Effect of Various Pretreatment for Extracting Intracellular Lipid from Nannochloropsis oculata under Nitrogen Replete and Depleted Conditions

Microalga is one of the most compelling microbial biomasses for biodiesel production. Various pretreatment processes, namely, enzyme treatment, lysis by acid, ultrasonicator, microwaves, autoclave, and 40% NaCl, for nitrogen replete and depleted algal cultures of Nannochloropsis oculata had been carried out to check the most feasible and effective technique to disrupt cells for procuring lipids, for which concentrations were determined. Fatty acid composition, essential functional groups, and cell disruption were analyzed by GC-MS, FT-IR Spectroscopy, and Nile Red fluorescent microscopy, respectively. The present investigation showed that lipid yield was higher in nitrogen depleted cells than that in normally nourished cells. GC-MS revealed the presence of major fatty acids—palmitic, oleic, stearic, arachidic, lauric, and linoleic acids. Highest efficiency was found when cells were pretreated using acid for 3 h. The lipid content was calculated as 33.18% and 54.26% for nitrogen rich cells and nitrogen starved cells, respectively. This work thus aided in identifying the most eligible pretreatment process to avail lipids from cells, to convert them to eco-friendly and nonpolluting biodiesel.

[1]  H. Oh,et al.  Comparison of several methods for effective lipid extraction from microalgae. , 2010, Bioresource technology.

[2]  Xuewu Zhang,et al.  Biodiesel Production by Microalgal Biotechnology , 2018, Renewable Energy.

[3]  Rupam Kataki,et al.  Microalgae Chlorella as a potential bio-energy feedstock , 2011 .

[4]  W. Omar,et al.  Responses of Tetraselmis sp . and Nannochloropsis sp . Isolated from Penang National Park Coastal Waters , Malaysia , to the Combined Influences of Salinity , Light and Nitrogen Limitation , 2022 .

[5]  Arief Widjaja,et al.  Study of increasing lipid production from fresh water microalgae Chlorella vulgaris , 2009 .

[6]  Fan Yang,et al.  Enzyme-assisted extraction of lipids directly from the culture of the oleaginous yeast Rhodosporidium toruloides. , 2012, Bioresource technology.

[7]  Yung-Chuan Liu,et al.  Continuous Production of Lipase-Catalyzed Biodiesel in a Packed-Bed Reactor: Optimization and Enzyme Reuse Study , 2010, Journal of biomedicine & biotechnology.

[8]  Imogen Foubert,et al.  Optimization of an Analytical Procedure for Extraction of Lipids from Microalgae , 2012 .

[9]  Teodoro Espinosa-Solares,et al.  Macroscopic mass and energy balance of a pilot plant anaerobic bioreactor operated under thermophilic conditions , 2006, Applied biochemistry and biotechnology.

[10]  Gang Li,et al.  Effects of inorganic carbon concentration on carbon formation, nitrate utilization, biomass and oil accumulation of Nannochloropsis oculata CS 179. , 2012, Bioresource technology.

[11]  Farid Chemat,et al.  Combined Extraction Processes of Lipid from Chlorella vulgaris Microalgae: Microwave Prior to Supercritical Carbon Dioxide Extraction , 2011, International journal of molecular sciences.

[12]  G. Ahlgren,et al.  NITROGEN LIMITATION EFFECTS OF DIFFERENT NITROGEN SOURCES ON NUTRITIONAL QUALITY OF TWO FRESHWATER ORGANISMS, SCENEDESMUS QUADRICAUDA (CHLOROPHYCEAE) AND SYNECHOCOCCUS SP. (CYANOPHYCEAE) , 2003 .

[13]  Chun-Chong Fu,et al.  Simultaneous estimation of chlorophyll a and lipid contents in microalgae by three-color analysis. , 2008, Biotechnology and bioengineering.

[14]  Clemens Posten,et al.  Composition of Algal Oil and Its Potential as Biofuel , 2012 .

[15]  S. Renganathan,et al.  Optimization and kinetic studies on algal oil extraction from marine macroalgae Ulva lactuca. , 2012, Bioresource technology.

[16]  R. Ruoff,et al.  Lipid analysis of Neochloris oleoabundans by liquid state NMR , 2010, Biotechnology and Bioengineering.

[17]  S. Elumalai,et al.  Optimization of abiotic conditions suitable for the production of biodiesel from Chlorella vulgaris , 2011 .

[18]  Wen‐Teng Wu,et al.  Hydrolysis of microalgae cell walls for production of reducing sugar and lipid extraction. , 2010, Bioresource technology.

[19]  K. Row,et al.  Optimization of enzymatic extraction of polysaccharides from some marine algae by response surface methodology , 2012, Korean Journal of Chemical Engineering.

[20]  U. Pick,et al.  Kinetic anomalies in the interactions of Nile red with microalgae. , 2012, Journal of microbiological methods.

[21]  Ganti S. Murthy,et al.  Enzymatic Degradation of Microalgal Cell Walls , 2009 .

[22]  W. L. Zemke-White,et al.  Acid lysis of macroalgae by marine herbivorous fishes : effects of acid pH on cell wall porosity , 2000 .

[23]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[24]  Catherine A. Carmichael,et al.  Age of nitrogen deficient microalgal cells is a key factor for maximizing lipid content. , 2012 .

[25]  Elba Pinto da Silva Bon,et al.  Evaluation of Chlorella (Chlorophyta) as Source of Fermentable Sugars via Cell Wall Enzymatic Hydrolysis , 2011, Enzyme research.

[26]  W. J. Dyer,et al.  A rapid method of total lipid extraction and purification. , 1959, Canadian journal of biochemistry and physiology.

[27]  B. Cheirsilp,et al.  Effect of nitrogen, salt, and iron content in the growth medium and light intensity on lipid production by microalgae isolated from freshwater sources in Thailand. , 2011, Bioresource technology.

[28]  Razif Harun,et al.  Influence of acid pre-treatment on microalgal biomass for bioethanol production , 2011 .

[29]  T. Volova,et al.  Effect of Nitrogen Limitation on the Growth and Lipid Composition of the Green Alga Botryococcus braunii Kutz IPPAS H-252 , 2005, Russian Journal of Plant Physiology.

[30]  Song Xue,et al.  Increased lipid production of the marine oleaginous microalgae Isochrysis zhangjiangensis (Chrysophyta) by nitrogen supplement. , 2011, Bioresource technology.

[31]  K. Vanselow,et al.  TFA and EPA Productivities of Nannochloropsis salina Influenced by Temperature and Nitrate Stimuli in Turbidostatic Controlled Experiments , 2010, Marine drugs.

[32]  Xiao-Jun Ji,et al.  Disruption of Chlorella vulgaris Cells for the Release of Biodiesel-Producing Lipids: A Comparison of Grinding, Ultrasonication, Bead Milling, Enzymatic Lysis, and Microwaves , 2011, Applied biochemistry and biotechnology.

[33]  Zhang Yu-zhong,et al.  Developments in oil extraction from microalgae , 2012 .