Revised Assignment 1

UNIVERSITAS INDONESIA
PRODUCTION OF GLUCONIC ACID FROM SUGARCANE JUICE BY

AEROBIC SUBMERGED FERMENTATION USING ASPERGILLUS
NIGER
Revised Assignment 1
GROUP 05
GROUP PERSONNEL:
ALWENDO GUNAWAN (1506723452)
DIMAS RAHADI PITOYO (1506673473)
SAMSON PATAR SIPANGKAR (1506723774)
WIDA ADELIA PUTRI (1506673290)
YAZIDIE RIZQI ISNAINDI (1506728434)
CHEMICAL ENGINEERING DEPARTMENT

ENGINEERING FACULTY
UNIVERSITAS INDONESIA
DEPOK
OKTOBER 2018
EXECUTIVE SUMMARY
Gluconic acid is a chemical commodity with many applications in food and

chemical industries. This compound is industrially produced from glucose whereby
gluconic acid is derived by chemical or microbial oxidation. The glucose itself is
chosen to be sourced from sugar cane juice using fermentation process. In general,
gluconic acid and is used as a food additive and as acid regulators in food
formulations, pharmaceuticals, and cleaning products for industries. In addition,
gluconic acid is also used as a supplement to prevent calcium deficiency, iron and
as a buffer salt. Different gluconic acid salts have different applications according
to their respective properties.
The most well-known industries in Indonesia gluconic acid is food and
cosmetics. The world's need for gluconic acid continues to increase along with the
development of its user industries. The trade value of organic acids estimated will
reach $25 million in 2024. Until recently, Indonesia's demand for gluconic acid is
still fulfilled through the import channel, considering that currently there is no
producing plant in Indonesia gluconic acid.
Based on data from various sources, it is noted that gluconic acid plant can
utilize sugarcane juice from fermentation plant surroundings in East Java Province.
In 2013, the production of sugarcane reached 2.55 million tons and increased 0.86%
in 2014 to be 2.58 million tons. While in 2015, sugarcane production decreased by
1.57% (2.53 million tons with the production in East Java was 1.24 million tons
(48.75% of overall sugarcane production in Indonesia).
Gluconic acid is produced from the glucose by conversion using
fermentation process. In moving toward sustainable and clean technology regime,
chemical and biochemical process industries are gradually adopting bio-based
production strategies rather than pure chemical technologies. The gluconic acid will
be produced as a food grade product and targeted the industry of pharmaceuticals,
food, and beverages.
This production strategies consist of plenty process alternatives on every
process in fermenting gluconic acid. Five processes will be considered to make the
production as economic and as environmentally safe as possible. These processes
ii Universitas Indonesia
include reaction process, glucose pre-treatment process, incubation process,
purification process, and drying process. After making consideration parameter and
giving score to each parameter, one best process is selected from few alternatives
available for each process. Fermentation reaction using Aspergillus niger as the
fermentative agent is used for the reaction of the process. Pre-treatment process
chosen is pH regulation, sucrose hydrolysis, carbon filtration, and sterilization. In
incubation process, the medium chosen is PDA (Potato-Dextrose-Agar) due to its
short inoculation time and better quality even though it is a bit costly.
Microfiltration and Nanofiltration membranes are used in purification process due
to its lower energy consumption, sustainable processing, simpler operation and
relatively easy scale-up provisions. In drying process, spray dryer is used because
it is possible to make powdered product from its usage. For fermentating agent,
bacteria, and Method, Aspergillus niger, Gluconobacter oxydans, and Submerge,
respectively is used in this Gluconic acid fermentation.
The mass and energy balance calculation were done by using simulation
program in SuperPro. SuperPro was used because of its advantages which can
proceed and provide bio-solid material properties such as biomass. Based on the
simulation, the Gluconic acid that can be produced is about 13.33 ton per day (3200
ton per year), converted from (9.6) ton per day of sugar cane juice from Malang
region with assumption the effective working day is (240) days a year. With the
presence of gluconic acid, it is expected that it will reduce the number of imports
carried out by Indonesia which reduce national gluconic acid needs with the aim of
increasing the economy of our country by exporting gluconic acid to the worldwide.
The operational conditions - in the simulators - are obtained from many sources
such as paper, book, and trial-error to get the highest gluconic acid yield.
After doing several calculations using SuperPro, the flowrate of glucose
inputted to the production is 400 kg/h. the glucose will be reacted with oxygen from
the air to produce gluconic acid. The flowrate of gluconic acid produced is 525.84
kg/h. The conversion efficiency calculated is 57.65%. the yield is 1.0612 kg
gluconic acid per kg glucose. The energy consumption needed for every product is
5838.2 kWh per 1 kg gluconic acid. The production of gluconic acid is estimated
will diminish the number import which will help the economy of Indonesia.
iii Universitas Indonesia

LIST OF CONTENT
LIST OF CONTENT ............................................................................................iv

LIST OF TABLES ................................................................................................. v
LIST OF FIGURES ..............................................................................................vi
CHAPTER 1 INTRODUTION ............................................................................. 1
1.1 Background ...................................................................................... 1
1.2 Review of Literature ........................................................................ 2
1.2.1 Fermentation .................................................................... 2
1.2.2 Gluconic Acid .................................................................. 6
1.2.3 Glucose ............................................................................ 9
1.3 Market Analysis ............................................................................. 12
1.4 Raw Material Analysis .................................................................. 14
1.4.1 Glucose .......................................................................... 14
1.4.2 Fermentation Agent ....................................................... 17
1.5 Plant Location Analysis ................................................................. 19
CHAPTER 2 PROCESS SELECTION ............................................................. 22
2.1 Raw Material Selection.................................................................. 22
2.1.1 Sugarcane ....................................................................... 23
2.1.2 Carbohydrate-Containing Waste ................................... 23
2.2 Process Synthesis............................................................................ 23
2.2.1 Process Production Method Selection ........................... 24
2.2.2 Selection of Fermentation Process................................. 25
2.2.3 Fermentation Agent ....................................................... 27
2.2.4 Process Selection ........................................................... 29
2.2.5 Process Description ....................................................... 34
2.3 Block Flow Diagram ...................................................................... 38
2.4 Process Flow Diagram ................................................................... 39
CHAPTER 3 MASS AND ENERGY BALANCE ............................................ 40
3.1 Mass Balance .................................................................................. 44
Table 3.1. Mass Balance of Overall Process .......................................... 44
Table 3.1. Mass Balance of Overall Process (continued) ...................... 45
3.2 Energy Balance............................................................................... 47
3.3 Product Conversion Efficiency ..................................................... 47
3.4 Product Yield.................................................................................. 48
3.5 Energy Consumption of Unit Product ......................................... 48
CHAPTER 4 CONCLUSION ............................................................................. 49
REFERENCE ....................................................................................................... 50
iv Universitas Indonesia
LIST OF TABLES
Table 1.1. General Characteristics of Gluconic Acid .............................................. 7

Table 1.2. Physical Properties of Glucose ............................................................. 10
Table 1.3. Sugar Content of Selected Common Plant Foods (g/100g) .................. 11
Table 1.4. Calculation of Production Capacity ...................................................... 14
Table 2.1. Parameter of Scoring for Fermentation Method Selection ................... 26
Table 2.2. Scoring for Fermentation Method Selection ......................................... 27
Table 2.3. Parameter of Scoring for Fermentation Agent Selection ...................... 28
Table 2.4. Scoring for Fermentation Agent Selection ........................................... 28
Table 2.5. Parameter of Scoring for Reaction Method Selection .......................... 30
Table 2.6. Scoring for Fermentation Reactor Selection ......................................... 30
Table 2.7. Parameter of Scoring for Incubation Method Selection ....................... 31
Table 2.8. Scoring for Incubation Method Selection ............................................. 32
Table 2.9. Parameter of Scoring for Fermentation Reaction Method Selection .... 33
Table 3.1. Mass Balance of Overall Process.......................................................... 44
Table 3.2. Energy Balance of Overall Process ...................................................... 47
v Universitas Indonesia
LIST OF FIGURES
Figure 1.1. Chemical Structure of Gluconic Acid ................................................... 6

Figure 1.2. Indonesia Sugarcane Plantation Area Maps ........................................ 15
Figure 1.3. Sugarcane Production Development ................................................... 16
Figure 1.4. Plant Location...................................................................................... 20
Figure 1.5. Actual Area of The Plant ..................................................................... 21
Figure 2.1. Black Box Diagram ............................................................................. 24
Figure 2.2. Fermentation Reaction of Gluconic Acid Production ......................... 36
Figure 2.3. Block Flow Diagram ........................................................................... 38
Figure 3.1. SuperPro Simulation for Pre-Treatment Process ................................. 41
Figure 3.2. SuperPro Simulation for Fermentation and Purification Process ........ 42
Figure 3.3. SuperPro Simulation for Drying Process ............................................ 43
vi Universitas Indonesia
CHAPTER 1
INTRODUTION
1.1 Background
In moving toward sustainable clean technology regime, chemical and
biochemical process industries are gradually adopting bio-based production
strategies rather than pure chemical technologies. However, bio-based production
alone cannot ensure eco-friendliness of a process technology as downstream
separation–purification of the products often involves quite a few energy-intensive
steps that may require harsh chemicals also. This is where membrane-based
purification schemes are stepping in. In this context, development of a clean
technology for production of gluconic acid assumes significance. Being a
multifunctional organic acid, the gluconic acid (GA) has wide applications in food,
pharmaceutical and chemical industries.
Pentahydroxycaproic acid, ("# $%& '( ) or gluconic acid, naturally occur in
plants, fruits, wine, honey, rice, meat, vinegar and other natural sources. The alkali
salt of gluconic acid such as calcium gluconate or sodium gluconate are extensively
used in chemical, pharmaceutical, food, beverages and construction industries
being a multifunctional carbonic acid. Due to multiple applications of metal
glutamates in different industries, the demand of gluconic acid is steadily increasing
globally. By virtue of low toxicity, low corrosiveness and high capability of
forming water-soluble complexes with divalent and trivalent metal ions, sodium
gluconate has been designated as GRAS (generally recognized as safe) by the US-
FDA (United States-Food and Drug Administration). Some specific properties of
gluconic acid permits its wide applications in preventing the deposition of milk
stone or in removing it in dairy industry and in preventing cloudiness in beverages.
Gluconic acid is also used as an additive in cement to control the setting and
increase the strength and water resistance. This organic acid can produce and
improve the taste and can form complexes with metal ions in various foods. It is
also used as water conditioner by removing alkali and protein films. The non-
corrosiveness nature of this acid may be used as gentle metal cleaning agent where
cleaning and degreasing of ferrous and non-ferrous metal ions like Ca2+, Fe2+,
1 Universitas Indonesia
2
Al3+ and other heavy metals is possible. Sodium gluconate is used in the shampoo
and toothpaste as a chelator. In dishwashing and laundry detergent application, this
acid can replace toxic phosphates which are harmful to the bio-environment. It can
be used with artificial sweetener. For its non-corrosiveness nature, gluconic acid is
added in low concentration in water recirculation systems such as cooling towers
and heat exchangers. Gluconic acid finds application in tanning, textile industry and
in fermented tea for its anti-microbial activity.
In last 20 years, demand of gluconic acid has increased steadily reaching
60,000 tons per year. Market researcher Global Industry Analysts predicts that the
global market for organic acids will amount to EUR 1 billion by 2016, driven by
increasing demand in developing economies, stable demand for meat and meat
products from the developed economies and, a growing global population. And
some industry sources consider this figure as conservative. Use of gluconic acid
and its derivatives is currently restricted in many cases because of high prices which
is about USD 1.2–8.5/kg due to use of glucose as substrate and the system specific
requirements during fermentation. However, its increasing demand in different
industries has spurred interest in the development of an effective and economically
viable system for gluconic acid production.
1.2 Review of Literature
This report will mainly discuss about Fermentation, Glucose and Gluconic
Acid. Therefore, in this sub-chapter are some theories about it such as what is
Fermentation, Fermentation method and agent, what is glucose, the properties,
occurrence and also its application.
1.2.1 Fermentation
Fermentation is a metabolic process that consumes sugar in the absence of
oxygen. The products are organic acids, gases, or alcohol. It occurs in yeast and
bacteria, and also in oxygen-starved muscle cells, as in the case of lactic acid
fermentation. The science of fermentation is known as zymology.
1.2.1.1 Fermentation Method
There are so many ways to ferment but we have shrunk to 2 sections which
is Solid State Fermentation and Submerged Fermentation.
a) Solid-State Fermentation
Universitas Indonesia
3
Solid State Fermentation has been widely used and has gained renewed
attention as an alternative for submerged fermentation. It provides the opportunity
for economical production of fermented food, amino acids and flavor inducing
compounds by inducing the natural growth on water- insoluble solid support in the
absence or near absence of free liquid medium. SSF is better than submerged culture
due to less variation of osmotic pressure, DO concentration, availability of water
and nutrients. Buckwheat substrate was used as SSF medium for the production of
spores of Aspergillus niger and these entrapped spores were used for the
bioconversion of glucose to gluconic acid with conversion rate of 1.06 g per mass
of glucose without any nitrogen source.
b) Aerobic Submerged Fermentation
Submerged fermentation is a process involving the development of
microorganisms in a liquid broth. This liquid broth contains nutrients and it results
in the production of industrial enzymes, antibiotics or other products. The process
involves taking a specific microorganism such, as fungi, and placing it in a small
closed flask containing the rich nutrient broth. A high volume of oxygen is also
required for the process. The production of enzymes then occurs when the fungi
interact with the nutrients on the broth resulting in them being broken down. At
industrial level this production of yeasts has become a major output of
microbiological industries as a result of improved fermentation technologies.
Fermentation in industries is carried out using fermenters which are large vessels
which can store huge volumes. In an effort to reduce nitrogen and carbon levels,
microorganisms secrete enzymes in the selected medium. There are two common
methods by which submerged fermentation takes place; they are batch-fed
fermentation and continuous fermentation. In batch-fed fermentation sterilized
growth nutrients are added to a culture. It is most common in bio-industries as it
occurs during the growth of bio-mass in the fermenter. It helps raise the cell density
in the bioreactor and it is typically highly concentrated to stop dilution. The rate of
growth in the culture is maintained by adding nutrients, this also reduces the risk of
overflow metabolism. An open system is constructed for continuous fermentation.
Then sterilized liquid nutrients are slowly and continuously added to the
bioreactor at the same rate at which the converted nutrient solution is being
4
recovered from the system. This results in a steady-rate production of the

fermentation broth. In order to maintain a successful fermentation, certain variable
must be monitored, for example, temperature, pH, as well as oxygen and carbon
dioxide levels.
1.2.1.2 Fermentation Agent
Fermentation agent that is potential for converting glucose to gluconic acid
are bacteria and fungi species. Bacteria that commonly used for gluconic acid
production are Pseudomonas fluorescens and Gluconobacter oxydans. Meanwhile,
the filamentous fungi that usually used is Aspergillus niger.
a) Pseudomonas fluorescens
Pseudomonas fluorescens is a member of the fluorescent pseudomonad group
and (unlike P. aeruginosa) has generally been regarded to be of low virulence and
an infrequent cause of human infection. However, it has been reported to cause
infections such as blood transfusion-related septicemia, catheter-related bacteremia,
and peritonitis in peritoneal dialysis patients.
Pseudomonas fluorescens is a rod-shaped aerobic, non-lactose-fermenting,
Gram-negative bacterium. It can survive and replicate in moist reservoirs, and as a
result, nosocomial outbreaks often lead to the investigation of water sources.
Optimal growth generally occurs at lower temperatures than those for P. aeruginosa,
which can make identification difficult at the standard microbiology laboratory
incubation temperature of 37°C. It can grow at temperatures as low as 4°C,
temperatures at which blood products, distilled water, and disinfectants provide the
ideal environment for proliferation.
b) Gluconobacter oxydans
Gluconobacter oxydans, previously known as Acetobacter suboxydans, are
Gram-negative rod or oval shaped bacteria ranging from about 0.5 to 0.8mm x to
4.2mm. The name oxy from Gluconobacter oxydans is Latin for 'sharp' and 'acidic',
and dans is 'giving'. They tend to have a small genome size because of their limited
metabolic abilities. These abilities include partially oxidizing carbohydrates and
alcohols through the process of oxidative fermentation, and they can be used for
synthesis of Vitamin C, D-gluconis acid and ketogluconic acids. G. oxydans are
found in flowers, fruits, garden soil, alcoholic beverages, cider, and soft drinks
5
because they contain strains capable of growing in high concentrations of sugar

solutions and low pH values (optimal pH for growth is 5.5-6.0). Although they are
able to grow in extreme conditions, their growth rate is slow, and the concentration
of mature cells are low. The importance of G. oxydans is its ability to incompletely
oxidize carbon substrates such as D-sorbitol, glycerol, D-fructose, and D-glucose
for the use in biotechnological instruments. Conversion of D-glucose into 2,5-
diketogluconic acid is mediated by membrane bound NADP + -independent three
dehydrogenases [glucose dehydrogenase (GDH), gluconate dehydrogenase
(GADH) and 2-ketogluconate dehydrogenase (2KGADH)]
c) Aspergillus Niger
Aspergillus niger forms easily recognized black or dark brown colonies, as do
the closely related species A. carbonarius, which produces conidia 7–10 µm in
diameter and A. awamori, which produces finely roughened conidia. A. awamori is
used in food fermentations and is perhaps a domesticated form of A. niger. A. niger
and closely related species (A. carbonarius, A. awamori, A. aculeatus and A.
japonicus) all belong in Aspergillus Section Nigri.
Aspergillus niger is more prevalent in warmer climates, both in field situations
and stored foods. The black spores apparently provide protection from sunlight and
UV irradiation, providing a competitive advantage in such habitats. A. niger is very
frequently isolated from sun-dried products such as vine fruits where it may produce
ochratoxin A.
Aspergillus niger is among the most common fungi isolated from nuts (peanuts,
pecans, pistachios, hazelnuts, walnuts, kola nuts, coconut and copra). Cereals and
oilseeds are also frequent sources, especially maize, but A. niger can be isolated
from almost any type of stored commodity. Dried, smoked and cured fish and meat
products are other common sources.
Growth temperatures for Aspergillus niger are minimum, 6–8 °C, maximum,
45–47 °C and optimum 35–37 °C. A. niger is a xerophile: reported germination at
0.77 aw at 35 °C. A. niger is able to grow down to pH 2.0 at high aw. Until relatively
recently, Aspergillus niger was regarded as a benign fungus, and has been widely
used in food processing.
6
1.2.2 Gluconic Acid

Gluconic acid is produced from glucose through a simple dehydrogenation
reaction catalysed by glucose oxidase. Oxidation of the aldehyde group on the C-1
of b-D-glucose to a carboxyl group results in the production of glucono-d-lactone
and hydrogen peroxide. Glucono-d-lactone is further hydrolyzed to gluconic acid
either spontaneously or by lactone hydrolyzing enzyme, while hydrogen peroxide
is decomposed to water and oxygen by peroxidase. The conversion process could
be purely chemical. However, the most commonly involved method is the
fermentation process.
Figure 1.1. Chemical Structure of Gluconic Acid

(Source: Ramachandran, S. et al., 2006)
1.2.2.1 Properties of Gluconic Acid

Gluconic acid is a noncorrosive, nonvolatile, non-toxic, mild organic acid.
It imparts a refreshing sour taste in many food items such as wine and fruit juices,
Sodium gluconate has a high sequestering power. It is a good chelator at alkaline
pH; its action is comparatively better than EDTA, NTA and other chelators.
Aqueous solutions of sodium gluconate are resistant to oxidation and reduction at
high temperatures. It is an efficient plasticizer and a highly efficient set retarder. It
is easily biodegradable (98% at 48 hours). It has an interesting property of inhibiting
bitterness in foodstuffs. Concentrated gluconic acid solution contains certain
lactone structures (neutral cyclic ester) showing antiseptic property. The
7
characteristics are described in Table 1.1.

Table 1.1. General Characteristics of Gluconic Acid
Noncorrosive, mildly acidic, less irritating,

Nature nonodorous, nontoxic, easily
biodegradable, nonvolatile organic acid
Relative molecular mass 196.16
Chemical formula C6H12O7
Synonym 2,3,4,5,6-pentahydroxyhexanoic acid
pKa 3.7
Melting Point (50% solution) Lower than 12 °C
Boiling Point (50% solution) Higher than 100 °C
Density 1.24 g/mL

Appearance Clear to brown
Solubility Soluble in water

Sourness Mild, soft, refreshing taste
Degree of sourness (sourness
of citric acid is regarded as 29 - 35
100)
(Source: Ramachandran, S. et al., 2006)
1.2.2.2 Gluconic Acid Occurrence

Gluconic acid is abundantly available in plants, fruits and other foodstuffs
such as rice, meat, dairy products, wine (up to 0.25 %), honey (up to 1 %), and
vinegar. It is produced by different microorganisms as well, which include bacteria
such as Pseudomonas ovalis, Acetobacter methanolicus, Zymomonas mobilis,
Acetobacter diazotrophicus, Gluconobacter oxydans, Gluconobacter suboxydans,
and Azospirillum brasiliense. Furthermore, gluconic acid could produce by fungi,
such as Aspergillus niger, Penicillium funiculosum, P. variabile, P. amagasakiense,
and various other species such as Gliocladium, Scopulariopsis, Gonatobotrys,
Endomycopsis, also yeasts such as Aureobasidium pullulans (formerly known as
Dematium or Pullularia pullulans). Ectomycorrhizal fungus Tricholoma robustum,
which is associated with the roots of Pinus densiflora, was found to synthesise
8
gluconic acid.
1.2.2.3 Application of Gluconic Acid
Gluconic acid is a mild organic acid, which finds applications in the food
industry. As stated above, it is a natural constituent in fruit juices and honey and is
used in the pickling of foods. Its inner ester, glucono-d-lactone imparts an initially
sweet taste which later becomes slightly acidic. It is used in meat and dairy products,
particularly in baked goods as a component of leavening agent for preleavened
products. It is used as a flavouring agent (for example, in sherbets) and it also finds
application in reducing fat absorption in doughnuts and cones. Foodstuffs
containing D-glucono-d-lactone include bean curd, yoghurt, cottage cheese, bread,
confectionery and meat.
In general, gluconic acid and its salts are used in the formulation of food,
pharmaceutical and hygienic products. They are also used as mineral supplements
to prevent the deficiency of calcium, iron, also as buffer salts. Different salts of
gluconic acid find various applications based on their properties. Sodium salt of
gluconic acid has the outstanding property to chelate calcium and other divalent and
trivalent metal ions. It is used in the bottle washing preparations, where it helps in
the prevention of scale formation and its removal from glass. It is well suited for
removing calcareous deposits from metals and other surfaces, including milk or
beer scale on galvanised iron or stainless steel. Its property of sequestering iron over
a wide range of pH is exploited in the textile industry, where it prevents the
deposition of iron and for desizing polyester and polyamide fabrics. It is also used
in metallurgy for alkaline derusting, as well as in the washing of painted walls and
removal of metal carbonate precipitates without causing corrosion. It also finds
application as an additive to cement, controlling the setting time and increasing
the strength and water resistance of the cement. It helps in the manufacture of frost
and crack resistant concretes. It is also used in the household cleaning compounds
such as mouthwashes.
Calcium gluconate is used in pharmaceutical industry as a source of calcium
for treating calcium deficiency by oral or intravenous administration. It also finds a
place in animal nutrition. Iron gluconate and iron phosphogluconate are used in iron
therapy. Zinc gluconate is used as an ingredient for treating common cold, wound
9
healing and various diseases caused by zinc deficiencies such as delayed sexual
maturation, mental lethargy, skin changes, and susceptibility to infections.
1.2.3 Glucose
Glucose is a simple sugar with the molecular formula "# $%& '# . Glucose is
the most abundant monosaccharide, a subcategory of carbohydrates. Glucose is
mainly made by plants and most algae during photosynthesis from water and
carbon dioxide, using energy from sunlight. There it is used to
make cellulose in cell walls, which is the most abundant carbohydrate. In energy
metabolism, Glucose is the most important source of energy in all organisms.
Glucose for metabolism is partially stored as a polymer, in plants mainly
as starch and amylopectin and in animals as glycogen. Glucose circulates in the
blood of animals as blood sugar. The naturally occurring form of glucose is D-
glucose, while L-glucose is produced synthetically in comparably small amounts
and is of lesser importance.
1.2.3.1 Glucose Chemical Properties
With six carbon atoms, it is classed as a hexose, a subcategory of the
monosaccharides. D-Glucose is one of the sixteen aldohexose stereoisomers.
The D-isomer, D-glucose, also known as dextrose, occurs widely in nature, but
the L-isomer, L-glucose, does not. Glucose can be obtained by hydrolysis of
carbohydrates such as milk sugar, (lactose), cane sugar (sucrose), maltose, cellulose,
glycogen, etc. It is commonly commercially manufactured from cornstarch by
hydrolysis via pressurized steaming at controlled pH in a jet followed by further
enzymatic depolymerization. Unbonded glucose is one of the main ingredients
of honey. All forms of glucose are colorless and easily soluble in water, acetic acid,
and several other solvents. They are only sparingly soluble in methanol and ethanol.
Glucose is a monosaccharide with formula "# $%& '# or H-(C=O)-
(CHOH)5-H, whose five hydroxyl (OH) groups are arranged in a specific way
along its six-carbon back. Glucose is usually present in solid form as
a monohydrate with a closed pyran ring (dextrose hydrate). In aqueous solution, on
the other hand, it has an open-chain to a small extent and is present predominantly
as α- or β-pyranose, which partially mutually merge by mutarotation. From aqueous
solutions, the three known forms can be crystallized: α-glucopyranose, β-
10
glucopyranose and β-glucopyranose hydrate. Glucose is a building block of the

disaccharides lactose and sucrose (cane or beet sugar), of oligosaccharides such as
raffinose and of polysaccharides such as starch and amylopectin, glycogen or
cellulose. The glass transition temperature of glucose is 31℃ and the Gordon-
Taylor constant (an experimentally determined constant for the prediction of the
glass transition temperature for different mass fractions of a mixture of two
substances) is 4.5. Glucose is no toxic chemical compound and have highly
combustible. This compound also finely dispersed particles that become explosive
when exposed to air.
1.2.3.2 Glucose Physical Properties
The physical properties of glucose can be used to characterize matter and
energy also their interactions. The phase of glucose can be solid or liquid depending
from the source. This compound has a sweet taste, but not have an odor. Glucose is
soluble in water and acetic acid. All forms of glucose are colorless also clear. The
another of glucose physical properties are described in Table 1.2.
Table 1.2. Physical Properties of Glucose
Property Value Unit

Δ, -° -793.74 kJ/mole
Δ, $°/01 -1035.02 kJ/mole
2345$° 19.93 kJ/mole
2$°607 117.51 kJ/mole
logPoct/wat -3.379 -
AB 6631.37 kPa
DEFGH 844.48 K
DIJHK 419 K
DB 1034.02 K
D,L1 423.00 ± 3.00 K
MN 0.46 m3/kg mole
Density 1.54 g/cm³
Weight 180.16 g/mole
Source: chemeo.com
11
1.2.3.3 Sources of Glucose

Most dietary carbohydrates contain glucose, either as their only building
block (as in the polysaccharides starch and glycogen), or together with another
monosaccharide (as in the hetero-polysaccharides sucrose and lactose). Unbounded
glucose is one of the main ingredients of honey. The sources of glucose are
described in Table 1.3.
Table 1.3. Sugar Content of Selected Common Plant Foods (g/100g)

Sucrose
Total Fructose/ as a %
Total Free Free
Food Item Carbo- Sucrose Glucose of
Sugars Fructose Glucose
hydrate Ratio Total
Sugars
Apple 13.8 10.4 5.9 2.4 2.1 2.0 19.9
Apricot 11.1 9.2 0.9 2.4 5.9 0.7 63.5
Banana 22.8 12.2 4.9 5.0 2.4 1.0 20.0
Fig, dried 63.9 47.9 22.9 24.8 0.9 0.93 0.15
Grapes 18.1 15.5 8.1 7.2 0.2 1.1 1
Navel
12.5 8.5 2.25 2.0 4.3 1.1 50.4
orange
Peach 9.5 8.4 1.5 2.0 4.8 0.9 56.7
Pear 15.5 9.8 6.2 2.8 0.8 2.1 8.0
Pineapple 13.1 9.9 2.1 1.7 6.0 1.1 60.8
Plum 11.4 9.9 3.1 5.1 1.6 0.66 16.2
Beet, Red 9.6 6.8 0.1 0.1 6.5 1.0 96.2
Carrot 9.6 4.7 0.6 0.6 3.6 1.0 77
Red Pepper,
6.0 4.2 2.3 1.9 0.0 1.2 0.0
Sweet
Onion,
7.6 5.0 2.0 2.3 0.7 0.9 14.3
Sweet
Sweet
20.1 4.2 0.7 1.0 2.5 0.9 60.3
Potato
Yam 27.9 0.5 traces traces traces N/A traces
0.2 – 0.2 –
Sugarcane - 13–18 11–16 1.0 high
1.0 1.0
0.1 – 0.1 –
Sugar Beet - 17–18 16–17 1.0 high
0.5 0.5
Corn, Sweet 19.0 6.2 1.9 3.4 0.9 0.61 15.0
Source: Journal
12
1.2.3.4 Glucose Function

Glucose can form from formaldehyde under abiotic conditions, so it may
well have been available to primitive biochemical systems. Probably more
important to advanced life is the low tendency of glucose, by comparison to other
hexose sugars, to non-specifically react with the amino groups of proteins. This
reaction (glycation) reduces or destroys the function of many enzymes. The low
rate of glycation is due to glucose's preference for the less reactive cyclic isomer.
Nevertheless, many of the long-term complications of diabetes (e.g., blindness,
kidney failure, and peripheral neuropathy) are probably due to the glycation of
proteins or lipids. In contrast, enzyme-regulated addition of glucose to proteins by
glycosylation is often essential to their function.
Glucose is a ubiquitous fuel in biology. It is used as an energy source in
most organisms, from bacteria to humans. Use of glucose may be by either aerobic
or anaerobic respiration (fermentation). Carbohydrates are the human body's key
source of energy, through aerobic respiration, providing approximately 4
kilocalories (17 kilojoules) of food energy per gram. Breakdown of carbohydrates
(e.g. starch) yields mono- and disaccharides, most of which is glucose. Through
glycolysis and later in the reactions of the Citric acid cycle (TCAC), glucose is
oxidized to eventually form CO2 and water, yielding energy, mostly in the form of
ATP. The insulin reaction, and other mechanisms, regulate the concentration of
glucose in the blood. A high fasting blood sugar level is an indication of prediabetic
and diabetic conditions.
Use of glucose as an energy source in cells is via aerobic or anaerobic
respiration. Both of that starts with the early steps of the glycolysis metabolic
pathway. The first step of this is the phosphorylation of glucose by hexokinase to
prepare it for later breakdown to provide energy. The major reason for the
immediate phosphorylation of glucose by a hexokinase is to prevent diffusion out
of the cell. The phosphorylation adds a charged phosphate group, so the glucose 6-
phosphate cannot easily cross the cell membrane. Irreversible first steps of a
metabolic pathway are common for regulatory purposes.
1.3 Market Analysis
Gluconic Acid (also called as Sodium Gluconate) can be used in food,
13
pharmaceutical products, and chemical industries which may contribute towards

industry growth. Currently, food industry consists of the largest market share due
to its high use as an acidulant in the meat processing industry and recognized as a
generally permitted food additive. In medicinal application in pharmaceuticals, the
high demand comes up from calcium therapy and treatment to become the fastest
growing segment in the application type.
Based on H.A. El-Enshasy (2005) data journal, demand of gluconic acid has
increased steadily reaching 60,000 tons per year in last 20 years. The growth of the
gluconic acid industry in Indonesia especially from fermentative methods is
expected to grow up rapidly in line with the growing need for carbonic acid as an
additive in food and beverages industry. Gluconic acid primarily occurs and imparts
a refreshing sour taste in many food items such as honey, fruit juice, wine, meat,
and dairy products. It is placed in beverages such as lemonade for acidification, as
it is a fairly weak acid that has a neutral taste. Breweries and creameries tend to use
them to prevent calcification during the processing of beer and milk. It also is
included with processed vegetables and fruits, along with their additives and for
preparations.
Gluconic Acid dominates the market of organic acids due to its application
in various fields and produced 60,000 tons world-wide annually. It is available in
the market as 50% aqueous solution (by mass) and 99% purity crystal powder.
Indonesia is one of country which has been importing gluconic acid for several
years because no industry in Indonesia produced gluconic acid commercially.
Based on Foreign Trade Statistical Bulletin by BAPPENAS in December 2013,
Indonesia has been importing 7,329,946 kilograms of gluconic acid, salts and esters
cumulatively since 2013. The capacity for only gluconic acid is estimated around
30% from overall import capacity.
-O4NPQRN SNRT "UVUNRWX = 30% ] 7,329,946 de
-O4NPQRN SNRT "UVUNRWX = 2,198,983.8 de
-O4NPQRN SNRT "UVUNRWX = 2,199 WPQQi5
Based on the import data and world-wide annually supply, the production capacity
estimated by projecting the variable with the growth using CAGR value in 2013 –
2017 in the amount of 5.5% and CAGR value in 2018 – 2024 in the amount of 7.2%
14
based on Global Market Insight Report. The calculation shown in Table 1.4.
Table 1.4. Calculation of Production Capacity

Year Growth Capacity (tons)
2013 2199
2014 2320
2015 5.5% 2448
2016 2582
2017 2724
2018 2920
2019 3131
2020 3356
2021 7.2% 3598
2022 3857
2023 4134
2024 4432
From the calculation above, the production capacity of gluconic acid in

2019 is estimated 3,131 tons and calculated the growth annually until 2024 with
CAGR value.
1.4 Raw Material Analysis
The main ingredients to produce gluconic acid with fermentative methods
are glucose and fermentation agent. Glucose can be obtained from high
carbohydrate-containing waste or natural plant such as sugarcane. Fermentation
agent commonly uses fungi or bacteria as the living agent to ferment the glucose.
1.4.1 Glucose
Glucose is the essential raw material in gluconic acid production. The
gluconic acid is usually produced by microbial oxidation of glucose. Enzymatic
conversion of glucose into gluconic acid can be achieved through a simple
dehydrogenation reaction using glucose oxidase as the enzyme.
Glucose could be obtained in many materials such as sugarcane, starch,
potatoes, fruits and also from industrial waste that contain high carbohydrate. At
this section, the glucose sources will be explained more from sugarcane and some
15
example of high carbohydrate containing waste which have a great quantity to

produce high amount of gluconic acid.
1.4.1.1 Sugarcane
Sugarcane as the raw material of sugar industry is one of the commodities
that has strategic role in economic sector of Indonesia. Sugarcane has been
producing in several areas of Indonesia such as North Sumatera, South Sumatera,
Lampung, West Java, Central Java, Yogyakarta, and East Java. Based on
Indonesian Sugarcane Statistics in 2015, East Java takes the first place for the
widest sugarcane plantation area in the amount of 207.15 thousand hectares or the
percentage of 45.4 from overall sugarcane plantation area in Indonesia in 2015.
Another area that also has wide plantation area are Lampung (27.34%), Central Java
(10.33%), and South Sumatera (4.88%) based on BPS Report in 2015. The average
production of sugarcane production shown in Figure 1.2.
Figure 1.2. Indonesia Sugarcane Plantation Area Maps

(Source: Badan Pusat Statistik, 2015)
The development of sugarcane production in Indonesia over the past 3 years

is fluctuating based on Badan Pusat Statistik in 2015. In 2013, the production of
sugarcane reached 2.55 million tons and increased 0.86% in 2014 to be 2.58 million
tons. While in 2015, sugarcane production decreased by 1.57% (2.53 million tons).
The production of sugarcane for the last 5 years can be seen in Figure 1.3. In 2015,
16
the production in East Java was 1.24 million tons (48.75% of overall sugarcane
production in Indonesia) and Lampung was 756,55 thousand tons (29,85% of
overall sugarcane production in Indonesia).
Figure 1.3. Sugarcane Production Development

(Source: Badan Pusat Statistik, 2015)
Due to highly supply of sugarcane in Indonesia, the plantation has been

exporting the sugarcane and reached the volume of 537,57 thousand tons
(US$ 66.42) then creates a fluctuation during 2013 – 2015 as shown in Figure 1.3.
The raw sugarcane has been importing to 23 countries in 2015. The five most
importer countries were Philippines (190.89 thousand tons), South Korea (151.36
thousand tons), Japan (40.98 thousand tons), Taiwan and Vietnam (15.3 thousand
tons each). Based on the explained data, sugarcane has a great possibility to be used
as the raw material of gluconic acid production due to the availability of sugarcane.
1.4.1.2 Carbohydrate Containing Waste
Due to the main raw material of gluconic acid is glucose which is a simple
type of carbohydrate, the industry could utilize the high carbohydrate containing
waste such as paper waste and potato waste. These materials would be a cheaper
raw material as substrate.
Paper waste is one of the cellulolytic biomasses targeted to be recycled
(Ikeda, Y. et al., 2006) because it is a cause of environmental problems in Indonesia.
In 2016, paper waste reached 10% of 175 tons per day based on Media Indonesia
Report, 2016. Paper waste has been studied previously in bioconversion by
17
filamentous fungi and also as an alternative way to produce gluconic acid. Ikeda,
Y. et al. used paper waste hydrolysate, compared the gluconic acid yield and
production rate to those obtained with a glucose medium in a flask and bioreactor
using the filamentous fungus Aspergillus niger. The study has been used
saccharified solution of waste paper with glucose concentration adjusted to 50–100
g/L for bioconversion with Aspergillus niger. These previous study shows that
paper waste is a potential material role as glucose to produce gluconic acid.
Potato waste (potato pulp) is an agricultural waste product that is obtained
from potato starch production, and it contains cellulose, starch, and some proteins
(Jiang, Yi et al., 2017). This pulp is usually buried as feed or waste disposal,
resulting in soil and water pollution and a problem with low waste utilization
(Mayer, F. 1997). Based on Kementerian Pertanian report in 2017, the consumption
of potato reached more than one million thousand per year and the waste is
estimated 10% from the consumption which have the value around 100.000 kg/year.
Potato waste is significantly different from conventional agricultural lignocellulosic
biomasses, such as wheat straw, rice straw, because of its loose and hydrated
structure as well as its low lignin content (Delgado R. et al., 2009). These properties
of potato waste make it easier to hydrolyze into sugar so that it is a suitable resource
for producing biochemicals, such as gluconate, gluconic acid, lactic acid,
bioethanol. Based on the previous study by Jiang, Yi et al., the enzymatic
hydrolysates of the potato pulp were fermented into 81.4 g/L gluconic acid by
Gluconobacter oxidans with the result of the overall conversion yield from glucose
to gluconic acid was 94.9%, and the productivity was 4.07 g/L/h. This study shows
that potato waste is also another potential carbohydrate containing waste for
gluconic acid industry.
1.4.2 Fermentation Agent
Fermentative process and catalytic oxidation are found to be the two basic
approaches in production of gluconic acid. Thus, in manufacturing gluconic acid,
fermentation is most widely acceptable and widely practiced method. Fungus,
Aspergilus niger and bacteria Gluconobacter oxydans are widely used micro-
organisms for fermentation.
1.4.2.1 Aspergilus niger
18
Aspergilus niger as one of the fermentative agents which produces all the
enzyme required for the conversion of glucose to gluconic acid. The most important
things when a plant is producing a product using living things is by knowing the
ideal and critical condition which will be used to control the system of the bioreactor
in the plant. Aspergillus nigger grows optimally in temperature of 35 – 37 C, with
the minimum temperature at 6 – 8 C, and maximum temperature at 45 – 47 C in
aerobic environment.
Aspergillus niger should be activated before the reaction start called as
incubation process. Incubation process provide a condition for the filamentous
fungi to inoculate its spore inoculum. Driouch, H. et al. (2010) has been inoculate
Aspergillus niger by growing thawn spores from the maintenance culture at 30 °C
for 3 days on Potato Dextrose Agar (PDA). Spores are harvested as spore
suspension from the plate into 20 mL 0.9 % NaCl solution, which is spreaded onto
the plate. After filtration (Miracloth, 25 µm pore size, CalBioChem, Darmstadt,
Germany) the spore concentration is determined photometrically then make a
calibration between spore number and optical density to precisely inoculate with a
well-defined spore concentration. Cultivations for enzyme production are
typically inoculated to an initial spore concentration of 106/ml.
Another study also revealed Aspergillus niger culture activation in inoculum
in fermentation media containing molasse which shows the best result in the
concentration of 30% at 30˚C for 4 days. The spore of mutant A. niger from slant
were harvested in 5 ml of sterile 50 mM sodium phosphate buffer (pH 6.8)
containing 0.1% presterilized Tween 80. The spores in inoculums were maintained
at 108 -1010 spores per ml (Nilesh, K. et al., 2012).
Aspergillus niger is the most applicable fungi that usually used for
converting glucose to gluconic acid. Besides, these filamentous fungi are easily
obtained and available in powder and granule which have purity of 97% – 99%.
1.4.2.2 Gluconobacter oxydans
Gluconobacter oxydans is a genus of bacteria which also has the ability for
converting glucose to gluconic acid. Gluconobacter oxydans is an aerobe bacterial
which has oxygen as a terminal electron acceptor. The highest growth rate occurs
at temperatures between 25 ˚C – 30˚C and it could not withstand high temperatures
19
above 37 ˚C (Comprehensive Macrobial Resource). Gluconobacter oxydans was

grown in mannitol medium (Buchert and Viikari, 1998) or ethanol medium
(Mostafa et al., 2002) which were adjusted to pH 5.0 or pH 6.0 by hydrochloric acid
on a shaker at 130 rpm.
Gluconic acid is derived from glucose by a simple oxidation reaction.
Bacterial gluconic acid production has limited success at industrial scale as the
oxidation proceeds with the secondary reaction leading to oxogluconic acids.
Gluconobacter oxydans oxidizes glucose via two alternative pathways. The first
pathway requires an initial phosphorylation followed by oxidation via the pentose
phosphate pathway. The second is the direct glucose oxidation pathway, which
results in the formation of gluconic acid and ketogluconic acid (A. Stadler-Szoke,
et al., 1980).
Gluconobacter oxydans converts glucose to gluconic acid and subsequently
to 2-keto-D-gluconic acid (2-KGA) and 5-keto-D-gluconic acid (5-KGA) by
membrane-bound periplasmic pyrroloquinoline quinone-dependent and flavin-
dependent dehydrogenases. Based on the study by Elfari, M. et al. 2005,
Gluconobacter oxydans converted about 11% of the available glucose to 2-KGA
and 6% to 5-KGA, with growing cells and improved growth under defined
conditions. The 2-KGA is a by-product that highly disadvantageous for an
industrial application of Gluconobacter oxydans so it will need more improvements
to reduce the production of 2-KGA. The yield of the production of 5-KGA without
improvement is around 45%. Meanwhile, under improved growth conditions,
Gluconobacter oxydans converted the available glucose almost completely 84%
into 5-KGA.
1.5 Plant Location Analysis
Plant location is one of the important things in plant design consideration,
because it will be related to the operation process in a plant. To choose the right
location, there are several factors that must be considered. These factors will affect the
continuity of the production of the plant in the future. We choose Malang, as our plant
location. The selection of the plant location is determined by various factors of
considerations, such as:
1. Proximity to Raw Material
20
Based on the sugarcane plantation distribution area in Indonesia that have been
explained, the highest production of sugarcane is East Java which is located
specifically in Kedungkandang, Malang. The raw material is raw sugarcane from
sugarcane plantation which located behind the area of the selected plant area. The
location of this factory is close to raw materials so that it could reduce distribution
and logistics or transportation costs to buy sugarcane juice.
Figure 1.4. Plant Location

(Source: maps.google.co.id)
2. Proximity to Markets
Target of the gluconic acid markets are pharamauticals, food, and beverages
industry in Indonesia especially in East Java area such as PT. Gunung Batu and PT.
Golden Harvestindo which is not yet fulfilled the need of gluconic acid. The market
also will be reached to another industry all over Indonesia to replace the need which
has been imported before based on BAPPENAS Report.
3. Available Size in Location
Due to our plant location in industrial area, we have to consider our plant size and
compromise it with the available lot sizes in the area. Our plant sizes is depend on the
equipment’s sizes and layout, so we need to consider the equipment specification and
its layout before choosing the suitable lot for our plant.
4. Infrastructure and Transportation Facility
Our plant location is in Mayjen Sungkono street, Kedungkandang, Malang, East
21
Java. Where the selection of the area is based on the proximity to major roads, Ports,
Electricity Short Villages, and close to Raw Material, so that will facilitate the
distribution of our factory products to the consumer.
Figure 1.5. Actual Area of The Plant

(Source: maps.google.co.id)
CHAPTER 2
PROCESS SELECTION
2.1 Raw Material Selection

Renewable carbon resources like sugarcane juice and starch waste may be
used as carbon source. But different raw material will need different pre-treatment
process, so it can enter the main process. Some kind of raw material will be selected
to find the most suitable raw material to produce the specified gluconic acid. The
selection also considers the ease and the simplicity of the pre-treatment the raw
material needed.
Sugar cane may be used as carbon source without the necessity of much pre-
treatment, saccharification and liquefaction. The use of sugarcane juice has been
encouraged for the production of gluconic acid through fermentation with suitable
micro-organisms. Gluconic acid which obtain from fermentation of sugarcane juice
has the highest concentration among all the available raw material, about 76.3g/L.
This type of carbon sources has been readily accepted by food industries and the
consumers as raw materials and is derived from biological origin. Sugarcane juice
is also easily available throughout the year in Indonesia.
Waste paper is one of the cellulolytic biomasses targeted to be recycled
because it is a cause of environmental problem. Moreover, when paper materials
are recycled, they are usually turned into lower grade paper products. One way of
using waste paper is to decompose it to reducing sugars and to convert the sugars
to gluconic acid. Waste paper has a great potential as the raw material of gluconic
acid production for it is has low price and easy to get. But the number of waste
paper production in Indonesia is only about 900.000 ton and it is already been used
in another sector. Furthermore, the number of this waste is decreasing annually
because the use of paper in working space is replaced. This waste is not eligible to
be the raw material of gluconic acid production for its not sustain.
Waste potato such as potato pulp is an agricultural waste product that is
obtained from potato starch production, and it contains cellulose, starch, and some
proteins. Potato pulp is signiﬁcantly diﬀ erent from conventional agricultural
lignocellulosic biomasses, such as corn stove, and so on, because of its loose and
23
hydrated structure as well as its low lignin content. These properties of potato pulp
make it easier to hydrolyze into sugar (Jiang. 2018). But, usually buried as feed or
waste disposal for the processing costs of this waste is high. In Indonesia the waste
potato potential represented from the number, annually Indonesia could produce 1.3
million ton of potato (Kementrian Pertanian, 2016). Thus, potato pulp is a suitable
resource for producing gluconic acid, but it will involve more complex raw material
pretreatment.
From explanation above, the raw material that will be used in the production
of gluconic acid is sugarcane. Sugar cane is used because this source of carbon has
simpler pre-treatment process and been readily accepted by the food industries.
Also, sugarcane juice is easily available in Indonesia for it is annually produced
about 2.4 million ton (Kementrian Pertanian, 2016)
2.1.1 Sugarcane
Sugarcane may be used as carbon source without the necessity of much pre-
treatment, saccharification and liquefaction. The use of sugarcane juice has been
encouraged for the production of gluconic acid through fermentation with suitable
micro-organisms. Gluconic acid which obtain from fermentation of sugarcane juice
has the highest concentration among all the available raw material, about 76.3g/L.
This type of carbon sources has been readily accepted by food industries and the
consumers as raw materials and is derived from biological origin. Sugarcane juice
is also easily available throughout the year in Indonesia
2.1.2 Carbohydrate-Containing Waste
Based on the raw material analysis, the raw material that will be used in the
production of gluconic acid is sugarcane. Sugarcane is used because this source of
carbon has simpler pre-treatment process and been readily accepted by the food
industries and the consumers as raw materials. Also, sugarcane juice is also easily
available in Indonesia for Indonesia annually produce about 2.4 million tons of
sugarcane (Kementerian Pertanian, 2016).
2.2 Process Synthesis
There are mainly three different methods for the commercial production of
gluconic acid like; chemical methods, enzymatic catalysis and fermentation. Each
method consists of three main process, those are: raw material preparation; glucose
24
to gluconic acid reaction, and purification. Each method is used in production of

gluconic acid based on the product specification that is required and the raw
material that is used. In this plant design, gluconic acid product specification that
will be produced is 3131 tons/year and the raw material that will be used is
sugarcane juice.
There are plenty of alternative process from each step in each methods of
production of gluconic acid. The methods and the process steps then will be
analyzed and selected to find the most suitable production process to produce the
xspecified gluconic acid.
Figure 2.1. Black Box Diagram
2.2.1 Process Production Method Selection

There are mainly three different methods for the commercial production of
gluconic acid like; There are mainly three different methods for the commercial
production of gluconic acid like; chemical methods, enzymatic catalysis and
fermentation process where specific micro-organisms are grown in medium
containing glucose and other ingredients.
2.2.1.1 Fermentation method
Fermentation method for the production of gluconic acid from glucose is
the easiest and economically most excellent process known abroad (Pal, 2016).
Thus, in manufacturing gluconic acid, fermentation is most widely acceptable and
widely practiced method. However, it has also many disadvantages such as
difficulties in the separation of microbes and product, control of byproduct
formation and disposal of waste water.
2.2.1.2 Chemical method
Chemical method such as chemical oxidation of glucose by hypochlorite
solution and electrolytic oxidation of glucose solution in presence of bromide are
adopted in industrial scale production of gluconic acid. Several studies have been
reported on heterogeneous catalysis of glucose to produce gluconic acid by using
25
Pt, Pd or Au nanoparticles as catalysts and oxygen from air as oxidant under mild
conditions. Although the conversion is a simple one-step process, the chemical
method is not at all favored due to its limited selectivity towards gluconic acid,
environmental toxicity and biological hazards; whereas the rapid increase in the
cost of electrical power in the recent years has turned electrolytic process costly.
Different oxidizing agents are used in this process, but still the process appears to
be costlier and less efficient than fermentation method.
2.2.1.3 Enzymatic method
Enzymatic method is developed for the commercial production of gluconic
acid. Various enzymatic processes have been described in the technical
bibliography, aiming in the replacement of commercial fermentation processes
suffering under difficulties including the removal of glucose and separation of
gluconic acid from fermentation broths. However, enzymatic production of
gluconic acid is economically unfeasible for large-scale industrial production due
to enzyme’s instability and resulting very high cost.
From explanation above, the gluconic acid production plant will use
fermentation method for it is the most reliable method for large-scale production.
This method is also the most economically method among other methods. For the
weakness of this method such as difficulties in the separation, new superior
fermentation processes system has been developed. Membrane-based separation
processes are considered as viable alternative to some traditional separation
processes. This system can largely overcome these problems due to high cell
density and continuous removal of products and ensures reduced energy
consumption as it involves no phase change (except pervaporation).
2.2.2 Selection of Fermentation Process
The plant will produce gluconic acid through fermentation. Based on the
mode of '& supply, two types of culture conditions have been defined for GA
fermentation: including aerobic submerged fermentation (ASF) and solid-state
fermentation (SSF).
2.2.2.1 Solid-State Fermentation
Solid-state fermentation (SSF) is a process of growth of microorganism in
an SSF solid medium (substrates). The main advantage of using these substrates is
26
that nutrient-rich waste materials can be easily recycled as substrates. SSF is better
than submerged culture due to less variation of osmotic pressure, DO concentration,
availability of water and nutrients. In this fermentation technique, the substrates are
utilized very slowly and steadily, so the same substrate can be used for long
fermentation periods. Hence, this technique supports controlled release of nutrients.
However, it cannot be used in fermentation processes involving organisms that
require high aw (water activity), such as bacteria.
2.2.2.2 Aerobic Submerged Fermentation
Aerobic Submerged Fermentation (ASF) is a process of growth of
microorganism in a free-flowing liquid medium (substrates). This fermentation
technique is best suited for microorganisms such as bacteria that require high
moisture content. An additional advantage of this technique is that purification of
products is easier. However, oxygen supply in the center of large pellets sometimes
lack enough oxygen supply, leading to autolysis of the fungus or bacteria, which
further negatively affects fermentation. Submerged fermentation is usually
employed for commercial production of gluconic acid which is sensitive to process
control parameters, demands investment and is an energy intensive process.
Shown in the Table 2.1 below is the scoring for selection process based on
the comparison of fermentation time, yield, and applicability.
Table 2.1. Parameter of Scoring for Fermentation Method Selection
Weight Score
Parameter
(%) 3 2 1
Fermentation time
10 < 19 19-22 >22
(h/batch)
Yield (using
25 > 90% 70-90% 50%-70%
Sugarcane juice)
DO concentration 15 1 vvm 2-3 vvm 4 vvm
High
Medium Low because
because no
because need need complex
Effectivity 30 need next
some next step purification’s
step
purification step
purification
Applicability 20 Industrial Pilot Research
27
For purpose of process selection, the weight factor of the criteria was
determined subjectively by our team from our rational intent. The total weight
factor has to be 100% and it is normally used to assign the weight factor value of
each criteria. Processes are rated through a comparison to the reference design
process. Representative ratings or ranks are valued from 1 to 3, indicating much
worse to much better process that the reference, this would be explained here after.
Once all the concepts are rated, a total score for each design concept is calculated.
The calculation is shown in Table 2.2 below.
Table 2.2. Scoring for Fermentation Method Selection

Type of Fermentation
Aerobic Submerged Solid State
% Fermentation Fermentation (SSF)
Parameter
weight (ASF)
rank weight score rank weight
score
Fermentation time
15 2 0.3 2 0.3
(h/batch)
Yield (using
20 2 0.4 3 0.6
Sugarcane juice)
DO concentration 15 2 0.3 3 0.45
Effectivity 30 2 0.6 1 0.3
Applicability 20 3 0.6 2 0.4
Total 100 2.2 2.05
The table shows that each fermentation process has advantage and
disadvantage, but one alternative has to be chosen. From scoring in the Table 2.2,
Submerged fermentation is chosen as the fermentation process in the production of
gluconic acid. ASF is the most reliable method for large-scale production for it is
the most used method in the production of gluconic acid. Also, this technique has
easier products purification.
2.2.3 Fermentation Agent
Based on the literature, there are a few microorganisms that has been used
for gluconic acid production. Each one of them has their own benefits and
challenges. To produce gluconic acid in the most economical way, which
fermentation agent used should be considered. To choose which one is the best
28
fermentation agent to be used, there are few parameters to be considered which are
related to the fermentation process and ability for the conversion of glucose to
gluconic acid. The parameters are tabulated in Table 2.3 below.
Table 2.3. Parameter of Scoring for Fermentation Agent Selection
Weight Score
Parameter
(%) 3 2 1
Operability (max.
20 ≥ 40 ˚C 30 – 40˚C < 30˚C
temperature)
Yield (%) 25 > 90% 70% – 90% 50%-70%
Targeted
products and Only targeted No products
Overall Product 25 by-products products are were
are advantageous advantageous
advantageous
Not many
Easily No market
markets
Availability 30 obtained in provided the
provided the
market materials
materials
Table 2.4. Scoring for Fermentation Agent Selection

Fermentation Agent
Gluconobacter
Aspergillus niger
Parameter % weight oxydans
weight
rank weight score rank
score
Operability (max.
20 3 0.60 2 0.40
temperature)
Yield (%) 25 3 0.75 2 0.50
29
Table 2.4. Scoring for Fermentation Agent Selection (continued)

Fermentation Agent
Gluconobacter
Aspergillus niger
Parameter % weight oxydans
weight
score
Overall Product 25 2 0.50 1 0.25
Availability 30 3 0.90 2 0.60
Total 100 2.75 1.75
The table shows that each fermentation process has advantage and
disadvantage, but one alternative has to be chosen. From scoring in the Table 2.4,
Aspergillus niger is chosen as the fermentation agent in the production of gluconic
acid. Aspergillus niger has a good temperature operability and easily obtained in
the market. These filamentous fungi also could create high yield value and
advantageous by-product which also has a market value.
2.2.4 Process Selection
The process of the gluconic acid production is being selected to choose more
efficient and economical decision. The selection including reaction process,
2.2.4.1 Reaction
Conversion of glucose into gluconic acid with Aspergillus niger as a
fermentation agent can be done in reaction process. In the fermentation process of
raw materials with fermentation agent, that can be use by three methods, there are
Solid State Fermentation (SSF), Immobilization, and Aerobic Submerged
Fermentation. However, the conversion of gluconic acid from glucose could not be
done by Solid State Fermentation, because this method can only be used if the
medium and raw materials in solid phase, while the glucose is liquid phase.
Fermentation with Immobilization can be assumed as a Packed Bed Reactor (PBR)
and fermentation with Aerobic Submerged Fermentation can be assumed as a
reactor in general, there are batch reactor or continuous reactor with stirred (CSTR).
To choose which one is the best reactor to be used, there are few parameters to be
considered which are related to the fermentation process and ability for the
conversion of glucose to gluconic acid. The parameters are tabulated in Table 2.5
below.
30
Table 2.5. Parameter of Scoring for Reaction Method Selection
Weight Score
Parameter
(%) 3 2 1
More than 10 2 – 10 ton per Lower than 2
Product Capacity 20
ton per day day ton per day
Yield (%) 25 > 90% 70% – 90% 50%-70%
Reaction Capacity 15 >1.1 L/hour 0.9-1.1 L/hour <0.9 L/hour
Less
Need more
fermentative
Amount of the fermentative
10 agent (not In the middle
fermentative agent agent (easily to
easily to
saturated)
saturated)
Can recycle
Can recycle
the product, Cannot recycle
Product Recycle 15 the product
but not the product
efficiently
efficient
Investment Investment
One of
cost and cost and
Cost of Reactor investment or
15 operational operational
Manufacture operational
cost are cost are
cost is cheaper
cheaper expensive
Table 2.6. Scoring for Fermentation Reactor Selection
Fermentation Reactor
Batch CSTR PBR
Parameter % weight
weight weight weight
rank rank rank
score score score
Product Capacity 20 3 0.60 3 0.60 2 0.40
31
Table 2.6. Scoring for Fermentation Reactor Selection

Fermentation Reactor
Batch CSTR PBR
Parameter % weight
weight weight weight
rank rank rank
score score score
Yield (%) 25 3 0.75 3 0.75 2 0.50
Reaction Capacity 10 3 0.30 2 0.20 2 0.20

Amount of the
15 2 0.30 3 0.45 3 0.45
fermentative agent
Product Recycle 15 2 0.30 3 0.45 3 0.45
Cost of Reactor
15 3 0.45 2 0.30 2 0.30
Manufacture
Total 100 2.70 2.75 2.30
From the scoring table of fermentation reactor selection, all of the reactor
has advantage and disadvantage, but one alternative has to be chosen. The result of
the scoring method is Continuous Stirred Tank Reactor (CSTR) that chosen to be
fermentation reactor. Furthermore, CSTR also appropriate to use membrane for
separation (purification) of gluconic acid.
2.2.4.2 Incubation
Based on the literature, there are 2 (two) types of incubation methods that
has been used for gluconic acid production. The first type is using Potato Dextrose
Agar (PDA) as the first media of strainer before cultivation process, and the other
type is inoculation without PDA but directly uses molasse to activate the spore.
Each one of them has their own benefits and challenges. To produce gluconic acid
in the fastest way, which incubation method used should be considered. The
parameters are tabulated in Table 2.7 below.
Table 2.7. Parameter of Scoring for Incubation Method Selection
Weight Score
Parameter
(%) 3 2 1
Inoculation time
30 <5 5–7 >7
(days)
Rotation (rpm) 20 50 – 150 150 – 200 > 200
Nutrients needs 20 < 5 types 5 – 10 types > 10 types
Spore (spore/ml) 30 > 1010 105 - 1010 < 105
32
Table 2.8. Scoring for Incubation Method Selection
Type of Fermentation
Incubation with Incubation without
Parameter % weight PDA cultivation PDA cultivation
weight
score
Inoculation time
30 3 0.9 2 0.6
(days)
Rotation (rpm) 20 2 0.4 2 0.4
Nutrients needs 20 3 0.6 2 0.4
Spore (spore/ml) 30 3 0.9 2 0.6
Total 100 2.8 2.0
From the scoring table of fermentation reactor selection, all of the reactor
has advantage and disadvantage, but one alternative has to be chosen. The result of
the scoring method is incubation with Potato Dextrose Agar (PDA) that chosen to
be the incubation method. Although using PDA would be a little bit costly, but the
running time of the incubation will be reduced and faster.
2.2.4.3 Purification
To determine which type of purification that will be used, the basic concepts
of purification itself must be understood. There are few criteria to be considered in
order to decide which type is the best Microfiltration membranes, Nanofiltration
membranes and Absorption The steps of process selection are determining the
parameters to compare some the alternatives, determining scoring parameter which
consists of weight and rank for each parameter, and doing scoring based on scoring
parameter where the process with the highest score will be chosen for purification
process in the plant.
33
Table 2.9. Parameter of Scoring for Fermentation Reaction Method Selection

No Parameter Weight (%) Score Description Score
1 Maturity 20% Frequent usage 1-3
2 Flexibility 20% Easy to shape 1-3
3 Operability 20% Complex to simple usage 1-3
4 Capital Expense 20% High to low cost 1-3
5 Availability 20% Easy to find 1-3
a. Maturity
Microfiltration membrane has been frequently commercialized among all
purification for its sustainable processing, while absorption is still at the research
stage and there is no plant that has been using this technology.
b. Flexibility
This membrane covers a wide range application of this purification process.
the process is free of additives. The choice of membrane, and thus the pore size,
depends on the purification purpose, which can be organic removal or softening of
Gluconic Acid.
c. Operability
Membrane based system has the advantages of lower energy consumption,
sustainable processing, simpler operation and relatively easy scale-up provisions.
Being selective in nature, NF membrane is the most prominent candidate for
recovery of GA due to their negative charge and low molecular weight while using
absorption needs reagents that meet the specification to absorb the GA product and
disrupt the purification process.
d. Capex
Capital expense is divided into two majors, they are fixed capital investment
and working capital investment. The construction cost of microfiltration membrane
is cheaper than the cost of absorption due to the installment and also the
maintenance of absorption.
e. Availability
Availability of the two types of purification are easily found in markets. But
it is easier to find membranes these days due to its modern technologies than
34
absorption. Based the scoring of the purification processes, we choose

Microfiltration and Nanofiltration as our process to purify Gluconic Acid.
2.2.4.4 Drying Process
The market target is in the form of powder not liquid. In order to change the
form, the solution needs to be dried. The process chosen for drying the solution is
using spray drier. Spray drier is used because it dries the solution rapidly only using
hot air. Also, because spray drier used atomizer in form of sprayer. It makes powder
form possible.
2.2.5 Process Description
Based on the raw material selection explained, the main source of glucose
will be obtained from sugarcane juice. Sugarcane juice obtained by grinding
sugarcane stalk that is come from sugarcane field near the plant location. The
filtered sugarcane juice yield from crushing the sugarcane is about 50%
(Chauan,2002) and it contains approximately 78 to 86% water, 10 to 20% sucrose,
1 to 2% reducing sugar, 0.3 to 0.5% ash and between 0.5 and 1.0% of nitrogen
compounds. The pH of the sugarcane juice varies between 5.2 and 6.8 (Corazza et
al., 2001). Sugarcane juice then will be processed even more to attain desired
specification. Processes such as, heat sterilizing, pH stabilizing, clarifying, and
evaporating next will be done.
After extracting the sugarcane juice from sugarcane stalk, the sugarcane
juice then will be mixed with HCl to hydrolyze the sucrose become sucrose and
fructose. This step is needed to increase the medium’s glucose concentration and
also the fermentation agent (A. niger) can only digest sucrose and fructose become
gluconic acid. Before entering the hydrolysis reactor, the sugarcane-HCl mixture is
heated first to attain the reaction temperature (80oC) then will be reacted in the
CSTR reactor. Even though the mixture has been heated before the entering reactor,
after the thermal hydrolyzing process the mixture still needs more heat sterilizing
process to make sure that no other microorganisms in the mixture. The present of
other microorganisms in medium will be competitor to the fermentation agent (A.
niger) and reduce the production of gluconic acid. After passing the heat sterilizer,
some nutrients (KH2PO4 and MgSO4) will be added to the mixture then the mixture
will enter the pH stabilizing process. In this process, NaOH will be added to the
35
mixture respectively to adjust the pH value of sugarcane juice because the pH of

sugarcane juice increases after the thermal hydrolysis reaction. The pH stabilizing
process will produce salt. The salt will next be separated from the juice by entering
the clarifying process then the mixture will enter the evaporation process to reduce
the water content of the juice. The final yield of sugarcane juice is 61.4 g/L glucose.
To meet the demand of production (400kg/h of glucose), the total of sugarcane stalk
that will be need is 4000kg/day.
After pre-treatment process that produce glucose from sugarcane juice, the
next process to create gluconic acid is fermentation reaction. In fermentation
reaction process, the glucose reacted with fermentative agent which is Aspergillus
niger. Aspergillus niger should be activated before the reaction start called as
incubation process. Incubation process provide a condition for the filamentous
fungi to inoculate its spore inoculum.
Based on Purane, K. et al. (2012), Aspergillus niger NCIM 530 derived from
wild type Aspergillus niger NCIM 530 was provided by NCL Pune, India. During
the study, this strain was maintained on agar slant of molasses medium at 30°C for
4 days. The spore of mutant A. niger from slant were harvested in 5 ml of sterile 50
mM sodium phosphate buffer (pH 6.8) containing 0.1% pre-sterilized Tween 80.
The spores in inoculums were maintained at 108 -1010 spores per ml. It was
inoculated in erlenmeyer flasks containing spore germination medium with
following composition: glucose 5%, di-ammonium phosphate 0.2%, MgSO4,
0.25%, KH2PO4 0.1% (pH 5.5). The inoculated medium was put on orbital shaking
incubator at 28°C with 180 rpm for 48 hrs.
To react fermentation process, the plant uses Continuous Stirred Tank
Reactor (CSTR) that continuously processes the products. Furthermore, the reaction
needs peanut oil as a nutrient of fermentative agent also append sodium hydroxide
(NaOH) to make the product in pH 6.8. The method of fermentation reaction is use
Aerobic Submerged Fermentation, so this process use air to react glucose with
fermentative agent. The reaction to produce gluconic acid is as follows:
2"# $%& '# + '& → 2"# $%& '( (-O4NPQRN UNRT)
Feed of reactor is sugarcane juice in α-D-glucose. The Aspergillus niger as
36
fermentative agent can produce mutarotase enzyme to create l-D-glucose from m-

D-glucose. After that, Aspergillus niger also produce glucose oxidase to create D-
glucono-n-lactone and D-glucono-o-lactone from previous form of glucose. Last,
the fermentative agent changes D-glucono-o-lactone to gluconic acid as a product
that needed. Beside produce gluconic acid with fermentation of glucose, the reactor
also processes some recycle product from purification, that does not accumulate
absolutely. Feeds of recycle product are microbial cell, unconverted carbon source,
and water & micro-nutrients recycle product. These recycle product will reacted
with another feed in CSTR, there are glucose and Aspergillus niger. So, the product
of gluconic acid has higher of yield and maximum conversion.
Figure 2.2. Fermentation Reaction of Gluconic Acid Production

(Source: Pal, P. et al., 2016)
The CSTR as a fermentor has temperature of operation condition, that work

o
in 37 C also has pressure that work in at atmospheric pressure (approximately 1
atm). This reactor has design capacity that approximately up to 50,000 L. As a
continuous reactor, the feed of glucose to reacted in CSTR has mass flowrate 263.94
kg/hour and the feed of Aspergillus niger as fermentative agent has mass flowrate
258 kg/hour. To produce gluconic acid, the reaction also needs nutrients that has
mass flowrate 32.09 kg/hour also need water to be solvent that has mass flowrate
24,457 kg/hour. This reactor can produce gluconic acid up to 17,958 kg/hour.
In purification process, Microfiltration and Nanofiltration membranes is
used. With 0.45 µm polyvinylidene fluoride (PVDF) pore size for microfiltration
membrane and 0.22 µm Nylon pore size for nanofiltration membrane, five thin film
composite polyamide membranes of each filtration were procured and tested for
37
product separation and concentration. The microfiltration of fermentation broth was

done to recycle the biomass.
There are 3 filtrations will be applied in this purification, 1 microfiltration
and 2 nanofiltrations. The first microfiltration is to filtrate the microbial cells and
to flow it back to reactor to be re-used. The second filtration which is nanofiltration
is to send back unconverted carbon source to reactor to have it processed. The last
one is nanofiltration which the function is to filter water and micronutrients, and
have it back recycled to reactor.
The concentrated gluconic acid solution from purification process is then
dried using spray drier. The solution is pumped from the harvesting tank into spray
drier tower to be dried using hot air. The air comes from ambient air that is
compressed and and then heated using electric heater. The solution is atomized
using a sprayer on the top of spray drier column and then contacted with hot air.
The solution is rapidly dried and become powder. The powder then collected and
then cooled using electric cooler to be stored in product tank
2.3 Block Flow Diagram
Figure 2.3. Block Flow Diagram
2.4 Process Flow Diagram
Figure 2.4. Process Flow Diagram
CHAPTER 3
MASS AND ENERGY BALANCE
From process flow diagram in previous chapter, the next step is calculating
mass and energy balance for this plant, and the energy balance is calculated for a day.
The calculation uses two simulators such as SuperPro. The urge to use SuperPro
simulator is because the lack of UniSim itself which cannot defines fermentation
process and its material stream.
Figure 3.1. SuperPro Simulation for Pre-Treatment Process
Figure 3.2. SuperPro Simulation for Fermentation and Purification Process
Figure 3.3. SuperPro Simulation for Drying Process
3.1 Mass Balance
Table 3.1. Mass Balance of Overall Process
Pre-Treatment
Stream Unit Glucose Sucrose Fructose HCl KH2PO4 MgSO4 Air A. niger NaOH NaCl Water GA
MX-101 0.02 0.07 0.01 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.00 0.00
HX-101 0.02 0.07 0.01 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.00 0.00
R-101 0.02 0.07 0.01 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.00 0.00
ST-101 0.06 0.00 0.45 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.99 0.00
MX-102 0.06 0.00 0.05 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.00 0.00
In
R-102 0.06 0.00 0.05 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.00 0.00
CL-101 0.06 0.00 0.05 0.00 0.00 0.00 0.00 0.00 0.00 0.00 90.49 0.00
EV-101 0.06 0.00 0.45 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.00 0.00
EV-102 0.06 0.00 0.45 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.25 0.00
EV-103 0.06 0.00 0.45 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.13 0.00
MX-101 0.02 0.07 0.01 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.00 0.00
Out HX-101 0.02 0.07 0.01 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.00 0.00
R-101 0.06 0.00 0.45 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.99 0.00
Table 3.1. Mass Balance of Overall Process (continued)
ST-101 0.06 0.00 0.05 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.99 0.00
MX-102 0.06 0.00 0.05 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00
R-102 0.06 0.00 0.05 0.00 0.00 0.00 0.00 0.00 0.00 0.00 90.49 0.00
Out CL-101 0.06 0.00 0.45 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.00 0.00
EV-101 0.06 0.00 0.45 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.25 0.00
EV-102 0.06 0.00 0.45 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.13 0.00
EV-103 496.2 0.00 373.3 0.00 0.15 0.04 0.00 0.00 0.00 0.00 519.17 0.00
Fermentation & Purification
FR-101 0.10 0.00 0.90 0.00 0.00 0.00 0.67 0.89 0.00 0.00 0.71 0.41
MF-101 0.04 0.00 0.90 0.00 0.00 0.15 0.00 0.89 0.00 0.00 23.25 0.48
In
MF-102 0.00 0.00 0.01 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.71 0.38
MF-103 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.57 0.30
FR-101 0.04 0.00 0.90 0.00 0.00 0.00 0.67 0.89 0.00 0.00 0.71 0.48
Out
MF-101 0.00 0.00 0.01 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.71 0.38
Table 3.1. Mass Balance of Overall Process (continued)
MF-102 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.57 0.30
Out
MF-103 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.11 0.06
Drying
GV-101 0.87 0.00 1.38 0.00 0.11 0.41 0.00 0.00 0.00 0.00 933.77 525.84
SR-101 0.87 0.00 1.38 0.00 0.11 0.41 0.00 0.00 0.00 0.00 933.77 525.84
HX-101 0.00 0.00 0.00 0.00 0.00 0.00 4076.77 0.00 0.00 0.00 452.97 0.00
In
HX-102 0.87 0.00 1.38 0.00 0.11 0.41 0.00 0.00 0.00 0.00 27.82 525.84
G-101 0.00 0.00 0.00 0.00 0.00 0.00 4473.33 0.00 0.00 0.00 497.03 0.00
FD-101 0.00 0.00 0.00 0.00 0.00 0.00 4473.33 0.00 0.00 0.00 497.03 0.00
GV-101 0.87 0.00 1.38 0.00 0.11 0.41 0.00 0.00 0.00 0.00 933.77 525.84
SR-101 0.87 0.00 1.38 0.00 0.11 0.41 4076.77 0.00 0.00 0.00 1386.75 525.84
HX-101 0.00 0.00 0.00 0.00 0.00 0.00 4076.77 0.00 0.00 0.00 452.97 0.00
Out
HX-102 0.87 0.00 1.38 0.00 0.11 0.41 0.00 0.00 0.00 0.00 27.82 525.84
G-101 0.00 0.00 0.00 0.00 0.00 0.00 4473.33 0.00 0.00 0.00 497.03 0.00
FD-101 0.00 0.00 0.00 0.00 0.00 0.00 4473.33 0.00 0.00 0.00 497.03 0.00
47
3.2 Energy Balance
Table 3.2. Energy Balance of Overall Process
Pre-Treatment
Equipment Input (kWh/h) Output (kWh/h) Requirement (kWh/h)
Mixer 1 249.09 249.09 0.00
Heater 249.09 348.62 -99.53
Reactor 1 348.62 379.33 -30.71
Sterilizer 379.33 351.96 27.37
Mixer 2 353.16 353.16 0.00
Reactor 2 354.36 354.35 0.01
Clarifier 354.35 254.13 100.22
Evaporator 1 253.19 4866.78 -4613.59
Pump 1 115.56 115.60 -0.04
Evaporator 2 115.60 859.15 -743.55
Pump 2 67.38 67.40 -0.02
Evaporator 3 67.40 439.17 -371.77
Pump 3 43.28 43.23 0.05
Fermentation & Purification
Reactor 0.06 0.07 -0.01
Pump 1 0.06 0.06 0.00
Filtration 1 0.06 0.06 0.00
Pump 2 0.036 0.036 0.00
Splitter 0.024 0.024 0.00
Filtration 2 0.036 0.036 0.00
Pump 3 0.029 0.029 0.00
Filtration 3 0.029 0.029 0.00
Drying
Valve 47.65 47.65 0
Drier 536.11 205.97 330.14
Compressor 1 41.8 66.89 -25.09
Storage 66.89 66.89 0
Heater 66.89 488.46 -421.57
Cooler 15.38 5.49 9.89
Total 3725.46 9563.66 -5838.20
3.3 Product Conversion Efficiency

The reactant will be converted to gluconic acid. According to overall mass
balance, the product conversion efficiency is:
48
$%&' ()*+,-. ()*+,-/+

ŋ = ×100%
)%0 $%./)&%1 ,2/+
525.84 AB/ℎ
ŋ789:; = ×100%
9120.96 AB/ℎ
ŋGHIJK = LM. NL%
3.4 Product Yield
Yield based on gluconic acid obtained divided by the glucose needed,
respectively, is:
$UV XP98YT:R
O&/1+P8Q:R ST UV SP98WT: = ×100%
$UZ[\SQ: T::R:R
0.06331 AB/ℎ
O&/1+P8Q:R ST UV SP98WT: = ×100%
0.05966 AB/ℎ
O&/1+P8Q:R ST Q89[;89:R Q9:8^ = _`N. _a%
3.5 Energy Consumption of Unit Product

The total required energy shown in Table 3.2 is -5838.2 kWh/hour. The
energy required to get the gluconic acid is:
b'/)Bc d/e,&)/+ 5838.2 Agℎ/ℎ) Agℎ

= = 11.1
f)*+,-. f)*+,-/+ 525.84 AB/ℎ) AB ()*+,-.
CHAPTER 4
CONCLUSION
Based on the report explained, therefore can be concluded:

1. Indonesia's demand for gluconic acid is still fulfilled through the import
channel, considering that currently there is no producing plant in Indonesia
gluconic acid.
2. Our plant uses sugar cane juice as raw material, and located in Malang, East
Java, using Aspergillus niger as fermentative agent of gluconic acid
conversion with aerobic submerged fermentation method.
3. The flowrate of sugar cane juice is 9120.96 kg/h and it will produce 525.84
kg/h of gluconic acid, based on SuperPro Simulation and Calculation.
4. The main products produced by our plant is gluconic acid with the purity of
95% - 99%.
5. The production processes which has been selected from process selection
are incubation process, fermentation reaction process, membrane
purification, and drying process.
6. The product conversion efficiency is 57.65% and the product yield based on
gluconic acid is 106.12 %.
7. Producing gluconic acid will diminish the number import which will help
the economy of Indonesia.
50
REFERENCE
A.P. Joshi, et al.: Continuous Production of Gluconic Acid by Aspergillus niger:

Immobilized on a Cellulosic Support: Study of Low pH Fermentative
Behaviour of Aspergillus niger. Process Biochemistry. 317-325 (1999)
Chemical and Physical Properties of Glucose. 2018. [ONLINE] Available
at: http://www.edinformatics.com/math_science/science_of_cooking/glucos
e.htm. [Accessed 9 September 2018].
Glucose (CAS 50-99-7) - Chemical & Physical Properties by Cheméo.
2018. [ONLINE] Available at: https://www.chemeo.com/cid/33-990-
6/Glucose. [Accessed 10 September 2018].
Lantero et al.: Process for the Preparation of Gluconic Acid and Gluconic Acid
Produced Thereby. United State Patent. (2015).
Mustafa Elfari et al.: A Gluconobacter Oxydans Mutant Converting Glucose
Almost Quantitatively to 5-keto-D-gluconic acid. Applied genetics and
Molecular Biotechnology. (66) 668-674 (2005).
Nilesh K. Purane et al.: Gluconic Acid Production from Golden Syrup by
Aspergillus niger Strain using Semiautomatic Stirred-Tank Fermenter.
Journal of Microbial & Biochemical Technology. (2012).
Parimal Pal, Ramesh Kumar, and Subhamay Banarjee: Manufacture of Gluconic
Acid: A Review Towards Process Intensification for Green Production.
Chemical Engineering and Processing: Process Intensification. (104) 160-171
(2016).
Petra Dersch et al.: Optimized Bioprocess for Production of Fructofuranosidase by
Recombinant Aspergillus niger. Biotechnological Products and Process
Engineering. (87) 2011-2024 (2010).
S. Ramachandran et al.: Gluconic Acid: A Review, Food Technol. Biotechnol. 44
(2) 185–195 (2006)
The Glucosee Molecule - Chemical and Physical Properties. 2018. World of
Molecules. [ONLINE] Available
at: https://www.worldofmolecules.com/foods/glucose.htm. [Accessed 10
September 2018].

Revised Assignment 1

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Revised Assignment 1

Uploaded by

Copyright:

Available Formats

UNIVERSITAS INDONESIA

PRODUCTION OF GLUCONIC ACID FROM SUGARCANE JUICE BY

CHEMICAL ENGINEERING DEPARTMENT

Gluconic acid is a chemical commodity with many applications in food and

iii Universitas Indonesia

LIST OF CONTENT ............................................................................................iv

Table 1.1. General Characteristics of Gluconic Acid .............................................. 7

Figure 1.1. Chemical Structure of Gluconic Acid ................................................... 6

recovered from the system. This results in a steady-rate production of the

because they contain strains capable of growing in high concentrations of sugar

1.2.2 Gluconic Acid

Figure 1.1. Chemical Structure of Gluconic Acid

1.2.2.1 Properties of Gluconic Acid

characteristics are described in Table 1.1.

Noncorrosive, mildly acidic, less irritating,

Density 1.24 g/mL

Solubility Soluble in water

1.2.2.2 Gluconic Acid Occurrence

glucopyranose and β-glucopyranose hydrate. Glucose is a building block of the

Table 1.2. Physical Properties of Glucose

Property Value Unit

1.2.3.3 Sources of Glucose

Table 1.3. Sugar Content of Selected Common Plant Foods (g/100g)

1.2.3.4 Glucose Function

pharmaceutical products, and chemical industries which may contribute towards

Table 1.4. Calculation of Production Capacity

From the calculation above, the production capacity of gluconic acid in

example of high carbohydrate containing waste which have a great quantity to

Figure 1.2. Indonesia Sugarcane Plantation Area Maps

The development of sugarcane production in Indonesia over the past 3 years

Figure 1.3. Sugarcane Production Development

Due to highly supply of sugarcane in Indonesia, the plantation has been

above 37 ˚C (Comprehensive Macrobial Resource). Gluconobacter oxydans was

Figure 1.4. Plant Location

Figure 1.5. Actual Area of The Plant

2.1 Raw Material Selection

to gluconic acid reaction, and purification. Each method is used in production of

Figure 2.1. Black Box Diagram

2.2.1 Process Production Method Selection

Table 2.1. Parameter of Scoring for Fermentation Method Selection

Table 2.2. Scoring for Fermentation Method Selection

Table 2.3. Parameter of Scoring for Fermentation Agent Selection

Table 2.4. Scoring for Fermentation Agent Selection

Table 2.4. Scoring for Fermentation Agent Selection (continued)

Table 2.5. Parameter of Scoring for Reaction Method Selection

Table 2.6. Scoring for Fermentation Reactor Selection

Table 2.6. Scoring for Fermentation Reactor Selection

Reaction Capacity 10 3 0.30 2 0.20 2 0.20

Table 2.7. Parameter of Scoring for Incubation Method Selection

Table 2.8. Scoring for Incubation Method Selection

Table 2.9. Parameter of Scoring for Fermentation Reaction Method Selection

absorption. Based the scoring of the purification processes, we choose

mixture respectively to adjust the pH value of sugarcane juice because the pH of

2"# $%& '# + '& → 2"# $%& '( (-O4NPQRN UNRT)

Feed of reactor is sugarcane juice in α-D-glucose. The Aspergillus niger as

fermentative agent can produce mutarotase enzyme to create l-D-glucose from m-

Figure 2.2. Fermentation Reaction of Gluconic Acid Production

The CSTR as a fermentor has temperature of operation condition, that work

product separation and concentration. The microfiltration of fermentation broth was

Figure 2.3. Block Flow Diagram

Figure 2.4. Process Flow Diagram

Table 3.1. Mass Balance of Overall Process

$%&' ()+,-. ()+,-/+