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Name/s (Optional): Nur Fatihah binti Mohd Arnawi
Student/s ID: 0318345
Programme : Biotechnology
Email (Individual/Group Leader): fatihah2312@gmail.com

Contact No (Individual/Group Leader) : 012-3760100

Module code and title: SCT 60203 : Bioprocess Technology

Module Lecturer: Dr Lai Zee Wei
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Due date:24th June 2016

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15

GRADE
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16

INTRODUCTION
AJINOMOTO Malaysia Berhad or previously known as AJI-NO-MOTO Monosodium Glutamate
Producer

is

Japanese

based

company

that

being

established

in

Malaysia

on

1961( AJINOMOTO 2006) . One of achievement that they achieved throughout this century,
Ajinomoto promoted the new sensory taste which known as UMAMI, which was discovered by
Kikunae Ikeda. He had succeeded in isolating, purifying and identifying the principal component
of umami from Konbu seaweed. He patented his discovery on the umami taste, and
collaborated with Saburosuke Suzuki who was an entrepreneur to start the industrial production
of monosodium glutamate which is in the form of sodium salt (Sano 2009).
Glutamate is type of amino acid that can be derived either chemically or biology pathway.
Generally, the application of glutamate is famously known as flavor enhancing in cooking as it
provides a unique taste (Gul etal. 2012). Even though, glutamate had been approved by FDA
with their GRAS status but there are minority of public criticized this usage of glutamate due to
some rumors about the side effect such as may elicit asthma, migraine headache and Chinese
Restaurant Syndrome (CRS) as there are no consistent clinical data to support this claim (Jinab
& Hajeb 2010). Besides, the consumption of glutamate may beneficial to human metabolism as
glutamate is a specific precursor for amino acid synthesis such as arginine, proline and for
tripeptide glutathione in the small intestinal mucosa. Glutathione have important role in
protection of the guts from peroxide damage and dietary toxin). In addition, the consumption of
glutamate may be used for other metabolic purpose such as protein synthesis, energy
metabolism, and ammonia fixation or as reused transmitter (Beyreuther et al. 2007). Other than
focusing in producing monosodium glutamate, Ajinomoto also manufacture various type of
amino acid that being uses in different fields such as food, pharmaceutical and bio-fine
(AJINOMOTO 2006).

As a pioneer Japanese company in Malaysia, their target is to be a group of companies that

contributes to human health globally by continually creating unique value to benefit customers
(citation). Along with the vision, bellows are three missions that always be their guideline to be a
No.1 company in amino acid manufacturing:

To become a global group of food companies centered on the worlds

No. 1 seasoning business.

To become a global group of amino science companies that

contributes to humankind with the worlds No. 1 amino acid technology.

To become a group of health-promoting companies with a scientific

approach to good taste and health

(AJINOMOTO 2006)

DISCUSSION
During ancient time, the production of glutamate was using conventional method where using
Domyoji game vessel. Previously, Ikeda proposed source of carbohydrate is wheat gluten
instead of cassava or sugarcane because consist of high content of L-glutamic acid. The gluten
was separated from wheat flour by washing the starch from dough. Then, the gluten was
transferred to pottery vessels with addition of hydrochloric acid and heated for 20h. The product
of this process were filtered to eliminate a black residue which called as humus as resulted from
the reaction of amino acids with carbohydrates to be concentrated. The concentrated of this
product, were allowed to be crystallized. The crystallization of L-glutamic acid hydrolysate is
also known as partial purification as the salt crystal possesses high selectivity against other
amino acid. However, this conventional method is considered as hazardous and not safe due to
exposure towards hydrogen chloride gas which can corrosive may cause irritation (Sano 2009).
Due to this limitation, Ajinomoto Company projected a new economical technique termed as
fermentation. In this method, glutamate is produced with the aid of microbe with selected strains
in large scale rather than chemical synthesis. According to Ajinomoto (2014), the sugarcane or
tapioca is chosen for the raw materials as source of carbohydrates. The selection of the starch
because of Ajinomotos principle which to promote the use of sustainable pro and supply of
amino acids using local agricultural raw materials like cassava and sugarcane that considered
as one of major crop in this tropic country. Hence, Ajinomoto also applied bio-cycle as their
fermentation plants in worldwide, hence practicing sustainable procurement of agriculture raw
materials and supporting local agriculture industry along increase the productivity of local
farmers. Besides, all the by product from this process were not being wasted, they are recycled
as organic fertilizer to the crops.
On the first stages, the cassavas were undergoing pre-treatment for example being washed and
chopped into small pieces as to increase the surface area. Then, the starch is extracted by
flushing the pulp with the root juice. The crude starch suspension is separated to milk and juice
on hydrocyclones. The concentrated crude starch milk is further processed with multistage to
produce pure concentrated starch slurry (International Starch Institute, 2014). This form of
starch was being added into the bioreactor as carbon sole for microorganism such as
Corynebacterium species (Shyamkumar etal. 2014). Addition of -amylase will enhance the
fermentation by hydrolyzing the long polysaccharide chain to shorter chain. Subsequently, the

microbe will metabolize the starch using the Glutamate dehydrogenase or by glutamate
synthase pathways as shown in figure 1.

Figure 1: Metabolic pathway of glutamate production by selected bacteria (Gul etal. 2012).
After the pre-treatment, the selected strain of bacteria which in this glutamate manufacture by
Ajinomoto, the stock of Corynebacterium glutamicum are used being culture into the shake flask
cultures. The resulting cells are transferred and grown into the small seed culture tank. Then, it
was used for inoculating to a larger tank. The intermediate seed culture volumes are varies in
size but commonly in the range of 200L-1000 L,10,000 L-20,000 L and 50 000 L- 500 000 L
respectively. This process is very crucial and each condition is being controlled carefully for
example cell density, nutrient composition, temperature, pH, aeration rate, agitation rate and
sugar flow rate. All these control need to be consistent throughout the entire process. Oleic acid
is being introduced at the beginning of the fermentation as to encourage the excretion of
glutamate from the Corynebacterium. Overall, in the entire process, pH is maintained at 7.8
(Herman 2003).
The bacteria culture was being incubated for 14 hours, and temperature of the culture rose from
32C to 38C. The sugar (cassava source) which that already being treated during pre-treatment
process being fed in the fermenter in fed batch mode for 36 hours. There are approximately
about 160g of glucose per liter being added into the fermentation. The amount of glucose is
being monitored in order to prevent from the crab-tree effects. The glutamate concentrations
are monitored within the interval as to meet the required concentration and being controlled by
specific probe. After completing all the process, the broth is pumped out from the bioreactor for
the recovery process in order to get the pure glutamate (Tien nd). During purification and
recover process, ion chromatography is used as it offers fast results and low operational cost
compared to other purification method (Nampoothiri & Pande 1999). The whole process of
fermentation can be illustrated as in figure 2.

Figure 2 : the overall illustration of monosodium glutamate fermentation ( Ajinomoto 2016).

For the industrial production of L-glutamate, the process is under control of stirred tank
bioreactor. The most concern in producing glutamate in the large scale is the cost; this stirred
tank bioreactor offers low operation and capital cost compared to other bioreactors. The use of
stirred tank bioreactor also provide advantages in producing more glutamate in one time as the
working volume of this reactor can be up to more than 500 000 L. Besides, the sterilization
method is easily understood and relatively easy to implement by the workers for example using
steam to sterilized the bioreactor (William 2002).
Usually, the production of glutamate is operated under fed batch mode where it allows addition
of sugar during fermentation process. As during initial process, the concentration of sugar will be
high as to supply enough carbon sources for the selected strain of bacteria. However, the
addition of high level of sugar in the beginning will contribute to other problem such as
incomplete oxidation of the sugar to lactic acid which eventually can influence the yield of
desired product. This situation is known as crab-tree effect (Hermann 2003). The other issue of
using fed batch fermentation as primary bioreactor instead of batch fermentation because of the
substrate inhibition. The Corynebacterium will metabolized the all the nutrient inside the
fermenter and produce the glutamate in form of glutamic acid. Subsequently, if the
concentration of glutamic acid is too high, the yield of glutamate produced will be reduced as the
glutamic acid will inhibit the growth of Corynebacterium. This situation is known as substrate
inhibition (Anschau, Santos & Alegre 2013). By implementing the fed batch fermentation, the
risk of these two problems can be overcome because the adding of glucose accordingly may
reduce the formation of high concentration of glutamate acid and low concentration of glucose
will not causing the crab-tree effect.

Last but not least, for industrial production glutamate there are certain requirements need to be
considered as to maximize the production of glutamate. Hence, it is very important to keep in
mind the glutamate has to be excreted by microorganism in excessive manner and have
extracellular properties. Therefore, there are many ways as to increase the production of
glutamate. Commonly, the glutamate producers are biotin auxotroph where it required addition
nutrient for some specific metabolic pathway. The dependence of biotin subsequently will
reduce the yield of glutamate. Therefore, industries use alternative ways by applying the biotin
deficient medium as it founded that this kind of medium helps to trigger the production of
glutamate. Biotin is a one of vitamin that acts as cofactor that essential for carboxylase activity.
Example of enzyme is acetyl-CoA carboxylase which enables converting Acetyl-CoA to malonylCoA during fatty acid biosynthesis (Cao, Duan & Shi 2013). Research reported that
approximately 30% of the phospholipid membrane loses their structure during biotin starvation
or addition of surfactant (Kimura 2002). In addition, mycolic acid content of the outer layer
began to decrease under this situation which lead to glutamate over-production was induced
(Hashimoto et al. 2006). Fundamentally, the membrane of microorganism will be more
permeable due to alteration if membrane structure and thus will trigger the secretion of
glutamate Other than that, for nitrogen sources Ammonia and ammonium sulfate are used.
Thus, ammonia also can act to control pH during fermentation. In order to minimized the cost
without lowering the quality, cheap sources of vitamins and other nutrients for instance corn
steep liquor, which is a by-product of cornstarch manufacture which rich with amino acid, nucleic
acids, mineral and vitamin (Sano 2009 ).

CONCLUSION
As for conclusion, AJINOMOTO has positioned itself as one of the most popular brand by
serving consumer with high quality products as with it is status as pioneer company in glutamate
manufacture. For further in manufacturing monosodium glutamate (MSG) suitable condition
such as the media selection, pH, temperature, bacterial strain, and operation need to be
optimized as to maximize the production of monosodium glutamate. Therefore, by taking up
these criteria as consideration will reduce the operational cost which one of the major concern in
the industrial sector as to exploit profit with minimum capital and operational cost. For the
further development with the aid of research, this industry can use another strain of bacteria
such as MNZ of Lactobacillus plantarum as reported by Zareian etal (2012) where this strain
have capability to produce high amount of glutamic acid comparative with the existing bacterial
strain which is Corynebacterium glutamicum. Finally, there are still ongoing research needs to
be done as to enhance the effectiveness of glutamate production along to maintain the brands
name of AJINOMOTO as the precious company in this sector.

1920 words including citation

REFERENCES
Ajinomoto Group, 2014, Promoting the Sustainable Use of Food Resources, Ajinomoto Group
Sustainability

Report,

viewed

on

17th

June

2016,

<https://www.ajinomoto.com/en/activity/csr/pdf/2014/021-028.pdf>
AJINOMOTO, 2006, The Ajinomoto Group: About Us, viewed 19th June 2016, <
http://www.ajinomoto.com.my/about-ajinomoto-group/>
Anschau, A, Santas, LO, Alegre, RM, 2013, A Cost Effective Fermentative Production of
Glutathione by Saccharomyces cerevisiae with Cane Molasses and Glycerol, Brazilian Archives
of Biology and Technology, Vol. 56, No. 5, pp 849-857.
Beyreuther, K, Biesalski,HK, Fernstorm, JD, Grim, P, Hammes, WP & Heinemann, U, 2007,
Consensus meeting: Monosodium Glutamate: An Update. European Journal of Clinical
Nutrition, Vol 61, No. 3, pp 304313.
Cao, Y, Duan, Z, & Shi, Z, Effect of biotin on transcription levels of key enzymes and glutamate
efflux in glutamate fermentation by Corynebacterium glutamicum ,

World J Microbiology

Biotechnology, Vol.30, pp461-468

Gul, H, Shah, AH & Younis, N, 2012, Fermentative Production of Glutamate by Newly Isolated
Soil Bacteria, International Journal of Pharmaceutical & Biological Archives, Vol 3, No. 6,pp
1368-1376.
Hashimoto, K, Kawasaki, H, Akazawa, K,Nakamura, J Asakura, Y , Kudo, T, Sakuradani, E,
Shimizu, S , Shimizu, S, & Nakamatsu, T, 2006, Changes in composition and content of mycolic
acids in glutamate overproducing Corynebacterium glutamicum . Biosci Biotech Biochem,
Vol.70, No.1, pp 2230
Hermann, T, 2003, Industrial production of amino acids by coryneform bacteria , Journal of
Biotechnology, Vol 103, pp 155-172.
Jinab, S, & Hajeb, P, Glutamate: Its applications in food and contribution to health, Elsevier,
Vol 55, pp 1-10.
Kimura, E, 2002, Triggering mechanism of L-glutamate overproduction by DtsR1 in coryneform
Bacteria , J.Biosci Bioeng , Vol. 94, No.6, pp 545-551

Nampoothiri, KM, Pande, A , 1999, Fermentation and Recovery of L-Glutamic Acid from
Cassava Starch Hydrolysate by Ion-Exchange Resin Column , Revista de Microbiologia , Vol
30, pp 258-264.
Sano, C, 2009, History of Glutamate Production, The American Journal of Clinical Nutrition,
Vol 90, pp 728s- 732s , viewed on 20th June 2016 , < http://www.ajcn.nutrition.org>
Shyamkumar, R , Moorthy, IMG, Ponmurugan, K & Baskar, R , 2014, Production of L-glutamic
Acid with Corynebacterium glutamicum (NCIM 2168) and Pseudomonas reptilivora (NCIM
2598): A Study on Immobilization and Reusability , Avicenna J Med Biotech , Vol. 6, No.3, pp
163-168
Tien,

KC,

nd,

Amino

Acid

Production,

viewed

on

15th

June

2016,

<

https://www.academia.edu/4936314/Amino_Acid_Production>
William, JA, 2002, Keys to Bioreactors Selection, Bioreaction, viewed on 20th June 2016 <
www.cepmagazine.org>
Zareian, M, Ebrahimpour,A, Abu Bakar, F, Mohamed, AKS, Forghani, B, Ab Kadir, MS & Saari, N
, A Glutamic Acid-Producing Lactic Acid Bacteria Isolated from Malaysian Fermented Foods ,
International Journal of Molecular Sciences, Vol. 13, pp 5482-5497.