biodiesel microalgae chart diesel | Biodiesel | Diesel Fuel

Bioresource Technology 97 (2006) 841–846

Biodiesel production from heterotrophic microalgal oil
Xiaoling Miao
a

a,b

, Qingyu Wu

a,*

Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, PR China b Department of Biological Sciences, Ningde Teachers College, Fujian, Ningde 352100, PR China Received 6 September 2004; received in revised form 14 February 2005; accepted 6 April 2005 Available online 4 June 2005

Abstract The present study introduced an integrated method for the production of biodiesel from microalgal oil. Heterotrophic growth of Chlorella protothecoides resulted in the accumulation of high lipid content (55%) in cells. Large amount of microalgal oil was efficiently extracted from these heterotrophic cells by using n-hexane. Biodiesel comparable to conventional diesel was obtained from heterotrophic microalgal oil by acidic transesterification. The best process combination was 100% catalyst quantity (based on oil weight) with 56:1 molar ratio of methanol to oil at temperature of 30 °C, which reduced product specific gravity from an initial value of 0.912 to a final value of 0.8637 in about 4 h of reaction time. The results suggested that the new process, which combined bioengineering and transesterification, was a feasible and effective method for the production of high quality biodiesel from microalgal oil. Ó 2005 Elsevier Ltd. All rights reserved.
Keywords: Biodiesel; Transesterification; Microalgal oil; Heterotrophic; Chlorella protothecoides; Acid catalyst

1. Introduction Fatty acid methyl esters originating from vegetable oils and animal fats are known as biodiesel. Biodiesel fuel has received considerable attention in recent years, as it is a biodegradable, renewable and non-toxic fuel. It contributes no net carbon dioxide or sulfur to the atmosphere and emits less gaseous pollutants than normal diesel (Lang et al., 2001; Antolin et al., 2002; Vicente et al., 2004). There are four primary ways to make biodiesel, direct use and blending, microemulsions, thermal cracking (pyrolysis) and transesterification (Ma and Hanna, 1999). The most common way is transesterification as the biodiesel from transesterification can be used directly or as blends with diesel fuel in diesel engine (Peterson et al., 1991; Zhang et al., 2003). Biodiesel, primarily rapeseed methyl ester, has been in commercial use as

*

Corresponding author. Tel./fax: +86 10 62781825. E-mail address: qingyu@tsinghua.edu.cn (Q. Wu).

an alternative fuel since 1988 in many European countries (Lang et al., 2001). However, in spite of the favourable impact that its commercialization could provide, the economic aspect of biodiesel production prevents its development and large-scale use, mainly due to the high feed cost of vegetable oil (Antolin et al., 2002; Lang et al., 2001). Biodiesel usually costs over US$0.5/l, compared to US$0.35/l for normal diesel (Zhang et al., 2003). Exploring ways to reduce the high cost of biodiesel is of much interest in recent biodiesel research, especially for those methods concentrating on minimizing the raw material cost. Microalgae have been suggested as very good candidates for fuel production because of their advantages of higher photosynthetic efficiency, higher biomass production and faster growth compared to other energy crops (Milne et al., 1990; Ginzburg, 1993; Dote et al., 1994; Minowa et al., 1995). Heterotrophic growth of some microalgae has been used for efficient production of biomass and some metabolites such as lipid (Shi et al., 2000, 2002; Wen et al., 2002), which can reduce

0960-8524/$ - see front matter Ó 2005 Elsevier Ltd. All rights reserved. doi:10.1016/j.biortech.2005.04.008

10 g/l glucose was added to the basal medium and the glycine was reduced to 0. flash point. protothecoides results in high production of biomass and accumulation of high lipid content in cells. protothecoides resulted in not only the disappearance of chlorophyll in cells (Fig. The molecular weight of the oil was calculated from saponification and acid value. In the present study. For the heterotrophic growth of C. Chlorella protothecoides is a microalgae that can be photoautotrophically or heterotrophically grown under different culture conditions. Experiments were designed to determine effections of catalyst quantity. heterotrophic growth of C.2. which was .97 mg KOH/g). 1992). Biodiesel yield (wt.3. cooled.1. Heterotrophic microalgal oil As shown in Fig. 2. Aeration was provided by bubbling air at regular pressure. A standard reaction mixture consisted of oil. 2004).4.. solidifying point and heating value were measured. Al-Widyan and Al-Shyoukh. Q. The specific gravity of the upper layer was precisely measured. most of the research has concentrated on biodiesel production from vegetable oil such as soybean oil. 70:1 and 84:1) and three different temperatures (30. sunflower oil. According to the methods of Al-Widyan and Al-Shyoukh (2002) and Siler-Marinkovic and Tomasevic (1998). Lipid compounds (referred as microalgal oil) was prepared by pulverization of heterotrophic cell powder in a mortar and then extraction with n-hexane.. Analysis of biodiesel from microalgal oil The properties of biodiesel such as density. 1.. 2. The characteristics of acidic transesterification of microalgal oil and the properties of the biodiesel were investigated. 60% and 100% H2SO4 based on oil weight). Reaction conditions Preliminary experiments indicated that the basic catalyst (alkali catalyst) was not suitable for the transesterification of microalgal oil probably because of the high acid 3. The elemental compositions of biodiesel were determined by a CE-440 elemental analyzer. cold filter plugging point. Miao. 2. 2002). protothecoides. Each 9.. more the reaction was completed (Al-Widyan and Al-Shyoukh. 1994). and washed with petroleum ether and then washed with hot water (50 °C) until the washing were neutral. Methods 2. Lower values of the specific gravity indicated that more of the heavy glycerine was removed. In order to investigate the changes of the product specific gravity with reaction time. aliquots of samples were taken from the reaction mixture at specified times and centrifuged to obtain the upper layer. Texas. palm oil and rapeseed oil (Siler-Marinkovic and Tomasevic. The saponification (189. The alga was grown autotrophically and axenically in batch cultures under 26 °C (±1) with continuous illumination at intensities of 40 lmol mÀ2 sÀ1. and left to separated into two layers in a separating funnel. 2002.01 mol) of microalgal oil was used in all experiments. the acid catalyst (sulfuric acid) was used in the research.20% (Table 1). 30:1. Results and discussion 3. 2001.12 g (about 0.1. Acidic transesterification of microalgal oil The acid catalyzed transesterification were carried out in flasks and heated to the reaction temperature on a shaker or constant temperature bath. However. There is no information available regarding the production of biodiesel from heterotrophic microalgal oil. molar ratio of methanol to oil.97 mg KOH/g) of the microalgal oil were determined according to the method of Vicente et al. viscosity.. The reaction mixture was heated for the specified period. 45:1. Lipid content in heterotrophic cells reached as high as 55. Heterotrophic growth of C.842 X. Therefore. methanol and concentrated sulfuric acid. 50%. value of microalgal oil (8. The culture medium and method were as described before (Wu et al. USA). four levels of the catalyst quantity (25%. 2. Wu / Bioresource Technology 97 (2006) 841–846 the cost of microalgal biomass production and microalgal oil production. 56:1.1 g/l. time and the temperature in reaction on the yield and properties of biodiesel product. The biodiesel product was obtained by evaporating the ether solution.3 mg KOH/g) and acid value (8.%) relative to the weight of microalgal oil was estimated. 1A) but also accumulation of high lipid content in cells. 2002). The average molecular weight of the oil was 933. Antolin et al. six levels of molar ratio of methanol to oil (25:1. 1998. Lang et al. 50 and 90 °C) were used in the experiments. Microalgae and sample preparation The strain of C. The details of the culture of heterotrophic cells were reported in our previous research (Wu et al. protothecoides were measured as described before (Miao et al. The main chemical components of autotrophic and heterotrophic C. protothecoides was provided by the Culture Collection of Algae at the University of Texas (Austin. The upper oil layer (biodiesel) was separated. an integrated method for biodiesel production from heterotrophic microalgal oil by acidic transesterification was developed. Cells were harvested by centrifugation and washed with distilled water and then dried by a freeze dryer. (2004).

10 55.61 0 20 A 40 60 80 Temperature (oC) 25% ‘Catalyst quantity’ 50% 60% 100% 100 0.865 0.20 ± 0. linoleic acid.28 15. protothecoides under differential interference microscopy.2.16 10. 50 °C and 90 °C. 60% and 100%.88 0.28 ± 0. esterification and gas chromatographical analysis (Wu et al.87 0. protothecoides results in high production of biomass and accumulation of high lipid content in cells. green) and heterotrophic (right. heterotrophic growth can be performed in conventional microbial bioreactors.39 ± 0. 5 h of reaction time. protothecoides cells.17 5.) The yield and the specific gravity of biodiesel product under different temperature and catalyst quantity (on oil basis) are shown in Fig. (For interpretation of the references in colour in this figure legend. 1. Biodiesel. 0. which were mainly lipid compounds (referred as oil). primarily made from vegetable oil is being commercialized in many countries (Lang et al.20 ± 0.36 ± 0. Q.. the reader is referred to the web version of this article. Temperature: 30 °C.86 0 20 40 60 80 Temperature (oC) 100 B Fig. Effect of temperature and catalyst quantity Fig.57%) (Table 1).42 ± 0.. protothecoides under autotrophic (left. In addition.93 ± 0. Developing high lipid content microalgae or Ôbioengineering microalgaeÕ would be a new and the promising way for biodiesel production in the future.26 14. which will improve the yield of biomass and reduce the cost of microalgal biomass production and microalgal oil production. Large amount of microalgal oil was efficiently extracted from these heterotrophic cells by using n-hexane.57 ± 0. .14 6. It is important as the low cost is desirable for commercial application in biodiesel production. (C) The cells of heterotrophic C. 2. protothecoides were full of lipid vesicles.65 HC 10.96 ± 0. At different levels of catalyst 25% ‘Catalyst quantity’ 50% 60% 100% 100 Biodiesel yield (wt. The fatty acid composition of the oil mainly composed oleic acid. cetane acid by hydrolysis. C) Cells of autotrophic and heterotrophic C.02 11. Miao. Wu / Bioresource Technology 97 (2006) 841–846 843 However. 1C). Reaction conditions: 30:1 molar ratio of methanol to oil. (B. which could be easily observed under differential interference microscopy (Fig.89 0. protothecoides is a microalgae that can be photoautotrophically or heterotrophically grown under different culture conditions. 50%.875 0. The influence of catalyst quantity and temperature on the yield (A) and the specific gravity (B) of biodiesel product. (B) Almost no lipid vesicles were observed in autotrophic C.43 ± 0.62 ± 0. 2001).X. The lipid-soluble compounds from the autotrophic cells appeared in a blackish green with chlorophyll and carotenoid as the major components.64 ± 0. 1992).885 0. 2.05 5. The heterotrophic cells were full of lipid vesicles. protothecoides Component (%) Protein Lipid Carbohydrate Ash Moisture Others AC 52.895 Specific gravity 0. Heterotrophic growth of C.%) 80 60 40 20 0 Table 1 Contents of the main chemical components of cells of autotrophic (AC) and heterotrophic (HC) C. The catalyst quantity based on oil weight: 25%. C.04 10. the high cost of biodiesel due to the high cost of vegetable oil prevents its large-scale use and development. whereas the lipid-soluble compounds from the heterotrophic cells appeared in a state of light yellow grease.04 1. 160 rpm. yellow) culture conditions. 3.9 about four times that in autotrophic cells (14. (A) Growth of the cells of C.

At level of 100% catalyst quantity. and the values of product specific gravity were 0.89 0. 0. 100% catalyst quantity and the temperature of 30 °C were used in the experiments of the effects of molar ratio of methanol to oil on the yield and the specific gravity of biodiesel product. As reported by Al-Widyan and Al-Shyoukh (2002) and Siler-Marinkovic and Tomasevic (1998). it took about 5–7 h to reach the asymptotic A 0. The presence of excess methanol in the transesterification process is essential for it is responsible for breaking the glycerine–fatty acid linkages (Al-Widyan and Al-Shyoukh. In the present work. good quality of biodiesel could be obtained in the presence of 100% acid catalyst at a higher temperature. values of product specific gravity (Fig. they accounted for 68% and 63%.86 0 1 2 3 4 5 6 7 8 Reaction time (h) Fig. the process combination of 100% catalyst quantity with temperature of 30 °C would be best. 4).3. At the same temperature. Miao. low values of specific gravity were obtained at levels of 100% catalyst quantity (Fig. This is probably because that high temperature and high concentration of H2SO4 could burn some of oil.878 and 0. the product specific gravity decreased with reaction time in a roughly exponential fashion (Fig. Fig. 7 h of reaction time. respectively. the lowest value of specific gravity (0. high catalyst concentrations (2. Therefore. 2B). The values of the product specific gravity at these two molar ratio levels were as low as 0. 2B). an excess of methanol in large quanti- . whereas the yield was lowest at these levels.91 0.91 30:1 ‘Molar ratio’ 45:1 56:1 Specific gravity 0. However. The biodiesel product yields at temperature of 30 °C and 50 °C were 56% and 58% respectively (Fig. 3A). 160 rpm. there were only slight difference in the yields of biodiesel and their specific gravity at temperature of 30 °C and 50 °C.873 and 0. Q. Production cost in this case is lower than that when H2SO4 was used at high temperature for a long time. 2002). Reaction conditions: 30 °C.878). 4).%) Molar ratio Specific gravity 0. On economic basis. 2A). High yields of biodiesel were obtained at molar ratio levels of 45:1 and 56:1 (Fig.87 0. It may be noticed that at any given level of methanol. 2A).25 M or 100% H2SO4) could produce biodiesel with low specific gravity in much short time. 3. Therefore. Wu / Bioresource Technology 97 (2006) 841–846 100 80 60 40 20 0 25:1 30:1 45:1 56:1 70:1 84:1 quantity. The molar ratio of methanol to oil at level of 56:1 resulted in the lowest terminal product specific gravity and in the shortest reaction time (4 h). In spite of this. The yield (A) and the specific gravity (B) of biodiesel product with different molar ratio of methanol to oil.92 0.87 0.88 0. The level of 30:1 also gave the low product specific gravity (0.90 Biodiesel yield (wt. above conditions were applied to the following experiments. high yields of biodiesel products were all obtained at 50 °C (Fig. 2B). whereas low values of specific gravity were all obtained at 90 °C (Fig.862) was obtained at 90 °C in the presence of 100% catalyst quantity.875 respectively (Fig. Reaction conditions: 30 °C. 3.88 0.90 0. 4. 3B).86 25:1 30:1 45:1 56:1 70:1 84:1 B Molar ratio Fig. 100% catalyst quantity based on oil weight. Effect of molar ratio of methanol to oil From the results presented in Fig. which cause low yield of biodiesel product. 160 rpm. 100% catalyst quantity based on oil weight.89 0.844 X. The specific gravity of the product varied with reaction time.864 respectively (Fig. 3 shows the product yield and specific gravity under different molar ratio of methanol to oil after 7 h of reaction time. the changes of the product specific gravity with reaction time were further investigated from these three molar ratio levels as shown in Fig. 2. 4. At 45:1 and 30:1 levels. The variation of product specific gravity with reaction time under different molar ratio of methanol to oil.

Heterotrophic growth of C. Energy 3. The biodiesel from microalgal oil showed much lower cold filter plugging point of À11 °C in comparison with the diesel fuel (Table 2). Bioresour. Therefore. solidifying point and heating value were determined. Castano. which reduced product specific gravity from an initial value of 0. The physical and fuel properties of biodiesel from microalgal oil in general were comparable to those of diesel fuel.I.K. Conclusions The present study introduced an integrated method for the production of biodiesel from heterotrophic microalgal oil. B. T.86–0. 4.0 (Max À6. Wu / Bioresource Technology 97 (2006) 841–846 Table 2 Comparison of properties of biodiesel from microalgal oil and diesel fuel and ASTM biodiesel standard Properties Density (kg/l) Viscosity (mm2/s. N. 1999. cold filter plugging point. It was also supported by National Key research plan 2004BA411B05 from Chinese Ministry of Science and Technology.. Al-Widyan and Al-Shyoukh.4. Acknowledgements This research was supported by NSFC project 40272054 and NSFC key project of 40332022 to Q.912 to a final value of 0. Renew.X. 1994. Dote.9 3.81 Diesel fuela 0. Y. 2001. Fuel 73. S.A. 3. which combined bioengineering and transesterification.2 115 À12 À11 0. Bioresour. 111–114. The biodiesel from heterotrophic microalgal oil could be a competitive alternative to conventional diesel fuel.. Technol. Large amount of microalgal oil was efficiently extracted from these heterotrophic cells by using n-hexane. Antolin et al. cSt at 40 °C) Flash point (°C) Solidifying point (°C) Cold filter plugging point (°C) Acid value (mg KOH/g) Heating value (MJ/kg) H/C ratio a 845 Biodiesel from microalgal oil 0. M... 1999...N. 2002. 2004) and biodiesel from microalgal oil and ASTM biodiesel standard is shown in Table 2.838 1. Biodiesel comparable to conventional diesel was obtained from heterotrophic microalgal oil by acidic transesterification. 2002. Al-Shyoukh. Recovery of liquid fuel from hydrocarbon-rich microalgae by thermochemical liquefaction. Briceno. Miao. Ma. The best combination of factors was 100% catalyst quantity (based on oil weight) with 56:1 molar ratio of methanol to oil at temperature of 30 °C. Hertz.V. viscosity. X. winter max < À15 Max 0. 249– 252. 53–62. amount of lipid in cells. the properties of biodiesel such as density. Ramirez.8637 in about 4 h of reaction time. Antolin.. 2002).. A. C. Technol. Tinaut. Dalai.864 5. protothecoides is a microalgae that can be photoautotrophically or heterotrophically grown under different culture conditions. S. ties slowed down the separation of the two phases produced as in the case of the levels of 70:1 and 84:1 molar ratio.5 – – The data about diesel fuel were taken from published literature as indicated in the text. Lang..B. Biodiesel production: a review.I. flash point. 2002.7) Max 0. Hanna. Good quality of biodiesel could be obtained in the presence of 100% acid catalyst (on oil basis) at high temperature. 83... Experimental evaluation of the transesterification of waste palm oil into biodiesel. Sawayama. Liquid fuel (oil) from halophilic algae: a renewable source of non-polluting energy. The results suggested that the new process. F.. Wu. Yokoyama.20%. Inoue.. Bakhshi. Reaney.. F. Vicente et al. Developing high lipid content microalgae or Ôbioengineering microalgaeÕ for biofuels production would be becoming a new and the promising way in biofuels production in the future. References Al-Widyan. C. Perez.. Bioresour.1 75 À50 to 10 À3.374 41 1. A. 2001.0 Min 100 – Summer max 0. M.. G. A comparison of these properties of diesel fuel (Ma and Hanna.8637 in about 4 h of reaction time. Ginzburg. 2002..Z..5–5. protothecoides resulted in the accumulation of a large . 80.. P. 70. Minowa. 1–15..912 to a final value of 0. 1993. Bioresour. 1855–1857. V. A. 85. M. Technol...O. Lang et al.J. Y.. Preparation and characterization of bio-diesels from various bio-oils.. the best process combination of acidic transesterification of microalgal oil was 100% catalyst quantity with 56:1 molar ratio of methanol to oil at temperature of 30 °C. Optimisation of biodiesel production by sunflower oil transesterification.9–4. Q.. could be a feasible and effective method for the production of high quality biodiesel from heterotrophic microalgal oil. 253–256. Most of these parameters comply with the limits established by ASTM related to biodiesel quality (Antolin et al. Characteristics of biodiesel from microalgal oil To assess the potential of biodiesel as a substitute of diesel fuel.5 40–45 1. It reduced product specific gravity from an initial value of 0. Lipid content in heterotrophic cells reached as high as 55. S. Technol.81 ASTM biodiesel standard 0.

Sheng. Wu / Bioresource Technology 97 (2006) 841–846 Siler-Marinkovic. Korus. 855– 863. D. Minowa. G. Sci.Y. 711–716.A. Miao. N. Chen.. M. Integrated biodiesel production: a comparison of different homogeneous catalysts systems. Zhang.. 1995... Technol. (in Chinese) 3. Shi. Q. R. M.Y... Wu. 1447–1453.Y.. Y.. Q. Process design and technological assessment.846 X... Okakurat... Tomasevic. Aracil. 1–16. Feldman. 1998. with shapeselective zeolites.. Wen. Q. Fast pyrolysis of microalgae to produce renewable fuels. Transesterification of sunflower oil in situ. M. 312–318. Appl. Sci. X. Biotechnol.. Anal. X.. Pyrolysis 71 (2). M. 2004.Y. Miao.. 92. Biomass 21. Appl.L. Fuel 74. . T.Y. C. Y.. Y... Bioresour. S. F. J... S. Sheng. Wu. C. J. Oil production from algal cells of Dunaliella tertiolecta by direct thermochemical liquefaction. 1. F.L. S. Fuel 77 (12). Agric. A. Catalytic conversion of microalgae and vegetable oils to premium gasoline. Nagle. 1991. X. Chen. X. New discoveries in study on hydrocarbons from thermal degradation of heterotrophically yellowing algae. Fu.. Yin.. Martinez. J.D. 1990. 2003. Eng. T. Zhang. 723–727.Y. 2002.L.. Yang. Heterotrophic production of biomass and lutein by Chlorella protothecoides on various nitrogen sources. 1389–1391. S.A.. Bioresour.. D. McLean. Kishimoto. Jiang. J.. Milne. Auld. Shi. High-yield production of lutein by the green microalga Chlorella protothecoides in heterotrophic fed-batch culture.. Prog. Technol.M. Dube. 1992. 27. 37. Q. Vicente... Jiang.. Wu. 2002. Prog.M. High cell density culture of the diatom Nitzschia laevis for eicosapentaenoic acid production: fedbatch development.. 18. Peterson. M. Yokoyama. Process Biochem. Biodiesel production from waste cooking oil. 219–232.Y. 435–440. 1735–1738. Nat... 89. G. Chen. G. 2004..M. Kates.W. Yin.. Batch type transesterification process for winter rape oil. R. 326–335. T.. A comparative study of gases generated from stimulant thermal degradation of autotrophic and heterotrophic Chlorella. Evans. Z. 1994.M.. Fu. 297–305. Enzyme Microb. Technol. 7 (6). China (B) 37. F. 2000..Y.J..

Sign up to vote on this title
UsefulNot useful