CELL BIOLOGY FOR ENGINEERS
CHEMICAL AND BIOCHEMICAL PROPERTIES
The table below shows the chemical and biochemical properties of each element.
Table Error! No text of specified style in document..1
biochemical properties of each elemet
Name Chemical properties
Lactose -Soluble in water
-It has high melting point which
is at 202 ℃
Ammonium Nitrate -Soluble in cold water, hot water.
Soluble in acetone. Partially
soluble in methanol, insoluble in
diethyl ether.
-Reactive with reducing agents,
combustible materials, organic
materials, metals, alkalis.
Carbon dioxide - Linear covalent molecule.
-Colorless, odorless gas that can
be formed by the body and is
necessary for the respiration
cycle of plants and animals.
Eicosapentaenoic acid - carboxylic acid with a 20-
carbon chain and five cis double
bonds; the first double bond is
located at the third carbon from
the omega end.
Pythium irregulare It has the decompose waste
substance to be used as carbon
source such as glycerol and corn
–steep liquour.
Source:PubChem,1 December 2018
Chemical and
Biochemical properties
-Sweeten food, beverages,
medications, etc., such as sugar,
saccharine or other low-calorie
synthetic products.
-Substances that are energetically
unstable and can produce a sudden
expansion of the material, called an
explosion, which is accompanied by
heat, pressure and noise.
-added to the soil to supply nutrients
or to make available nutrients
already present in the soil, in order
to increase plant growth and
productivity.
-colorless, odorless, incombustible
gas resulting from the oxidation of
carbon.
-Eicosanoids are chemical
messengers derived from 20-carbon
polyunsaturated fatty acids that
play critical roles in immune and
inflammatory responses.
It secrete significant amount of
enzyme to the media (>80g/L)
which the temperature is optimally
kept around 37℃. Thus, it is still a
dominant production host due to it
can achieve yield up to 80-86 g/L
TAXONOMY OF SELECTED CELL

The table below shows the taxonomy and characteristics of Pythium irregulare.

Table Error! No text of specified style in document..2 Taxonomy of Pythium

Irregulare
Properties Explaination
Domain Eukarya
Kingdom Chromista
Phylum Oomycoota
Class Oomycetes
Order Pythiales
Family Pythiacceace
Genus Pythium
Species Irregulare
Cell Reproduction Hyphae and asexual spores
Colony Forms colonieswith a moderate amount of aerial mycelium when
grown on cornmeal agar. Its hyphae can be up to 6 μm wide with
terminal and intercalary seldom produced. Spores are spherical
with a 10–20 μm diameter. The hyphal swellings can be globose,
obovate, limoniform or of irregular shape, with terminal and
intercalary up to 25 μm in diameter. The morphology of oogonia
change from globose to irregular, while intercalary is about 18.5
μm.

Source : (Wu et al. 2013)

SELECTION OF CELLS

There are a few types of cells that can be used to produce EPA has been selected and compared.
The comparison has been tabulated into table below.

Table Error! No text of specified style in document..3 Types of cells producing EPA
Strain Concentration of EPA Culture time Lactose ultilization
(mg/l) (days) (g/l)
Pythium 22 5 9.3
irregulare
Thraustotheca 9 6 6.0
clavata
Conidiobolus 5 6 5.9
obscurus

Source:(Gandhi & Weete 2018)

We choose Pythium irregulare as our type of fungi because it produces the optimum production
of desired product and it takes the shortest time to incubate compared to the other yeast. The
longer the time taken to produces the product, the higher the cost. We found that Pythium
irregulare is a filamentous fungi, that is capable of withstanding stressful conditions and has
a high fermentation efficiency, rapid growth, effective sugar use, the ability to produce and low
levels of oxygen, osmotolerance, thermotolerance, , which are fundamental to its industrial
usefulness, ( Andrietta,2007). Other than that, Pythium irregulare has been widely used
because it can be grown and manipulated easily.Moreover,Pythium irregulare has high rate
lactose of consumption as its carbon source.P. irregulare is the best overall candidate for future
study because it produced the highest concentration of EPA and lactose appeared to be used
for dry weight and EPA production. (Cheng et al. 1999)

CULTIVATION AND PROPAGATION OF THE CELLS

Pythium irregulare is a soil borne oomycete plant pathogen.In nature, Pythium irregulare has
been isolated from habitats such as soil and roots of the infected plants. It can primarily be
found in sugar rich environments, which are ideal for fermentation and reproduction, ( NC
Stated University,2007 ). There are the physical and chemical factors which influence cell
growth and the productivity such as PH, temperature and substrate concentration which act
as carbon source(lactose).

Pythium irregulare can tolerate extreme temperatures although it may lose its viability with
time. For Pythium irregulare , its population will grow the most at the optimum temperature
(25°C to 27°C), and also the population growth at temperatures below the optimum will be
higher than the population growth at temperatures above the optimum.

Most of the fungi grows effectively in a small range of pH values. The optimum pH of Pythium
irregulare is 6.5 to 7.In the optimum pH, there will be the highest Pythium irregulare
population growth. (Brien et al. 1993)

In an enzyme-catalysed reaction, the rate increases in direct proportion to the substrate

concentration. The optimum lactose concentration of Pythium irregulare is 8%. In the optimum
lactose concentration, will occur the highest fungi growth. In the lactose concentration below
of the optimum there will be much lower growth, whereas in the absence of lactose there will
be almost no growth.(Gandhi & Weete 2018; Wu et al. 2013)
TYPE OF CARBON SOURCE AND NITROGEN SOURCE

Carbon and nitrogen metabolism is linked, requiring shared regulation between these two
elements. During cell cutivation and fermentation, we preferred sweet whey pemeate as a
source of lactose for Pythium irregulare.Since fungi are universal decomposers,they prefer to
use complex carbon source such as sweet whey permeate. Lactose is a favored carbon and
energy source in fungi. It provides faster growth rate and higher yield of EPA compared to
other sugars.

While, for nitrogen source we preferred to use ammonium nitrate. Ammonium nitrate
will limit the uptake of harmful elements such as chloride into large quantities and it can be
readily absorbed by the plant and do not need to undergo any further conversion, as is the case
with other nitrogen source before plant uptake, ( Mengel and Kirkby, 1987 ).

The nutritional composition used was prepared in 250 mL Erlenmayer flasks with 100
mL of sterilized medium (15 min 121 °C) composed of 150 g/L sweet whey permeate, , 5 g/L
ammonium chloride, 1 g/L magnesium sulphate and 1 g/L potassium chloride. The yeast grown
in Potato Dextrose Agar (PDA) slant for 24 h at 32 °C was resuspended in sterilized water and
inoculated at a 10% concentration of total volume of the fermentation medium. The flasks were
incubated in a shaker for 24 h at 32 °C and 150 rpm, (Braz J Microbiol,2013)