Growth Media for Enzymes and

Starter Cultures in Halal Perspective

Dr. Ali Salim Fanous

Halal Control, EU, European Inspection-and Certification Body for Halal-Food

Germany

The First Gulf Conference on Halal Industry and its Services24-26 January 2011

Salmiyah, State of Kuwait-Holiday Inn Hotel, Al

البيئة المغذية ل�نزيمات

والبادئات من منظور الح�ل

فانوسمليعلي س. د

، خدمات الح�ل في أوروبا، ألمانيا)ا�تحاد ا وروبي( كونترولح�ل

مؤتمر الخليج ا�ول لصناعة الحل وخدماته2011يناير 24-26

دولة الكويت–السالمية –فندق ھوليدي إن

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What are the stages of biotechnology?

• Ancient Biotechnology

• early history as related to food and shelter, including domestication

• Classical Biotechnology

• built on ancient biotechnology

• fermentation promoted food production

• medicine

• Modern Biotechnology

• manipulates genetic information in organism

• genetic engineering

What are starter cultures ?

Starter cultures are those microorganisms that are used in the

production of cultured dairy products such as yogurt and cheese.

A starter culture can provide particular characteristics in a more

controlled and predictable fermentation. The primary function

of lactic starters is the production of lactic acid fromlactose.

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Application of starter cultures in food

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Production of starter cultures

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1. Preparation of growth media

Production of starter cultures

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1. Preparation of growth media

Production of starter cultures

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2. Cultivation of bacteria (fermentation)

Production of starter cultures

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2. Cultivation of bacteria (fermentation)

Production of starter cultures

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3. Concentration of bacteria and freezing the pellets

Production of starter cultures

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Production of starter cultures

4. Freeze drying

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• Slimy structure e.g. yogurt

• Bad flavours and taste e.g. cheese

• Excessive gas production

• Sensory deprivation

Trouble-Shooting Fermentation

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Fish blood is a component of growth media

The potential production of fish blood is about 12,000 metric

tons in 2006. Salmon blood, for example, contains easily

digested proteins and a high concentration of poly-

unsaturated Omega-3 fatty acids.

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Application of fish blood in food

• Fish blood the key to low-fat ice cream!

The blood protein originally comes from

the eel-like ocean pout fish. This is added

to the ice cream and lowers the

temperature at which ice crystals form

and the shape they take up.(www.dailymail.co.uk/health/article-392219/Fis).

• Direct supplements to food, as iron or binding agents.

• Fish blood can be used as a component of growth

media e.g. starter culture.

Halal status of Fish blood

قال � تعالى سفوح( ما على طاعم يطعمه إ)# أن يكون ميتة أو دما م# أو لحم خنزير ا قل )# أجد في ما أوحي إلي# محر#

ه رجس أو فسقا أھل# لغير � به فمن اضطر# غير باغ و) عاد فإن# رب#ك غفور ر# ) ) 145(يم ح فإن#]سورة ا نعام[

“He hath forbidden you only carrion, and blood, and swine flesh, and that

which hath been immolated to (the name of) any other than Allah. But he who

is driven by necessity, neither craving nor transgressing, it is no sin for him. Lo!

Allah is Forgiving, Merciful” (2:173).

:عن النبي صلى � عليه وسلم قال , عن عبد � بن عمر رضي � عنه قال )والكبد والطحال , السمك والجراد , أحلت لنا ميتتان ودمان (

Allah’s Messenger (peace be upon him) said: “Made lawful for you are the

flesh of two dead animals and two types of blood. As for the two dead

animals, they are the locust and the fish. And as for the two types of blood,

they are the blood of the spleen and the liver.”

The Use of Fungi and Microorganism

in the Production of Food Products

Micro-organisms are today widely used in the manufacture

of food and drink, acting primarily as production agents

rather than as principal raw materials.

They play an important role in the conversion of ingredients

into food, for example, in the preparation of bread, cheese,

yogurt and alcoholic drinks.

Yeasts and other fungi play a critical role in drug production,

food processing, bio-control agents, as well as enzyme

biotechnology.

Production of Amino acids

Corynebacterium glutamicum, is a Gram-positive soil bacterium, which

is used for the industrial production of different amino acids, mainly L-

glutamate and L-lysine, and of nucleotides. One of the most important

organisms in biotechnology, Corynebacterium glutamicum is currently

used to produce 2 million tons of amino acids.

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Ingredients of BHI

Pancreatic Digest of Casein

Brain Heart Infusion (swine)

Peptic Digest of Animal

Tissue

Sodium Chloride

Disodium Phosphate

Glucose

BHI Broth is used for the cultivation

of a wide variety of microorganisms,

including bacteria, yeasts and molds.

Brain Heart Infusion Broth (BHI)

Production of Amino acids

No Material Source

1 Beef extract Extracted from beef

2 Peptone Hydrolysis of protein from meat, casein, or gelatin using

acid of enzymes

a. Acid-hydrolyzed

casein

Hydrolysed protein from casein using acid for hydrolysis

example: casamino acids (Difco); Hy-case (Sheffield)

b. Enzymatic digest

of casein

Hydrolysed casein using enzymes for hydrolysis

Example: Casitone (Difco); Tryptone; trypticase peptone

c. Enzymatic digest of

meat protein

Hydrolysed protein from meat using enzymes for

hydrolysis.

Bactopeptone proteos pepton

d. Enzymatic digest of

plant (soy) protein

Hydrolysed protein from plant (soyben) using enzymes

for hydrolysis. Example: Soyapeptone

3 Mixed hydrolisate Biosate (yeast extract and casein digest); Trypton

(mixed-enzymatic source)

4 Yeast extract Extracted from yeast

5 Agar Carbohydrate from seaweed

Critical materials in growth media

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Production of Amino acids

Metabolic Engineering, manipulation of pathways within an organism to

optimize the production of a compound

Done by turning off particular genes, either through mutation or

deletion Products are also gained by altering the microbe’s environment

Metabolically

Engineered Cell

Cheap Carbon Source Industrial Chemicals

Chemical Precursors

Biopolymers

VitaminsAmino Acids

Production of Amino acids

Glucose

NH4+

Pyruvate

L-lactate

D-Alanine

L-Alanine100%L-Alanine

NAD+

NADH

NADH

NAD+

L-LDH

AlaOH

NADH NAD+

Racemase

The microbe is forced to produce alanine at higher than normal amounts

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Production of citric acid

Most sodas and soft drinks contain citric acid as a main

ingredient. Citric acid is also used in other drinks, many candies,

canned goods, baked goods, etc. It is too expensive to isolate the

citric acid from citrus fruits so it is produced in large-scale

fermentation vats utilizing Aspergillus niger

Why are enzymes important in the food industry?

Enzymes are highly specialized proteins that catalyze

specific biochemical reactions.

Advantages

• Natural, Nontoxic

• Catalyze specific reactions

• Active under mild conditions

• Active at low concentrations

• Can control rate of reaction

• Can be inactivated

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Uses of Enzymes in Food Industry

Food industry utilizes a variety of enzymes for processing of various

foods, e.g., production of various types of syrups from starch or

sucrose (a- and β-amylases, glucamylase, pullulanase, invertase, and

glucose isomerase), meat/protein processing using proteases,

removal of glucose and or molecular oxygen (O2) using glucose

oxidase and catalase, use of lactase in dairy industry and use of

enzymes in fruit juice.

saccharose is broken down by the enzyme invertase in glucose and fructose

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Application Enzymes used Uses

Food processing

Amylases catalyze the release

of simple sugars from starch

Amylases from fungi

Proteases

Production of sugars from starch, such as in making high-

fructose corn syrup. In baking, catalyze breakdown of

starch in the flour to sugar.

Biscuit manufacturers use them to lower the protein

level of flour.

Baby foods Trypsin To predigest baby foods.

Fruit juices Cellulases, pectinases Clarify fruit juices

Dairy industry Rennin, Microbially produced

enzyme

Lipases

Lactases

Manufacture of cheese, used to hydrolyze protein.

production of Roquefort cheese to enhance the ripening

of the blue-mould cheese.

Break down lactose to glucose and galactose.

Meat tenderizers Papain To soften meat for cooking.

Starch industryAmylases, amyloglucosideases and

glucoamylases

Glucose isomerase

Converts starch into glucose and various syrups.

Converts glucose into fructose in production of high

fructose syrups These syrups have enhanced sweetening

properties and lower calorific values than sucrose for the

same level of sweetness.

Uses of Enzymes in Food Industry

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α-acetolactate bacteria Removes bitter substances

decarboxylase from beer

α -amylase bacteria Converts starch to simple sugar

Catalase fungi Reduces food deterioration

Chymosin bacteria or fungi Clots casein to make cheese

β-glucanase bacteria Improves beer filtration

Glucose isomerase bacteria Converts glucose to fructose

Glucose oxidase fungi Reduces food deterioration

Lipase fungi Oil and fat modification

Maltogenic amylase bacteria Slows staling of breads

Pectinesterase fungi Improves fruit juice clarity

Protease bacteria Improves bread dough

structure

xylanase (hemicellulase) bacteria or fungi Enhances rising of bread dough

Production of Enzymes

Enzyme Name GE Organism Use (examples)

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Enzymes produced by genetically modified microorganisms

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Advantages of using (GMMs) for the production of enzymes

There are several advantages of using genetically modified

microorganisms GMMs for the production of enzymes, including:

• It is possible to produce enzymes with a higher specificity and

purity.

• It is possible to obtain enzymes which would otherwise not be

available for economical, occupational health or environmental

reasons.

• For enzymes used in the food industry particular benefits are for

example a better use of raw materials (juice industry), better keeping

quality of a final food and thereby less wastage of food (baking

industry) and a reduced use of chemicals in the production process

(starch industry)

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HALAL CONTROL

Thank you for your attention

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