ENZYMES in Food ENZYMES in Food ENZYMES in Food ENZYMES in Food

Processing Processing Processing Processing

Useful links for Useful links for ENZYMES ENZYMES
d d in Food Processing in Food Processing
http://www.amfep.org/ http://www.amfep.org/index.html index.html
http://www.efsa.europa.eu/ http://www.efsa.europa.eu/
http://www foodnavigator com/ http://www foodnavigator com/Product Product Categories Categories//Cultures Cultures enzymes enzymes yeast yeast http://www.foodnavigator.com/ http://www.foodnavigator.com/Product Product--Categories Categories//Cultures Cultures--enzymes enzymes--yeast yeast
Enzyme Enzyme database database yy
http://www.brenda http://www.brenda- -enzymes.org/ enzymes.org/
http://www.expasy.ch/ http://www.expasy.ch/enzyme enzyme/ /
http://www.enzyme http://www.enzyme- -database.org/ database.org/
http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intenz intenz//
http://www biochem ucl ac uk/ http://www biochem ucl ac uk/bsm bsm//enzymes enzymes// http://www.biochem.ucl.ac.uk/ http://www.biochem.ucl.ac.uk/bsm bsm//enzymes enzymes//
http://metacyc.org/ http://metacyc.org/
P ti f P ti f Properties of enzymes Properties of enzymes
l l Control ripening. Control ripening.
Cause food spoilage (rotting). Cause food spoilage (rotting). p g g p 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 foods storage stability of foods storage stability of foods. storage stability of foods.
P ti P ti Properties Properties
U d f f t ti i f d U 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 d E i I d Enzymes in Industry Enzymes in Industry
Distribution of enzymes by substrate Distribution of enzymes by substrate
Protein hydrolyzing Protein hydrolyzing 59% 59%
Carbohydrate hydrolyzing Carbohydrate hydrolyzing 28% 28% Carbohydrate hydrolyzing Carbohydrate hydrolyzing 28% 28%
Lipid hydrolyzing Lipid hydrolyzing 3% 3%
Speciality (analytical, pharma, research) 10% Speciality (analytical, pharma, research) 10%
US Market US Market
Bakery Bakery
US$ 210m 5 US$ 210m 5--7% pa 7% pa
Beverage Beverage
US$ 130m, 4 US$ 130m, 4--6% pa 6% pa
Dairy Dairy
US$ 195m, 3 US$ 195m, 3--5% pa 5% pa
Fats Fats & Oils & Oils
US$ 30m, 10 US$ 30m, 10--12% pa 12% pa
Culinary Culinary
US$ 30m, 5 US$ 30m, 5--6% pa 6% pa $ , $ , pp
Meat and others ( Meat and others (incl. food protection) incl. food protection)
US$ 30m 10 US$ 30m 10--12% pa 12% pa US$ 30m, 10 US$ 30m, 10 12% pa 12% pa
A f l tit A f l tit A few large competitors A few large competitors
Novozymes (all applications) Novozymes (all applications) y pp y 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 tit th tit other competitors other competitors
Advanced Enzymes Advanced 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 Biochemicals Biochemicals, is headquartered , is headquartered
in California in the US) in California in the US) f ) f )
http://www.ingredientsnetwork.com/profile/full/advanced http://www.ingredientsnetwork.com/profile/full/advanced--enzymes enzymes- -
food food--enzymes enzymes yy
produces more than $100 million worth of produces more than $100 million worth of probiotics probiotics 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 pp
engineers, 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 non Founded in 1977, the is a non- -profit European industry association profit European industry association
The main objectives of AMFEP are The main objectives of AMFEP are::
to to provide provide aa common common basis basis for for representing representing the the interests interests of of its its members members both both
t d t d th th i tit ti i tit ti ff th th EE U i U i ll ll ti l ti l th iti th iti toward toward the the institutions institutions of of the the European European Union Union as as well well as as national national authorities authorities; ;
to to represent represent the the interests interests of of its its members members in in international international organisations organisations; ;
to to assure assure a a free free flow flow of of information information between between its its members members on on developments developments
related related to to the the regulatory regulatory status status of of enzymes enzymes in in the the EU EU;; related related to to the the regulatory regulatory status status of of enzymes enzymes in in the the EU EU;;
to to inform inform its its customers customers and and other other interested interested parties parties on on the the efficacy efficacy and and safety safety
aspects aspects of of its its enzyme enzyme products products; ;
to to defend defend and and promote promote the the products products of of the the enzyme enzyme industry industry..
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 non other non-- food industries, excluding enzymes for pharmaceutical and diagnostic food industries, excluding enzymes for pharmaceutical and diagnostic
use. use.
Amfep fruitfully co Amfep 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.html http://www.amfep.org/index.html
AMFEP members AMFEP members
Enzymes REACH Consortium (ERC) Enzymes REACH Consortium (ERC)
Industrial Industrial enzymes enzymes are are used used in in aa variety variety of of applications applications some some of of which which require require their their registration registration Industrial Industrial enzymes enzymes are are used used in in aa variety variety of of applications, applications, some some of of which which require require their their registration registration
under under the the so so- -called called REACH REACH Regulation* Regulation* ..
Following Following aa six six- -month month month month pre pre- -registration registration phase phase full full registration registration starts starts on on December December 2010 2010 or or
later later depending depending on on tonnage tonnage later, later, depending depending on on tonnage tonnage..
REACH REACH requires, requires, subject subject to to certain certain exceptions, exceptions, multiple multiple registrants registrants of of the the same same substance substance to to share share
data data and and jointly jointly submit submit part part of of the the relevant relevant registrations registrations..
Manufacturers Manufacturers and and importers importers of of enzymes, enzymes, recognizing recognizing that that enzyme enzyme registration registration under under REACH REACH will will
benefit benefit from from being being discussed discussed and and coordinated, coordinated, have have created created the the Enzymes Enzymes REACH REACH Consortium Consortium (ERC) (ERC)
with with the the overall overall purpose purpose of of facilitating facilitating aa smooth smooth REACH REACH implementation, implementation, hereunder hereunder by by creating creating
effective effective data data sharing sharing processes processes and and aa reduction reduction of of associated associated costs costs.. effective effective data data sharing sharing processes processes and and aa reduction reduction of of associated associated costs costs..
ERC ERC intends intends to to discuss discuss and and develop develop a a harmonized harmonized approach approach to to aspects aspects such such as as nomenclature nomenclature and and
classification, classification, sameness, sameness, read read--across across and and data data waiving waiving..
The The approach approach will will be be based based on on the the common common features features that that are are shared shared by by all all enzymes enzymes..
Unlike Unlike many many chemicals, chemicals, enzymes enzymes are are aa unique unique set set of of substances substances that that have have predictable predictable properties properties.. For For
instance instance all all enzymes enzymes are are classified classified as as respiratory respiratory sensitizers sensitizers regardless regardless of of their their biological biological source source instance, instance, all all enzymes enzymes are are classified classified as as respiratory respiratory sensitizers sensitizers,, regardless regardless of of their their biological biological source source
and and catalytic catalytic activity activity..
http://www.enzymes http://www.enzymes- -reach.org/index.html reach.org/index.html
*Regulation *Regulation (EC) (EC) No No 1907 1907//2006 2006 of of the the European European Parliament Parliament and and of of the the Council Council of of 18 18 December December
2006 2006 concerning concerning the the Registration, Registration, Evaluation, Evaluation, Authorisation Authorisation and and Restriction Restriction of of Chemicals Chemicals
(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 members Full members which are Manufacturers Importers and/or Only which are Manufacturers Importers and/or Only -- Full members Full 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 members Associate 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 Pre contributing 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 Skaskevitch Mr. Youri Skaskevitch
Telephone: +32 (0)2 740 29 62 Telephone: +32 (0)2 740 29 62
EE il il di @ di @ hh EE--mail: mail: coordinator@enzymes coordinator@enzymes- -reach.org reach.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 Report A Global Market Report Industrial Enzymes Industrial Enzymes -- A Global Market Report A 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 Reactions Enzymatic Reactions
High fructose corn syrup: $ 1 billion
Aspartame: $ 850 million Aspartame: $ 850 million
Acrylamide: $ 300 million
Acrylamide (or acrylic amide) Acrylamide (or acrylic amide)
chemical compound with the chemical formula C chemical compound with the chemical formula C
33
HH
55
NO. NO.
white white odourless odourless 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 acrylonitrile acrylonitrile by by nitrile nitrile
hydratase hydratase..
Acrylamide Acrylamide also occurs in many cooked starchy foods. also occurs in many cooked starchy foods. Acrylamide Acrylamide 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 acrylamide acrylamide in the heating process in the heating process
was shown to be temperature was shown to be temperature- -dependent dependent). ).
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 poliacrilammidi poliacrilammidi 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 Cl I d t i l E Cl Industrial Enzyme Classes Industrial Enzyme Classes
Commodity enzymes Commodity enzymes Commodity enzymes Commodity 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. $5 Low cost (e.g. $5- -40 per kg) 40 per kg)
Low profit margins Low profit margins
Speciality enzymes Speciality enzymes
Low volume (g Low volume (g kg) kg)
High purity High purity
High cost ($5 High cost ($5 10,000 per g) 10,000 per g)
High profit margins High profit margins High profit margins High profit margins
Important Factors in Using Enzymes p 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 processing g 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 enzymes Industrial enzymes Industrial enzymes Industrial enzymes
Food processing Food processing p g p g
Amylases in bread Amylases in bread--making making
Lipases in flavor development Lipases in flavor development
P t s s i h s ki P t s s i h s ki Proteases in cheese making Proteases in cheese making
Pectinases in clarifying fruit juices Pectinases in clarifying fruit juices
Textiles Textiles Textiles Textiles
Cellulases in treating denim to generate stone Cellulases in treating denim to generate stone--washed washed
texture/appearance texture/appearance
Grain processing Grain processing
Conversion of corn starch to high fructose syrups Conversion of corn starch to high fructose syrups
I d t i l I d t i l Industrial enzymes Industrial enzymes
F d F d Feed enzymes Feed 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 management Waste management
(cellulase, xylanase, (cellulase, xylanase,
phytase) phytase)
Lipases as drain Lipases as drain cleaning cleaning
gg
Di i Di i
Lipases as drain Lipases as drain--cleaning cleaning
agents agents
Reporter enzymes (alkaline Reporter enzymes (alkaline
Diagnostic enzymes Diagnostic 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 t A li ti i f d i d t Applications in food industry Applications in food industry
b 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
cheese cheese
li i li i Applications Applications
Gl Gl id id Glucose Glucose oxidase oxidase: :
desugaring desugaring 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.
Pectinases Pectinases: :
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 g p q y g
products products
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 production Renin (chymosin): cheese production Renin (chymosin): cheese production Renin (chymosin): cheese production
Applications Applications
Flavorases Flavorases: restoration and enrichment : restoration and enrichment Flavorases Flavorases: 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 cannned cannned goods or addition of goods or addition of alliinase alliinase of of cannned cannned goods or addition of goods or addition of alliinase alliinase
to convert to convert alliin alliin of garlic into garlic oil. of garlic into garlic oil.
L'alliina un amminoacido non proteinogeno ed un solfossido naturale costituente
dell'aglio fresco, insieme a cicloalliina, isoalliina e metiina. La sua biosintesi trae origine
dall'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 nei
sulfonati, in primis allicina passando per intermedi sulfenici, La trasformazione avviene
anche 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 l A l Amylases Amylases
l h l h l l Alpha Alpha amylase amylase
cuts 1,4 bonds cuts 1,4 bonds
Yields Yields dextrins dextrins and oligosaccharides and oligosaccharides Yields Yields dextrins dextrins and oligosaccharides and oligosaccharides
Beta Beta- -amylase amylase maltose units only maltose units only
Combo Combo produces almost all maltose produces almost all maltose Combo Combo -- produces almost all maltose produces almost all maltose
Gluco Gluco- -amylase amylase glucose glucose
Pullunase Pullunase Pullunase Pullunase --
Cuts beta Cuts beta - - 1,6 linkages 1,6 linkages
Corn starch processing 1 Corn starch processing 1
Maize grain
Corn steep liquor
Endosperm
Edible oil
Oil meal
Germ
Hulls
Gluten
Starch
Industrial and
food uses
Short chain
Corn syrups
F d dditi
Short chain
dextrins (foods)
Maltose syrups
Corn syrups
Ethanol
Food additives
High fructose syrups
Ethanol
composto da due polimeri: l'amilosio (che ne costituisce circa il 20%) e l'amilopectina (circa
l'80%). In entrambi i casi si tratta di polimeri del glucosio, che si differenziano l'uno dall'altro
per 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 it unit di glucosio sono legate tra loro con legami glicosidici (14) (tra il sito 1 di una unit e
quello 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 polimero
ramificato che presenta catene di base di struttura simile all'amilosio che si dispongono a ram f cato ch pr s nta cat n as struttura s m a am os o ch s spongono a
formare una struttura ramificata; ogni 24-30 unit di glucosio si innestano catene laterali
attraverso 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 q
Il processo di gelatinizzazione modifica la digeribilit dell'amido: la forma cristallina
difficilmente attaccabile dagli enzimi (quali - e -amilasi), mentre quella gelatinizzata lo di
pi in conseguenza della disorganizzazione strutturale avvenuta.
--amilasi amilasi ((endoamilasi endoamilasi))
appartenente appartenente alla alla classe classe delle delle idrolasi idrolasi che che appartenente appartenente alla alla classe classe delle delle idrolasi, idrolasi, che che
catalizza catalizza l' l'endo endo--idrolisi idrolisi dei dei legami legami 1 1,,4 4----D D- -
glucosidici glucosidici in in oligosaccaridi oligosaccaridi ee polisaccaridi polisaccaridi glucosidici glucosidici in in oligosaccaridi oligosaccaridi ee polisaccaridi polisaccaridi
contenenti contenenti tre tre residui residui oo pi pi..
L'enzima L'enzima agisce agisce in in modo modo casuale casuale su su amido, amido,
glicogeno glicogeno e e molecole molecole ad ad esse esse correlate correlate.. Il Il termine termine si si
riferisce riferisce alla alla configurazione configurazione degli degli zuccheri zuccheri prodotti prodotti (che (che presentano presentano infatti infatti una una
configurazione configurazione ), ), ee non non al al tipo tipo di di legame legame glucosidico glucosidico che che viene viene scisso scisso.. gg ), ), pp gg gg
Le Le alfa alfa--amilasi amilasi producono producono prevalentemente prevalentemente Le Le alfa alfa amilasi amilasi producono producono prevalentemente prevalentemente
destrine destrine
--amilasi amilasi ((esoamilasi esoamilasi) )
d ll d ll ll d ll d ll d l d l hh ll ll dd enzimi enzimi della della classe classe delle delle idrolasi, idrolasi, che che catalizzano catalizzano la la digestione digestione
dell'amido, dell'amido, ossia ossia l'idrolisi l'idrolisi dei dei legami legami ( (1144))----D D- -glucosidici glucosidici nei nei
polisaccaridi, polisaccaridi, rimuovendo rimuovendo di di fatto fatto le le unit unit di di maltosio maltosio dalle dalle estremit estremit pp
non non riducenti riducenti delle delle catene catene
agiscono agiscono sull'amido sull'amido sul sul glicogeno glicogeno ee sui sui poli poli ed ed oligosaccaridi oligosaccaridi correlati correlati agiscono agiscono sull amido, sull amido, sul sul glicogeno glicogeno ee sui sui poli poli-- ed ed oligosaccaridi oligosaccaridi correlati, correlati,
producendo producendo --maltosio maltosio attraverso attraverso una una inversione inversione..
La La lettera lettera si si riferisce riferisce all'iniziale all'iniziale configurazione configurazione anomerica anomerica dello dello zucchero zucchero libero libero rilasciato, rilasciato, e e non non
alla alla configurazione configurazione del del legame legame idrolizzato idrolizzato alla alla configurazione configurazione del del legame legame idrolizzato idrolizzato..
sono sono pi pi selettive selettive rispetto rispetto alle alle alfa alfa- -amilasi, amilasi, le le quali quali "tagliano" "tagliano" a a caso caso la la
molecola molecola; ; sono sono utilizzate utilizzate per per produrre produrre maltosio maltosio durante durante il il processo processo di di
""mashing mashing" " (o (o "ammostamento") "ammostamento")
AA differenza differenza delle delle alfa alfa- -amilasi, amilasi, che che sono sono presenti presenti nei nei vegetali vegetali e e in in
gran gran parte parte degli degli animali, animali, le le beta beta- -amilasi amilasi sono sono presenti presenti solo solo nei nei batteri batteri
ii l i l i i li i li ee in in alcuni alcuni animali animali
Production of High Fructose Corn Syrups
from Starch
Corn Starch Slurry (30-35% DS( dissolved solids), pH 6.0-
from Starch
y ( ( ), p
6.5, Ca
2+
50 ppm)
Liquefaction
Thermostable -Amylase
Gelatinization (105C 5 min) Gelatinization (105 C, 5 min)
Dextrinization (95C, 2h)
Liquefied Starch DE (dextrose equivalent) 10-15
Saccharification
Glucoamylase
(60C, pH 4.0-4.5, 24-72 h)
Glucose Syrups DE 95-96
Isomerization
Glucose isomerase
(pH 7.5-8.0, 55-60C, 5 mM Mg
2+
)
High Fructose Corn Syrups (42% fructose) g y p ( )
Production of Glucose from Starch Production 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 93
cooking/ -amylase cooking/ 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 l T m m l n st h n l s n st h n l s Termamyl Termamyl on starch granules on starch granules
Termamyl Termamyl is an is an --amylase (cleaves amylase (cleaves
--1 1--4 4 glucosidic glucosidic 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 amylose amylose and and
amylopectin amylopectin chains more accessible chains more accessible
Maltose concentration
Termamyl Termamyl is sufficiently stable at is sufficiently stable at
high temperatures if short reaction high temperatures if short reaction
times are used times are used times are used times 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 glucose of maltose to glucose
Amyloglucosidase Amyloglucosidase is not as is not as thermostable thermostable
as as Termamyl Termamyl (temperature must be (temperature must be
d d) d d) reduced) reduced)
Amyloglucosidase Amyloglucosidase has a pH optimum of has a pH optimum of
6 5 ( 6 5 (T l T l t ti ll t t ti ll t 6.5 ( 6.5 (Termamyl Termamyl operates optimally at operates optimally at
8.5): pH must be reduced 8.5): pH must be reduced
R ti ki ti l R ti ki ti l Reaction kinetics are slower Reaction kinetics are slower
Long incubations result in Long incubations result in caramelization caramelization
f th f th h id h id lti i d t lti i d t of the of the saccharides saccharides - - resulting in product resulting in product
loss and increase in impurities loss and increase in impurities
Enzyme step 3: Conversion of Enzyme step 3: Conversion of
l f l f glucose to fructose glucose to fructose
Fructose Fructose is is much much sweeter sweeter than than glucose glucose; ; it it can can be be used used
t i t i tt ii f d t ff f d t ff dd ii as as a a sweetening sweetening agent agent in in foodstuffs, foodstuffs, and and is is more more
profitable profitable than than glucose glucose
The enzyme The enzyme xylose xylose isomerase isomerase will convert glucose to will convert glucose to
fructose, in an equilibrium reaction fructose, in an equilibrium reaction
Glucose Glucose Fructose Fructose
A 50:50 mixture of A 50:50 mixture of glucose:fructose glucose:fructose is sold as high is sold as high gg gg
fructose syrup (HFS) fructose syrup (HFS)
Xylose Xylose (glucose) (glucose) isomerase isomerase is much less is much less thermostable thermostable, , Xylose Xylose (glucose) (glucose) somerase somerase s much less s much less thermostable thermostable, ,
and inhibited by Ca ions. and inhibited by Ca ions.
Enzymes in Processing and food Enzymes in Processing and food
Storage Storage
P l h l P l h l id id f it t f it t Polyphenol Polyphenol oxidase oxidase fruit storage fruit storage
Amylase Amylase DE Starches DE Starches
Protease Protease rennin/ rennin/chymosin chymosin and and ficin ficin (beer (beer
clarifier) clarifier)
Lipase Lipase hydrolytic rancidity hydrolytic rancidity
Lipoxidase Lipoxidase oxidizes fats oxidizes fats pp
Muscle Muscle tendrizer tendrizer bromelin bromelin from from pinepapple pinepapple
E ti b i E ti b i Enzymatic browning Enzymatic browning
Phenolic substances Phenolic substances from brown to black from brown to black
pigments pigments p g p g
Enzymes Enzymes + S + S -- Brown color Brown color ((melanosis melanosis))
Polyphenol Polyphenol- -oxidase oxidase (Cu (Cu
++
dependent) dependent) yp yp ( u ( u p ) p )
EC EC. 1.14.18.1 . 1.14.18.1
Need Oxygen and tissue damage Need Oxygen and tissue damage Need Oxygen and tissue damage Need 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 enzymes Inhibition of enzymes
S lfit S lfit t ith t ith ii t t t t Sulfite Sulfite reacts with reacts with quinone quinone to prevent to prevent
further chemical steps further chemical steps
HH i ( it i id ) i ( it i id ) pH pH-- vinegar (citric acids) vinegar (citric acids)
Sodium Sodium
h t h h t h t h h t // b t b t / it t / it t hexametaphosphate hexametaphosphate//ascorbate ascorbate/citrate /citrate
EDTA (binds copper of PPO) EDTA (binds copper of PPO)
Sugar (limit oxygen diffusion) Sugar (limit oxygen diffusion)
Vacuum package Vacuum package p g p g
Cysteine Cysteine
Blanching Blanching Blanch ng Blanch ng
Irradiation Irradiation
Color and bleach Color and bleach
l d l d bb l l l l hh h h h h ff Colored Colored substances substances contain contain molecules molecules with with chromophores chromophores, , areas areas of of
the the molecule molecule that that have have double double bonds bonds between between carbon carbon atoms atoms or or
oxygen oxygen atoms atoms.. A A good good example example is is beta beta carotene, carotene, and and that that section section yg yg gg pp
goes goes into into more more detail detail on on how how molecules molecules become become colored colored..
Bleaches Bleaches attack attack these these chromophores chromophores in in one one of of two two ways ways..
Oxidizing Oxidizing bleaches bleaches like like sodium sodium hypochlorite hypochlorite break break the the molecules molecules at at
the the double double bond bond.. This This results results in in either either a a shorter shorter molecule molecule that that does does
not not absorb absorb visible visible light light or or aa molecule molecule whose whose chromophore chromophore is is either either not not absorb absorb visible visible light, light, or or aa molecule molecule whose whose chromophore chromophore is is either either
shorter shorter or or non non--existant existant.. AA shorter shorter chromophore chromophore will will absorb absorb light light of of
aa shorter shorter wavelength wavelength than than visible visible light light (such (such as as ultraviolet ultraviolet light), light), and and
so so does does not not appear appear colored colored so so does does not not appear appear colored colored..
Reducing Reducing bleaches bleaches such such as as lemon lemon juice juice (in (in combination combination with with sunlight) sunlight)
or or sulfur sulfur dioxide dioxide convert convert the the double double bonds bonds in in the the chromophore chromophore into into or or sulfur sulfur dioxide dioxide,, convert convert the the double double bonds bonds in in the the chromophore chromophore into into
single single bonds, bonds, eliminating eliminating its its ability ability to to absorb absorb visible visible light light. .
Sometimes Sometimes the the reaction reaction is is reversible, reversible, where where oxygen oxygen in in the the air air reacts reacts
with with the the molecule molecule to to repair repair the the chromophore chromophore and and the the stain stain returns returns with with the the molecule molecule to to repair repair the the chromophore chromophore,, and and the the stain stain returns returns..
UUUses Uses
B k f B k f In Baking to increase fermentation rate In Baking to increase fermentation rate
Corn syrup Corn syrup y p y p
Liquid center chocolates Liquid center chocolates
Clarification of Apple juice Clarification of Apple juice Clarification of Apple juice Clarification of Apple juice
Li d E t Li d E t Lipase and Esterase Lipase and Esterase
FF d d FF d d d d FFA are produced FFA are produced may cause rancidity may cause rancidity
Form mono and Form mono and diglycerides diglycerides g y g y
But in Cheese it is desirable But in Cheese it is desirable
In seeds it is destroyed by heat In seeds it is destroyed by heat In seeds it is destroyed by heat In 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 i T il ki f b b i Tailor making of cocoa butter substitute Tailor making of cocoa butter substitute
I bili d E I bili d E Immobilized Enzymes Immobilized Enzymes
E l b d E l b d Enzyme in solution can be used once Enzyme 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
continuously continuously
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
crosslinked crosslinked
They are more heat stable pH is shifted They are more heat stable pH is shifted They are more heat stable, pH is shifted They are more heat stable, pH is shifted
O id d t O id d t Oxidoreductase Oxidoreductase
Gl Gl dd Glucose Glucose oxidase oxidase
From From Aspergillus Aspergillus niger niger and and Penicillium Penicillium notatum notatum
Used in removal of glucose and oxygen Used in removal of glucose and oxygen
HH
22
OO
22
is produced and is destroyed by is produced and is destroyed by
22 22
p y y p y y
Catalase Catalase
HH
22
OO
22
is used sometime to is used sometime to pasterurize pasterurize milk milk HH
22
OO
22
is used sometime to is used sometime to pasterurize pasterurize milk milk
and the excess is removed by and the excess is removed by Catalase Catalase
Other applications Other applications
Aldehyde Aldehyde dehydrogenase dehydrogenase Aldehyde Aldehyde dehydrogenase dehydrogenase
Unsaturated FA in Soy produce Unsaturated FA in Soy produce hexanal hexanal (bean like (bean like
flavor) flavor)
Butanediol Butanediol Dehydrogenase Dehydrogenase
Diacetyl Diacetyl formed during beer production formed during beer production
T l t i T l t i Transglutaminase Transglutaminase
crosslinking crosslinking enzyme enzyme lysine and lysine and glutamic glutamic acid acid
Naringinase Naringinase Naringinase Naringinase
hydrolyzes bitter hydrolyzes bitter narigin narigin to to naringenin naringenin