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A Newly Isolated Green Alga, Scenedesmus Acuminatus, from Thailand with Efficient Hydrogen Production

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A Newly Isolated Green Alga, Scenedesmus Acuminatus, from Thailand with Efficient Hydrogen Production 1270 Chiang Mai J. Sci. 2017; 44(4) Chiang Mai J. Sci. 2017; 44(4) : 1270-1278 http://epg.science.cmu.ac.th/ejournal/ Contributed Paper A Newly Isolated Green Alga, Scenedesmus acuminatus, from Thailand with Efficient Hydrogen Production Yuwalee Unpaprom [a], Rameshprabu Ramaraj [b, c] and Kanda Whangchai* [d] [a] Biotechnology Program, Faculty of Science, Maejo University, Chiang Mai, Thailand. [b] School of Renewable Energy, Maejo University, Chiang Mai, Thailand. [c] Energy Research Center, Maejo University, Chiang Mai, Thailand. [d] Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, Thailand. *Author for correspondence; e-mail: [email protected]; [email protected] Received: 3 March 2015 Accepted: 11 September 2015 ABSTRACT Biological H2 production is considered the most environmentally friendly route of producing H2, fullling the goals of recycling renewable resources and producing clean energy. It has attracted global attention because of its potential to become an inexhaustible, low cost, renewable source of clean energy and appears as an alternative fuel. This paper presents laboratory results of biological production of hydrogen by green alga was isolated from freshwater fish pond in Sansai, Chiang Mai province, Thailand. Under light microscope, this green alga was identified as belonging to the genus Scenedesmus and species S. acuminatus. The successful culture was established and grown in poultry litter effluent medium (PLEM) under a light intensity of 37.5 μmol-1m2 sec-1 and a temperature of 25°C. The nutrient requirements and process conditions that encourage the growth of dense and healthy algal cultures were explored. The highest H2 was produced when cultivated cells in PLEM for 21 hours under light and then incubated under anaerobic adaptation for 4 hours. Keywords: Scenedesmus acuminatus, poultry litter effluent, biohydrogen 1. INTRODUCTION Algae, the major biomass of living one of the best strategies of CO2 fixation organisms in marine and freshwater [1], are and biomass production [4]. There are the most important CO2 fixer, the primary several reasons for this approach: (i) the best producer in aquatic ecosystem through growth rate among the plants, (ii) low impacts the photosynthesis into biomass to fulfill on world’s food supply, (iii) specificity for the responsible duty of CO2 [2, 3], and play CO2 sequestration without gas separation to a crucial niche of CO2 bio-fixation of the save over 70% of total cost, (iv) excellent ecosystem [3]. treatment for combustion gas exhausted At present algae is broadly recognized as with NOx and SOx, (v) high value of algae Chiang Mai J. Sci. 2017; 44(4) 1271 biomass including of feed, food, ammonium nitrogen, nitrate, nitrite and which nutrition, pharmaceutical chemicals, are suitable for growing algae. In addition, fertilizer, aquaculture, biofuel, etc [5-7]. pathogens in the effluent were eliminated by As efficient photosynthetic organisms, anaerobic digestion particular after two-stage microalgae have unique advantages in (thermophilic and mesophilic) digestion [17]. capturing solar energy to generate reducing Therefore, digested effluent as medium of equivalents and converting atmospheric algal culture could be an alternative to CO2 to organic molecules [9]. Microalgae using costly mineral medium which is not also have special advantages in ability to environmentally sustainable. adapt to various stressful environments, In addition, microalgae grown under non-requirement of agricultural land and heterotrophic condition (using organic carbon) so on [4]. Microalgae’s high ability to could have more potential to produce use inorganic nutrients (nitrogen and hydrogen than in autotrophic condition phosphorous) from wastewater makes them (inorganic carbon). In heterotrophic condition, a useful bioremediation tool in waste water high biomass is achieved. Moreover, in treatment process. Many green microalgae heterotrophic mode of cultivation organic species are commonly used in the wastewater wastes, as cheaper carbon sources can be treatment system due to their high tolerance used. Unlike autotrophic condition, light is to soluble organic compounds [10-12]. also not required in heterotrophic cultivation Algae can be successfully cultivated in [18]. Therefore, heterotrophic cultivation wastewaters. It can be potentially a sustainable can be cheaper than autotrophic cultivation. growth medium for the algal feed stock One of the advantages of green algae [12, 13]. Use of microalgae in treatment and for producing hydrogen is their ability recycling of waste water has attracted a great to grow under photoautotrophic and deal of interest because of excessive biomass photoheterotrophic condition. There is little generation at cheaper cost without extra input information available for undertaking of nutrients such as inorganic fertilizer intensive algal production by using digested (chemical medium) [14]. Microalgae can use poultry effluent as a nutrient re-source. inorganic carbon (CO2) and organic carbon The present article reports simultaneous algae source (glucose, mannitol, acetate, sucrose). growth, biomass production and waste Cultivation of microalgae in swine wastes, recycling with the green microalga S. acuminatus dairy manure, and other animal residues has from the poultry litter waste water. been reported by several authors [14-16]. At present, the potential of microalgae More importantly, there were no bacteria as a source of renewable energy has received and pathogens found on aerated manure considerable interested in biofuel such as when algae are grown. The algae culture biodiesel, bioethanol, biogas, biohydrogen carried out in different types of reactors, and bio-oils [17, 18]. H2 is a valuable energy flasks and plastic bags are common practice carrier, an important feedstock to the chemical with mineral medium as nutrient sources. industry, and useful in detoxifying a wide Kumar et al. [17] stated digested piggery range of water pollutants. Compared to fossil effluent could be an alternative nutrient source fuels as traditional energy sources, hydrogen for mass algal production since anaerobically is a promising candidate as a clean energy treated animal waste contains nutrients such carrier in the future because of its higher as phosphorous, nitrogen species including, heating values 141.6 MJ kg-1, or 12.6 MJ m-3 1272 Chiang Mai J. Sci. 2017; 44(4) [8, 9]. Heat and water are the only products 10 days with cool white fluorescent light of combustion of H2 with no releasing using the same light intensity. of CO2 into the atmosphere [19, 20]. The single colonies on agar were picked Thus, hydrogen is a clean, renewable, and up and cultured in liquid BBM medium, non-polluting fuel. Previous reports showed and the streaking and inoculation procedure that photosynthetic microorganisms, was repeated until pure cultures were cyanobacteria and green algae, can generate obtained. The purity of the culture was hydrogen from solar energy and water monitored by regular observation under [17-20]. microscope. The isolated microalgae were The algal cultivation process was focused identified microscopically using light on in this study for reducing the cost. microscope with standard manual for algae Cultivation of microalgae in swine wastes, [23,24]. dairy manure, and other animal residues has been reported by several literatures 2.2 Inoculums Preparation [10-17, 20, 21]; by using Scenedesmus sp., which Isolated and purified microalgae is part of our ecosystem and is very accessible, (S. acuminatus) were inoculated in 250-ml a more environmental friendly and renewal Erlenmeyer flasks containing 125 ml culture fuel can be produced. In this study, we medium (BBM). Flasks were placed on a examined a newly isolated species of green reciprocating shaker at 120 rpm for 7 d at microalga S. acuminatus for biohydrogen room temperature of 25±1 °C. Light was production applied with inexpensive poultry provided by cool white fluorescent lamps at litter effluent medium (PLEM). an intensity of 37.5 μmol-1m2 sec-1. The algae culture was then transferred to 500-ml 2. MATERIALS AND METHODS Erlenmeyer flasks containing 450 ml. 2.1 Isolation and Identification of Microalgae 2.3 Preparation of Poultry Litter Effluent Microalgae were collected by plankton Medium net (20-μm pore size) from freshwater fish The anaerobically digested poultry litter pond (18°55′4.2′′N; 99°0′41.1′′E) at a effluent (PLEM) raw was collected from the location near Maejo University, Sansai, chicken farm at Mae Fak Mai, Chiang Mai Thailand. The collected samples were province, Thailand (18°97′20.01′′N; samples of about 5 ml were inoculated into 98° 97′99.41′′E) and transported to the 5-ml autoclaved Bold Basal Medium (BBM) Maejo University laboratories in 10 L plastic [22] in 20-ml test tubes and cultured at room containers and stored in a cold room temperature (25 °C) under 37.5 μmol-1m2 maintained at 4°C. Pre-treatment was sec-1 intensity with 16:8 h photoperiod for carried out by sedimentation and filtration 10 days. After incubation, individual colonies with a filter cloth to remove large, non-soluble were picked and transferred to the same particulate solids. After filtration the substrate media for purification in 250 mL conical was autoclaved for 20 min at 121°C, after flask. The culture broth was shaken manually which the liquid was stored at 4 °C for for five to six times a day. The pre-cultured 2 days for settling any visible particulate samples were streaked on BBM medium- solids and the supernatant was used for enriched agar plates and cultured for another microalgae growth studies. Chiang Mai J. Sci. 2017; 44(4) 1273 2.4 Growth Conditions and Measurements electron impact, EI, 70 eV with HP 5973 mass The isolated green alga was grown in selective detector and a molecular sieve 450 ml Erlenmeyer flasks each containing 5A 60/80 mesh packed column using a 450 ml of poultry litter effluent medium thermal conductivity detector). The injector (PLEM) for the heterotrophic growth and detector temperatures were kept at condition. It was cultivated at room 100°C whereas the oven temperature was temperature for 3 days under fluorescent maintained at 50°C.
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