ENZYMES in Food ENZYMES in Food ENZYMES in Food ENZYMES in Food ProcessingProcessingProcessingProcessing

Useful links forUseful links for ENZYMES ENZYMES d d in Food Processingin Food Processing

http://www.amfep.org/http://www.amfep.org/index.htmlindex.html

http://www.efsa.europa.eu/http://www.efsa.europa.eu/

http://www foodnavigator com/http://www foodnavigator com/ProductProduct CategoriesCategories//CulturesCultures enzymesenzymes yeastyeasthttp://www.foodnavigator.com/http://www.foodnavigator.com/ProductProduct--CategoriesCategories//CulturesCultures--enzymesenzymes--yeastyeast

EnzymeEnzyme databasedatabaseyyhttp://www.brendahttp://www.brenda--enzymes.org/enzymes.org/http://www.expasy.ch/http://www.expasy.ch/enzymeenzyme//http://www.enzymehttp://www.enzyme--database.org/database.org/http://www.ebi.ac.uk/http://www.ebi.ac.uk/intenzintenz//http://www biochem ucl ac uk/http://www biochem ucl ac uk/bsmbsm//enzymesenzymes//http://www.biochem.ucl.ac.uk/http://www.biochem.ucl.ac.uk/bsmbsm//enzymesenzymes//http://metacyc.org/http://metacyc.org/

P ti f P ti f Properties of enzymesProperties of enzymesl l Control ripening.Control ripening.

Cause food spoilage (rotting).Cause food spoilage (rotting).p g gp g g Responsible for changes in flavor, color, Responsible for changes in flavor, color,

texture and nutritional properties.texture and nutritional properties.texture and nutritional properties.texture and nutritional properties. Can be inactivated by heat to extend Can be inactivated by heat to extend

storage stability of foodsstorage stability of foodsstorage stability of foods.storage stability of foods.

P tiP tiPropertiesPropertiesU d f f t ti i f dU d f f t ti i f d Used for fermentation purposes in foods.Used for fermentation purposes in foods.

Can be immobilized to a surface of a Can be immobilized to a surface of a b th i t bj t i t t b th i t bj t i t t membrane or other inert object in contact membrane or other inert object in contact

with the food being processed.with the food being processed.C b t t d d ifi d t hi h C b t t d d ifi d t hi h Can be extracted and purified to a high Can be extracted and purified to a high degree.degree.

E i I dE i I dEnzymes in IndustryEnzymes in Industry

Distribution of enzymes by substrateDistribution of enzymes by substrate Protein hydrolyzingProtein hydrolyzing 59%59%

Carbohydrate hydrolyzingCarbohydrate hydrolyzing 28%28% Carbohydrate hydrolyzingCarbohydrate hydrolyzing 28%28%

Lipid hydrolyzingLipid hydrolyzing 3%3%

Speciality (analytical, pharma, research) 10%Speciality (analytical, pharma, research) 10%

US MarketUS Market BakeryBakery

US$ 210m 5US$ 210m 5--7% pa7% pa

BeverageBeverageUS$ 130m, 4US$ 130m, 4--6% pa6% pa

DairyDairyUS$ 195m, 3US$ 195m, 3--5% pa5% pa

Fats Fats & Oils& OilsUS$ 30m, 10US$ 30m, 10--12% pa12% pa

CulinaryCulinaryUS$ 30m, 5US$ 30m, 5--6% pa6% pa$ ,$ , pp

Meat and others (Meat and others (incl. food protection) incl. food protection) US$ 30m 10US$ 30m 10--12% pa12% paUS$ 30m, 10US$ 30m, 10 12% pa12% pa

A f l titA f l titA few large competitorsA few large competitors

Novozymes (all applications) Novozymes (all applications)y ppy pp DSM (most applications) DSM (most applications) Chr Hansen (dairy enzymes only) Chr Hansen (dairy enzymes only) Chr. Hansen (dairy enzymes only) Chr. Hansen (dairy enzymes only) AB Enzymes (some applications) AB Enzymes (some applications)

th titth titother competitorsother competitors Advanced EnzymesAdvanced Enzymes was founded in 1989 as a biotechnology company was founded in 1989 as a biotechnology company

(sister company, (sister company, Specialty Enzymes and Specialty Enzymes and BiochemicalsBiochemicals, is headquartered , is headquartered in California in the US)in California in the US)f )f )

http://www.ingredientsnetwork.com/profile/full/advancedhttp://www.ingredientsnetwork.com/profile/full/advanced--enzymesenzymes--foodfood--enzymesenzymesyy

produces more than $100 million worth of produces more than $100 million worth of probioticsprobiotics and enzymes every and enzymes every year.year.yy

As well as products for the baked goods and fruit juice processing As well as products for the baked goods and fruit juice processing sectors, sectors, ,,

Advanced Enzymes is based in Mumbai, India and has its Advanced Enzymes is based in Mumbai, India and has its main main manufacturing manufacturing plant in Nasik. Its workforce includes chemical plant in Nasik. Its workforce includes chemical gg ppengineers, food technologists, microbiologists, biochemists and engineers, food technologists, microbiologists, biochemists and biotechnologists.biotechnologists.

Association of Manufacturers and Association of Manufacturers and F mul t s f En m P du ts (AMFEP)F mul t s f En m P du ts (AMFEP)Formulators of Enzyme Products (AMFEP)Formulators of Enzyme Products (AMFEP)

Founded in 1977, the is a nonFounded in 1977, the is a non--profit European industry association profit European industry association

The main objectives of AMFEP areThe main objectives of AMFEP are::

toto provideprovide aa commoncommon basisbasis forfor representingrepresenting thethe interestsinterests ofof itsits membersmembers bothbotht dt d thth i tit tii tit ti ff thth EE U iU i llll ti lti l th itith ititowardtoward thethe institutionsinstitutions ofof thethe EuropeanEuropean UnionUnion asas wellwell asas nationalnational authoritiesauthorities;; toto representrepresent thethe interestsinterests ofof itsits membersmembers inin internationalinternational organisationsorganisations;; toto assureassure aa freefree flowflow ofof informationinformation betweenbetween itsits membersmembers onon developmentsdevelopmentsrelatedrelated toto thethe regulatoryregulatory statusstatus ofof enzymesenzymes inin thethe EUEU;;relatedrelated toto thethe regulatoryregulatory statusstatus ofof enzymesenzymes inin thethe EUEU;; toto informinform itsits customerscustomers andand otherother interestedinterested partiesparties onon thethe efficacyefficacy andand safetysafetyaspectsaspects ofof itsits enzymeenzyme productsproducts;; toto defenddefend andand promotepromote thethe productsproducts ofof thethe enzymeenzyme industryindustry..

Members of AMFEP produce and sell enzymes for use in food, feed, detergents and Members of AMFEP produce and sell enzymes for use in food, feed, detergents and other nonother non-- food industries, excluding enzymes for pharmaceutical and diagnostic food industries, excluding enzymes for pharmaceutical and diagnostic use. use.

Amfep fruitfully coAmfep fruitfully co--operates with European institutions and partner associations in operates with European institutions and partner associations in the EU, Japan and USA.the EU, Japan and USA.

http://www.amfep.org/index.htmlhttp://www.amfep.org/index.html

AMFEP membersAMFEP members

Enzymes REACH Consortium (ERC)Enzymes REACH Consortium (ERC)IndustrialIndustrial enzymesenzymes areare usedused inin aa varietyvariety ofof applicationsapplications somesome ofof whichwhich requirerequire theirtheir registrationregistrationIndustrialIndustrial enzymesenzymes areare usedused inin aa varietyvariety ofof applications,applications, somesome ofof whichwhich requirerequire theirtheir registrationregistrationunderunder thethe soso--calledcalled REACHREACH Regulation*Regulation* ..

FollowingFollowing aa sixsix--monthmonth monthmonth prepre--registrationregistration phasephase fullfull registrationregistration startsstarts onon DecemberDecember 20102010 ororlaterlater dependingdepending onon tonnagetonnagelater,later, dependingdepending onon tonnagetonnage..

REACHREACH requires,requires, subjectsubject toto certaincertain exceptions,exceptions, multiplemultiple registrantsregistrants ofof thethe samesame substancesubstance toto sharesharedatadata andand jointlyjointly submitsubmit partpart ofof thethe relevantrelevant registrationsregistrations..

ManufacturersManufacturers andand importersimporters ofof enzymes,enzymes, recognizingrecognizing thatthat enzymeenzyme registrationregistration underunder REACHREACH willwillbenefitbenefit fromfrom beingbeing discusseddiscussed andand coordinated,coordinated, havehave createdcreated thethe EnzymesEnzymes REACHREACH ConsortiumConsortium (ERC)(ERC)withwith thethe overalloverall purposepurpose ofof facilitatingfacilitating aa smoothsmooth REACHREACH implementation,implementation, hereunderhereunder byby creatingcreatingeffectiveeffective datadata sharingsharing processesprocesses andand aa reductionreduction ofof associatedassociated costscosts..effectiveeffective datadata sharingsharing processesprocesses andand aa reductionreduction ofof associatedassociated costscosts..

ERCERC intendsintends toto discussdiscuss andand developdevelop aa harmonizedharmonized approachapproach toto aspectsaspects suchsuch asas nomenclaturenomenclature andandclassification,classification, sameness,sameness, readread--acrossacross andand datadata waivingwaiving..

TheThe approachapproach willwill bebe basedbased onon thethe commoncommon featuresfeatures thatthat areare sharedshared byby allall enzymesenzymes..

UnlikeUnlike manymany chemicals,chemicals, enzymesenzymes areare aa uniqueunique setset ofof substancessubstances thatthat havehave predictablepredictable propertiesproperties.. ForForinstanceinstance allall enzymesenzymes areare classifiedclassified asas respiratoryrespiratory sensitizerssensitizers regardlessregardless ofof theirtheir biologicalbiological sourcesourceinstance,instance, allall enzymesenzymes areare classifiedclassified asas respiratoryrespiratory sensitizerssensitizers,, regardlessregardless ofof theirtheir biologicalbiological sourcesourceandand catalyticcatalytic activityactivity..

http://www.enzymeshttp://www.enzymes--reach.org/index.htmlreach.org/index.html

*Regulation*Regulation (EC)(EC) NoNo 19071907//20062006 ofof thethe EuropeanEuropean ParliamentParliament andand ofof thethe CouncilCouncil ofof 1818 DecemberDecember20062006 concerningconcerning thethe Registration,Registration, Evaluation,Evaluation, AuthorisationAuthorisation andand RestrictionRestriction ofof ChemicalsChemicals(REACH)(REACH)..

Enzymes REACH Consortium (ERC)Enzymes REACH Consortium (ERC)The Enzyme REACH Consortium operates with two kinds of membership:The Enzyme REACH Consortium operates with two kinds of membership:

-- Full membersFull members which are Manufacturers Importers and/or Only which are Manufacturers Importers and/or Only -- Full membersFull members, which are Manufacturers, Importers and/or Only , which are Manufacturers, Importers and/or Only Representatives of one or more enzymes, who are subject to the Representatives of one or more enzymes, who are subject to the registration requirements pursuant to REACH;registration requirements pursuant to REACH;

-- Associate membersAssociate members, which are Downstream Users, Distributors, , which are Downstream Users, Distributors, industry associations, manufacturers not established within the European industry associations, manufacturers not established within the European Union, and/or third party holding information on enzyme Substances Union, and/or third party holding information on enzyme Substances Union, and/or third party holding information on enzyme Substances Union, and/or third party holding information on enzyme Substances (Data Holder), who are not subject to the registration requirements (Data Holder), who are not subject to the registration requirements pursuant to REACH but who have an identified and reasonable interest in pursuant to REACH but who have an identified and reasonable interest in contributing to the achievement of the Precontributing to the achievement of the Pre--consortium goals.consortium goals.gg gg

Inquiries regarding membership of the Enzyme REACH Consortium can be Inquiries regarding membership of the Enzyme REACH Consortium can be made to its coordinator:made to its coordinator:

Mr. Youri SkaskevitchMr. Youri SkaskevitchTelephone: +32 (0)2 740 29 62Telephone: +32 (0)2 740 29 62EE il il di @di @ hhEE--mail: mail: coordinator@enzymescoordinator@enzymes--reach.orgreach.org

Industrial Enzyme Market

Annual Sales: $ 1.6 billion

Food and starch processing: 45%Detergents: 34%Textiles: 11%Textiles: 11%Leather: 3%Pulp and paper: 1.2%

Industrial Enzymes Industrial Enzymes -- A Global Market ReportA Global Market ReportIndustrial Enzymes Industrial Enzymes -- A Global Market ReportA Global Market Report

BizAcumen, Inc. BizAcumen, Inc.

N mb 1 2009 N mb 1 2009 238 P s 238 P s November 1, 2009 November 1, 2009 -- 238 Pages 238 Pages --

US $ 2,995.00 US $ 2,995.00

World Market for Some Products of Enzymatic ReactionsEnzymatic Reactions

High fructose corn syrup: $ 1 billionAspartame: $ 850 millionAspartame: $ 850 millionAcrylamide: $ 300 million

Acrylamide (or acrylic amide)Acrylamide (or acrylic amide) chemical compound with the chemical formula Cchemical compound with the chemical formula C33HH55NO. NO. white white odourlessodourless crystalline solid, soluble in water, ethanol, ether and chloroform. crystalline solid, soluble in water, ethanol, ether and chloroform.

is prepared on an industrial scale by the hydrolysis of is prepared on an industrial scale by the hydrolysis of acrylonitrileacrylonitrile by by nitrilenitrilehydratasehydratase..

AcrylamideAcrylamide also occurs in many cooked starchy foods. also occurs in many cooked starchy foods. AcrylamideAcrylamide was was accidentally discovered in foods in April 2002 by scientists in Sweden when they accidentally discovered in foods in April 2002 by scientists in Sweden when they found the chemical in starchy foods such as potato chips French fries and found the chemical in starchy foods such as potato chips French fries and found the chemical in starchy foods, such as potato chips, French fries and found the chemical in starchy foods, such as potato chips, French fries and bread that had been heated (production of bread that had been heated (production of acrylamideacrylamide in the heating process in the heating process was shown to be temperaturewas shown to be temperature--dependentdependent).).

una potente neurotossina e nella forma non polimerizzata, facilmente una potente neurotossina e nella forma non polimerizzata, facilmente assorbita attraverso la pelle. Una volta avvenuta la gelificazione perde la sua assorbita attraverso la pelle. Una volta avvenuta la gelificazione perde la sua pericolosit in quanto non pi assorbita.pericolosit in quanto non pi assorbita.

Le Le poliacrilammidipoliacrilammidi reticolate possono assorbire acqua, formando dei gel con reticolate possono assorbire acqua, formando dei gel con svariate propriet (possono ad esempio essere impiegate nella realizzazione di svariate propriet (possono ad esempio essere impiegate nella realizzazione di l i bid ll l f i ll l i bid ll l f i ll lenti a contatto morbide o come supporto nelle elettroforesi o nella lenti a contatto morbide o come supporto nelle elettroforesi o nella cromatografia). cromatografia).

I d t i l E ClI d t i l E ClIndustrial Enzyme ClassesIndustrial Enzyme Classes Commodity enzymesCommodity enzymes Commodity enzymesCommodity enzymes

High volume (tonnes p.a)High volume (tonnes p.a) Low purity (but not necessarily so)Low purity (but not necessarily so) Low purity (but not necessarily so)Low purity (but not necessarily so) Low cost (e.g. $5Low cost (e.g. $5--40 per kg)40 per kg) Low profit marginsLow profit margins

Speciality enzymesSpeciality enzymes Low volume (g Low volume (g kg)kg) High purityHigh purity High cost ($5 High cost ($5 10,000 per g)10,000 per g) High profit marginsHigh profit margins High profit marginsHigh profit margins

Important Factors in Using Enzymesp g y

Reactions possible that are not possible using chemistry

Specificity of reaction including substrate specificity, positional specificity, stereo-specificity

Allows milder process conditions e.g. temperature, pH, sterility etc.

Reduces number of process steps required

Eliminates the need to use organic solvents in processingg p g

Immobilization of enzyme to allow its reuse or continuous use

Use of enzymes in combination with other separate chemical steps

Genetic engineering to improve enzymes Genetic engineering to improve enzymes

Industrial enzymesIndustrial enzymesIndustrial enzymesIndustrial enzymes Food processingFood processingp gp g

Amylases in breadAmylases in bread--makingmaking Lipases in flavor developmentLipases in flavor development

P t s s i h s kiP t s s i h s ki Proteases in cheese makingProteases in cheese making Pectinases in clarifying fruit juicesPectinases in clarifying fruit juices

TextilesTextiles TextilesTextiles Cellulases in treating denim to generate stoneCellulases in treating denim to generate stone--washed washed

texture/appearancetexture/appearance

Grain processingGrain processing Conversion of corn starch to high fructose syrupsConversion of corn starch to high fructose syrups

I d t i l I d t i l Industrial enzymesIndustrial enzymesF d F d Feed enzymesFeed enzymes Enzymes to assist in the Enzymes to assist in the

digestibility of animal feeds digestibility of animal feeds ( ll l l ( ll l l

Waste managementWaste management

(cellulase, xylanase, (cellulase, xylanase, phytase)phytase)

Lipases as drainLipases as drain cleaning cleaning gg

Di i Di i

Lipases as drainLipases as drain--cleaning cleaning agentsagents

Reporter enzymes (alkaline Reporter enzymes (alkaline Diagnostic enzymesDiagnostic enzymes Reporter enzymes (alkaline Reporter enzymes (alkaline phosphatase, glucose phosphatase, glucose oxidase, oxidase, --glucosidase) and glucosidase) and g )g )diagnostic enzymes (DNA diagnostic enzymes (DNA polymerase)polymerase)

A li ti i f d i d tA li ti i f d i d tApplications in food industryApplications in food industryb h d b h d Carbohydrases: Carbohydrases:

production of corn syrups from starch production of corn syrups from starch ( l l ) ( l l ) (glucoamylase); (glucoamylase);

conversion of cereal starches into fermentable conversion of cereal starches into fermentable i lti b i di till i lti b i di till sugars in malting, brewing, distillery, sugars in malting, brewing, distillery,

baking industry (amylase).baking industry (amylase).P d i (b li P d i (b li Proteases: meat tenderizers (bromelin, Proteases: meat tenderizers (bromelin, papain, ficin)papain, ficin)

Lipases: Flavor production in chocolate and Lipases: Flavor production in chocolate and cheesecheese

li ili iApplicationsApplicationsGl Gl idid Glucose Glucose oxidaseoxidase: : desugaringdesugaring of eggs, flour and potatoes to prevent of eggs, flour and potatoes to prevent

browning;browning;gg preparation of salad dressings. preparation of salad dressings.

PectinasesPectinases: : l ifi ti f f it j i i f i ld f j i l ifi ti f f it j i i f i ld f j i clarification of fruit juices; increase of yield of juice clarification of fruit juices; increase of yield of juice

from grapes and other products; from grapes and other products; removal of excess pectin from juices before removal of excess pectin from juices before

concentration.concentration.

A li ti td A li ti td Applications contd. Applications contd. bl h f fl bl h f fl Lipoxygenase: bleaching of flours.Lipoxygenase: bleaching of flours.

Phosphatase: quality testing of food Phosphatase: quality testing of food p q y gp q y gproductsproducts

Phenol oxidase: imparts the characteristic Phenol oxidase: imparts the characteristic Phenol oxidase imparts the characteristic Phenol oxidase imparts the characteristic dark hue to tea, cocoa, coffee and raisins.dark hue to tea, cocoa, coffee and raisins.

Renin (chymosin): cheese productionRenin (chymosin): cheese production Renin (chymosin): cheese productionRenin (chymosin): cheese production

ApplicationsApplications FlavorasesFlavorases: restoration and enrichment : restoration and enrichment FlavorasesFlavorases: restoration and enrichment : restoration and enrichment

of flavor by addition of enzyme of flavor by addition of enzyme preparations to food products e g fresh preparations to food products e g fresh preparations to food products e.g. fresh preparations to food products e.g. fresh corn enzyme extracts to improve flavor corn enzyme extracts to improve flavor of of cannnedcannned goods or addition of goods or addition of alliinasealliinaseof of cannnedcannned goods or addition of goods or addition of alliinasealliinaseto convert to convert alliinalliin of garlic into garlic oil.of garlic into garlic oil.

L'alliina un amminoacido non proteinogeno ed un solfossido naturale costituentedell'aglio fresco, insieme a cicloalliina, isoalliina e metiina. La sua biosintesi trae originedall'amminoacido cisteina.

Alliina ed isoalliina sottoposte ad azione enzimatica da parte dell'alliinasi (alliina liasi)Alliina ed isoalliina, sottoposte ad azione enzimatica da parte dell alliinasi (alliina liasi),contenuta nel sistema di vacuoli cellulari della stessa pianta vengono convertite neisulfonati, in primis allicina passando per intermedi sulfenici, La trasformazione avvieneanche solo per schiacciamento degli spicchi, e produce il caratteristico odore. Il composto poco stabile, e per prevenirne l'ossidazione va mantenuto a 4 C.

A lA lAmylasesAmylasesl h l h l l Alpha Alpha amylase amylase cuts 1,4 bondscuts 1,4 bonds Yields Yields dextrinsdextrins and oligosaccharidesand oligosaccharides Yields Yields dextrinsdextrins and oligosaccharidesand oligosaccharides

BetaBeta--amylase amylase maltose units onlymaltose units only Combo Combo produces almost all maltoseproduces almost all maltose Combo Combo -- produces almost all maltoseproduces almost all maltose GlucoGluco--amylase amylase glucoseglucose PullunasePullunase PullunasePullunase --

Cuts beta Cuts beta -- 1,6 linkages 1,6 linkages

Corn starch processing 1Corn starch processing 1Maize grain Corn steep liquor

EndospermEdible oilOil meal

Germ

Hulls

Gluten

Starch Industrial andfood uses

Short chain

Corn syrups F d dditi

Short chain dextrins (foods)

Maltose syrups Corn syrups

Ethanol

Food additives

High fructose syrupsEthanol

composto da due polimeri: l'amilosio (che ne costituisce circa il 20%) e l'amilopectina (circal'80%). In entrambi i casi si tratta di polimeri del glucosio, che si differenziano l'uno dall'altroper la struttura. L'amilosio un polimero lineare che tende ad avvolgersi ad elica, in cui le

it di l i l t t l l i li idi i (1 4) (t il it 1 di itunit di glucosio sono legate tra loro con legami glicosidici (14) (tra il sito 1 di una unit equello 4 dell'unit successiva). L'elica costituita da 6 molecole di glucopiranosio (glucosio)per spira, stabilizzate da legami idrogeno come nel DNA. L'amilopectina invece un polimeroramificato che presenta catene di base di struttura simile all'amilosio che si dispongono aram f cato ch pr s nta cat n as struttura s m a am os o ch s spongono aformare una struttura ramificata; ogni 24-30 unit di glucosio si innestano catene lateraliattraverso legami (16).

H si t t l i s l bilit i t t bi t i l s l bili i Ha quasi totale insolubilit in acqua a temperatura ambiente; una parziale solubilizzazione possibile attraverso un aumento della temperatura, e porta alla formazione di un gel.

Il processo influenzato dall'origine botanica dell'amido e dalla quantit di acqua.p f g q qIl processo di gelatinizzazione modifica la digeribilit dell'amido: la forma cristallina difficilmente attaccabile dagli enzimi (quali - e -amilasi), mentre quella gelatinizzata lo dipi in conseguenza della disorganizzazione strutturale avvenuta.

--amilasi amilasi ((endoamilasiendoamilasi))appartenenteappartenente allaalla classeclasse delledelle idrolasiidrolasi checheappartenenteappartenente allaalla classeclasse delledelle idrolasi,idrolasi, chechecatalizzacatalizza l'l'endoendo--idrolisiidrolisi deidei legamilegami 11,,44----DD--glucosidiciglucosidici inin oligosaccaridioligosaccaridi ee polisaccaridipolisaccaridiglucosidiciglucosidici inin oligosaccaridioligosaccaridi ee polisaccaridipolisaccaridicontenenticontenenti tretre residuiresidui oo pipi..

L'enzimaL'enzima agisceagisce inin modomodo casualecasuale susu amido,amido,glicogenoglicogeno ee molecolemolecole adad esseesse correlatecorrelate.. IlIl terminetermine sisiriferisceriferisce allaalla configurazioneconfigurazione deglidegli zuccherizuccheri prodottiprodotti (che(che presentanopresentano infattiinfatti unaunaconfigurazioneconfigurazione ),), ee nonnon alal tipotipo didi legamelegame glucosidicoglucosidico cheche vieneviene scissoscisso..gg ),), pp gg gg

LeLe alfaalfa--amilasiamilasi produconoproducono prevalentementeprevalentementeLeLe alfaalfa amilasiamilasi produconoproducono prevalentementeprevalentementedestrinedestrine

--amilasi amilasi ((esoamilasiesoamilasi) ) d lld ll ll d lld ll d ld l hh ll ll ddenzimienzimi delladella classeclasse delledelle idrolasi,idrolasi, cheche catalizzanocatalizzano lala digestionedigestione

dell'amido,dell'amido, ossiaossia l'idrolisil'idrolisi deidei legamilegami ((1144))----DD--glucosidiciglucosidici neineipolisaccaridi,polisaccaridi, rimuovendorimuovendo didi fattofatto lele unitunit didi maltosiomaltosio dalledalle estremitestremitppnonnon riducentiriducenti delledelle catenecatene

agisconoagiscono sull'amidosull'amido sulsul glicogenoglicogeno ee suisui polipoli eded oligosaccaridioligosaccaridi correlaticorrelatiagisconoagiscono sull amido,sull amido, sulsul glicogenoglicogeno ee suisui polipoli-- eded oligosaccaridioligosaccaridi correlati,correlati,producendoproducendo --maltosiomaltosio attraversoattraverso unauna inversioneinversione..LaLa letteralettera sisi riferisceriferisce all'inizialeall'iniziale configurazioneconfigurazione anomericaanomerica dellodello zuccherozucchero liberolibero rilasciato,rilasciato, ee nonnonallaalla configurazioneconfigurazione deldel legamelegame idrolizzatoidrolizzatoallaalla configurazioneconfigurazione deldel legamelegame idrolizzatoidrolizzato..

sonosono pipi selettiveselettive rispettorispetto allealle alfaalfa--amilasi,amilasi, lele qualiquali "tagliano""tagliano" aa casocaso lalamolecolamolecola;; sonosono utilizzateutilizzate perper produrreprodurre maltosiomaltosio durantedurante ilil processoprocesso didi""mashingmashing"" (o(o "ammostamento")"ammostamento")

AA differenzadifferenza delledelle alfaalfa--amilasi,amilasi, cheche sonosono presentipresenti neinei vegetalivegetali ee iningrangran parteparte deglidegli animali,animali, lele betabeta--amilasiamilasi sonosono presentipresenti solosolo neinei batteribatteri

ii l il i i lii liee inin alcunialcuni animalianimali

Production of High Fructose Corn Syrupsfrom Starch

Corn Starch Slurry (30-35% DS( dissolved solids), pH 6.0-

from Starchy ( ( ), p

6.5, Ca2+ 50 ppm)Liquefaction

Thermostable -AmylaseGelatinization (105C 5 min)Gelatinization (105 C, 5 min)Dextrinization (95C, 2h)

Liquefied Starch DE (dextrose equivalent) 10-15Saccharification

Glucoamylase(60C, pH 4.0-4.5, 24-72 h)

Glucose Syrups DE 95-96Isomerization

Glucose isomerase(pH 7.5-8.0, 55-60C, 5 mM Mg2+)

High Fructose Corn Syrups (42% fructose)g y p ( )

Production of Glucose from StarchProduction of Glucose from Starch____________________________________________________________Liquefaction Saccharification DE Glucose_____________________________________________________________Acid Acid 92 85

Acid Glucoamylase 95 91

Acid/-amylase Glucoamylase 96 92

-Amylase/High pressure Glucoamylase 97 93cooking/ -amylasecooking/ amylase

-Amylase (thermostable) Glucoamylase 97 94

-Amylase (thermostable) Glucoamylase 97-98.5 95-97.5____________________________________________________________

Enzyme step 1: Enzyme step 1: Action Action of of T m m lT m m l n st h n l s n st h n l sTermamylTermamyl on starch granules on starch granules

TermamylTermamyl is an is an --amylase (cleaves amylase (cleaves --11--4 4 glucosidicglucosidic bonds in starch)bonds in starch)Hi h t t ds st h Hi h t t ds st h High temperature expands starch High temperature expands starch granules, making granules, making amyloseamylose and and amylopectinamylopectin chains more accessiblechains more accessible Maltose concentration

TermamylTermamyl is sufficiently stable at is sufficiently stable at high temperatures if short reaction high temperatures if short reaction times are usedtimes are usedtimes are usedtimes are used

Starch hydrolysis is a batch process Starch hydrolysis is a batch process (the enzyme is not (the enzyme is not reused)reused) Amylase activity

0 10(minutes)( )

Enzyme step 2: Enzyme step 2: Conversion Conversion of maltose to glucoseof maltose to glucose

AmyloglucosidaseAmyloglucosidase is not as is not as thermostablethermostableas as TermamylTermamyl (temperature must be (temperature must be

d d)d d)reduced)reduced) AmyloglucosidaseAmyloglucosidase has a pH optimum of has a pH optimum of

6 5 (6 5 (T lT l t ti ll t t ti ll t 6.5 (6.5 (TermamylTermamyl operates optimally at operates optimally at 8.5): pH must be reduced8.5): pH must be reducedR ti ki ti lR ti ki ti l Reaction kinetics are slowerReaction kinetics are slower

Long incubations result in Long incubations result in caramelizationcaramelizationf th f th h idh id lti i d t lti i d t of the of the saccharidessaccharides -- resulting in product resulting in product

loss and increase in impuritiesloss and increase in impurities

Enzyme step 3: Conversion of Enzyme step 3: Conversion of l fl fglucose to fructoseglucose to fructose

FructoseFructose isis muchmuch sweetersweeter thanthan glucoseglucose;; itit cancan bebe usedusedt it i tt ii f d t fff d t ff dd iiasas aa sweeteningsweetening agentagent inin foodstuffs,foodstuffs, andand isis moremore

profitableprofitable thanthan glucoseglucose The enzyme The enzyme xylosexylose isomeraseisomerase will convert glucose to will convert glucose to

fructose, in an equilibrium reactionfructose, in an equilibrium reactionGlucose Glucose '' FructoseFructose

A 50:50 mixture of A 50:50 mixture of glucose:fructoseglucose:fructose is sold as high is sold as high gg ggfructose syrup (HFS)fructose syrup (HFS)

XyloseXylose (glucose) (glucose) isomeraseisomerase is much less is much less thermostablethermostable, , XyloseXylose (glucose) (glucose) somerasesomerase s much less s much less thermostablethermostable, , and inhibited by Ca ions.and inhibited by Ca ions.

Enzymes in Processing and food Enzymes in Processing and food StorageStorage

P l h lP l h l idid f it tf it t PolyphenolPolyphenol oxidaseoxidase fruit storagefruit storage Amylase Amylase DE StarchesDE Starches Protease Protease rennin/rennin/chymosinchymosin and and ficinficin (beer (beer

clarifier)clarifier) Lipase Lipase hydrolytic rancidityhydrolytic rancidity LipoxidaseLipoxidase oxidizes fatsoxidizes fatspp Muscle Muscle tendrizertendrizer bromelinbromelin from from pinepapplepinepapple

E ti b iE ti b iEnzymatic browningEnzymatic browning Phenolic substances Phenolic substances from brown to black from brown to black

pigmentspigmentsp gp g Enzymes Enzymes + S + S -- Brown color Brown color ((melanosismelanosis)) PolyphenolPolyphenol--oxidaseoxidase (Cu(Cu++ dependent)dependent)ypyp ( u( u p )p )

ECEC. 1.14.18.1. 1.14.18.1 Need Oxygen and tissue damageNeed Oxygen and tissue damage Need Oxygen and tissue damageNeed Oxygen and tissue damage Present in foods Present in foods Banana, Apples, Pear, Banana, Apples, Pear,

Peaches Tea leaves Coffee beans Peaches Tea leaves Coffee beans Peaches, Tea leaves, Coffee beans Peaches, Tea leaves, Coffee beans (desirable).(desirable).

I hibiti f I hibiti f Inhibition of enzymesInhibition of enzymesS lfit S lfit t ith t ith ii t t t t Sulfite Sulfite reacts with reacts with quinonequinone to prevent to prevent further chemical stepsfurther chemical stepsHH i ( it i id )i ( it i id ) pHpH-- vinegar (citric acids)vinegar (citric acids)

Sodium Sodium h t h h th t h h t // b tb t / it t/ it thexametaphosphatehexametaphosphate//ascorbateascorbate/citrate/citrate

EDTA (binds copper of PPO)EDTA (binds copper of PPO) Sugar (limit oxygen diffusion)Sugar (limit oxygen diffusion) Vacuum packageVacuum packagep gp g CysteineCysteine Blanching Blanching Blanch ng Blanch ng IrradiationIrradiation

Color and bleachColor and bleachl dl d bb l ll l hh h hh h ff ColoredColored substancessubstances containcontain moleculesmolecules withwith chromophoreschromophores,, areasareas ofof

thethe moleculemolecule thatthat havehave doubledouble bondsbonds betweenbetween carboncarbon atomsatoms ororoxygenoxygen atomsatoms.. AA goodgood exampleexample isis betabeta carotene,carotene, andand thatthat sectionsectionygyg gg ppgoesgoes intointo moremore detaildetail onon howhow moleculesmolecules becomebecome coloredcolored..

BleachesBleaches attackattack thesethese chromophoreschromophores inin oneone ofof twotwo waysways..

OxidizingOxidizing bleachesbleaches likelike sodiumsodium hypochloritehypochlorite breakbreak thethe moleculesmolecules atatthethe doubledouble bondbond.. ThisThis resultsresults inin eithereither aa shortershorter moleculemolecule thatthat doesdoesnotnot absorbabsorb visiblevisible lightlight oror aa moleculemolecule whosewhose chromophorechromophore isis eithereithernotnot absorbabsorb visiblevisible light,light, oror aa moleculemolecule whosewhose chromophorechromophore isis eithereithershortershorter oror nonnon--existantexistant.. AA shortershorter chromophorechromophore willwill absorbabsorb lightlight ofofaa shortershorter wavelengthwavelength thanthan visiblevisible lightlight (such(such asas ultravioletultraviolet light),light), andandsoso doesdoes notnot appearappear coloredcoloredsoso doesdoes notnot appearappear coloredcolored..

ReducingReducing bleachesbleaches suchsuch asas lemonlemon juicejuice (in(in combinationcombination withwith sunlight)sunlight)oror sulfursulfur dioxidedioxide convertconvert thethe doubledouble bondsbonds inin thethe chromophorechromophore intointooror sulfursulfur dioxidedioxide,, convertconvert thethe doubledouble bondsbonds inin thethe chromophorechromophore intointosinglesingle bonds,bonds, eliminatingeliminating itsits abilityability toto absorbabsorb visiblevisible lightlight..SometimesSometimes thethe reactionreaction isis reversible,reversible, wherewhere oxygenoxygen inin thethe airair reactsreactswithwith thethe moleculemolecule toto repairrepair thethe chromophorechromophore andand thethe stainstain returnsreturnswithwith thethe moleculemolecule toto repairrepair thethe chromophorechromophore,, andand thethe stainstain returnsreturns..

UUUsesUses B k f B k f In Baking to increase fermentation rateIn Baking to increase fermentation rate

Corn syrupCorn syrupy py p Liquid center chocolates Liquid center chocolates Clarification of Apple juiceClarification of Apple juice Clarification of Apple juiceClarification of Apple juice

Li d E tLi d E tLipase and EsteraseLipase and EsteraseFF d d FF d d d d FFA are produced FFA are produced may cause ranciditymay cause rancidity

Form mono and Form mono and diglyceridesdiglyceridesg yg y But in Cheese it is desirableBut in Cheese it is desirable In seeds it is destroyed by heatIn seeds it is destroyed by heat In seeds it is destroyed by heatIn seeds it is destroyed by heat 1,3 Specific enzymes (position on glycerol) 1,3 Specific enzymes (position on glycerol)

T il ki f b b iT il ki f b b i Tailor making of cocoa butter substituteTailor making of cocoa butter substitute

I bili d EI bili d EImmobilized EnzymesImmobilized EnzymesE l b d E l b d Enzyme in solution can be used onceEnzyme in solution can be used once

It can be fixed on a carrier so can be used It can be fixed on a carrier so can be used continuouslycontinuously

It can be bound, adsorbed, entrapped or It can be bound, adsorbed, entrapped or It can be bound, adsorbed, entrapped or It can be bound, adsorbed, entrapped or crosslinkedcrosslinked

They are more heat stable pH is shiftedThey are more heat stable pH is shifted They are more heat stable, pH is shiftedThey are more heat stable, pH is shifted

O id d tO id d tOxidoreductaseOxidoreductaseGl Gl dd Glucose Glucose oxidaseoxidase From From AspergillusAspergillus nigerniger andand PenicilliumPenicillium notatumnotatum Used in removal of glucose and oxygenUsed in removal of glucose and oxygen

HH22OO22 is produced and is destroyed by is produced and is destroyed by 22 22 p y yp y yCatalaseCatalase

HH22OO22 is used sometime to is used sometime to pasterurizepasterurize milk milk HH22OO22 is used sometime to is used sometime to pasterurizepasterurize milk milk and the excess is removed by and the excess is removed by CatalaseCatalase

Other applicationsOther applications AldehydeAldehyde dehydrogenasedehydrogenase AldehydeAldehyde dehydrogenasedehydrogenase

Unsaturated FA in Soy produce Unsaturated FA in Soy produce hexanalhexanal (bean like (bean like flavor) flavor)

ButanediolButanediol DehydrogenaseDehydrogenase DiacetylDiacetyl formed during beer productionformed during beer production

T l t iT l t i TransglutaminaseTransglutaminase crosslinkingcrosslinking enzyme enzyme lysine and lysine and glutamicglutamic acidacid

NaringinaseNaringinase NaringinaseNaringinase hydrolyzes bitter hydrolyzes bitter nariginnarigin to to naringeninnaringenin