Journal of Life Sciences 9 (2015) 229-233
doi: 10.17265/1934-7391/2015.05.006
The Effect of Salinity on Growth, Dry Weight and Lipid Content of the Mixed Microalgae Culture Isolated from Glagah as Biodiel Substrate
Eko Agus Suyono1, Winarto Haryadi2, Muhammad Zusron1, Matin Nuhamunada1, Sri Rahayu1 and Andhika Puspito Nugroho1
1. Faculty of Biology, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia
2. Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia Received: May 9, 2015 / Accepted: May 30, 2015 / Published: May 30, 2015.
Abstract: Microalgae u photosynthesis to convert solar energy into chemical energy, such as lipid and they can be a replacement for oil-bad fuels. They are among the fastest growing plants in the world, and about 50% of their weight is oil. This lipid oil can be ud to make biodiel. Unfortunately, there are only some of potential strains isolated from Indonesia and most of the biodiel productions a
re usually using a single strain. Then, although they are rich of oils, their biomass productivity is still low. Salinity treatment can be ud to increa their biomass as well as their lipid content. Therefore, the rearch aim was to study the effect of salinity on the growth, dry weight and lipid content of mixed microalgae isolated from Glagah, Yogyakarta. The mixed microalgae were cultured in 3NBBM medium with different salinities or types of water (a water, brackish water, and fresh water). The cultures were incubated at light intensity 3,000 lux under dark:light exposure of 12:12 h for 7 days. The number of cells was counted every 24 h with a Haemocytometer, and the biomass was calculated bad on the dry weight. The lipid content was measured on days 0, 3, and 7 using NR (Nile Red) staining, and then the amount of lipid was analyzed using a fluorescence microscope and measured with CellProfiler 2.0 software. The highest dry weight and lipid content were found in awater medium, they accounted for 3.42 mg/mL and 13.58% at day 7, respectively. Whereas, the highest number of cells was found in freshwater medium, this was 9.8 × 106 cells/mL.
Key words: Salinity, growth, dry weight, lipid content.
1. Introduction
The worldwide increa of human population and transportation has generated greater energy cons
umption of petroleum fossil fuels that lead energy crisis becau of depleting fossil fuel rerves [1]. Therefore, for solving the issue above, a renewable energy as an alternative resource should be developed. Microalgae-bad biodiel production could be a potential source for the future renewable energy [2]. Microalgae as a potential candidate for biodiel production has generated significant interest [3, 4], becau the organism is the most efficient biological
Corresponding author: Eko Agus Suyono, rearch field: algal biotechnology and engineering. E-mail: *****************.id.producer of oil and biomass source due to u photosynthesis to convert solar energy and combine water with fixing CO2 into chemical energy, such as lipid and it can be a replacement for oil-bad fuel [5, 6].
英语绘本推荐Currently, biodiel production were dominated by commercial single strains microalgae [3]. Most of the commercial single cell cultivations of microalgae have low biomass production and lipid content due to various problems such as culture condition and susceptibility to contaminant [7]. Tho problems can be overcome through the arch and lection of the local microalgae strains for biodiel production which have highest growth rate, biomass productivity,
and lipid content [8, 9]. Furthermore, the u of mixed
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The Effect of Salinity on Growth, Dry Weight and Lipid Content of the Mixed Microalgae
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culture of the lected local strains shows that cells grow faster and high biomass yield becau the cells are capable for utilizing organic carbon sources optimally [10].
Even so, optimization technology is commonly done by regulating environmental conditions and culture medium [11]. By adjusting the salinity was reported able to increa biomass production, such as in Scenedesmus almeriensis culture [12] and Scenedesmus sp. culture [13]. Moradi and Ismail [14] also reported that Botryococcus braunii adapted to grow in low salinity was able to increa the biomass production, hydrocarbon, fat, carbohydrate, and carotenoids. Microalgae have a respon against increa of salinity and osmotic stress on the environment by accumulating small molecules components for osmoregulation [6]. Hu [15] and Rao et al. [16] describe that the increasin
g of salinity leads to slight increa in the total lipid content of algae, but excessive salinity gives an negative effect on growth due to salt stress caus microalgae tend colonies-form in the growth pha, inhibit the photosynthesis and decrea the growth rate. Therefore, it is important to study the effect of different salinities in culture medium of the mixed microalgae culture isolated from Glagah cell on growth, dry weight, and lipid content for biodiel substrate.
2. Materials and Methods
2.1 Algal Cultures
The mixed microalgae culture isolate Glagah were obtained from Glagah beach in the coastal area of South Yogyakarta-Indonesia. Microalgae samples were isolated in Laboratory of Biotechnology, Faculty of Biology, UGM by using the microcapillary pipette method and rial dilution method [17]. The isolated microalgae was not identified yet, however, according to morphological characters, they consisted of three genus, there were Chlorella, Scenedesmus [18], and Nannochloropsis [19].
The strains were cultivated in modified a water medium of f/2 for lection. Then, survived strains were cultivated in modified 3NBBM medium + vitamins medium [20] They were grown in 500 mL glass bottles and incubated at light intensity of 3,000 lux under dark:light exposure of 12:12 h for 7 d
ays at 18-25 °C.
2.2 Cultivation Condition with Different Salinity
梦到别人死了是什么意思For all experiments, the microalgae were grown in modified 3NBBM medium + vitamin in 500 mL glass bottle under condition described above. The effect of salinity was investigated at 3NBBM + vitamins diluted in fresh water, a water, and brackish water (mixture of fresh water:a water (1:1)) with three replications. The cultures were inoculated into glass bottles of different salinity medium with ratio 50 mL stock culture and 200 mL the medium.
2.3 Determination of Cells Growth
To compare cell growth in different salinity, cells were counted using a light microscope and Haemocytometer Neubauer 1 mm every 24 h. Sample was shaken to homogenize, then 900 μL of sample put into microtube 2 mL mixed with 100 μL of alcohol 70% for fixing. Number of cells was calculated as follows:
Cell number = cell count × 5 × 104 cell/mL
2.4 Determination of Algal Biomass
The biomass production was measured bad on dry weight of the culture for ven days. Sample culture (2 mL) was transferred into microtube 2 mL. Sample was centrifuged at 3,300 rpm for 10 min. Supernatant was discarded, then washed using distillate water. Cell suspensions on micro tube bottom were dried in the incubator oven at 34 °C until having constant weight.
2.5 Determination of Lipid Content by Nile Red Staining
Lipid content was measured by using Nile Red
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The Effect of Salinity on Growth, Dry Weight and Lipid Content of the Mixed Microalgae
Culture Isolated from Glagah as Biodiel Substrate
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(9-(Diethylamino)-5H–benzophenoxazin-5-one) staining [21]. Nile red staining was conducted to detect intracellular lipid droplets [22]. The cultured cells (1 mL) were collected into micro tube and 0.01 mg/mL then was added with Nile Red for staining. Stained microalgae cells were obrved by F北京长城介绍
luorescent Microscope to get the lipid fluorescence. The lipid fluorescence from neutral lipid will be smeared yellow-orange [21, 23]. Then, lipid fluorescences were quantified using image analysis software CellProfiler 2.0 as an intracellular lipid droplet [24].
3. Results and Discussion
The comparison of, cell growth, dry weight, cell quota and lipid content in mixed cultures of microalgae isolated from Glagah, Yogyakarta using various salinities of medium in bacth cultures were investigated. Tho data are prented in Figs. 1a-1d.
As can be en from Fig. 1a., the microalgae were growth better in fresh water medium as similar to their habitat. The highest number of cells was found in freshwater medium reached 9.8 × 106 cells/mL at day 7, followed by brackish water and a water treatments. However, the highest total dry weight was found in a water treatment accounted for 3.42 mg/mL, followed by brackish water and fresh water. Both a water and brackish water treatments reached a pick at day 5, but fresh water treatment was at a pick at day 4. (Fig. 1b). It can be assumed that microalgae growth in fresh water medium were stimulated to proliferate their cells rather than construct their cell walls. The fresh water nutrient absorbed by microalgae cells did not ud much to construct the cell wall, but to
stimulate cell division. Then, their cells became smaller and their cell walls were relatively thinner and lighter. On the other hand, a water nutrient provided more and suitable compounds for producing carbohydrate stored in the cell walls as llulo and hemillu, so that the total dry weight was higher.
The trend of cell quota (dry weight per cells) of each treatment (Fig. 1c) was different from total dry weight (Fig. 1b). The highest cell quota was in the brackish water treatment at day 5 with reached 3.79 ng/cell, followed by a water treatment and fresh water treatment, they accounted for 1.52 ng/cell and 0.17 ng/cell, respectively (Fig. 1c). Therefore, the total dry weight in a water was the highest, but the dry weight per cells in brackish water was the highest. This happened becau the cell division in brackish water was growth very slow (Fig. 1a).
Lipid contents of the microalgae in all treatments tend to increa during their growth. The highest lipid content of the microalgae was found in awater treatment at day 7 accounted for 13.58%, followed by brackish water treatment (5.86%), and the lowest was found in fresh water treatment (0.88%) (Fig. 1d). Increasing of lipid content was caud by the accumulation of lipid under stress conditions. The results were similar to the rearch done by Takagi et al. [25] in Dunaliella cells. This was also confirmed by Hu [15] that the increa of salinity could accelerate the total lipid conten
t in microalgae cells. Under high salinity condition, the microalgae were stimulated to enhance their lipid production as osmoprotectant which necessary to protect them from salt stress [26]. High salinity water might effect on intracellular osmolarity due to hypertonic medium condition over the microalgae cells. Then, it caud relea of water inside of the microalgae cells into the environment. The mechanisms for encountering salinity stress were regulated by producing lipid in large quantities to prevent the escape of water from microalgae cells. So, the high amounts of lipid lead to enhancing the microalgae dry weight. Therefore, the high salinity treatment could be ud as method to increa the biomass productivity and stimulate lipid production of microalgae for biodiel substrates.
4. Conclusions
Different salinity have potential for increa the
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The Effect of Salinity on Growth, Dry Weight and Lipid Content of the Mixed Microalgae
Culture Isolated from Glagah as Biodiel Substrate
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(a) (b)
(c) (d)
Fig. 1 Growth (A), dry eight (B), cell quota (C) and lipid content (D) of the mixed microalgae cultures i
solated from Glagah were cultured in fresh water ().
number of cells, dry weight and lipid content on the
mixed microalgae cultures isolates from Glagah. The
highest dry weight and lipid content were found in
awater medium, they accounted for 3.42 mg/mL and
13.58% at day 7, respectively. Whereas, the highest
number of cells was found in freshwater medium, this
was 9.8 × 106 cells/mL.
Acknowledgments
All authors would like to sincerely acknowledge to
Directorate General of Higher Education, Ministry of
Education and Culture, Indonesia for funding this
rearch.
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