.g:fruit inclusion in yoghourt, cider…)

59 presentation enzymes TROPICAL FRUITS With a high level in hemicellulase, PECLYVE PINEAPPLE allows to reduce viscosity of the juice To make a total clarification of cloudy juice from tropical fruit, PECLYVE TROPICA is well adapted.

60 presentation enzymes PECLYVE OLIVA During maceration step, PECLYVE OLIVA allows to reduce the emulsifying phase. Emulsion with enzymes Emulsion without enzymes Oil

61 presentation enzymes RED FRUITS & VEGETABLE Red fruit mash is particularly acid and PECLYVE FR is well adapted for acid phase. Vegetable have a high content in cellulose and pectins and it is why our PECLYVE LI is specially formulated for vegetables

62 presentation enzymes WARNING The dosage of all these enzymes must be very precise. If too much enzymes is added, it will lead to a liquefaction and the filtration will not be able anymore. Due to its experience Lyven is ready to help you to find the best dosage to use.

63 presentation enzymes ENZYMES IN BAKING

64 presentation enzymes BAKING PRINCIPLES BREAD: Food made from dough of flour or meal and usually raised with yeast or baking powder and then baked 3 elements are imperatives: –WATER: Allows to give a network of gluten –YEAST: Allows to give CO2 and flavour molecules producing from flour components –CO2 is trapped in gluten network and gives bread volume

65 presentation enzymes FLOUR COMPOSITION COMPONENT% STARCH60-72% DIFFERENT OSES1-2% PROTEINS8-12% LIPIDS1,2-1,4% MINERALS0,5-0,6 HEMICELLULOSE2-3% WATER13-16%

66 presentation enzymes BREAD PROCESS MOULDING STAGE PROVING STAGE MIXING STAGE DIVIDING STAGE BAKING STAGE Flour, Water, Ingredient (salt, fat..) enzymes

67 presentation enzymes Enzymes in Baking AMYLASE HEMICELLULASE GLUCOSE-OXYDASE PROTEASE PHOSPHOLIPASE & LIPASE

68 presentation enzymes AMYLASES Starch is composed of amylose and amylopectine Amylase: Enzyme that liquefy starches and convert them to maltose (sugar) and dextrins. AMYLOSE AMYLOPECTINE Fermentable Sugars :amylase

69 presentation enzymes LYVEN Amylases Panlyve Amy : Fungal Alpha-Amylase Increase bread volume by faciltating yeast activity : Increase colorness of the crust Panlyve AR : Bacterial Alpha-Amylase Retard the staling of baked goods Avoid starch retrogradation

70 presentation enzymes Correlation between Falling Number and Amylase dose Examples: If the falling number is 350 seconds, FAU per 100 kg of flour should be added. If after malt addition, the falling number is 300 seconds, FAU per 100 kg of flour should be added. Falling Number FAU (Fungal Amylase Unit): Amylase activity Unit

71 presentation enzymes HEMICELLULASE Hemicellulase (xylanase): Enzyme that breaks down hemicellulose. Hemicellulose catagorizes a variety of complex polysaccharides chain (mainly pentosans). Within hydrolising pentosans, xylanases give more free water and increase the dough workability and bread volume

72 presentation enzymes LYVEN Xylanase Panlyve HCB increases bread volume and gives better color and structure to the crumb.

73 presentation enzymes GLUCOSE-OXIDASE Glucose-Oxidase: converts glucose to gluconic acid and hydrogen peroxyd and will lead to molecule oxidation. GOX GLUCOSE GLUCOSE **+ H SH HS Gluten proteins SS Di-sulfure bonds created

74 presentation enzymes LYVEN Glucose-Oxidase Panlyve GO: Fungal Glucose Oxidase Well adapted for weak flour Increase tenacity and decrease elasticity of the dough Partials ascorbic acid replacer Well adapted to deep-frozen raw dough Panlyve GO should be avoided in dough with butter (rancid flavour)

75 presentation enzymes PROTEASE Protease: Hydrolysis of the protein chains of the gluten and the formation of fragments of smaller size S S S S protease

76 presentation enzymes LYVEN Protease Panlyve BI: Bacterial protease Replacer of chemical reducers (metabisulfite…) Specially adapted for use in the manufacture of Biscuits and crackers

77 presentation enzymes LIPASE & PHOSPHOLIPASE Enzyme that catalyzes the hydrolysis of triglycerides to yield glycerol and fatty acids lipase Hydrophobic partHydrophilic part EMULSIFYING E472, E471, E481 … AIR AMIDON GLUTEN triglycerides

78 presentation enzymes LYVEN Lipase & Phospholipase Panlyve TDR: Enzymatic complex with lipase & phospholipase Replacer of Chemical emulsifier as Datem or SSL. Increasing bread volume Giving a thiner crumb

79 presentation enzymes SPECIAL FORMULATION Panlyve BR: Alpha-Amylase+Hemicellulase+Glucose oxidase –Bromate replacer –Improvement of machenability and strength of the dough –Improvement of crispiness of the crust Panlyve NPB Alpha-Amylase+Protease –Dough relaxer (actives on gluten) –Gives expanding dough –Well adapted for strong flour

80 presentation enzymes SPECIAL FORMULATION Panlyve AG: Fungal amylase+amyloglucosidase –Gives directly glucose and maltose –Increases the yeast activity –Increase the colour of the bread Panlyve ASN: Alpha-Amylase+Hemicellulase –Improves softness and fineness of the crumb –Extends the shelf-life of the bread

81 presentation enzymes SPECIAL FORMULATION Panlyve B2G Alpha-amylases+hemicellulases –Specially adapted for retarder proven processes –Specially adapted for deep freezing of unbaked doughs

82 presentation enzymes SPECIAL FORMULATION Panlyve B2S Alpha-amylase + specific hemicellulases –Specially recommended for good quality flours –Improves crumb softness and crust aspect Panlyve XT Alpha-Amylase+endo-xylanase –Well adapted on rye flour and short fermentation process –Convenient for low hydrated dough (biscuit)

83 presentation enzymes Enzymes dosage should be adapted to the quality of the flour. Due to his good knowledge in Flour and Baking, LYVEN is ready to help you in the choice of enzymes and dosages.

84 presentation enzymes ENZYMES IN BEER

85 presentation enzymes BEER It is a fermented beverage obtained from malted cereals (barley, wheat, corn…) water, yeast and hops. For several reasons, malted cereals could be replaced by cereals like wheat, oats… Enzymes will attack the cereal in order to help yeast during the fermentation

86 presentation enzymes Brewing Flowchart Adjuncts Cooker Mash Tub Lauter Tub Copper Fermentation Beer Tank FILTRATION Fermentation & Filtration Brewhouse Malt Yeast

87 presentation enzymes AMYLASES Starch is composed of amylose and amylopectine Amylase: Enzyme that liquefy starches and convert them to maltose (sugar) and dextrins. AMYLOSE AMYLOPECTINE Fermentable Sugars :amylase

88 presentation enzymes Lyven Amylases in Brewing Amylyve UHT: Bacterial Alpha-Amylase –Heat Stable enzymes –Liquefaction of the starch at high temperature Amylyve A30: Bacterial Alpha-Amylase –Liquefaction of starch at intermediate temperature Amylyve AN2300L:Fungal Alpha-Amylase –Maltose and maltotriose forming effect Amylyve AG400L: Fungal Glucamylase –Transforms dextrins into glucose

89 presentation enzymes Brewing Flowchart Adjuncts Cooker Mash Tub Lauter Tub Copper Fermentation Beer Tank FILTRATION Fermentation & Filtration Brewhouse Malt Yeast Amylyve UHT Amylyve A30 Amylyve AG 400L Amylyve AN 2300L

90 presentation enzymes GLUCANASE An enzyme that act on beta glucans, a type of gum found in the endosperm of unmalted barley, oatmeal, and wheat. Beta-glucans Beta-glucanase Dextrins

91 presentation enzymes LYVEN Glucanase in Brewing GLUCALYVE PB 750 : Fungal Beta Glucanase –Heat stable –Reduces viscosity of the wort BREWLYVE AXC 1500L: Fungal xylanase –Acts on the pentosane –Completes action of Glucalyve PB750 FILTERLYVE 1500L: Fungal Beta Glucanase –Improve filterability of beer –Efficient at the pH of beer

92 presentation enzymes Brewing flowchart Adjuncts Cooker Mash Tub Lauter Tub Copper Fermentation Beer Tank FILTRATION Fermentation & Filtration Brewhouse Malt Yeast Glucalyve PB 750 Brewlyve AXC 1500L Filterlyve 1500L

93 presentation enzymes PROTEASE An enzyme that hydrolyzes (breaks down a bond and adds water) proteins, especially to peptides S S S S protease

94 presentation enzymes LYVEN Protease in Brewing Brewlyve NP900: Bacterial Protease –Improves the content of peptides and amino- acid in the wort –Gives more FAN (Free Amino Acid) to the yeast Lypaine 6500: Vegetal Protease –Prevent hazing of beer kept in cold storage

95 presentation enzymes Brewing Flowchart Adjuncts Cooker Mash Tub Lauter Tub Copper Fermentation Beer Tank FILTRATION Fermentation & Filtration Brewhouse Malt Yeast Brewlyve NP 900 Lypaine 6500

96 presentation enzymes ENZYMES IN ANIMAL FEED

97 presentation enzymes Enzymes using in Animal Feed Enzymes in animal feed will allow to higher absorption of nutritional particles and to a better digestibility of feedingstuff, leading to an increasing growth of the animals Three enzymes are mainly used in Animal Feed: –XYLANASE –GLUCANASE –PHYTASE

98 presentation enzymes Animal Feed Definitions Feed Conversion Ratio: FCR=Quantity of feed used/ Quantity of meat produced Apparent Metabolisable Energy: AME=General Energy (GE) of feed-Energy Contained in Excretion (ECE) AME ECE TE

99 presentation enzymes Non-Starch Polysaccharides Non-Starch Polysaccharides (NSP) are present in cereal based diets: –Traps of valuable nutrients in the cells of vegetable feedstuffs –Increase the viscosity of the digesta in the intestine.

100 presentation enzymes Non-Starch Polysaccharides (NSP) rice maize sorgho wheat rye barley AME/GE %NSP

101 presentation enzymes FEEDLYVE AXC - contains xylanase as the main enzymatic activity, which degrade pentosan contained in cereal like wheat and rye * fungal xylanase from Trichoderma longibrachiatum * xylanase concentration is described in units of AXC - contains main side activities (B glucanase, pectinases,cellulases...) Feedlyve AXC is available - in 2 different forms : liquid and powder (Feedlyve AXC L and Feedlyve AXC P) -in different concentration : -Feedlyve AXC 200 contains at least 200 u AXC / g -Feedlyve AXC 400 contains at least 400 u AXC / g -Feedlyve AXC 1500 contains at least 1500 u AXC / g -Feedlyve AXC 6000 contains at least 6000 u AXC / g -Feedlyve AXC contains at least u AXC / g

102 presentation enzymes AME with FEEDLYVE AXC Apparent Metabolisable Energy Roslin Institute, Scotland Results AME levels, in MJ / kg %

103 presentation enzymes FCR with FEEDLYVE AXC in broilers

104 presentation enzymes FEEDLYVE AGL - contains endo glucanase as the main enzymatic activity which degrade glucans contained in cereal like barley * fungal B glucanase from Aspergillus Niger * B glucanase concentration is described in units of AGL - contains main side activities (xylanase, pectinases,..) Feedlyve AGL is available - in 2 different forms : liquid and powder (Feedlyve AGL L et Feedlyve AGL P) -in different concentration : -Feedlyve AGL 200 contains at least 200 u AGL / g -Feedlyve AGL 400 contains at least 400 u AGL / g -Feedlyve AGL 1500 contains at least 1500 u AGL / g -Feedlyve AGL 6000 contains at least 6000 u AGL / g -Feedlyve AGL contains at least u AGL / g

105 presentation enzymes AME with FEEDLYVE AGL Apparent Metabolisable Energy Roslin Institute, Scotland Results AME levels, in MJ / kg %

106 presentation enzymes FCR with FEEDLYVE AGL in Broilers

107 presentation enzymes PHYTATES Oilseed (soya…) and Cereals (wheat…)contain 90% of their phosphorus locked up in phytate which is almost indigestible to monogastric animals. The unabsorbed phytate passes through the gastrointestinal tract, elevating the amount of phosphorus in the manure, leading to environmental problem. O P O-O- OH -O-O O P O-O- -O-O O P O-O- -O-O O P O-O- -O-O O P O-O- -O-O + + Furthermore, Other nutrients are linked with phytates unavalaible for the intestinal absorption. Phytates minerals

108 presentation enzymes FEEDLYVE PHY 5000 FEEDLYVE PHY 5000 is a phytase which will release the phophorus and the minerals allowing the absorption by the intestinal cells. O P O-O- OH -O-O O P O-O- -O-O O P O-O- -O-O O P O-O- -O-O O P O-O- + + FEEDLYVE PHY 5000 Ca STARCH FEEDLYVE PHY 5000

109 presentation enzymes FEEDLYVE PHY 5000 for broilers

110 presentation enzymes Legislation in Feed Enzymes European authorisation : xylanase : definitive authorisation in chicken (3 trials already done) definitive authorisation in turkey (3 rd trial running in spring) B glucanase : definitive authorisation in chicken (3 trials already done) definitive autorisation in turkey (2nd trial running in spring, 3 rd trial in late summer) For other country than european countries, used should be adapted to the legislation. LYVEN plans : new segments (mainly pig) to think for new enzymes activities (alpha galactosidase, cellulases, proteases, amylases for ex)

111 presentation enzymes Enzymes shelf-life All the Lyven enzymes get only Best Before Date. It means that after this date the activitycould be lower than the specification.. This Best Before Date last for 1 year for the liquids and 2 years for the powders

112 THANK YOU FOR YOUR ATTENTION


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