Food analysis

Explaining how the safety of food from a genetically engineered food product is evaluated.
Source of novel protein
A food or a protein can either be allergenic,less commonly allergenic or unknown allergenic potential.
Allergenic food in plants includes;peanuts,soybeans treenuts and wheat. In animals the allergenic foods
includes milk eggs and fish.
If a gene is obtained from a known allergenic source, a careful assessment must be conducted to ensure
that the gene of interest does not encode an allergen particuraly if gene is expressed in edible portion of
genetically modified organism.
Level of expression of the protein.
In most casses ,most introduced proteins are expressed in low levels in genetically modified
organism,but enough to provide the function which they are introduced for. The threshold dose exist
below which the allergenic individuals will not affected by the food.
Sequence homology to known allargen.
A comparison of an amino acid sequence homology of the novel protein to the amino acid of known
allergens is done. This is a useful approach in the determination of allergenic potential.The amino acid
of many allergens is known and curated.
Function charactaristics of the novel proteins
the pathogenesis related protein of several different types are prominently involved.if the novel protein
introduced into genetically modified organismfall into functional categories that contains known
allargens.great caution must be excercised in assessing potontial allergenicityof these particular protein.
Immunoreactivity with serum IgE.
If the gene is derived from a known allergenic source an assessment of the immunoreactivity of the
novel protein with IgE antibodies from the sera of the individual allergenic to the source material must
be conducted.IgE binding could be used to determine whether IgE patient sera react with novel protein
or the extract of the novel food.
Physiochemical stability of the novel protein.
To become allergenic .a protein must react in the intestinal tract in a form that is enough to provoke the
immune system.
In a case where the protein is digested under simulated gastric and intertinal digestive model, allargenic
prospect is unlikely.
2.Discuss molecular engineering methods and procedures for improving the catalyst performance
of industrial enzymes.
Rational design
detailed information about structure and mechanism is required to redesign the nature of catalyst.
Although this information is not available the increasing growth protein structure database is helping in
the lack of this information Because new enzymes are evolved by minor modification of the active site
structures, homology-driven experiment are done to the binding site to fit the different substrate and
new catalytic residues to modify functions and mechanism.
Random design
The development of evolutionary design methods using random mutagenesis,gene recombination,and
high-throughput screening is one of the key factor contributing to the expanding application of
biocatalyst in industrial processing
Random redesign techniques are being used currently used to generate enzyme with improved
properties such as activity and stability at different PH values and temperatures,altered substrates
specificity,novel substrate specificity and activity as well ass increased biological activity of biological
molecules.
3.discuss about regulation measures of enzymes used in food industry
Replacing of dry enzymes preparation by liquid preparation.This reduces the chance of aerosal
formation during handling.
Encapsulating and granulating dry enzyme preparation,
dissolving any waste enzyme in water before disposal into the sewage system'
4.Discuss the production and application of amino acids in the food industry
Amino acids can be produced by the following methods

 extraction from natural sources

 chemical synthesis

 fermentation

 enzymatic catalytic
Extraction from natural source involves the following procedure
1. hydrolysis with aqueous acid
2. capture of the amino acid by passing of the hydrolysate over a strong acid ion exchange resin
3. washing of the resin with water
 4. elution with aqueous ammonia to free the amino acid
5. collection of the amono acid as fraction .The method is much economic for the production of
(S)-tyrosine and (R-R)-cystine.
Chemical synthesis
The method heve an advtage that it can be carried out on a very large scale and on a continous
way.even thogh the method typically give a racemic mixture of the enantiometric forms of amino acids.
Fermentation method
Althogh it is possible to prepare any natural amino acid by fermentation,the special mutant that allow
production to be done on large scale has been developed only for the preparation of (S)-lysine and (S)-
glutamic acid. In the process,cane raw sugar or starch are used as a source of carbon. Ammonia
provides source of nitrogen and oxygen is provided by passing compressed air into the fermentation
mixture.
Method one
fermentation process for the production of lysine makes the use of a pair of E.coli mutant .normal
E.coli can synthesize its own lysine from carbohydrate and ammonia but the first mutant lacked the
alpha E-diamino-pimelic carboxylase.
Method two (single stage fermentation)
This process makes the use of a mutant of corynobacterium glutamicum in which feedback mechanism
of product inhibition are overcomed. Mollasses is the most common carbon source,that contains
enough biotin to provide more than 30 microgram per litre needed to suppress the excretion of glutamic
acid .the final concentration of lysine is about 60g/l, and the fermentation cycle takes about 48-92 hrs.
Enzyme synthesis method
pure enzyme are used in this process.example in (S) -aspatic acid was produced mainly by the enantio-
selective enzyme-catalysed,addition of ammonia to fumaric acid .
5 Explain how microbial stains are developed for enzyme production.
Objectives
The organism should grow on an inexpensive medium and give constant high yield within a short time
Secodary enzyme activities and the content of metabolites in the fermentation broth should be
minimal.
The recovary of the enzyme should be simple andinexpensive and lead to products the can be handled
safely and of acceptable appearance.
The process must be safe to the person in the production plant and the effluent from the plant should
not disterb the environment.
Steps in the development of reccombinant strain
1. development of the recipient/host organism
2. construction of the expression vector.
3. Transformation of the host strain.
4. Identification of the best reccombinant strain.
5. Addition improvement
6. characterization of the produced strains.
Most of the host strains for food-processing enzyme have been derived from a reratively small number
of bacteria and fungal spices primarily Bacillus subtils B.lichenifomis A.nigar and A.oryzae.
Bacterial host strain,
the host are B. subtilis and B.lichenifomis.B.sus\btilis is used for the production of alpha-amylase and
proteases.this strain have an advantage because they secrete enzyme directly to the fermentation
medium.
E.coli
E.coli K-12 has been used to produce chymosin.
Fungal host strain
this includes Aspagillus nigar and A.oryzae. A oryzae has been used for many years as a source of koji
and for production of soy sauce,miso and rice wine sake.
A.nigar is commonly used for the production of citric acid.
6.Describe the criteria for lactic acid bacteria for process development ane explain why the
criteria are important.
The need for the organism to be a homo-fermenter meaning that it will only produce one product ie
lactic acid
the type of the lactic acid required.there exist three enzymes which are responsible for the different
type of lactic acid.
A steriospecific l(+)-lactate dehydrogenase
Astereospecific L(-) lactic dehydrogenation
Alactate racemes