PENGARUH VINASSE DALAM BERBAGAI KONDISI

TROFIK TERHADAP LAJU PERTUMBUHAN DAN KADAR

METABOLIT Euglena sp. DENGAN AERASI CO2

Tesis

Untuk Memenuhi Sebagian Persyaratan

Mencapai Derajat Master of Science
Program Studi Magister Biologi

Disusun oleh
Maya Cindiati
21/490262/PBI/01809

PROGRAM STUDI BIOLOGI

PROGRAM PASCASARJANA FAKULTAS BIOLOGI
UNIVERSITAS GADJAH MADA
YOGYAKARTA
2024
 THE EFFECT OF VINASSE UNDER VARIOUS TROPHIC
CONDITIONS ON THE GROWTH RATE AND METABOLITE
CONTENTS OF Euglena sp. WITH CO2 AERATION
by
Maya Cindiati
21/490262/PBI/01809
Faculty of Biologi, Gadjah Mada University, 55231

Abstract

Euglena sp. can accumulate valuable metabolites in its biomass for use in diverse
industries. However, due to the high production costs and low biomass content, this
research explores the utilization of carbon sources as a cost-effective, abundant,
and high-nutrient cultivation medium obtained from vinasse waste and CO2. This
research aimed to determine the effect of vinasse waste on cultivation in various
trophic conditions with CO2 aeration on the growth rate and metabolite levels of
Euglena sp. The parameters measured in this study were optical density (OD) and
cell density, which were measured every day; biomass, carbohydrate, lipid, protein,
chlorophyll, carotenoid, and paramylon levels were measured every three days
during the 18-day observation period using specific methods, while cell size and
morphology were measured at the beginning and end of cultivation. The
concentration of vinasse waste used in this research was 4%. Data analysis used
one-way ANOVA followed by the Duncan test with a significance level of 95%. The
results showed that heterotrophic treatment produced the best growth rate, with
cell density reaching 30.7 x 106 cells/mL on the day 13 and OD of 1.036 on the day
11; Biomass, carbohydrate, protein, and paramylon levels reached 0.832 g/L, 0.275
g/L, 0.168 x 10-1 g/L, and 0.513 g/L respectively on the day 12. Meanwhile, the
positive control showed the highest levels of lipid and chlorophyll a, reaching 0.273
g/L and 3.332 x 10-3 g/L on day 12, and chlorophyll b and carotenoids reached
2.991 x 10-3 g/L, and 1.288 x 10-3 g/L respectively on day 18. The aspect ratio value
showed that the control treatment produced Euglena sp. spindle and elongated-
shaped; in the heterotrophic and mixo-heterotrophic treatments, the cells were
spherical and spindle-shaped, while in the mixotrophic treatment, there were
Euglena sp. with spherical, spindle, and elongated cells. Based on the one-way
ANOVA test, adding vinasse waste from various trophic modes significantly
affected growth rate and metabolite levels. This research shows that vinasse waste
can be used as an alternative medium to increase the growth rate and metabolite
accumulation in Euglena sp. However, the chosen trophic conditions must be
adjusted to the objectives of the cultivation.

Keywords: Carbon Carbon dioxide, Euglena sp., Metabolites, Trophic conditions,

Vinasse waste.

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 SUMMARY

THE EFFECT OF VINASSE IN VARIOUS TROPHIC

CONDITIONS ON GROWTH RATE AND METABOLITE
CONTENTS OF Euglena sp. WITH CO2 AERATION.

Maya Cindiati
21/490262/PBI/01809

Euglena sp. can accumulate functional metabolites such as chlorophyll

pigments (Gissibl et al., 2019), carotenoids, lipids, proteins, polysaccharides
(Rammuni et al., 2019; Tang et al., 2020) vitamins, minerals and high antioxidants
(Khavari et al., 2021) in its biomass which can be utilized in various industries such
as the food industry, nutraceuticals (Udayan et al., 2017) biofuels, biofertilizers,
hydrocarbons (Tyoker and Chetty, 2021), feed, pigments, polymers (Perez-Garcia
et al., 2011) supplements, enzymes, waste processing (Yuosuf et al., 2020),
agriculture (Renuka et al., 2018), and cosmetics (Fu et al., 2019). However, due to
the high production costs and low biomass content, this research explores the
utilization of carbon sources as a cost-effective, abundant, and high-nutrient
cultivation medium obtained from vinasse waste and CO 2.
This research aimed to determine the effect of vinasse waste on cultivation
in various trophic conditions with CO 2 aeration on the growth rate and metabolite
levels of Euglena sp. The parameters measured in this study were optical density
(OD) and cell density, which were measured every day; biomass, carbohydrate,
lipid, protein, chlorophyll, carotenoid, and paramylon levels were measured every
three days during the 18-day observation period using specific methods, while cell
size and morphology were measured at the beginning and end of cultivation. Optical
density was measured using a spectrophotometer, cell density was measured with
the cell count method (Punchard et al., 2001) using a Haemocytometer, biomass
was measured using the gravimetric method (Olguin et al., 2015) by weighing the
dry weight, while the metabolite content measured included protein. using the
Bradford method (Bradford, 1976), lipids using the Bligh & Dryer method (Bligh
& Dyer, 1959), carbohydrates and paramylon using the Phenol Sulfuric Acid

98
method (Dubois et al., 1956), pigments such as chlorophyll, carotenoids, using the
spectrophotometric method using formula for determining the amount of
chlorophyll (a and b) (Ritchie et al., 2006), and carotenoids (Strickland and Parsons,
1972). The concentration of vinasse waste used in this research was 4%. Data
analysis used one-way ANOVA followed by the Duncan test with a significance
level of 95%.
The results showed that heterotrophic treatment produced the best growth
rate, with cell density reaching 30.7 x 106 cells/mL on the day 13 and OD of 1.036
on the day 11; Biomass, carbohydrate, protein, and paramylon levels reached 0.832
g/L, 0.275 g/L, 0.165 g/L, and 0.513 g/L respectively on the day 12. Meanwhile,
the positive control showed the highest levels of lipid and chlorophyll a, reaching
0.273 g/L and 3.332 x 10 -3 g/L on day 12, and chlorophyll b and carotenoids reached
2.991 x 10-3 g/L, and 1.288 x 10-3 g/L respectively on day 18. The aspect ratio value
showed that the control treatment produced Euglena sp. spindle and elongated-
shaped; in the heterotrophic and mixo-heterotrophic treatments, the cells were
spherical and spindle-shaped, while in the mixotrophic treatment, there were
Euglena sp. with spherical, spindle, and elongated cells.
Based on the one-way ANOVA test, adding vinasse waste from various
trophic modes significantly affected growth rate and metabolite levels. This
research shows that vinasse waste can be used as an alternative medium to increase
the growth rate and metabolite accumulation in Euglena sp. However, the chosen
trophic conditions must be adjusted to the objectives of the cultivation.

99
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