App of Biotranformation in Food Technology- Rupinder

8/6/2019 App of Biotranformation in Food Technology- Rupinder

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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..

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App of Biotranformation in Food Technology- Rupinder