Department of Biotechnology and Genetic Engineering

Islamic University, Kushtia-7003, Bangladesh

A Report On

Fermentation Technology & Distilled Beverages: Ethyl Alcohol from Molasses

and Its Distillation

(Carew and Co. Ltd. Darshana, Chuadanga, Bangladesh)

Course No: BT-412

Course Name: Industrial Tour

Submitted To
Final Year B.Sc Examination Committee-2020

Department of Biotechnology and Genetic Engineering,

Islamic University, Kushtia, Bangladesh

Submitted By-

Rizone Al Hasib
Roll: 1617016
Reg No: 1263
Session: 2016-2017

Submission Date:1 23 January, 2022

 INDUSTRIAL TOUR

TO

Carew and Co. (Bangladesh) Ltd. Darshana, Chuadanga.

PRESENTED BY

19th BATCH

DEPARTMENT OF BIOTECHNOLOGY & GENETIC ENGINEERING,

ISLAMIC UNIVERSITY, KUSHTIA-7003, BANGLADESH

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We are in front of Carew and Co. (Bangladesh) Ltd. Darshana, Chuadanga.

We are inside the distillery unit of Carew and Co. (Bangladesh) Ltd.

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 Table of Contents

Contents Page No

Acknowledgement i

Abstract 01

Introduction 02

Objectives of our Tour 03

Introduction of the Biotechnology wave 03-05

Brief description of Carew & Co (Bangladesh) Ltd 06

History 07

Visit-01 08-14

Visit-02 15-26

Conclusion 27

References 28

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5
 ACKNOWLEDGEMENT

At the outset I would like to express my whole hearted gratitude to the Almighty Allah for giving
me the strength and opportunity to this study tour report, manages each and everything soundly.
In reality, in order to do any type of employment or research in the biotechnological field, practical
expertise with biological processes is required in addition to sufficient academic understanding.
Keeping it mind, industrial tour or field work is included in the syllabus of our final year under the
course, BT-412. Undoubtedly, it is a timely and prudential step of our respectable teachers. I have
taken efforts in this project. It would not have been possible without the kind support and help of
many individuals and organizations. I would like to extend my gratefulness to all of them.
First and foremost, I am grateful to the Department of Biotechnology and Genetic Engineering for
allowing me to gain some practical experience. Bless to all of our honorable teachers, particularly
all members of the final year BSc (Honor's) examination committee, for their constant supervision
and for sharing important information regarding our visit. I am enormously glad to Prof. Dr. Md.
Rezuanul Islam, Prof. Dr. Md. Anwarul Haque, Prof. Dr. Md. Mizanur Rahman, Prof. Dr. Md.
Abul Kalam Azad, Associate Prof. Dr. A.K.M Nazmul Huda, Associate Professor Dr.Md. Khasrul
Alam and Assistant Prof. Md. Rezaul Karim for their guidance and industrial co-operation during
the visit. , I express my cordial gratefulness to my respectable teachers who presented us such a
tour of great consequence I must express my heartfelt thanks to the officers of the administration
chemist & worker of those industries for their act of kind cooperation that made us possible to
have a clear and accurate concept and to understand all the activities within those industries.
Nevertheless, I express our gratitude toward my families and friends for their kind co-operation
and encouragement which help me in completing this report.

Rizone Al Hasib
Department of Biotechnology and Genetic Engineering
Islamic University, Kushtia-7003, Bangladesh

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ABSTRACT
Biotechnology refers to the scientific and engineering principle to the processing of goods and
services. While technology generally aims to create tools to empower man, biotechnology aims to
change man himself, to better fit him to the world. In a changing climate, our prime need is to
introduce eco-friendly biotechnological approaches to address food security and safety,
biodiversity conservation and management of sustainable environment for the development of a
healthier, cleaner and greener nation. Carew and Company (Bangladesh) Ltd serves this
substantial purpose. It is a specialized institute for biotechnology research, technology transfer and
awareness creation on biotechnology products and processes. The Institute is entrusted with the
responsibility to develop eco-friendly and sustainable technology in environment, industry and
human health using modern biotechnology and to apply the benefits for the welfare of the nation.
It is committed to create a strong infrastructure both for research and commercialization ensuring
a steady flow of bioproducts, bioprocesses and new biotechnologies. These institutes will serve
the country as the coordinating canter of biotechnology research and promote awareness on
biotechnological innovations and technologies.

Keywords: Biotechnology, Carew and Company, Sugar, Molasses, Alcohol, Fermentation.

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INTRODUCTION
The goal of Biotechnology is obtain useful metabolic material. Biotechnology encompasses to
distinct phases: fermentation and product recovery. The word fermentation is derived from a Latin
verb “fervere" which means to boil conventionally. Fermentation means metabolism of larger
molecule for example carbohydrates into simple once under the influence of microorganisms or
enzymes. Fermentation simple means the production of alcohol.

Fermentation procedure must be developed for the cultivation of microorganisms under optimal
conditions and for the production of desired metabolites or enzymes by the microorganisms.
Product recovery or downstream processing involves the extraction than purification of Biological
product. Fermentation of ethyl alcohol from molasses is one of the oldest technologies of its kind.
Although this technology has been impractical for centuries sporadic research work are still being
carried out either to improve the yield of production or to employ new techniques for lowering
sown the production cost. Recently a fed-back culture technique has been reported by which
molasses has been added intermediately in the fermentation medium to increase the yield of
alcohol. In the present communication, a reverse technique entitled as a “Dilution of fermenting
wort” has been employed for fermentation of molasses. To determine the yield of alcohol relative
to the amount of residual sugar in the wort. For the first time, Louis Pasteur demonstrated the
fermentation of sugar by microorganisms and their regulation as well.

In the recent year a revolutionary success has been made for the production of alcohol on large
scale

I. Characterization of strains.
II. Improvement of potential mutant.
III. Their further use for mass production of chemicals.

It is the inherent ability of microorganisms that they utilize a variety of organic substrates and
results in wide ranges of products through metabolic processes. Owing to presence of Biochemical
diversity, microorganisms have been commercially exploited by fermentation industries.

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Objectives of Our Tour
In course of our academic study, fermentation is very popular process, we often heard of.
Fermentation is very important for all kind of biological system, and obviously our study tour to
Carew & Company (Bangladesh) Ltd. at Darshana, Chuadanga was involved with this primitive
process. The main objectives of our industrial tour are given below:

❖ To gain or achieve the practical experience of fermentation system;

❖ To know the present advancement of fermentation technology;
❖ To know how alcohol is produced by fermentation process;
❖ To know the difference between recombinant strain and mother strain in fermentation;
❖ To know the major types of products by molasses fermentation; and
❖ To know how to face major problems related to fermentation technology.

Whatever, our main purposes of this industrial tour were to understand the fermentation process
in a close view and to estimate the importance of fermentation technology in our daily life.

Introduction of the Biotechnology wave:

Biotechnology is the third wave in biological science and represents such an interface of basic and
applied sciences, where gradual and subtle transformation of science into technology can be
witnessed. Biotechnology is defined as the application of scientific and engineering principals to
the processing of materials by biological agents to provide goods and services. Biotechnology
comprises a number of technologies based upon increasing understanding of biology at the cellular
and molecular level. Biotechnology has applications in four major industrial areas, including
health care, agriculture, industrial uses of crops and other products like biofuels, biodegradable
plastics etc., and environmental uses. Moreover, biotechnology is not just a technological matter;
its development involves cutting-edge science, political, legal, and economic variables, and
external and internal negotiations and also to address the social and ethical challenges. A series of
derived terms have been coined to identify several branches of biotechnology namely Green, Red,
White, Gold etc.

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In a changing climate, our prime
need is to introduce eco-friendly
biotechnological approaches to
address food security and safety,
biodiversity conservation and
management of sustainable
environment for the development
of a healthier, cleaner and greener
nation. While technology
generally aims to create tools to
empower man, biotechnology
aims to change man himself, to
better fit him to the world. Simply
put, biotechnology is the
application of advances made in
the biological sciences, especially
involving the science of genetics
and its application.
Biotechnology has helped
improve food quality, quantity and processing. It also has applications in manufacturing, where
simple cells and proteins can be manipulated to produce chemicals.

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White biotechnology and Carew & Co (Bangladesh) Ltd industry:
White biotechnology, also known as industrial biotechnology, is biotechnology applied to
industrial processes. White biotechnology uses renewable resources such as carbohydrates from
cereals, corn and sugar beet or vegetable oils from sunflowers, rapeseed and oil palms. Increasing
efforts are also being made to use waste products as raw materials. An example is the designing
of organisms to produce a useful product.

Figure 01: Carew & Co (Bangladesh) Ltd Industry Functions Flowchart

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Brief description of Carew & Co (Bangladesh) Ltd
Carew and Company (Bangladesh) Ltd. were established under a private entrepreneur in 1938. At
that time a sugar factory, a distillery unit, and a drug factory started under it. After independence,
the Government of Bangladesh declared this institute as a state-owned institute. This large-scale
industrial complex comprises a sugar mill, distillery water, commercial farm, and organic
fertilizer. The land area of this is 3572 acres, whose 2450 acre is agricultural land. Here the main
product is a sugar produced from sugarcane. But after producing sugar from sugarcane a by-
product is found from which various products are produced. Among the produced goods from by-
products, domestic and foreign wine, vinegar, spirit, and organic fertilizer are noteworthy. Here
'Foreign liquor' or 'Foreign Wine' of nine brands of best quality is produced.

Carew & Co (Bangladesh) Ltd. is the only licensed distillery producing alcohol made from sugar
molasses in Bangladesh and is located at Darshana, Chuadanga, Khulna Division, Bangladesh. It
is the only distillery in Bangladesh owned by the Government of Bangladesh. It is located inside
Darsana Sugar Mills compound and is under the authority of Bangladesh Sugar and Food
Industries Corporation. It is one of the 15 sugar mills owned by Bangladesh Sugar and Food
Industries Corporation that makes a profit.

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 Figure 02: Molasses reserve tank

History
Carew & Co (Bangladesh) Ltd. traces its origin to the distillery established by the British
businessman John Maxwell in 1803. He built the distillery in Kanpur, Uttar Pradesh, British India.
The first distillery in the subcontinent. The distillery was successful and started making profits for
Maxwel. He hired Robert Russell Carew, spirits specialist. Carew was impressed by the result and
bought out the factory with two other investors. Carew hired his younger brother as the manager.
His brother died in the Indian Sepoy Mutiny in 1857.

After the mutiny, Carew reopened the distillery with the support of British Army. The company
became a joint-stock company in 1897. He opened branches in Asansol, Katni, and Darsana (East
Bengal, which became East Pakistan in 1947). The equipment in the factories were made in
Glasgow, Scotland. In 1971 East Pakistan became independent Bangladesh after the Bangladesh
Liberation War. The government of Bangladesh nationalised the distillery at Darsana in 1973
which became Carew & Co. (Bangladesh) Ltd.

The distillery is a profit-making company of the government and pays taxes to it as well. It has a
20 feet security wall and strong security system. It makes two kinds of liquor, domestic one using
indigenous recipes and foreign ones that do not use indigenous recipes. It has 200 distribution
agents and 13 country liquor distribution agents.

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 VISIT 01: Distillery Unit
Carew's distillery unit
The main product of Carew & Co is sugar. But after extracting sugar from sugarcane, various
products are produced as by-products, which include local liquor, foreign liquor, vinegar, spirits
and organic fertilisers. The company manufactures country spirit (CS), rectified spirit (RS), and
denatured spirit (DS), and two types of vinegar named malted vinegar and white vinegar. The
overall production capacity of the company's two distillery plants of CS, RS and DS is 1.35 crore
proof litres. The highest production so far was 52.76 lakh proof litres – half of the capacity
remaining unused as market demand was low.

The company markets liquor in 180ml, 365ml, and 750ml bottles. All designated warehouses of
the company have recently been placing higher demands, said officials, adding that the warehouses
in Dhaka and Srimangal, however, have outstripped others in this regard. According to officials,
one case contains 12 bottles of 750 ml, 24 bottles of 465 ml, and 48 bottles of 180 ml liquor.

Figure 03: Main gate of distillery plant

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 Figure 04: Fermentation House

High spirits
Entry into the distillery unit has to be authorized and strictly moderated. The first steps into the
premises are greeted by a pungent miasma of spirit. The walkways are interrupted by verdant
gardens, but the greenery does little to assuage the nostrils assailed by the smell of liquor.

The spirit is made from molasses, which are a by-product of sugar processing. Large industrial
vats are filled to the brim as workers in safety outfits oversee each individual step in the production
of the spirit. The distillery produces around five million litres of spirit ever year. The molasses are
processed into four different types of spirits: denatured spirit, rectified spirit, country liquor spirit
and extra neutral alcohol. The rectified spirit is used to distil whiskey. The denatured spirit acts as
solvent and fuel. The country liquor spirit is, as its name suggests, a local brew. The extra neutral
alcohol is used in the production of cosmetics, perfumes and white alcohol.

The rectified and country liquor spirits are fermented and added their respective flavours. The
fermentation process takes 72 hours. After the fermentation is complete, the liquor is bottled,
packaged and distributed over yet another 72 hours. They stand in neat rows of glassware ready to
be filled with alcohol. The bottles are organized in cases of nine litres. Workers haul cases to the
warehouse or directly to the waiting trucks to distribute them nationwide. Darsana Sugar Mill has
churned out a high volume of liquor ever since it was established in the late 19th century. It
currently produces 1,40,000 cases of foreign liquor annually.

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Products of Carew & Co (Bangladesh) Ltd
Carew has nine brands of "foreign liqueur"

1) Fine Brandy
2) Cherry Brandy
3) Yellow Label Malted Whiskey
4) Imperial Whiskey
5) Old Rum
6) Rosa Rum
7) Gold Riband Gin
8) Orange Curaçao
9) Tsarina Vodka (AKA Jorina Vodka)

Figure 05: Foreign liqueur brands of Carew

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1. Fine Brandy
In the field of oenology, the French term fine (high
quality) identifies and refers to high quality
brandy, usually of an Appellation d'Origine
Contrôlée (AOC), such as Cognac and Armagnac.
The varieties of fine include, Fine de Bordeaux,
Fine de Bourgogne, and Fine de la Marne.

2. Imperial Whiskey
Imperial Blue, abbreviated to IB and also known
as Seagram's Imperial Blue, is a brand of Indian
whisky, owned by Pernod Ricard, and launched in
1997. It is a blend of Indian grain spirits with
imported Scotch malts. It is commonly available in
750ml, 375ml and 180ml bottles, and also
available in 90ml bottles.

Figure 06: Fine Brandy and Imperial Whiskey

3. Cherry Brandy
Cherry brandy is actually defined as an unsweetened fruit brandy that has to be “distilled solely
from the fermented juice or mash of whole, sound, ripe fruit or from standard fruit wine”. Brandy
here is just a catch-all term for spirits distilled from fruit. Cherry 'brandy' is traditionally the largest
category of cherry liqueurs. A confusing term as most of these liqueurs are actually made by
macerating cherries in neutral spirit (vodka) rather than brandy. Some might also contain brandy,
but in most markets, this is not a legal requirement.

Cherry brandy liqueurs tend also to be flavored with spices such as cinnamon and cloves. Some
cherry liqueurs also include crushed cherry kernels to add a distinctive almond note to the finished
liqueur. When distilling cherries, it is important to use copper rather than stainless steel stills as
the copper not only helps produce a smoother distillate by removing sulphates but also helps
remove the cyanide produced when cherries are distilled

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4. Rosa Rum
Rosa Rum is a spirit made by distilling
fermented sugarcane product usually
molasses. It is produced from sugar
cane. Freshly cut cane is cut and
crushed among the rollers to extract the
juice. The residual pulp called the
bagasse is crushed several times to
extract all the juice.

5. Old Rum
Rum is a liquor made by fermenting
then distilling sugarcane molasses or
sugarcane juice. The distillate, a clear
liquid, is usually aged in oak barrels.
Most rums are produced in Caribbean
and North and South American
countries, but also in other sugar-
producing regions, such as the
Philippines and Taiwan.

Figure 07: Rosa Rum & Old Rum

Rums are produced in various grades. Light rums are commonly used in cocktails, whereas
"golden" and "dark" rums were typically consumed straight or neat, iced ("on the rocks"), or used
for cooking, but are now commonly consumed with mixers. Premium rums are made to be
consumed either straight or iced.

6. Orange Curaçao
Orange Curaçao is the original orange liqueur. It has a sweet, balanced orange flavor, but you can
also buy it in a “Dry Curaçao” style, which has a dryer finish. More common than the clear variety
is actually Blue Curaçao, bright blue in color and used in drinks like the Blue Hawaii and Blue
Lagoon

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7. Yellow Label Malted Whiskey
One of the Four Roses' blockbusters is the Original Bourbon, formerly known as the Yellow Label,
a fantastic NAS straight Bourbon. It's aged for around five years in oak barrels and supervised by
Master Distiller Brent Elliott.

8. Gold Riband Gin

Gold Riband gin, flavored,
distilled, colorless to pale yellow
liquor made from purified spirits
usually obtained from a grain mash
and having the juniper berry as its
principal flavouring ingredient. It
includes both the malty-flavored
and full-bodied Netherlands types
and the drier types, characterized
by distinct botanical flavouring,
produced in Britain and the United
States.

Figure 08: Yellow Malted Whiskey & Gold Riband Gin

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Carew’s vinegar
Vinegar manufactured by Carew and Company (Bangladesh) Ltd, a state-run sugar mill at
Darshana in Chuadanga under the Bangladesh Sugar and Food Industry Corporation, has hit the
market in the country to help the ongoing fight against coronavirus. Vinegar is made from a
mixture of acetic acid and water. Sugar or ethanol is converted into acetic acid through a process
of Gazan, called vinegar or vinegar. It has its uses all over the world. It is extremely effective in
removing excess flavor of fish or meat. Besides, vinegar is an important ingredient in various types
of cooking. In the foods such as soups or seed-made food, the use of vinegar is more commonly
seen. In addition to storing food, cooking vegetables, salad dressing is done with vinegar. It helps
to cut excess fat in the body. Each bottle of 450ml vinegar is selling at Tk 50 while the bottle of
750ml is at Tk 80,” he said, adding that they are capable of manufacturing 30,000 liters of vinegar
annually.

Carew & Co all set to produce affordable hand sanitizer

Carew and Company first introduced the Carew’s Hand Sanitizer to prevent coronavirus (COVID-
19) infection. It will be marketed under the name 'Carew's Hand Sanitizer'. Carew sanitisers consist
of 60 to 70 percent rectified spirit, glycerine, colour and fragrance. Expert chemists have been
involved with the production and this sanitiser can kill any virus instantly. Hand sanitisers can be
made with any kind of alcohol but normally it is made with isopropyl alcohol which is very scarce
now. Ethanol is a very good substitute and Carew & Co is the only source of the chemical in
Bangladesh.

Figure 09: Carew’s Vinegar & Hand Sanitizer

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 VISIT 02: Sugar Mill
Sugar production:
In the fiscal year of 2019-2020, more than 5 thousand (5,142) metric tons of sugar was produced
from sugarcane, of which about 4,683 (4,682.33) metric tons were in stock (unsold). Also, the lack
of raw materials resulted in not completing the capacity of Carew Sugar Mill, which increased the
production cost.

Figure 10: In front of sugar mill

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 Figure 11: Distillery laboratory

Fermentation substrate
Ethyl alcohol is produced from such organic materials that contain sugar or its precursor as
fundamental units. Sugarcane molasses, a by-product of sugar mill contains high % of sucrose &
Fructose.

Molasses is a dark color viscous liquid also known as mother liquor left after the crystallization of
cane sugar from cane juice still contains considerable amount of s sucrose. Black strap molasses
is concentrated cane juice from which no cane sugar has been extracted. Molasses is converted in
to ethyl alcohol by means of yeast, which contains enzyme invertase and zymase responsible for
fermentation. Another fermentable substrate are grapes, cereals, apple, fruit juice, pear & palmary,
and rice etc.

Principle
The hydrolysis of molasses by enzyme named invertase produced glucose & fructose. The
produced glucose is converted in to ethyl alcohol by an enzyme named zymase.

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Biochemistry of Alcoholic Fermentation
In 1815, Gay. Tussace formulated conversion of glucose to ethanol carbon dioxide (CO2). The
formula is given below.

C6H1206 — 2 C2H5OH + 2 CO2

Yeast (Saccharomyces cerevisiaea) converts di and oligosaccharides through EMP pathway in to
the pyruvate. In other microorganisms routes deviate from glucose - 6-phosphate to either pentose
phosphate pathway or ED- pathway. Even the fate of pyruvate defers in deferent alcoholic
producing microorganisms.

Figure 12: Alcohol Fermentation

Main Biochemical reaction

Main Reaction:

C12H22O4 + H2O → C6H12O6 (d-Glucose) + C6H12O6 (d-Fructose)

Zymase

C6H12O6 → 2C2H5OH + 2CO2; AH = 31.2 K Cal

Invertase

2C6H12O6 + H2O → ROH (alcohol) + RHO (Aldehyde)

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Raw Materials
▪ Molasses
▪ Enzyme (Invertase, Zymase) via yeast.
▪ H2SO4
▪ Urea.
▪ T.S.P.

Microorganisms used in alcohol production

There is a limited number of microorganisms which ferment carbohydrates (pentose or hexose
sugar) in to alcohols and yield some by-product. Some of the alcohol's producing organisms is...

Fungi: Aspergillus oryzae, Mucor sp, Endomyces lactis, S ellipsoideus, S saki & S cerevisiae etc.
(especially cerevisiae strain used in Carew & Co. Ltd.)

Properties of yeast's (Saccharomyces sp)

The origin of the word in many languages relates
primarily to its ability to ferment. The English
“yeast" and the Dutch ‘gist’ are derived from the
Greek term zestos which means boiled, a reference
to the bubbling foam cause by the evolution of
carbon dioxide. The German "hefe" and the
French "levure" both have their origins in verbs
meaning, "To raise" again referring to the
bubbling foam. Industrial terms such as "cultured,
true wild, top or bottom yeast” have little meaning
the botanical point of view. In fact, even from an
industrial view point they are confusing for a yeast
considered as a cultivated organism in one industry, for example a brewery yeast, may well to be
considered to be a wild yeast by bakers. From botanical point of view yeast are recognized 10 be
heterogeneous group. Botanists of the 19th century generally accepted the idea that yeasts belong
to the plant kingdom. At present, many taxonomists are in arrangement with the proceed by
Ainsworth in which the fungi are treated as a separate kingdom and the yeasts are included in the
division of eumycota:

• Yeasts are non-filamentous, unicellular fungi that are typically spherical, oval, and
elliptical.
• Yeasts cell wall components are chitin, cellulose hemicellulose.
• Yeasts are undergoing fission or budding,
• Yeasts are capable of facultative an aerobic growth.

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 • It is found in nature on ripe fruit.
• Under appropriate condition s cerevisiae forms “asci".
• Yeasts lack chlorophyll.
• Antibiotic susceptibility-resistant penicillin, tetracycline, chloramphenicol, sensitive to
griseofulvin.

Culture of Microorganism
Microorganisms isolated from deferent sources (soil, water & air) or genetic manipulation are
cultured on growth media. The growth media are supplemented with sources of carbon, nitrogen,
phosphorus, amino acids, trace-elements etc., sterilized and inoculated with specific
microorganism for the specific products. Some of the microbial cultures are described below:

Solid or semi-solid culture

Varying amount of agar amended with nutrient media gives solid or semi-solid phase. For the
research purpose, these types of media are used but they are generally avoided for microbial
product, because such media occupy space and create difficulty during harvesting.

Batch culture
In batch culture, growth phase of microorganisms passes through many stages. A microbe grows
in the medium until the nutrients are exhausted or toxic metabolites secreted by its reach to
inhibitory level.

Continuous Culture
A continuous culture is that where a steady exponential phase for growth of culture by addition of
fresh medium to the fermenter & Removal of spent medium at microbial biomass from it.

Fed batch culture

Basically, it is batch culture which is fed continuous with fresh medium without removal of
original culture medium from the fermenter. It results in continuous increase in volume of medium
in the fermenter.

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Fermenter
This is the simplest type of culture in which microorganisms grow in a vessel, known as a
fermenter or bioreactor. Hence it is also known as bioreactor; while designing a fermenter, several
criteria, which help in maximizing the yield, are taken in to the account. These are...

a) Long term operation in aseptic condition.

b) Adequate aeration and agitation.

c) PH control system.

d) Sampling facilities.

e) Minimum labor in operation, harvesting, cleaning & maintenance.

f) Temperature control system.

g) Minimum evaporation losses from fermenter.

h) Suitable for a Varity of processes.

Fermenter is provided with limited amount of medium containing all the nutrients at optimum
environmental conditions. Fermenter or bioreactor is used to provide an optimum environment in
which microorganism or enzymes can interact with a substrate & form the desired product.

Scale-up
The conversion of a laboratory procedure to an industrial process is termed scale-up. It is well
established in the field of industrial micro biology that a processes which works well at the
laboratory scale may work poorly or not at all when first attempted at large scale. It is generally not
possible to take fermentation conditions that have worked in the laboratory and blindly apply them
to industrial scale equipment. Scale-up is necessary in the following circumstances:

I. A new process is implemented in the plant.

II. Mutants with 10-20% greater yield are to be introduced in to large-scale production as soon
as possible.
III. Construction of a completely new fermentation plant, a rare occurrence.

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Material & Methods
Microorganism: A strain of Saccharomyces Cerevisiae: IMY obtained from the collection of
industrial microbiology section, IFST, B.C.S.I. R......, Dhaka was used.

It was maintained on a slant containing;

✓ Molasses 2%
✓ Yeast extract 1%
✓ Pentose 0.5%
✓ Agar 2%

The culture was propagated on the same solid medium in large culture flasks, where from the cell
were harvested in a saline solution & used to inoculate the fermentation wort. Yeast cells were
counted by a hemacytometer.

Composition of the wort

▪ Molasses dilutes to 10 to 18% sugars.
▪ Urea, 0.01%
▪ concentrated sulfuric acid 0.03%
▪ PH 4.8 to 5.0%.
▪ The final working volume each wort was 1 liter taken in a 10 liters flask and incubated at
370C for up to 42 hours. Number of yeast cells added to each wort were 5x108 /g sugar.

Dilution scheme
❖ Five deferent worts with sugar (Molasses) concentrations as 10, 12, 14 &
18% corresponding to 1.0(control) 1.2, 1.4, 1.6 & 1.8 strength were prepared. Multiple
dilutions were performed after 20, 12, 12, 10 hours of fermentation. Require a amount of
sterile water are added of them to make the final volume.
❖ Analysis of Molasses: total sugar in Molasses was estimated as invert sugar by Shaffer and
Hartman (1920) method and was found as 52% nitrogen and ass contents were found to be
0.29 &
❖ 8.1% respectively which were estimated by standard procedure (A.0.A.C official methods
of analysis 1965).
❖ Molasses, a by-product of sugar factory, usually contain 48% to 52% of total sugar.
Molasses is stored in large tanks from where it can be pumped in to the main distillery
building. It is diluted with water in such a proportion that the dilute Molasses called the
"wort" contains 10% to 12% of total sugar. This wort is then set for fermentation with the
help of yeast, which acting as a biocatalyst.

21
A quantity of about a gallon of yeast from the laboratory is brought to the yeast-developing plant
in the production floor where yeast is propagated, multiplied & thus made up its volume to 750-
800 gallon in three stages under sterile and pure condition. This contained is then transfer to open
- air pitching vat where in the volume is increased to 3000 gallon. This increasing volume are done
with the addition of wort, subsequently 3000 gallon is made up to 17000 gallon in fermentation
vat and kept for 25-30 hrs., for completion of the fermentation process.

➢ The step consists in diluting the Molasses to suitable concentration, because yeast does not
work in stone concentration of sugar, i.e. fermentation by yeasts interfered here for this
Molasses is diluted to a concentration up to this called mass, and presents the carbohydrate
ready for yeast inoculation.
➢ An ammonium salt and sulfuric acid are an added the one to furnish a nutritive constituent
deficient in Molasses, and the other the right environment pressure from 4-5 to facilitate
the ability of the selected yeast and to suppress the multiplication of wild yeast and bacteria.
➢ Magnesium sulfate is also added, when deficient, as is a small amount of phosphate.
➢ The addition of the nutrients, the pressure, the temperature, (76 F) and finally the clearing
and sterilizing of the yeast culture machine in readiness for the next batch.
➢ The temperature is maintained at 22-30° Cover 32-70 period rising about 350 C.
➢ Alcohol is for me le monosaccharaides, the sucrose is hydrolyzed D-glucose and fructose.
➢ In alcohol fermentation by yeast, this microorganism furnishes and organic catalyst or
enzyme," known as invertase which effects this hydrolysis. The east also produced another
more important enzyme zymase, which change the monosaccharaides into alcohol and
carbon dioxide.
➢ The alcohol is separated by distillation. It is pumped to the u per section, after passing
several heat exchangers passes down the spirit column, it is gradually losses its lighter
carrier’s protein, some residual sugar and vitamin products.
➢ The overhead containing alcohol, some water and the aldehyde passes through heat
exchanger to the partial condenser, which condenses sufficient of the vapors to afford a
reflux and also to strength then the vapors that do pass through to the condenser, where
about 50% alcohol containing the volatiles or aldehyde is condensed. This condensate is
conducted in to the aldehyde, column from which low boiling impurities or aldehydes are
separated as an overhead. The effluent liquor from part way down the aldehyde column
flows in to the rectifying column.
➢ In this third column the alcohol is brought to strength and finally purified. The more
volatile products, which may still contain a trace of aldehyde and some alcohol are totally
condensed and carried back to the upper part of the aldehyde still. Near the top of the
column 95-95.6 alcohol is taken off through a condenser for storage and sale.

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Estimation of ethanol
10ml of fermented broth wash diluted to 100ml with distilled water. From this, 10ml was distilled
and the distillate was absorbed in 10ml of potassium dichromate solution containing 5 6 ml
concentrated sulfuric acid. This distillate was then titrated against standard (0.1N) ferrous
ammonium sulfate solution using diphenylamine as an indicator.

𝑏𝑙𝑛𝑎𝑘 𝑟𝑒𝑎𝑑𝑖𝑛𝑔−𝑠𝑎𝑚𝑝𝑙𝑒 𝑟𝑒𝑎𝑑𝑖𝑛𝑔

Amount of alcohol v/v % = x dilution factor
𝑏𝑙𝑎𝑛𝑘 𝑟𝑒𝑎𝑑𝑖𝑛𝑔

Estimation of residual sugar

After fermentation is completed 10ml wort was centrifuged at 4000 rpm for at least 20minutes.
1ml of the supernatant solution was taken to determine the residual sugar (Shaffer and Hartman
1920).

Modern method of ethanol fermentation

Ethanol fermentation method: fermentation of ethanol is carried out in a large fermenter (size 1000
Ethanol to 1.5 million dm2). The inoculum of microorganism is maintained in fermenter at the
optimum growth conditions such as temperature, pH, oxygen and concentrations of carbohydrate,
the substrates.

Before starting the fermentation pure inoculum (starter... inoculum) of species of Saccharomyces
is prepared by inoculating the well-defined and sterilized medium. At the same time fermentation
medium formulated, sterilized and transferred to the sterile fermenter. Liquid medium in fermenter
is inoculated with a small amount of inoculum of yeast. Growth conditions of liquid broth are
maintained to provide optimum conditions such as temperature, pH, oxygen, etc. for the
production ethanol.

After inoculation at optimum growth conditions deferent periods the culture fluid is filtered when
growth of the microorganism is over. Consequently, the yeast cells (biomass) are separated from
the supernatant. From the supernatant products are recovered and purified. Commercial production
of alcohol varied in scale. Production & product recovery depend on

✓ Size of fermenter.
✓ Optimum culture conditions.
✓ Design for separation of solid & liquid.
✓ Culture medium.
✓ Potential microorganism.
✓ Joint efforts of microbiologists, geneticists, bio-chemists, engineers & chemists.

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Recovery of yeast
Yeast cells were separated from the fermented wort by centrifugation and purified by washing
three times with distilled water. It was then dried in an oven at 1000 C for 24 hours and weighed.

Preservation of yeast
The conventional methods of storage fail to prevent from losses caused by
➢ Attack of pathogens and pests.
➢ Climatic disorders.
➢ Natural disorders.
➢ Political and economic causes.
However, the conventional method could not save the viability of short lived seeds of economic
plants, for example- oil palm (Elaeis guiecmsis), rubber (Hevea brasilliensis) Citrus sp. and
Coffees sp. (Dodds and Roberts, 1985).

Especially the saccharomyces sp. is preserved by paraffin at 40 c in the laboratory of Carew and
co. ltd.

Process and quality control for fermentation

There are two lines of control, one is chemical control & other is process control. The chemical
control involves for a three-fold purpose.

❖ To guide the operation in progress to ensure the best practical results.

❖ To provide figures that will indicate the extent of the losses in the factory and at in
detecting these losses, and
❖ To accumulate data that will allow comparison of the work of the factory during
one period with that of other periods- day week, month, and year. An extension of the third
purpose is to supply figures for comparison with other factories owned by the same
company, with other factory in the same country, with factories throughout the world.

24
To obtain alcohol of good quality by fermentation process we should be
maintained or handles the following factors:
I. Temperature: most of the enzymes become inactive above 800 c and become very slow
below 200 c. So the temperature must be maintained between the above two temperature.
The optimum temperature of 30 400 c is most favorable.
II. Aeration: Fermentation process must be maintained in presence of air. The aeration rate
is 0.25-1. Ovum (air volume / liquid volume minute).
III. Concentration: An enzyme becomes inactive at high concentration of solution.
IV. Presence of nutrients: Certain in organic salt solution, which acts as a nutrient must be
added for good fermentation.
V. Absence of preservative: The preservatives, which destroy the ferment and retard the
fermentation, must be absent.
VI. Stirring: Fermentation process must be maintained the installation of a
continuous process.
VII. Pressure: In order to minimize the risk of contamination an over pressure of 0.2 - 0.5 bar
is used.

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Carew to produce organic fertilizer in a large scale
The state-owned distillery Carew & Company (Bangladesh) is going to produce its
environmentally-friendly organic fertilizer 'Sonar Dana' in a large scale in order to supply high
quality fertilizer to the farmer at a low price to increase agricultural production. "The state-owned
company has undertaken an initiative to increase the production and commercialization of the
fertilizer as per the direction of the Industries Ministry," according to an Industry Ministry press
release issued today. In this regard, Managing Director of Carew & Co Zahid Ali Ansari said that
farmers are facing loss due to low quality organic fertilizers in the market. The use of organic
fertilizers 'Sonar Dana' will increase productivity and quality of crops, he added.

Figure 13: Organic fertilizer of Carew & Company

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Conclusion
Industrial tour is part of our Honors’ final year academic course, it is very important for a person
who is destined to be biotechnologist or researcher of biological science. This type of tour increases
our practical knowledge. The reason behind this is, we have to live in a practical life where
something occurs in real, so if we don’t have enough knowledge to deal with the real-life
circumstances then we have nothing. However, by visiting Carew and Co. (Bangladesh) Ltd.
Darshana, Chuadanga, we learn something in real life which boost up ourselves to work on
biological science. It encourages us to learn something new and motivated to do something better,
because biological science is not a steady and constant science rather it’s an ever growing and
never-ending science which is still at its childhood. We should do more practical works like this
because it helps us to be creative, or conceive the fuel from which innovative something could be
done (who knows!). We always confined to our theoretical stage but it’s obviously needed to do
something in practical rather than reading. I really enjoyed this industrial tour, and learnt many
things which was slipped through from our course curriculum, or maybe I didn’t get it. I never
forget this lovely memory of my industrial tour. It still rejoices mine. Lastly, from my point of
view, industrial tour is very much important along with theoretical knowledge so that we could
see how theory works in real life and what it could be and I think this small practical knowledge
would be the greatest asset to our higher education, research or in our professional life.

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References

1.https://archive.dhakatribune.com/bangladesh/2017/08/01/carew-serving-liquor-since-1897.

2.https://www.thedailystar.net/news/bangladesh/news/ctg-rail-workers-threaten-go-strike-
demanding-overtime-2942986.

3. https://archive.dhakatribune.com/business/2017/08/01/carew-discreet-distillery.

4.https://www.tbsnews.net/economy/industry/carew-eyes-higher-output-meet-growing-liquor-
demand-348853.

5.https://bangladeshpost.net/posts/burden-of-losses-at-darshana-carew-farms-piles-up-day-by-
day-38629

6.https://www.tbsnews.net/economy/industry/carew-produce-organic-fertilizer-large-scale-
65461.

7.https://www.thedailystar.net/coronavirus-in-bangladesh-carew-co-all-set-produce-affordable-
hand-sanitiser-1883845#lg=1&slide=0

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