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Delivered to:
Dr. Ranjeeta Bhari
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Introduction
Historical background
Advantages
Applications In food industry Amino acids
Dextrins
Fructooligosaccharides
High fructose syrups Vitamins
Vinegar
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BiotranformationBiotranformation//BiocatalysisBiocatalysis
It is the use of biological agents to effect specific
change on compounds that are not part of their normalbiochemistry
Intact microbial cells
Plant cells
Isolated enzymes
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Sumerian and Babylonians were practicing
the brewing of beer before 6000 BC
Production of vinegar is reported in 2000 BC
Knowledge of production of alcohols and
organic acids were 1st reported in second
half of 19th century
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WHY BIOTRANSFORMATION
Simple to manipulate
Can work under mild reaction
conditionsHas high efficiency,
Is regio- and stereoselective
Is environment friendly
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Applications In food
industry
Amino acids
High fructose
syrups
Fructoo-
ligosaccharides
VitaminsVinegar
Dextrins
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Amino Acids
L-ASPARTIC ACID GLUTAMIC ACID
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Sodium aspartate is used
As seasoning in orange juice
Synthesis low calorie sweetner and in soft drink industry
The Enzyme responsible for the reaction isAspartase.This was thefirst enzyme isolated from the microbial cells and catalyses thesynthesis of amino acids.
Substrates used are: Fumaric acid and Ammonia.
Enzymatic reaction:
HO2CCH=CHCO2H + NH3 Aspartic acid
Aspartase
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Strain used: Brevibacterium flavum.
Cultured in a medium containing :
3.6% glucose and 0.5% L- Glutamic acid
Temperature: 30 C.
Incubation time: 4 days.
Enzyme: Aspartase.
Yield :4g/l.
However, Proteus vulgaris has high aspartase activity.
USEOF GLUCOSEAS CARBON SOURCEUSEOF GLUCOSEAS CARBON SOURCE
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USEOFFUMARIC ACID AS CARBON SOURCEUSEOFFUMARIC ACID AS CARBON SOURCE
Strain used: Bacillus megaterium.
Strain is pre-cultured with 0.5% Fumaric acid for 24 hrs(which is neutralized with NH3)
Addition to fermentative broth
Incubation for 72 hrs
Yield-80%
Pseudomonas fluorescens
yield
95% in 3 days.
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E. coli
However it is increased by the use of mutant E.coliKi-1023
derived from E.coliK-12.
0.5g dried E.colicells+100ml of 20% ammoniumfumarate
Incubated at 37C for 18 hrs (pH-7.2 - 7.4)
Yield 88.1%
Addition of surfactant such asAcetyl pyridinium chlorideandAcetyl methyl ammonium bromide released Asparticacid efficiently.
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GLUTAMIC ACID
Mostly produced as monosodium glutamate (MSG)
Microbial strains: Corynebacterium,Brevibacterium,Microbacterium and
Arthrobacter.
Culture conditions:
a) Carbon source- Glucose, Fructose, Maltose, Sucrose. Molassesand starch hydrolysates are also used.
b) Nitrogen source-Ammonium salts such as ammonium chlorideand ammonium sulphate .Urea is also used.
c) pH Control- gaseous ammonia is used to maintain the pH at 7-8.
d) Growth factors- Biotin
g)Temperature- 35C-38C
Yield 50-60 g/l
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CONTINUED..
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MONOSODIUM GLUTAMATEMONOSODIUM GLUTAMATE FLAVORFLAVOR
ENHANCERENHANCER
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ASCORBIC ACID (VITAMINASCORBIC ACID (VITAMIN--C)C)
L-ascorbic acid is used:-
in vitamin preparation
as an anti oxidant.
Reichstein-GrussnerSynthesis.
Consists of several chemical steps and one microbialconversion.
Oxidation stage from D-sorbitol to l-sorbose is carriedout byAcetobacter suboxydans in a submerged processat 30C-35C.
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REICHSTEINREICHSTEIN--GRUSSNER SYNTHESISGRUSSNER SYNTHESIS
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DEXT
RANS
DEXT
RANS
Homopolymer of -D-glucose units united by
-1,6- glycosidic linkages
At the branching positions -1,2; -1,3; and -
1,4 linkages also occurs.
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STRUCTURESTRUCTURE
linkages
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CONTINUED..
Dextrans are produced by a number of Grampositive and Gram negative bacteria likeAerobactersp., Streptococcus sp. andLeuconostocsp.
Commercially, it is mainly produced by Leuconostocsp. with sucrose as substrate.
Generally two species L. dextranicum and L.mesenteroides uses molasses as a substrate forthe large scale.
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GENERALLY TWO METHODS AREGENERALLY TWO METHODS AREUSED:USED:
Whole cell culture fermentation.
Enzymetic synthesis of dextran.
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ENZYMATIC SYNTHESISOFENZYMATIC SYNTHESISOF DEXTRANSDEXTRANS
Jeanes (1965) described a method for enzymatic dextransynthesis with L. mesenteroides NRRLB.512F. The mediumfor the initial stage of inoculum contains-
2.00% sucrose,
0.50% dipotassium acid phosphate,
0.50% yeast extract,
0.25% tryptone.
Bacterial cells are removed by centrifugation. Dextran isproduced enzymatically by adding sucrose to the cell-freeculture liquid at pH 5.0-5.2 and holding the temperature 250-300C.
Here the dextransucrase incorporate only glucose portion,and fructose remain in solution.
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APPLICATIONSOF DEXTRANAPPLICATIONSOF DEXTRAN
Dextran has found industrial applications in food,pharmaceutical and chemical industries asemulsifier, carrier and stabilizer.
In food industry dextran used as thickener forjam and ice cream.
It prevents crystallization of sugar, improves
moisture retention, and maintains flavour andappearance of various food items.
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FRUCTOOLIGOSACCHARIDESFRUCTOOLIGOSACCHARIDES
MICROORGANISMSArthrobactersp. Pseudomonas sp. Streptomyces rochei,Xanthomonas sp.Aspergillusficuum, Aspergillus fumigatus , Aspergillus niger, .Chrysosporium pannorum ,Penicillum sp, Kluyveromyces sp. Y-85
FOSs can be produced either from
-Sucrose by fructosyl transferase
-Inulin by endoinulinaseEndoinulinases act randomly and to yield FOSs containing a mixture of
-D-fru(21)-[-D-Fru-(21)-]n where n=1-9
and
-D-Glu-(12)-[-D-Fru-(21)-]n where n=2-9
Substrates used
pure inulinnaturally occurring inulin
Optimum conditions
Temperature 37-55 C
pH 6.0-7.0
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O
O
O
O
O
O
O
1
2
1
2
1
2
1
2
1
2
1
E- - l -(1 2)-[ F- - r -(1 2)-] n r n 2 -
i li s
F- - r -(2 1)-[ F- - r -(2 1)-] n r n 1 - 9
E- - l -(1 2)-[ F- - r -(2 1)-] n where n = 2 - 9
Sucr
1
2F
F
F
F
F
F
F
G
and
Elnga
tin
Deg
rada
tin
F-Fr ct f r anosidase
Fruct - li cchari
Inulin
Fruct - li acchari
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Applications of FOSs in Food Industry
Dietary fiber
Sugar substitute mainly in dairy and bakery products
Used in combination with high intensity sweetners toreplace sugars
Sweetening agent in light jam products Used in ice creams to replace all the sugar
Included in probiotic yoghurt and dairy drinks to producesynbiotic products
Application in confectionary, chocolate , dairy products &infant milk formulations
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HIGH FRUCTOSE SYRUPHIGH FRUCTOSE SYRUP
Aspergillus aureus,Botryosphaeria sp.
Chrysosporium pannorum,
Cladosporium phoenicis,
Fusarium oxysporum,F. roseum,
Penicillium glaucum,
Rhizopus sp.
K. fragilis,
K. lactis,
K. marxianus,
Kluyveromyces sp.
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Inulin
Major use in soft drink industries
In dairy products as bodying agent and to improve texture and
mouth feel
In confectionary for sweetness, grain control and humectancy
HFS (95%)Exoinulinase
Applications in food industriesApplications in food industries
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VINEGAR PRODUCTIONVINEGAR PRODUCTION
Conversion of ethyl alcohol to acetic acid
Acetobactersp. and Gluconobactersp.
Usual starting material is wine, beer or cider.
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CONTINUED..