Project of glucoamylase production by submerged cultivation of Aspergillus awamori. Курсовая работа (т). Химия.
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НАЦІОНАЛЬНИЙ АВІАЦІЙНИЙ УНІВЕРСИТЕТ
Проект виробництва глюкоамілази шляхом глибинного культивування
Aspergillus awamori. Відділення біосинтезу.
Тема курсової роботи:
Проект виробництва глюкоамілази шляхом глибинного культивування Aspergillus
awamori. Відділення біосинтезу
. Термін виконання
роботи: з 7.10.2011р. до 16.12.2011р.
мікроорганізм продуцент
виду Aspergillus awamori
підігрівач для поживного
середовища, що надходить у ферментер:
кількість поживного
середовища - 27 м 3
тривалість процесу
стерилізації - 3 год
опрацювання літературних
даних 7.10.11 - 21.10.11
розробка методики
визначення 22.10.11. - 4.11.11
написання основної
частини роботи 5.11.11. - 17.11.11
виконання креслень
18.11.11. - 2.12.11
- оформлення роботи та
її захист 2.12.11. - 16.12.11
4. Завдання видав (доцент,
к.т.н. Карпенко О.П.)
Голова комісії: д.б.н.
Гаркава К.Г. 16.12.2011 р.
Члени комісії: доцент,
к.т.н. Карпенко О.П. ___.
Ecological Safetyof
Biotechnologythe execution of yearly projectSuslova Violettatheme of course
work: Project of glucoamylase production by submerged cultivation of
Aspergillus awamori. Department of biosynthesis.
. The term of work
execution: from 7.10.2011- 16.12.2011
producer microorganism
Aspergillus awamori
Heater for nutrient
medium supplied to fermenter:
quantity of nutrient
medium - 27 m 3
duration of
sterilization process - 3 h
3. Stages of yearly
project creation
processing handling of
literature data 7.10.11 - 21.10.11
- elaboration of
determination method 22.10.11. - 4.11.11
writing of principal
part of the work 5.11.11. - 17.11.11
creation of drawings 18.11.11. - 2.12.11
- issuance of the work
and its defense 2.12.11. - 16.12.11
4. Task was given by (associate
professor Karpenko V.I..).
(sign of supervisor)
(full name of supervisor)
5. Took task for
execution_____.yearly project is defended with a mark__.of commission: Doctor
Garkava K.G. 6.12.2011.of commission: associate professor, Karpenko V.I..
note for the yearly
project “Project of glucoamylase production by submerged cultivation of
Aspergillus awamori. Department of biosynthesis” contains 54 pages, 23
references, 2 drawings, 6 figures, 5 tables, 3 appendixes.purpose of this work
is to investigate general method of producing enzyme glucoamylase by the most
suitable method and conditions of cultivation that is submerged
cultivation.method - literature data processing, description of apparatus and
of technological flowsheets of glucoamylase production in the department of
biosynthesis, drawing up the equipment scheme of glucoamylase biosynthesis by
Asp. awamori culture, calculation of heater for medium supplied to fermenter.
Пояснювальна записка до
курсового проекту на тему «Проект виробництва глюкоамілази шляхом глибинного
культивування Aspergillus awamori. Відділення біосинтезу» містить 54 сторінки, 23
літературних джерела, 2 креслення, 6 рисунків, 5 таблиць, 3 додатки.
Мета роботи полягає в
дослідженні загального методу виробництва ферменту глюкоамілази найбільш
придатним способом та умовами культивування , а саме глибинним культивуванням.
Метод дослідження:
обробка літературних даних, опис технологічної схеми виробництва глюкоамілази у
відділенні біосинтезу, креслення апаратурної схеми біосинтезу глюкоамілази
продуцентом Asp. awamori, розрахунок нагрівача для поживного середовища, що постачається у
ферментер.
.1 Characteristics of final product
.1.1 General notion about enzymes
.1.3 Characteristics of glucoamylase
`1.2 Characteristics of microorganisms
producers of glucoamylase. Aspergillus awamori
.1 Grounds of choosing technological
scheme
.2 Description of technological scheme
2.2.4 Processing of waste water and air
2.3 Description of equipment scheme.
Specification of equipment
. DESCRIPTION AND CALCULATION OF HEATER
FOR NUTRIENT MEDIUM
are biological catalysts
that bring about chemical changes in substances. With the development of the
science of biochemistry has come a fuller understanding of the wide range of
enzymes present in living cells and of their modes of action.enzymes, there can
be no life. Although enzymes are only formed in living cells, many can be
separated from the cells and can continue to function in vitro. This unique
ability of enzymes to perform their specific chemical transformations in
isolation has led to an ever-increasing use of enzymes in industrial and food
processes, in bioremediation, and in medicine, and their production is
collectively termed ‘enzyme technology’. Commercially produced enzymes will
undoubtedly contribute to the solution of some of the most vital problems with
which modern society is confronted, e.g. food production, energy shortage and
preservation, and improvement of the environment, together with numerous
medical applications.activity of an enzyme is due to its catalytic nature. An
enzyme carries out its activity without being consumed in the reaction, and the
reaction occurs at a very much higher rate when the enzyme is present. Enzymes
are highly specific and function only on designated types of compounds - the
substrates.is exoenzyme that attacks starch from the nonreducing end of
polysacharide chain and fully convert starch into glucose. Some general
properties at majority of glucoamylases of microbal origin are distinguished.
Glucoamylase is widely widespread in the nature. It is synthesized by many
microorganisms and forms in animal tissues, especially in a liver, kidney etc.
Glucoamylase is used preliminary for starch hydrolyses in beverages
production.use of microorganisms as a source material for enzyme production has
developed because of different reasons such as there is normally a high
specific activity per unit dry weight of product, seasonal fluctuations of raw
materials and possible shortages due to climatic change or political upheavals
do not occur, in microbes, a wide spectrum of enzyme characteristics, such as
pH range and high temperature resistance, is available for selection,
industrial genetics has greatly increased the possibilities for optimizing
enzyme yield and type through strain selection, mutation, induction and
selection of growth conditions and, more recently, by using the innovative
powers of gene transfer technology and protein engineering.task is to make the
best choice of microorganism for production of certain enzyme that gives the
highest yield and requires the cheapest raw materials.producers of amylolytic
enzymes most often use the molds of genera Aspergillus. Presently at the
industrial receipt of foods of hydrolysis of starch - decstrose, glucose and
fructose syrups, on the stage of saccharification mainly use glucoamylases of
producers, related to the species Asp. awamori, optimal conditions of action of
which рН 5.0 and
temperature 55 o С. Aspergillus are typical obligate aerobs, therefore they can
develop only on the surface of solid or liquid medium or in a liquid, aerated
enough medium.production of glucoamylase is actual problem nowadays because of
its ability to hydrolase the starch which then can be applied as low-price
glucose source for lots of industries. The purpose of this work is to
investigate general method of producing glucoamylase enzyme and to choose the
most optimal way of its production.
.1 Characteristics
of final product
Enzymes are biocatalysts
produced by living cells to bring about specific biochemical reactions
generally forming parts of the metabolic processes of the cells, they act as
catalysts in bringing about chemical changes in substances.are highly specific
in their action on substrates and often many different enzymes are required to
bring about, by concerted action, the sequence of metabolic reactions performed
by the living cell. All enzymes which have been purified are protein in nature,
and may or may not possess a nonprotein prosthetic group.occur in every living
cell, hence in all microorganisms. Each single strain of organism produces a
large number of enzymes, hydrolyzing, oxidizing or reducing, and metabolic in
nature. But the absolute and relative amounts of the various individual enzymes
produced vary markedly between species and even between strains of the same
species. Hence, it is customary to select strains for the commercial production
of specific enzymes which have the capacity for producing highest amounts of
the particular enzymes desired. Commercial enzymes are produced from strains of
molds, bacteria, and yeasts. [1]the development of the science of biochemistry
has come a fuller understanding of the wide range of enzymes present in living
cells and of their modes of action. Without enzymes, there can be no life.
Although enzymes are only formed in living cells, many can be separated from
the cells and can continue to function in vitro. This unique ability of enzymes
to perform their specific chemical transformations in isolation has led to an
ever-increasing use of enzymes in industrial and food processes, in
bioremediation, and in medicine, and their production is collectively termed
“enzyme technology”.activity of an enzyme is due to its catalytic nature. An
enzyme carries out its activity without being consumed in the reaction, and the
reaction occurs at a very much higher rate when the enzyme is present. Enzymes
are highly specific and function only on designated types of compounds - the
substrates.
Table
1. Application of enzymes in different industries
Lipases, proteases,
cellulases, amylases
Acids, alkali,
oxidizing agents, reducing agents
Starch (i.e. high
fructose, corn syrup, fuel ethanol, etc.)
Amylases,
pullulanases, glucose isomerases
Lower environmental
phosphate and waste (manure) levels
catalytic function of
the enzyme is due not only to its primary molecular structure but also to the
intricate folding configuration of the whole enzyme molecule. It is this
configuration which endows the protein with its specific catalytic function;
disturb the configuration by, for example, a change in pH or temperature, and
the activity can be lost.of their specificity, enzymes can differentiate
between chemicals with closely related structures and can catalyse reactions
over a wide range of temperatures (0-110 o C) and in the pH range
2-14. In industrial applications this can result in high-quality products,
fewer by-products and simpler purification procedures. Furthermore, enzymes are
non-toxic and biodegradable (an attractive ‘green’ issue) and can be produced
especially from microorganisms in large amounts without the need for special
chemical-resistant equipment.technology embraces production, isolation,
purification and use in soluble or immobilised form. [2]of microorganisms as a
source material for enzyme production has developed for several important
reasons:
(1) There is normally a
high specific activity per unit dry weight of product.
(2) Seasonal
fluctuations of raw materials and possible shortages due to climatic change or
political upheavals do not occur.
(3) In microbes, a wide
spectrum of enzyme characteristics, such as pH range and high temperature
resistance, is available for selection.
(4) Industrial genetics
has greatly increased the possibilities for optimizing enzyme yield and type
through strain selection, mutation, induction and selection of growth
conditions and, more recently, by using the innovative powers of gene transfer
technology and protein engineering.produced enzymes will undoubtedly contribute
to the solution of some of the most vital problems with which modern society is
confronted, e.i. food production, energy shortage and preservation, and
improvement of the environment, together with numerous medical applications.[3]
Enzymes are divided into
six main classes according to the type of reaction catalyzed. They are assigned
code numbers which contain four elements separated by points and have the
following meaning:
. the number first
indicates to which of the six classes the enzyme belongs,
. the second indicates
the subclass,
. the third number
indicates the sub-subclass, and
. the fourth is the
serial number of the enzyme in its sub-subclass.six classes are distinguished
in the following manner:
. Oxidoreductasesclass
encompasses all enzymes that catalyze redox reactions. The recommended name is
dehydrogenase whenever possible, but reductase can also be used. Oxidase is
used only when O 2 is the acceptor for reduction. The systematic name
is formed according to donor: acceptor oxidoreductase.
. Transferasescatalyze
the transfer of a specific group, such as methyl, acyl, amino, glycosyl, or
phosphate, from one substance to another. The recommended name is normally
acceptor group transferase or donor group transferase. The systematic name is
formed according to donor: acceptor group transferase.catalyze the hydrolytic
cleavage of C-O, C-N, C-C, and some other bonds. The recommended name often
consists simply of the substrate name with the suffix -ase. The systematic name
always includes hydrolase.
. Lyasescatalyze the
cleavage of C-C, C-O, C-N, and other bonds by elimination. The recommended name
is, for example, decarboxylase, aldolase, dehydratase (elimination of CO 2 ,
aldehyde, and water, respectively). The systematic name is formed according to
substrate group-lyase.
. Isomerasescatalyze
geometric or structural rearrangements within a molecule. The different types
of isomerism lead to the names racemase, epimerase, isomerase, tautomerase,
mutase, or cycloisomerase.
. Ligasescatalyze the
joining of two molecules, coupled with the hydrolysis of a pyrophosphate bond
in ATP or another nucleoside triphosphate.1983, the recommended name often
included synthetase, but the current recommendation is that names of the type
X-Y ligase be used instead, to avoid confusion with the name synthase (which is
not confined to enzymes of class 6). The systematic name is formed according to
X: Y ligase (ADP-forming). [4]
11 Oxidoreductases To catalyze oxidation
<#"555834.files/image001.jpg">
A mechanism of attack of
substrate with glucoamylase can be of two types: either one-chained or multiple
attack, and active center has ubcentered structure. Almost all glucoamylases
are glycoproteins, containing from 5 to 35% carbohydrates which consist of
olygo-, di- and monosaccharides. A carbohydrate component can be an integral
fragment or broken on individual compounds which attach to the protein through
a threonine and serine. [7]a rule, natural microorganisms form the complex of
amylolytic enzymes, able to hydrolyze plant substrates on the basis of starch
carbohydrates. This complex includes α-amylases, and glucoamylases hydrolyzing molecules of starch to
glucose and dextrins of different molecular mass; proteases, destroying the
proteins of raw material to amino acid, being a valuable nitrous feed for
yeasts; glucanases, which intensify the process of fermentative treatment of
raw material due to the hydrolysis of unstarch polysaccharidess; pectinases,
destroying a pectin, other enzymes of lytic action.nowadays polyenzymatic
preparations have different enzymatic composition and differentiate on the
level of activity of separate enzymes. On the whole the known polyenzyme
preparations with amylolytic activity need improvement of their functional
descriptions, that will promote efficiency of their industrial application.
Particular interest is presented by enzymatic preparations with the
overactivity of glucoamylase. [8]
.2 Characteristics of microorganisms producers of
glucoamylase. Aspergillus awamori
producers of amylolytic
enzymes most often use the molds of genera Aspergillus species oryzae, usamii,
awamori, batatae; of genera Rhizopus species delemar, fonkinsis, neveus,
tonnensis, japonicum, topnineusis, and also separate representatives of Neurospora
crassa and Mucor. Yeasts and yeast-like microorganisms of genera Candida,
Saccharomyces, Endomycopsis and Endomyces also able to synthesize the enzymes
of amylolytic action. Presently at the industrial receipt of foods of
hydrolysis of starch - decstrose, glucose and fructose syrups, on the stage of
saccharification mainly use glucoamylases of producers, related to the genus
Aspergilllus: Asp. niger. Asp. awamori, Asp. oryzae, optimal conditions of
action of which рН 5.0 and temperature 55 o С.
Aspergillus
are typical obligate aerobs, therefore they can develop only on the surface of
solid or liquid medium or in a liquid, aerated enough medium. Optimal
temperature for majority of Aspergillus 25-30 °С, for some it is to 35 °С. The majority of molds at surface cultivation can undergo
short-term increase of temperatures to 40 °C and even 45 °C without the
noticeable loss of enzymes activity. Optimal humidity of medium for them is
about 65 %.
The recombinant and
mutant strains of glucoamylase producers’ molds Aspergillus niger are known.
Such strains are described: Asp. niger, synthesizing 150 unit/ml of
glucoamylase; Asp. niger N 402, got on the basis of natural strain, contains 20
copies of gene of glucoamylase; Asp. niger B0-1. Asp. oryzae is a mutant that
synthesizes both glucoamylase and amylase. The use of recombinant strains is
related to the necessity of realization of permanent researches on maintenance
of strains in the stable and active state, by creation of the special
conditions of cultivations which not always are accessible at the industrial
conduct of process.of species Asp.awamori for the saccharification of
starch-containing raw material at the industrial receipt of dextroses, glucose
and fructose syrups, ethanol are known and widely used.enough in regard to the
synthesis of glucoamylase from the known mold Asp. awamori is Asp. awamori 466,
synthesizing 183 units/ml of enzyme at growth on medium with a corn-flour at
the use of saccharification by malt milk and malt mash with diammonium
phosphate during184 h of growth. Mycelium is strongly branched, with swelling,
septate; a diameter of hyphae is 10-12 μm, the form of
conidium is rounded cylindrical or irregular; diameter of conidium 4.4-6.4 μm, color - from
olive-yellow to darkly-olive.[Appendix. A1]descriptions: colonies on the Dox’s
agar of with a yeast extract, at 25 o С, have a diameter of a 70-71 mm/7days, radially grooved, surface
velvety, edge thin, a conidial area is an umber; an exudate absent, back is
dim-yellow.descriptions: conidial heads are spherical, disintegrating on
separate columns, conidiophores weakly tinctured in terminal part, apical
expansions are spherical 20-45 μm in a diameter, sterigmas are covered on all surface. Sterigmas
are mainly double-level, metulas 6-16 х 3,5 -7 μm . Conidium is spherical, 3,5-6 μm.culture of strain
assimilates glucose, saccharose, arabinose well, and poorly - maltose, lactose,
lactoglucose and ramnose. Starch hydrolyze to glucose. Well assimilates
ammoniacal salts of inorganic acids. It consumes a peptone, casein, amino acid,
peptonizes milk.disadvantages of the described strain is a necessity for the
receipt of high enough activity of cultural liquid, use of multiphase
preparation of inoculum and enriched cultural medium for the basic fermentation
process.[9]enzyme glucoamylase is commercially valuable biological product that
is widely used in food and agricultural industry, that is for beverages and
feed additives production. The most feasible and efficient method of this
enzyme production is microbial synthesis. According to reviewed literature the
best microorganism for glucoamylase production is mold Asp. awamori because of
its high activity for biosynthesis.
general notion
microorganism glucoamylase
.1
Grounds for choosing technological scheme
biotechnological
production of enzymes is realized by two methods - surface and submerged. The
first method, applied for cultivation of molds is characterized by development
of mycelium on a surface of solid or liquid substrate. The film of mycelium,
producing not only amylolytic enzymes but also organic acids, inactivating them
appears on liquid substrate, therefore solid substrates with the developed
surface - wheat bran, pellet of grains, potato fiber and others are used. Maximal
activity of enzymes is reached at cultivation of molds on wheat bran. The
pellet of grains is poor in nutrient substances, and activity of enzymes in the
cultures of molds, grown on it in 4-5 times lower, than on bran. The mature
culture of molds in result of bran particles covering with mycelium looks like
dense felt-like mass.[10]surface fermentation consists in growth of producer on
the surface of thin layer of solid loose medium. Submerged fermentation in a
liquid medium can be realized both in the conditions of batch process and with
the use of the flowing systems.surface fermentation for the receipt of inoculum
spore material is propageted by a superficial method or museum culture is grow
in the conditions of submerged liquid culture. Further inoculum is sent to the
stage of fermentation, which comes true on the surface of loose medium in
metallic trays or vertical perforated cuvettes. A culture develops on the
surface of solid loose medium, the basis of which is wheat bran, grain-growing
husk, being the source of growth substances. For loosening of medium the
arboreal sawdusts (5-10 %), and oat husk is added in brans. Mixture before
autoclaving is moistened to 20-40 % humidity and is acidified for the
improvement of sterilization conditions.medium a sterile termolabile
components, inoculum (0.02-0.1 % from mass of medium) are added, quickly mix
manually and lay out in trays with thickness 2-3 cm, which set in the
impermeable aerated chambers, preliminary sterilized. Initial humidity of
medium is 58-60 %, temperature of cultivation 28- 32°, duration of fermentation
about 36-48 hours. [7]
It allows organize the
centralized providing of biotechnological plants with the dry culture of molds
that is one of advantages of surface method of fermentation. A disadvantage of
surface method of cultivation is a necessity of setting of great number of
cuvettes, work with which it is difficult to mechanize. The prime price of
culture of mold-producer is high, thus mainly from the expense of plenty of
hand labour. Mechanization of process of cultivation is possible by creation of
continuous-action apparatus or cuvettes free vehicles with the vertical thick
layer of nutrient medium and intensive blowing of air through this
layer.submerged culture of microorganisms grow on a liquid nutrient medium at
the vigorous aeration in bottletight apparatus and in sterile conditions. A
process is fully mechanized. Sterility of submerged culture of microorganism-producer
of enzymes positively affects results. The next methods of submerged
cultivation are known: periodic, continuously-cyclic and
continuously-flowing.periodic method is characterized by irremovability of
nutrient medium in a fermenter, composition of which in the process of
development changes gradually. At continuously-cyclic method microorganisms,
located on immobile attachment in a fermenter, are washed by medium, flowing in
the reserved contour, to the complete consumption by them nutritives. Enriched
with nutritives medium during such cyclic fermentation is gradually exhausted;
at times medium stays in the area of reaction this process is more long, than
periodic.continuously-flowing method of cultivation of microorganisms is more
perfect. Essence of it consists in that microbial population develops in a
flowing nutrient medium. A method has two varieties: homogeneously-continuous
and gradient-continuous. In first case growth conduct in one fermenter; at
careful interfusion and aeration of medium the identical state of culture is
provided in all volume of liquid. In a fermenter continuously fresh medium is
supplied and from it continuously flows out an excess of cultural
liquid.continuous cultivation is carried out in the battery of fermenters,
connected by downpipes. The inoculated medium with large content of
carbohydrates and other components continuously flows from one fermenter in
other and also continuously flows out as the finished culture.continuous
cultivation in flowing mediums it is possible to grow microorganisms in
conditions optimal for their stages of development. Thus such important
factors, as concentration of nutritives, amount of products of exchange, рН, content of
dissolved oxygen, sharply changing at a periodic method of cultivations, are
maintained permanent on set level or change by worked out program. [12]
A nutrient medium for
fermentation is prepared based on physiological necessities of the used
microbial culture, and also from the type of aimed enzyme.synthesis of enzyme
in a submerged culture flows during a 3-4 days at the continuous supply of
sterile air, stabilizing of рН and temperatures of medium on strictly certain levels. The
insignificant changes of values of these parameters can cause the frequent
decline of fermentation activity. After completion of fermentation for
prevention of inactivation of enzymes cultural liquid is cooled and is directed
to down stream.
Basic difficulty in
realization of continuous cultivation is a large danger of infecting, and
necessity of frequent shutoff for realization prophylactic sterilization.
Table
3. Comparison of surface and submerged cultivations
Requires much space
for trays Requires much hand labor Uses lower pressure air blower Little
power requirement Minimum control necessary Little contamination problem
Recovery involves extraction with aqueous solution, filtration or
centrifugation, and perhaps evaporation and/or precipitation
Uses compact
fermenters Requires minimum of labor Requires high pressure air Needs
considerable power for air compressors and agitators Requires careful control
Contamination frequently a serious problem Recovery involves filtration or
centrifugation, and perhaps evaporation and/or precipitation
submerged cultivation
microorganisms develop in all volume of liquid nutrient medium. Because
majority of producers of enzymes is obligate aerobs, a medium is intensively
aerated. In microorganisms occures two indissolubly constrained processes that
is a synthesis of biomass and synthesis of enzymes. [1]
For the maximal
accumulation of enzymes certain composition of nutrient medium, providing of
air with Oxygen, timely taking off of metabolites and physiological heat,
optimal values of рН and temperatures is needed. A major condition is also sterility
of nutrient medium, supplied air, fermenters, pipelines and fittings., the best
method for the production of glucoamylase by cultivation of Asp.awamori is
submerged fermentation, because of easier controlling of parameters, minimal
requirements of hand labor, low cost of raw material and possibility of
sufficient providing medium with Oxygen due to requirements of aerobic culture.
.2 Description of the technological scheme
process of glucoamylase
production in department of biosynthesis include the next main stages:
Preparation of
equipmentsurface cultivation it is necessary sterilize an apparatus for
preparation of inoculum (capacities for inoculation, cuvettes, capacity for
water, for preparation of inoculum suspension, inoculums communications).
Sterilization of cuvettes and glassware in an inoculation department is
conducted by dry steam at a temperature 160 °C no less than 60 min Apparatus
and communications are sterilized by sharp steam at a temperature 105-120 °C
and excess pressure 0,05-0,1 МPа., especially inoculation boxing, is sterilized by irradiation by
means of the special bactericidal lamps. Sterilization of apparatus and
communications has significant mean at the submerged method of cultivation. The
most careful sterilization can not give an effect, if impermeability of
equipment is broken. [12]valves before setting check up by hydraulic
compression at pressure 0,3 МPа. Impermeability of connections is checked up at excess pressure
of steam 0,15- 0,2 МPа. The special attention is made to sterilization of apparatus and
communication for the serve of the defoamer. Sterilization of these knots is
conducted at 125-135°C during 1,5-2 h. On the stage of sterilization permanent
microbiological control of sterility of nutrient medium, air supplied to
fermenter, defoamer etc is conducted.process of fermentation for defoaming in
apparatus liquid defoamer is supplied. For receiving 0,05% emulsion of
defoamer, in a capacity bring in its concentrate, then dilute it to necessary
concentration. Emulsion of defoamer is sterilised in the special vehicle of
batch-type at temperature 123±2°C during 30 min in order to avoid bringing with
it infections to medium. After sterilization defoamer is cooled in the same
apparatus to temperature 30-32°C, then supply through a metering device in a
fermenter and inoculator.[7]a microbiological sterility check up the department
of sterilization, its walls and floor, apparatus, communications, and also
hands of workers.
The producer of
glucoamylase enzyme Asp.awamori is an aerobe, and for its normal development in
the process of cultivation it is necessary to give sterile air in a sufficient
amount. Especially high demands to sterility is required at preparation of air
for aeration of submerged culture.are a few methods of cleaning and
sterilization of air, based on two principles: killing of microorganisms and
their mechanical separation. Preparation of air for aeration is conducted as
follows:
cleaning air from rough
mechanical suspended particles (viscin filters)
preliminary conditioning
to the necessary temperature
thin cleaning of air
from microorganisms (head filter)
final clean
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