Chemical Engineering and Processing 81 (2014) 59–71

Contents lists available at ScienceDirect

Chemical Engineering and Processing:

Process Intensification
journal homepage: www.elsevier.com/locate/cep

Production and purification of glutamic acid: A critical review

towards process intensification
Ramesh Kumar, D. Vikramachakravarthi, Parimal Pal ∗
Environment and Membrane Technology Laboratory, Chemical Engineering Department, National Institute of Technology Durgapur,
Durgapur 713209, India

a r t i c l e i n f o a b s t r a c t

Article history: Amidst growing environmental awareness and stringent discharge regulations, chemical and allied pro-
Received 5 February 2014 cess industries are now desperately seeking replacement of the conventional, polluting processes by
Accepted 25 April 2014 clean and green processes. In this context, production and purification of amino acids like l-glutamic
Available online 6 May 2014
acid assumes significance. Concerned conventional process involves several steps like fermentation, cen-
trifugation, carbon adsorption, evaporation, crystallization, ion-exchange and so on to get glutamic acid
Keywords:
in desired concentration and purification. Despite its tremendous potential for large scale use in a wide
l-Glutamic acid
variety of applications, cost-effective production of high purity glutamic acid has remained a challenge
Membrane system
Green technology
for decades, mainly due to several downstream processing steps and the associated cost factors. With
Process intensification emergence of tailor-made membranes and modules, possibility of using membranes in downstream
purification of glutamic acid appears imminent with expectation of a turnaround in amino acid manu-
facturing industry. The present study through a brief yet comprehensive review of the very critical aspects
of glutamic acid production and purification, attempts to direct research efforts towards process inten-
sification encompassing the concepts of green processing, compact and flexible design with promise of
more economically attractive production with better quality.
© 2014 Elsevier B.V. All rights reserved.

1. Introduction acids in general is observed to double every decade with scope

Konbu which is a kelp-like seaweed and traditionally used in
Japan as flavour enhancer was identified as l-glutamic acid (GA)
[1]. This discovery led to industrial production of monosodium GA
by the Ajinomoto Company. Glutamic acid (GA) used to be pro-
duced in those days by acid hydrolysis of wheat gluten or soybean
protein. But in the subsequent years, l-glutamic acid-producing
micro-organisms were isolated [2] and research efforts culminated
in development of fermentative process for production of GA. Intro-
duction of fermentation technology provided a significant impetus
to the development of microbial production of primary metabo-
lites. Subsequently, in a series of research initiatives, attempts were
made to isolate wild strains or to derive genetic mutants for pro-
duction of amino acids. Thus amino acids are now commercially
produced by fermentation.
GA is one of the most important amino acid products with a wide
range of applications. Its salt-derivatives can be used as nutrition
elements and participate in body metabolism. Market for amino
∗ Corresponding author. Tel.: +91 9434788105; fax: +91 3432754078.
E-mail addresses: parimalpal2000@yahoo.com, ppal.nitdgp@gmail.com (P. Pal).
for exploitation of new uses of amino acids and progress in pro-
duction technology [3]. GA is used in seasoning throughout the
world [4]. It is also used as a starting material for synthesis of var-
ious kinds of speciality chemicals [5]. At the same time, glutamic
acid can improve the function of nervous centralize and cortical
brain for neurasthenia patients [6]. Poly glutamic acid (PGA) is
a naturally occurring anionic polymer that is biodegradable, edi-
ble, and non-toxic towards human and environment [7–10]. It
is a good candidate for various industrial applications including
thickener, bitterness reliving agent, cryoprotectant [11], drug car-
rier [12], curable biological adhesive [13], biodegradable fibres,
highly water absorbable hydrogels, biopolymer flocculants [14],
and heavy metals absorbers [15].
Development of innovative products and processes is a big chal-
lenge to the chemical and allied process industries all over the
world for survival in an era of emaciated profit margins amidst
highly globalized trade competition and fast growing environ-
mental constraints [16,17]. Thus process intensification through
revolutionary development of new products and processes that
ensure reduced material and energy consumption and reduced
environmental impacts while offering greater flexibility in scale of
operation are the need of the hour. Production of monomer grade

http://dx.doi.org/10.1016/j.cep.2014.04.012
0255-2701/© 2014 Elsevier B.V. All rights reserved.
60 R. Kumar et al. / Chemical Engineering and Processing 81 (2014) 59–71
glutamic acid which has traditionally been used in food, cosmetics,
medicine and agriculture has over the last few decades, attracted
attention of the world researchers. To separate amino acids from
fermentation broths, properties of the product, such as solubility,
molecular size, and affinity to adsorbent and charge characteristics
may be utilized. Membrane separation is expected to be one of the
most promising candidates in successfully separating target amino
acids from large amounts of other amino acids. Other methods
like electrodialysis that uses anion-exchange and cation-exchange
membranes has generally not been successful due to significant pH
changes in the feed solutions, being undesirable for enzyme reac-
tions when the electrodialysis separation process is incorporated
into the reactors. In addition, an anion-exchange membrane was
reported to be easily contaminated probably due to an oxidative
reaction of a sulfhydryl group present in the amino acids [18].
In many chemical, pharmaceutical, food and biotechnological
processes, the purification and recovery of amino acid plays
pivotal role. Nanofiltration(NF) is a relatively new class of the
pressure-driven membrane processes and is considered a viable
alternative to more traditional separation processes like extrac-
tion, ion-exchange, evaporation and distillation [19,20]. The major
technology barrier in cost-effective production of high purity GA is
its down-stream separation and purification. These are the fields
where membrane-based processes are stepping in. Being modu-
lar in design, membrane-based processes offer great flexibility in
scale of production depending on market demand. By virtue of high
selectivity, membranes can ensure high levels of separation and
purification. As membranes of chosen selectivity and permeability
can easily be integrated with conventional fermentor, membrane-
based processes permit simultaneous production and purification
in the same unit. This eliminates the need for separate purifi-
cation units and results in compact design with reduced capital
investment. Membrane-based separation and purification (barring
pervaporation) involves no phase change ensuring reduced energy
consumption. Thus such processes can meet all the goals of pro-
cess intensification. This paper first briefly discusses the traditional
downstream processes to highlight the major problems associ-
ated with these processes and then critically takes up review of
the developments in membrane-based processes. The objective
is to identify an environmentally benign, simple, economically
viable and continuous manufacturing scheme capable of producing
monomer grade GA with high productivity.
2. Upstream production of GA
2.1. Microbial strain
Table 1 shows the number of wild strain that have been isolated
as GA acid producing bacteria. Most of these GA producing bacte-
ria are gram positive, nonspore-forming, non-motile and require
biotin for growth. Table 2 shows the different authors got the
different yields of GA while using different processes and micro-
organisms.
Table 1
Different types of microbial strains producing l-glutamic acid.
Genus Species
Corynebacterium C. glutamicum, C. lilium, C. callunae, C. herculis
Brevibacterium B. divaricatum, B. aminogenes, B. flavum, B. lactofermentum,
B. saccharolyticum, B. roseum, B. immariophilum, B.
alanicum, B. aminoniagenes, B. thiogenitalis
Microbacterium M. salicinovolum, M. ammoniaphilum, M. flavum var.
glutamicum
Arthobacter A. globiformis, A. aminofaciens
2.2. Feedstock (carbon source)
Carbon sources like glucose, fructose, sucrose, maltose, ribose
or xylose can be utilized by the GA producing bacteria as the sub-
strate for cell growth and GA biosynthesis. A crude feedstock has
historically been avoided because high levels of extraneous mate-
rials can cause troublesome separation problems in the recovery
stage. In the late 1950, dextrose from corn starch was the most
commonly used feedstock [21]. Other feedstocks like concentrated
whey, hydrolysed potato, cellulosic material, sulphite liquor, and
molasses were also used [22–24]. Fermentation route from renew-
able resources like sugarcane juice with suitable microorganisms
has always been favoured to produce optically pure l-(+) glutamic
acid as a myriad of value-added products derived from biological
origin and are readily accepted by food industries and consumers
[25]. In addition to that sugarcane juice is easily available through-
out the year in some major sugarcane growing countries like India
and Brazil [26].
2.3. Others nutrients supplements
Addition of nutrient supplementation like yeast extract, pep-
tones and other micro-macro inorganic nutrients required for the
growth of micro-organism in fermentation broth, leads to increased
sugar utilization and reduced fermentation time but it also adds to
residual impurities in fermentation broth [27]. To produce pure and
monomer grade GA efficient separation of those impurities is essen-
tial. Ammonium ion and urea are detrimental to both cell growth
and product formation and its concentration in the medium must
be maintained at a low level. To neutralize the acid that is formed
during fermentation, calcium carbonate, calcium hydroxide and
gaseous ammonia are typically used.
2.4. Culture conditions optimization
Many techniques are available in the fermentation medium
designer’s toolbox for media optimization such as borrowing, com-
ponent swapping, biological mimicry, one-at-a-time, statistical and
mathematical techniques-experimental design and optimization,
artificial neural networks, fuzzy logic, genetic algorithms, con-
tinuous fermentation, pulsed batch and stoichiometric analysis
[28]. Each technique has its advantages and disadvantages, and
situations where they are best applied. While developing an indus-
trial fermentation, designing a fermentation medium is of critical
importance because medium composition can significantly affect
product concentration, yield and volumetric productivity.
The design and optimization techniques are mostly in the tra-
dition of Box and Wilson [29] which includes steepest ascent and
canonical analysis as components of response surface methodol-
ogy. During optimization process three phases of experimentation
can be distinguished: one for identifying important variables
(screening), two for optimization and three for central compos-
ite or Box–Behnken designs. In designing the perfect fermentation
medium, the first step is to integrate medium design and microbial
screening to assess as wide a range of microbial performance as
possible as shown in Fig. 1.
2.5. Microbial physiology and metabolic pathway of l-glutamic
acid fermentation
The biosynthesis of GA is an aerobic process requiring oxygen
throughout the fermentation (7 mg/L). Brevibacterium flavum, GA
producing bacteria accumulate lactic acid and succinic acid when
cultured under limited oxygen supply [30]. It was demonstrated
that the absence of ammonium ions, but with sufficient oxygen
supply, resulted in the accumulation of ␣-ketoglutaric acid in place
 R. Kumar et al. / Chemical Engineering and Processing 81 (2014) 59–71 61

Table 2
Micro-organisms used for the production of l-glutamic acid.

Micro-organism Substrate (carbon source) Yield (g/L) References

Micrococcus glutamicus/Corynebacterium glutamicum Glucose 30 [128]

Cephalosporium Distiller’s soluble, corn steep liquor 1 to 31/2 [129]
Bacillus strain 14B22 Glucose 12.5 mg/mL [130]
Arthrobacter globiformis Glucose 0.45 M per moles of glucose [131]
Brevibacterium divaricatum Hydrolysed grain sugars 41.5 g/L [132]
Brevibacterium Saccharide material 4 g/dl [133]
Micrococcus glutamicus Molasses 40 g/L [134]
Brevibacterium flavum ATCC No. 13826 Na/K acetate 15 g/L [135]
Brevibacterium roseum ATCC No. 13825 Acetic acid 14.8 g/L [135]
Brevibacterium lactofermentus ATCC No. 13869 Na/K acetate 14.3 g/L [135]
Corynebacterium acetoacidophilum ATCC No. 13870 Acetic acid 7.3 g/L [135]
Corynebacterium hydrocarboclastus (M-104) Glucose 6.3 g/L [136]
Corynebacterium Corn steep liquor 5 g/L [137]
Nocardia globerula ATTC15076 Hydro-carbon 4 g/L [138]
Bacillus thiogenitalis No. 653 Oleic acid 50 mg/mL [139]
Corynebacterium alkanolyticum No. 314 Glycerol 40 mg/mL [140]
Brevibacterium flavum (AJ 3612) Polyoxyethylene-sorbitan-mono-palmitate 52 mg/mL [141]
Brevibacterium lactofermentum 2256 Beet molasses 2 g/dL [142]
Brevibacterium divaricatum NRRL 2311 Ethanol 25 g/L [143]
Brevibacterium (mutant) Glucose 52 g/L [144]
Arthrobacter globiformis Glucose mineral salt 16.1 g/L [145]
Brevibacterium species Glucose (2%) 6.68 mg/mL [146]
Brevibacterium species Sugarcane baggase (10% glucose), dry baggase 80 mg/g [147]
Micrococcus glutamicus + Pseudomonas reptilivora Glucose 37.1 g/L [148]
Brevibacterium sp. DSM. 20411 Cassava starch hydrolysate 21 g/L [149]
Corynebacterium glutamicum 2262 Glucose (fed batch) 80 g/L [150]
Brevibacterium sp. Tc452 Glucose 41.42 g/L [151]
Brevibacterium divaricatum Cassava starch (sub. culture) 3.86% [152]
Corynebacterium glutamicum (ATCC 13032) Lime citrus aurantifoliaswingle + 2% glucose 13.7 g/L [153]
Brevibacterium roseum Glucose medium 0.5 g/L [154]
Corynebacterium glutamicum (CECT 690) Date waste juice 39.32 mg/mL [155]

of GA. When the pH controlling agent was switched from NH4 OH to
NaOH at the end of the growth phase, 18 g/L of ␣-ketoglutaric acid
was accumulated at a yield of 0.20 g/g substrate in 72 h cultivation
[31].
GA is converted into l-glutamine when the microbial culture
is used in presence of excess ammonium chloride at a weakly
acidic pH in the presence of zinc ions [32]. A key compound
controlling GA fermentation is biotin. As shown in Fig. 2, biotin
is a cofactor of acetyl-CoA carboxylase, the first enzyme in the
biosynthesis of oleic acid, and C16 –C18 saturated fatty acids inhibit
the biosynthesis of oleic acid by repressing acetyl-CoA carboxylase
[33,34]. The accumulation of GA is at a maximum when the biotin
concentration is suboptimal for maximum growth. GA producing
cells grown with limited biotin or grown with excess biotin and
treated with either penicillin or Tween-60 excreted intracellular
GA when washed with phosphate buffer. Fig. 3 shows the details
about the regulatory pathway for GA production. The properties of
␣-ketoglutarate dehydrogenase (KD) of GA producing bacteria are
favourable for the preferential synthesis of GA from ␣-ketoglutaric
Fig. 1. A co-ordination between medium design and microbes screening process
has promoted synergistic advantages as well as decreasing the chance of missing a
better performance system [28].
acid, preventing the further oxidation of ␣-ketoglutaric acid to
CO2 and H2 O via succinyl-CoA.
3. Traditional production process
Manufacturing methods of amino acids are categorized as (1)
extraction from hydrolysate of animal or plant protein, (2) fermen-
tation, and (3) enzymatic. Originally, GA used to be manufactured
by extraction from acid hydrolysate of plant protein. In the late
1950s, fermentation technology was established and was used for
commercial production of GA. This was the beginning of modern
amino acid production. Currently, most of the GA is produced by fer-
mentation. In last decade, GA industry developed rapidly, and the
quantum of GA production stood at 1.60 MT in China that is about
70% of the global production. More than 200,000 t of GA is pro-
duced every year. Usually, separation of glutamic acid hydrolysate
(GAH) from its sodium salt is carried out by isoelectric crystalliza-
tion method with or without prior removal of biomass present in
the fermentation broth. However, the presence of biomass reduces
the crystallization process and favours the formation of p-form
of crystal [35]. The fermentation broth still contains 1–2% of GAH
after separation in isoelectric supernatant [36]. The lower solubil-
ity of GAH in water compared to its salt is also a serious problem
for achieving high separation and recovery. Large amount of min-
eral acids and successive washing steps with water are required to
remove the salts formed.
All GA manufacturing industries have switched over to fer-
mentation based technology as presented in Fig. 4. One of the
methods for commercially producing glutamic acid relies on the
fermentation of relatively pure sugars with minimal amounts
of nitrogenous nutrients. After the fermenter broth is filtered,
activated vegetable carbon is used to bleach the calcium glutamate
for production of food grade acid. No activated carbon is used for
the technical grade. After that calcium glutamate is evaporated
to a 37% concentration at temperature around 70 ◦ C and 0.57 atm.
62 R. Kumar et al. / Chemical Engineering and Processing 81 (2014) 59–71
Fig. 2. Cell permeability of l-glutamic acid in relation to phospholipids contents in the membrane.
The next step is acidification with concentrated sulphuric acid
and the calcium sulphate precipitate is removed by continuous
filter and sent back for reuse. The filter acid is then treated with
activated carbon for bleaching. Glutamic acid is evaporated to con-
centrate it in 316 stainless steel evaporated. Heavy metals could
Fig. 3. Regulatory pathway for biosynthesis of glutamic acid.
be removed by ion exchange which may also remove other amino
acids. To get the higher grades of product the liquor is cooled,
crystallized and washed. Crystallization is presently performed in
a single unit and a stainless steel double effect evaporator is used.
Liquid–liquid extraction is another method to get purer form of GA
by using immiscible solvent. The extraction solvent should have
low water solubility, a high distribution coefficient for GA and a
low distribution coefficient for impurities such as residual sugars.
4. Immobilization of micro-organism for the production of
GA
Immobilization has been considered as one useful technique
in microbial production. The major advantages of this applica-
tion are the long-term utilization of biocatalysts and continuous
operation of stabilized systems which lead to reduction of cost of
bioprocessing. Disadvantages may also arise from the diffusional
barrier created by the immobilization matrix as well as the high
cell density. Another disadvantage has been the failure of efforts
to create a versatile, general matrix capable of holding a variety of
cells and functioning in differing bioprocesses. A number of amino
acids have been produced using this methodology. These include l-
aspartic acid [37], l-isoleucine [38], l-serine [39], l-lysine [40] and
l-glutamic acid [41]. Amin et al. [42] had studied the formation
of by-products during glucose conversion to glutamic acid using
Corynebacterium glutamicum immobilized in polyurethane foam.
Entrapment of protoplast of B. flavum in matrices of agar-acetyl cel-
lulose filtering is another attempt to produce GA [43]. Sunitha et al.,
1998, co-immobilized the whole cells of Micrococcus glutamicus and
Pseudomonas reptilivora for the higher yields of 37.1 kg/m3 of GA.
They found that the concentration of glucose, urea and biotin in the
production medium were proved to be the most suitable medium
constituents.
5. Limitations in conventional GA production
The traditional production process consists of a number of
downstream treatment schemes like precipitation, conventional
filtration, acidification, carbon adsorption, evaporation, etc. as
shown in Fig. 5. Thus the overall process plant scheme is quite com-
plex involving energy-intensive and expensive steps releasing huge
amount of wastewater and requiring relatively high manpower.
The other major drawbacks of the existing technology is substantial
 R. Kumar et al. / Chemical Engineering and Processing 81 (2014) 59–71
Fig. 4. Typical conventional fermentation-based glutamic acid production scheme.
demand for harsh chemicals like sulphuric acid, liquid ammonia
and other chemical supplemental compounds that work to the
overall economic disadvantage in production of glutamic acid (GA).
On the other hand, the GA extraction process also produces large
Fig. 5. Flow-sheet of conventional purification of glutamic acid.
63
amount of wastewater leading to environmental pollution [44,45].
Ion-exchange method is normally used for glutamic acid recovery
from fermentation broth without precipitation. The ion exchange
also involves use of large amount of acid and base to regenerate
ion-exchange resins. In addition to this, wastewater produced from
the process in reactivating and washing the ion-exchange resins
also causes a serious environmental pollution. Thus a new process
is urgently required which will be eco-friendly and economically
more attractive. In last two decades, efforts have been made by GA
enterprises and related scientific groups in solving the economic
and environmental problems related to glutamic acid production
[44,45]. Studies have revealed that the huge amount of wastewater
produced in GA extraction process is difficult to be handled with
the traditional end-of-pipe treatment method. During the past
half century, it has been recognized that end-of-pipe treatment is
hardly advisable in tackling environmental issues. Amidst tough
competition in the globalized market, cost of production needs to
be brought down significantly in the current regime of emaciated
profit margin in chemical and allied industries [46]. It has been
observed that there is no substitute to clean production technology
in matters of controlling pollution and bringing down cost [47,48].
6. Membrane processes
6.1. Concepts and principles
Membrane separation involves the use of a selective barrier
(membrane) to regulate the transport of substances, such as gases,
vapours and liquids at different mass transfer rates [49]. The rates
of mass transfer of different substances are controlled by the per-
meability of the barrier towards the feed components [49]. These
membranes can play effective role in downstream purification of
GA by microfiltration, ultrafiltration, nanofiltration, reverse osmo-
sis and electrodialysis membranes. Nanofiltration (NF) membranes
64 R. Kumar et al. / Chemical Engineering and Processing 81 (2014) 59–71
Fig. 6. Schematic diagram of a cross-flow membrane separation process.
have been developed recently and practiced for the separation
of small neutral and charged solutes in aqueous solutions. NF
membranes have two important features in their actual applica-
tions [50,51]. One is the intermediate molecular weight cut-off
(MWCO) between reverse osmosis (RO) membranes and ultrafil-
tration (UF) membranes, which ranges from 200 to 1000; the other
is salt rejection caused by the charge effect due to their materi-
als. They can be identified into the sieving (steric-hindrance) effect
and the Donann (electrostatic) effect from viewpoint of membrane
separation mechanism [52,53]. For continuous mode operation
of a fermentative process using renewal carbohydrate sources
for GA production, the components that need to be continuously
separated from fermentation broth are microbial cells, proteins,
nutrients (yeast extract, salts of ammonium, potassium, phospho-
rus, etc.), unconverted carbon sources, water and GA. Membranes
suffer from fouling by microbial cells and proteins, though extent
of fouling may be far less in nanofiltration/reverse osmosis mem-
branes compared to that in microfiltration membranes. However,
there are some particular membrane based modules which may
be operated long without much fouling like flat sheet cross-flow
types (Fig. 6). To separate the microbial cells for their subse-
quent recycling to the bioreactor to ensure high cell concentration
and thus high productivity, microfiltration membranes normally
used with high pore size (0.1–0.45 ␮m) are among the categories.
Nanofiltration membranes being in between reverse osmosis and
ultrafiltration membranes with average pore size less than 1 nm are
able to separate cells, proteins, nutrients, salts and unconverted car-
bon sources from GA fermentation broth. Reverse osmosis normally
known as nonporous membrane where separation is based on
solution diffusion mechanism can separate the same components
from fermentation broth as nanofiltration membranes do but it
required higher operating pressure than what is needed in nanofil-
tration [16]. Whereas ultrafiltration membranes with average pore
size less than that of the microfiltration membranes can retain
proteins along with the cells. Separation by microfiltration and
ultrafiltration membranes is based on size-exclusion and molecular
weight cut off (MWCO) value should be ensured [54]. Solutes hav-
ing larger molecular weight (MW) than the MWCO of a membrane
are rejected almost by the membrane and the ones having lower
MW than the MWCO of a membrane will permeate easily through
the membrane. That is so called the sieving effect. Thus, solutes
having different MWs can be separated based on sieving effect. The
Donnan effect of a membrane refers to the electrostatic interactions
between ions and the membrane. The membrane is charged and
mostly negatively charged since the thin films of NF membranes
are made of polyelectrolytes. Ions having the same sign of charge
as the membrane charged are excluded, and ions having the oppo-
site sign of charge can be attracted. Separation of electrolytes’ ions
having different signs and valences can be manipulated according
to the rejection differences by the membrane [52,53].
The modules of micro, ultra, nano or reverse osmosis mem-
branes can be coupled with fermenter permitting continuous
removal of acid from the broth and separation and recycle of cells,
nutrients and unconverted carbon sources. The type of membrane
used is the deciding factor for the separation and recycle of the
components. For example on use of microfiltration membrane,
microbial cells could be retained in the retentate side while per-
mitting acids, unconverted carbon sources, protein nutrients and
water to pass to the permeate side. If an ultrafiltration membrane
module is used in place of microfiltration module then proteins
along with cells get retained. This however, ensures continuous
removal of acids from the fermentor helping to arrest lowering of
pH. In such continuous process, pH adjustment may be redundant.
If nanofiltration or reverse osmosis membrane is used in place of
microfiltration then all the components barring acid solution are
retained.
6.2. Operating processes
For operation of membrane, a membrane module, pump, pres-
sure gauge, control valve, rotameter, etc. are required. The type
of membrane used in the module is the deciding factor for using
the type of pump for feeding. For microfiltration membrane,
low pressure (2–4 kgf/cm2 ) pump may be used. For an ultrafil-
tration module, a pump of 4–7 kgf/cm2 is used. Nanofiltration
module demands high pressure pump (5–15 kgf/cm2 ) whereas
reverse osmosis membrane system requires still higher pressure
(>20 kgf/cm2 ) for filtration. The recycling and permeation of the
components depends on the types of membranes used in the mod-
ule. The separation through electrodialysis membrane is based
upon electromigration of ions through a stack of cation and an
anion exchange membrane basically involves two steps – conven-
tional electrodialysis (CEP) and the bipolar electrodialysis (BED).
The first step (CEP) separates and concentrates organic acid salt
and the second step (BED) converts the salt form into acid form.
The next section reviews the developments in the first stage of
separation and purification by microfiltration and ultrafiltration
membranes and then moves over to nanofiltration, reverse osmosis
and electrodialysis.
6.3. Microfiltration and ultrafiltration of fermentation broth
Microfiltration and ultrafiltration of fermentation broth are
generally used for cell recycle and sometime for the recovery
of product. These may be applied to overcome the problem of
substrate-product inhibition during fermentation based batch pro-
duction process [55,56]. In addition to that the time loss for shut
down and start up after every batch of production is major con-
cerned. As the concentration of glutamic acid in the fermentation
cell goes up, microbial activities start getting reduced due to
increased difficulty of survival of microbes in low pH medium. Vari-
ation of pH-effects on bacterial growth results from the presence of
dissociated and undissociated of forms GA in fermentation broth.
The undissociated form is more inhibitory than dissociated form. At
low pH undissociated form dominates and at high pH complete dis-
sociated form of GA takes place. According to Milcent and Carrere
[57], pH is a key parameter for studying the membrane based sepa-
ration coupled to fermentation. Lowering of pH resulted in decrease
of flux and vice versa.
It was identified that critical fluxes were function of cross
velocity. Due to irreversible fouling resulting from adsorption of
molasses compounds used as a carbon source on the membrane
surface, cross flow velocity did not lead to an increase of flux. They
did not reuse the separated cells resulting in low productivity. To
ensure high productivity in a fermenter, two important tasks are
required at the outset, firstly, removal of acid (amino acid) from the
fermentor medium to maintain optimum pH and secondly, recy-
cle of the microbial cells at the late-logarithmic growth phase of
the microbial cells. For that membrane a separation unit should be
 R. Kumar et al. / Chemical Engineering and Processing 81 (2014) 59–71
coupled with a fermentor in an external unit permitting continu-
ous separation and removal of amino acids from the fermentation
broth preventing lowering of medium pH to inhibition level and
this simultaneously permits cell and recycle also. Taniguchi et al.
[58] achieved 29-fold increase (compared to the system without
filtration) by removing organic acid from the broth by a cross flow
filtration and by recycling the cells retained on the microfilter to the
fermentor. This ensured high cell concentration (81.5 g dry cells/L)
in the fermentor. Cell harvesting by microfiltration or ultrafiltra-
tion for its subsequent recycling leads to high cell concentration in
the fermentor but often excessive build up increases viscosity and
causes lowering of flux. To overcome this problem Crespo et al. [59]
suggested cell bleeding.
Membrane fouling is a major barrier in the way of effective use
of the MF/UF membrane. During the micro or ultra-filtration of
the fermentation broth, chances of membrane fouling by microbial
cells, proteins, etc. are obvious. If the membrane module operated
is in dead-end mode, concentration polarizations build up rapidly,
resulting in decreased flux. However, build-up of concentration
polarization is very much dependent on the mode of operation and
type of membranes. Chances of concentration polarization are dras-
tically reduced if the system is operated in a cross-flow module, due
to sweeping of the fluid on the membrane surface. To overcome the
problem of membrane fouling in microfiltration stage, Torang et al.
[60] suggested a shear-enhanced cross-flow ultrafiltration mod-
ule for separation of cells and proteins from fermentation broth.
However, this fouling problem can be significantly reduced with
use of the cross-flow module, as the fluid travels parallel to the
surface of the membrane imparting a sweeping action on the mem-
brane surface and thus leaving very little scope for the formation of
the concentration polarization layer. Fouling problem which is the
most serious obstacle in the application of membrane technology
to the fermentation broth filtration arises due to the formation of a
deposited layer, which has a harmful effect both on the permeation
rate and the ease of membrane cleaning, on the membranes during
ultrafiltration (UF) operation [61]. Although there are possibility to
reduce fouling and delay its onset by adopting hydrodynamic and
back-flushing technique [62,63], changing membrane material and
mode of operation [16], but it is impossible to eliminate the foul-
ing. The membrane manufacturers have recommended majority of
cleaning protocols which include a series of acid-alkali-cleaning
cycles depending on the feed processed and membrane material.
The main way to restore and maintain the permeability and selec-
tivity performance of a membrane system is periodical chemical
cleaning. Another factor is membrane materials which play vital
role in fouling problem. Like using ceramic membranes permitted
easy disinfection, but ceramic membranes suffer from quick foul-
ing and can retain only cells where unconverted substrate gets lost.
Polyamide membranes showed lower flux reduction than Polysul-
fone types but direct ultrafiltration of broth without microfiltration
resulted in quick reversible fouling as the system was operated at
cross flow velocity to protect the microbes from shear.
Hydrophobic (polyethersulfone) membrane (MWCO 25 kDa)
retained 100% protein but due to the blocking of the pores by
protein adsorbed on to the hydrophobic membrane surface the
flux was higher for hydrophilic (regenerated cellulose acetate)
membrane with MWCO of 20 kDa and flux of 12–85 L/(m2 h).
Hydrophilic membranes used had low protein binding tendency.
The chemical stability and as well as protein separation was bet-
ter for hydrophobic membrane so microfiltration or centrifugation
were suggested before the ultrafiltration to make the ultrafiltra-
tion step more efficient by avoiding the fouling of the membrane
by high molecular weight protein. The mechanism of flux decline
by the broth of MF/UF membranes are well understood and docu-
mented, there have been reported data on cleaning membranes
fouled during broth filtration. The formulation of an optimal
65
membrane-cleaning strategy through a systematic approach may
lead to important process improvements including optimized use
of chemicals (hence minimized environmental impact), reduced
loss of production time, improved permeate flux and quality con-
trol, and extended lifetime of the membranes which will add the
economy to the overall processes. By controlling tangential flow
in the PVDF microfiltration membrane (Millipore) attached with a
fermentor for downstream separation of cells, the problem of con-
centration polarization and fouling effect was controlled [64]. They
have claimed that membrane was operated 155 h with on-line ster-
ilization and cleaning of the membrane using NaOCl and distilled
water.
Moueddeb et al. [65] have designed a set-up in microfiltration
membrane bioreactor that consisted of two coaxial alumina tubes
having alpha alumina membrane (pore size: 2.0 × 10−7 m) on the
inner wall of the inner tube and on the outer wall of the outer tube
of the tubular membrane, aimed at total substrate conversion in
to product. The micro-organisms were fixed in the macroporous
support and confined in the annular space of two coaxial porous
tubes of a tubular membrane. The substrate solution was fed into
the reactor inner compartment whereas the liquid percolated in
the radial direction across the two membranes. The organic acid
was produced in the porous space between the two microfiltration
layers. Though the new design focused on elimination of bacterial
inhibition giving total substrate conversion, flux decreased rapidly
due to membrane plugging by microbes which could, however, be
reduced to some extent by sterilization of the ceramic membranes.
Feedback inhibition problem may be removed by direct removal
of product from the fermentation broth. Giorno et al. [66] inte-
grated one cross flow membrane module fitted with microfiltration
or ultrafiltration capillary membranes with a 2.5 L stirred cell. They
have also suggested that the ceramic membranes have the advan-
tage of easy disinfection in comparison to polymeric membranes.
Ultrafiltration membranes can retain both cells and proteins. Xavier
et al. [67] coupled ceramic ultrafiltration membrane to a fermentor
to separate both cells and proteins for their recycling and simulta-
neous removal of acid from the medium.
6.4. Electrodialysis
Electrodialysis (ED) is an electro-membrane process in which
ions are transported through ion permeable membranes from one
solution to another under the influence of potential difference
across the electrodes. Therefore, ED can separate selectively ions on
the basis of their charge [68,69]. The main industrial application of
this process is found in chlor-alkali, water purification, and indus-
trial effluent treatments [70,71]. The most recent application of
electrodialysis is in the separation and production of organic acids
from fermentation broths and enzymatically produced solutions
[72–74]. Electrodialysis (ED) and bipolar membranes electrodialy-
sis (BMED) as well as combination of ED and BMED have also been
widely used for desalination, concentration, separation and purifi-
cation in many fields. Desalination of brackish water [75,76] and
de-ashing of milk whey [77] are the main application areas of ED.
As fermentation based production requires constant maintenance
of near neutral pH for high productivity, this necessitates alkali in
most of the cases where product is acidic in nature. Traditionally,
GA is separated from fermentation broth by isoelectric crystalliza-
tion under low temperature with or without biomass. About 1–2%
residual GA still remains in isoelectric supernatant, after separa-
tion through crystallization. Ion-exchange process was used for
the removal of residual GA from isoelectric supernatant [78]. Sub-
stantial amount of acid and base is required in regeneration of
ion-exchange resins. This raises cost of recovery of glutamic acid
by ion-exchange resins. In addition, wastewater produced from the
process to reactivate and wash the ion-exchange resin will cause
66 R. Kumar et al. / Chemical Engineering and Processing 81 (2014) 59–71
serious environmental pollution and increase the burden on sub-
sequent wastewater treatment. Zhang et al. [36] have used two
electrodialysis processes: two-compartment bipolar membranes
electrodialysis and modified traditional electrodialysis, to recover
glutamic acid from isoelectric supernatant. Comparison of the two
electrodialysis processes indicated that the modified traditional
electrodialysis was more efficient method than two-compartment
BMED. Higher recovery ratio of glutamic acid and lower energy con-
sumption were obtained by the modified traditional electrodialysis.
Thus in the light of above facts, two routes of the membrane
technologies can be used for the recovery of GAH from GANa: (i)
the BMED process, to convert GANa into GAH and caustic soda
and their recovery; (ii) acidification of GANa by sulphuric acid and
subsequent separation of GAH and sodium sulphate by ED using
ion-exchange membranes. In recent years, ED processes are also
used in the production of organic acids, such as acetic acid, propio-
nic acid and lactic acid [79–82]. Due to its ions selective transport,
organic acid can be separated and concentrated. The problem of
disposal of by-product gypsum associated with conventional fer-
mentation process can be largely overcome through electrodialysis.
Datta et al. [83] studied the advantages associated with such elec-
trodialysis process. For maintaining pH, agents like NH4 OH or NaOH
used during fermentation can also be recycled by ED, which will
simultaneously reduce the environmental pollution [84,85].
Recently, bipolar membrane electrodialysis (BMED) has been
developed for the conversion of salts into corresponding acids and
bases [86,87]. The bipolar membranes are able to split water at high
applied potential, and generate the hydroxyl and hydrogen ions at
the anode and cathode side. Several studies have shown that, the
BMED process has economic potential for recovery of inorganic,
organic, and amino acids [88].
Energy consumption associated with ED processes is nor-
mally high and to make it economically attractive, attempts have
been made to increase energy efficiency. Shen et al. [89] car-
ried out experiments in the modified laboratorial electrodialyser
consisting of seven compartments with alternating cation- and
anion-exchange membranes, which was a self-made apparatus for
the separation of glutamine from its fermentation broth. The opti-
mized energy consumption was 1.95 kW h kg−1 sulphate under the
operation of current density of 204 A/m2 at a current efficiency of
81%. Kumar et al. [90] have developed an electro-membrane reac-
tor with four compartments (EMR-4) (anolyte, catholyte and comp.
1 and 2) based on in-house-prepared cation- and anion-exchange
membrane (CEM and AEM, respectively) to achieve separation and
recovery of glutamic acid (GAH) from its sodium salt by in situ ion
substitution and acidification.
Despite several studies have been undertaken to establish the
potential of bipolar electrodialysis as an efficient and eco-friendly
method of glutamic acid production, concentration and purifica-
tion, commercialization of bipolar membrane itself has been done
in very limited cases and electrodialysis fermentation (EDF) for
glutamic acid has hardly been commercialized. Due to the poor
conductivity of the organic phase power consumption in electro-
dialysis process is always high.
6.5. Nanofiltration and reverse osmosis
Purification and recovery of amino acids by NF is relatively new
class of the pressure-driven membrane processes and its appli-
cation for such purposes is a viable alternative over traditional
separation processes like extraction, ion-exchange, evaporation
and distillation. NF membranes have been developed recently and
adept for the separation of small neutral and charged solutes in
aqueous solution. The performance of a nanofiltration system in
terms of flux and retention of the target solutes will depend on
solution pH, cross-flow rate, membrane type, membrane module
and presence of impurities. Nanofiltration (NF) is widely used in
water softening, dye recovery, bio-product separation, desalina-
tion and wastewater treatment [91–95]. NF membranes can reject
salts due to charge effects since the separation layer is made of
polyelectrolytes [96]. Nanofiltration applied to the separation of
l-glutamine from fermentation broth in the place of traditional
ion-exchange technology not only can reduce acid and basic waste
treatment but also can prevent the conversion of l-Gln into l-Glu on
the ion-exchange resin in the ion exchange separation process [97].
Many studies have already been focused on the separation of
amino acids by NF membrane [98,99]. Tsuru et al. [98] firstly found
that several commercial polymeric NF membranes tested for sin-
gle amino acids and peptides at their pI (net charge zero) showed a
lower rejection than for amino acids and peptides with a net charge.
They also showed that the separation of amino acids and peptides
could be manipulated by modifying the pH value of the solution.
The rejection difference between charged and non-charged amino
acids was mainly explained by occurrence of the Donnan effect [98].
During the NF of a mixture of nine amino acids, acidic amino acids
(anions) are separated at pH < 3 and basic amino acids (cations)
are separated at pH > 9 [99]. Whatever the solution studied, charge
effects, repulsion of co-ions and attraction of counter-ions, more
than sieving effect, prevail in the behaviour of the solute [100]. NF
of amino acids was strongly influenced by the solute environment
and by hydrodynamic parameters [101]. The selectivity of the sepa-
ration of amino acids can be manipulated by modifying the charged
state of the amino acids by changing the pH value, or by modi-
fying the salt composition and concentration of the feed solution
[102,103]. Nanofiltration of some amino acids like l-phenylalanine
and l-aspartic acid aqueous solution were carried by Wang et al.
[104] by using two commercial NF membranes (ESNA2 and ES20).
They found that the rejections to l-Phe and l-Asp by ESNA2 mem-
branes are about 0 and 90% respectively at the pF value ranging
from 4 to 9, while ES20 membrane are almost 100% irrespective of
pH value. They have concluded that these two NF membranes are
possible to concentrate and separate l-Phe and l-Asp effectively by
choosing proper condition such as the pH value of solution.
Separation of l-glutamine (l-Gln) from Gln fermentation broth
by nanofiltration (NF) was investigated with changing the exper-
imental parameters such as transmembrane pressure, pH and
concentration of broth on the rejection of l-Gln and l-glutamate
(l-Glu) showed that NTR7450 was able to effectively separate
l-Gln and l-Glu when the appropriate conditions were chosen
[105]. Kovacs and Samhaber [106] have used the nanofiltration
membrane for the concentration of amino acid. To determine the
permeate flux and amino acid rejection from aqueous solutions
diprotic amino acids (l-glutamic and glycine) as a function of
increasing feed concentration and ionization state of the amino
acids, Kovacs and Samhaber [106] have used numerous polymeric
NF and tight UF membranes. The concentration of amino acids in the
whole range of their solubility was studied with a stepwise pH scan
ranging from 0 to −1 total net charge. Considerable higher rejection
and flux drop over the concentration was observed in higher pH
range, where amino acids are present in dissociated form. Mem-
branes with different types of active layer material show similar
concentration dependent tendency in the permeation behaviour.
This phenomenon can be explained by the dissociation dependency
of the osmotic pressure. In case of glutamic acid, at pH 8, where
net charge is −1, a less pronounced rejection drop (95–75%) was
measured over the concentration than close to its isoelectric point
(from 90 to 5%). McGregor [107] has used the thin film composite
reverse osmosis (RO) membrane to concentrate l-phenylalanine
from clarified bioreactor harvested media. He achieved the 100 g/L
concentration at pH 10 and 50 ◦ C with flux from 17 to 119 L/(m2 h).
He suggested that applications of RO are likely to be case spe-
cific.
 R. Kumar et al. / Chemical Engineering and Processing 81 (2014) 59–71
A combined process of nanofiltration and reverse osmosis was
developed by Li et al. [108] for separation and concentration of lac-
tic acid from cheese whey fermentation broth. Five NF membrane
(CK, DK, DL, HL, and GE) and two reverse osmosis membrane (DS 11
AG and ADF) were tested at different pressures. After nanofiltration
reverse osmosis was applied to concentrate lactic acid, 100% lactic
acid retention was achieved at 5.5 MPa pressure for ADF mem-
brane compared to DS11 AG membrane 96% at the same pressure.
Membranes suffered quick fouling in absence of prior filtration of
microbial cells.
6.6. Chemistry of NF membrane for separation of solute
Estimation and use of effective membrane charge density for
practical applications is rather complex and very few quantita-
tive description of the transport phenomena of amino acids has
been reported so far. Teixeira et al. [109] have demonstrated the
importance of divalent hardness (CaCl2 and MgSO4 ) on the NF per-
formance with pH in terms of both flux and retentions of ions by
using natural water. The steric hindrance and membrane solute
interactions are two major factors for separation of nanofiltration
membrane [110,111]. For the retention of uncharged molecules,
steric hindrance and non-electrostatic membrane–solute interac-
tions (e.g. Van-der-Waals forces) are mostly responsible, and their
transport takes place by convection due to a pressure difference and
by diffusion due to a concentration gradient across the membrane
[112,113]. Polarity decreases retention, which can be explained by
electrostatic interaction directing the dipole towards the mem-
brane [114]. Steric hindrance and electrostatic interactions are
responsible for separation for charged compounds [110]. In addi-
tion to that another important parameter in the transport process
through the membrane is the membrane charge along the surface
and through the pores [115]. When membrane surface in con-
tact with an aqueous solution acquires an electric charge like the
ionic surfactants, adsorption of polyelectrolytes or ions from the
solutions takes place along with dissociation of surface functional
groups [116]. To maintain the electroneutrality of the system, this
charging mechanism takes place on the exterior membrane surface
and on the interior pore surface of the membrane, because of the
distribution of ions in solution [115]. The ion separation resulting
from the electrostatic interactions between ions and membrane
surface charge is based on the Donnan exclusion mechanism [117].
In this mechanism the co-ions (which have the same charge of the
membrane) are repulsed by the membrane surface and to satisfy
the electroneutrality condition, an equivalent number of counter-
ions is retained which results in salt retention.
Retention of neutral solutes by size exclusion is dependent on
two parameters of membrane (effective pore radius, thickness–
porosity ratio) and also on stoke radius of solute. In addition to
the solute transport, membrane properties like pore radius (rp ),
effective thickness–porosity ratio (x/Ak) are determined by com-
parison and convergence between model and experiment rejection
data of sucrose, undissociated glutamic acid as neutral solutes.
Ionic rejection is mainly based on Donnan effect and dependent on
three parameters (effective pore radius, thickness–porosity ratio
and effective charge density of membrane).
6.7. Effect of pH of the solution on the NF membranes
performance
The osmotic pressure of feed solution containing amino acid is
greatly affected by the pH. Amino acids are ionizable compounds.
In neutral aqueous media, diprotic AA with non-ionizable residual
group is predominantly present in zwitterionic form as a result of
intermolecular proton transfer. If the pH of the solution is higher
than their isoelectric point, dissociation takes place, which results
67
in the anionic form. It has been reported in several studies, that
a considerably higher rejection can be observed for AA of anionic
form than for zwitterions [104]. As the amount of undissociated
glutamic acid and glutamate salts present in glutamic acid solution
depends on the pH equilibrium, the Henderson–Hasselbalch equa-
tion may be incorporated in model development to correlate the
pH effect on glutamic acid transport.
Apparent pore size of polyamide NF membranes can also vary
with solution pH. The protonation and deprotonation of the func-
tional groups present in the membrane surface and the molecules
in the solution, are very much dependent on pH over its range. This
will change the membrane charge and the size of the membrane
pores with consequences in the NF and UF performance [118]. At
the pore surface points of zero charge (isoelectric point), the mem-
brane functional group is minimal in charge and hence opens up,
as the absence of repulsion forces contribute to the widening of
the membrane pores. At high or low pH value, functional groups of
membrane polymers can dissociate and take on positive or nega-
tive charge functions. Interactions between these functions in the
membrane polymer reduce or close up membrane pores. At higher
pH values, addition of sodium hydroxide leads to an increase in
osmotic pressure and ionic strength, thus reducing the membrane
permeability and increasing rejection. Moreover, functional groups
such as carboxyl and hydroxyl groups present at the surface of the
membrane become deprotonated at high pH. High pH results in
an increased thickness of the double layer of the charged func-
tional groups over the surface of the membrane thus reducing the
apparent pore size and resulting in greater rejection of the charged
solutes [119]. In addition to that solution chemistry of natural
waters like pH, alkalinity, salinity, TDS and hardness of cations play
a significant effect on the membrane charge and on the character-
istics of the molecules in the solution. The NF membranes interact
with hardness cations during separation process, so they could
have a marked effect on fouling and NF performance [120]. Stream-
ing potential measurements along surface and through pores with
several electrolyte solutions at different pH help to investigate the
charge of the membrane surface and pore.
7. New approach: integrated membrane system
Membranes can be tailor-made in such a way that a high
degree of selectivity can be ensured, which in turn means that
very high degree of purity can be achieved during downstream
processing. Membrane fouling that is considered a major hindrance
in membrane separation can be overcome by using proper mod-
ule (flat sheet cross-flow membrane module). In the synthesis and
down-stream processing of amino acids (AA), the purification and
recovery is a challenge, where NF is a promising separation tool.
After fermentation, glutamate is the most abundant free amino acid
in bacterial cytoplasm and when micro-organism overproduces
glutamate in excess of their normal metabolic needs, it excretes
into culture broth. The major technology barrier in cost-effective
production of high purity of glutamic acid is its down-stream sep-
aration and purification from the fermentation broth. And this is
where, membrane-based processes are stepping in. Being modu-
lar in design, membrane-based processes offer great flexibility in
scale of production depending on market demand. By virtue of high
selectivity, membranes can ensure high levels of separation and
purification. As membranes of chosen selectivity and permeability
can easily be integrated with conventional fermenters, membrane-
based processes permit simultaneous production and purification
in the same unit. This eliminates the need for separate purifi-
cation units and results in compact design with reduced capital
investment. Membrane-based separation and purification (barring
pervaporation) involves no phase change ensuring reduced energy
68 R. Kumar et al. / Chemical Engineering and Processing 81 (2014) 59–71
consumption. Thus such processes can meet all the goals of process
intensification.
For downstream processing, conventional processes often
require many chemicals that may lead to environmental pollu-
tion. Many downstream units in conventional process are also
energy-intensive. Compared to such conventional processes, mem-
brane based processes involve very low energy consumption. With
integration of membrane modules with fermenter, continuous pro-
duction can be ensured with recycle of cell and unconverted carbon.
In such continuous mode of fermentation and purification, pro-
duction can be ensured without any pH adjustment. Recycling
of cells through the retentate of the microfiltration unit ensures
high sell concentration in the fermenter leading to high produc-
tivity. This may be a simple production scheme involving a few
steps only contrary to the use of a number of downstream treat-
ment steps of conventional production scheme. So many units in a
conventional process such as precipitation, filtration, acidification,
extraction, neutralization, carbon adsorption, crystallization and
evaporation as shown in Fig. 4 can be turned redundant. In conven-
tional production process, addition of lime for controlling pH leads
to production of calcium glutamate. Calcium glutamate is then sep-
arated from the microbial cells by filtration and, further purified
by activated carbon adsorption. In next phase, calcium glutamate
is evaporated and acidified by sulphuric acid to produce glutamic
acid. Conventional batch fermentation also suffers from low volu-
metric productivity due to both substrate and product inhibitions
in addition to involvement of high labour cost following shutdown
and start-up of such batch processes [64]. Over the last two decades,
several attempts have been made in this direction of integration of
traditional fermentor with membrane based separation and purifi-
cation. Literature shows that membrane integrated processes are
rarely used for the continuous production. In most of these stud-
ies, however, only a single stage of membrane separation has been
integrated with fermentor. In addition to that in these cases, stud-
ies have been conducted with finished raw materials like glucose.
Serious fouling is another problem associated with these mem-
brane modules used in these studies [16]. US Patent [121] claimed
to have developed a membrane-integrated process with the com-
bination of ceramic tubular UF module in the first stage for cell
separation and NF in the second stage for some organic acid separa-
tion. In very few studies, two-stage membrane separation has been
attempted. Gonzalez et al. [122] recovered lactic acid from ultrafil-
tered whey by two types of membranes – spiral wound DK 2540C
and tubular AFC80 nanofiltration membranes. Some authors have
produced organic acid by integrating membrane separation with
conventional fermentation process and have shown the possibility
of similar process intensification for many other chemical process
industries [123,124]. The situation improved when MF/UF of the
broth was done prior to nanofiltration or reverse osmosis. Two
stage novel membrane-integrated (micro and nano membrane) fer-
mentor under non-neutralizing conditions. Dey and Pal [125] have
used novel two stage membrane integrated system for the simul-
taneous production and purification of organic acid (lactic acid).
Membrane-integrated hybrid cell recycle bioreactors system
with continuous fermentation can mostly overcome these prob-
lems due to high cell density, much higher productivity and higher
acid concentration in a continuous process [16]. One of the most
vital criteria for continuous fermentation is steady state operation
with prolonged exponential growth phase and needs to be main-
tained with proper cell bleeding. The long term performance of
such a membrane-integrated system ensures a largely fouling-free
operation to ensure a desired flux for commercial viability when
operated by a properly selected membrane module as demon-
strated by Sikder et al. [126]. It can offer such fouling free long
hours of operation by virtue of sweeping flow of the fermentation
broth over the membrane.
To improve glutamic acid concentration as well as productiv-
ity, studies on multistage membrane cell recycle bioreactor may
be exploited. The productivity and yield are based on use of car-
bon source like glucose. Alternate renewal carbon source should
be searched to make the process more economic. To ensure long
term operation, selection of membrane module is also very impor-
tant, like involving expensive hollow fibre membrane modules
that accounted for 28% of the total capital cost. In this membrane
module, additional purification steps like precipitation through sul-
phuric acid, colour removal using adsorbent are necessary to get
the final product as pH adjustment is still done resulting in either
sodium or ammonium glutamate instead of glutamic acid directly.
Multivalent ions and disaccharides are rejected by nanofiltration
membranes [127]. Understanding these effects is essential to suc-
cessful modelling and scaling up of the process. Thus for better
understanding of the hydrodynamics and transport phenomena in
a more realistic setting pure glutamic acid can be directly obtained
instead of glutamic acid salt like sodium glutamate which domi-
nates most of the reported studies in the available literature.
8. Discussion
The present review study aimed at investigating the feasibility
of using nanofiltration for separation of unconverted sugars from
fermentation broth and purification of glutamic acid solution under
different operating conditions with provision of immediate sepa-
ration of glutamic acid from fermentation media eliminating the
need of addition of caustic solution for pH adjustment. Model solu-
tions were also filtered along with actual fermentation broth to
find out the impact of very low pH on the transport phenomena as
during actual fermentation without pH adjustment in a membrane-
integrated system, low pH regimes are likely to dominate.
9. Conclusion
It thus transpires that through the tireless efforts of early
researchers on membrane-based production schemes, world has
definitely moved towards a better process but serious attention still
needs to be paid to some areas to evolve a smaller, more compact,
more flexible, and less energy-intensive plant that could guarantee
large scale production of a highly demanding chemical product in
an environmentally benign process. Such a plant, in other words,
may be called to represent high degree of process intensification
which modern chemical process industries are desperately seek-
ing for their survival in highly competitive and environmentally
conscious world market. Selection of appropriate membranes as
well as modules in cell separation and product purification along
with provision of logical sequencing of operations are essential in
truly achieving such process intensification.
Acknowledgment
Authors are thankful to the Department of Science and Tech-
nology, Government of India for financial support under Start-Up
Research Grant for Young Scientist (SERB) (SB/FTB/ETA-59/2013).
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