甲酸处理对微藻油脂提取的影响

doi:10.7503/cjcu20140707

摘要/Abstract

摘要：

采用易于回收的有机酸处理微藻细胞, 以微藻Chlorella protothecoides为原料, 系统研究了甲酸处理对微藻油脂提取的影响. 结果表明, 甲酸能有效破碎微藻细胞, 最终油脂得率和脂肪酸甲酯(FAME)得率均与传统无机酸热法相当. 甲酸浓度、温度和处理时间对油脂提取有显著影响. 当甲酸浓度为88%, 液固比为6 mL/g, 100 ℃下处理30 min, Chlorella protothecoides油脂得率为49.2%, FAME得率高达92.1%. 甲酸处理提取到的微藻油脂脂肪酸组成与无机酸热法结果无明显区别. Chlorella protothecoides油脂主要含有5种脂肪酸, 其中C18:1含量最高. 实验还考察了甲酸处理对Nannochlorum sp和Nannochloropsis oceanic的适用性, 结果表明, 对于不同微藻原料, 甲酸处理提取到的微藻油脂得率和FAME得率均与无机酸热法结果相当.

关键词: 微藻, 甲酸, 油脂提取, 生物柴油

Abstract:

Developing an advanced cell treatment technology is very important for promoting microbial oils as the feedstock for biodiesel production. The traditional inorganic acid-heat treatment process has some problems such as difficulty in recycling inorganic acid, serious environmental pollution and so on. This paper explores using organic acid which is easy to recycle, to treat cells for lipid extraction from microalgae. Effect of formic acid treatment on lipid extraction from Chlorella protothecoides was investigated systematically. The results showed that formic acid was very effective in treating microalgae cells and the total lipid yield and fatty acid methyl esters(FAME) yield were comparable with those obtained from inorganic acid-heat treatment process. It was found that formic acid concentration, temperature, treatment time and liquid/solid ratio(l/s) had a significant influence on micro-algae lipid extraction. Under the optimum condition of formic acid concentration 88%, temperature 100 ℃, treatment time 30 min and ratio of liquid to solid 6:1, the total lipid yield and FAME yield reached 49.2% and 92.1%, respectively. There was no obvious difference in fatty acid composition in the microbial oils obtained from organic acid treatment and inorganic acid-heat treatment process. There were 5 kinds of fatty acid in the microbial oil from Chlorella protothecoides, among which the content of C18:1 was the highest. Then, the adaptation of formic acid treatment to different microalgae material was investigated. The results showed that the total lipid yield and FAME yield of Nannochlorum sp and Nannochloropsis Oceanic were comparable with those obtained from inorganic acid-heat treatment process.

Key words: Microalgae, Formic acid, Lipid extraction, Biodiesel

中图分类号:

TrendMD:

王松梅, 关鼎耀, 杜伟, 赵雪冰, 刘德华. 甲酸处理对微藻油脂提取的影响. 高等学校化学学报, 2015, 36(3): 499.

WANG Songmei, KUAN Dingyaw, DU Wei, ZHAO Xuebing, LIU Dehua. Effect of Organic Acid Treatment on Microalgae Lipid Extraction^†. Chem. J. Chinese Universities, 2015, 36(3): 499.

图/表 7

Table 1 Lipid extraction efficiency according to cell disruption method from Chlorella protothecoides*

Method	Bead-beating	Microwaves	Sonication	Steam explosion	HCl	Formic acid^*	No disruption
Total lipid yield(mass fraction, %)	28.9±3.1	6.0±0.5	15.4±1.1	7.4±0.8	47.2±2.6	46.7±0.8	5.7±0.4
FAME yield(mass fraction, %)	54.0±5.8	NA	27.1±1.9	NA	91.0±5.1	83.9±1.5	NA

Fig.1 Effects of formic acid concentrations on total lipid yield(a) and FAME yield(b) from Chlorella protothecoides Ratio of liquid to solid: 10 mL/g, temperature: 100 ℃, time: 30 min, stirring rate: 250 r/min.

Fig.2 Effect of temperature on total lipid yield(a) and FAME yield(b) from Chlorella proto-thecoides Mass fraction of formic acid: 88%, ratio of liquid to solid: 10 mL/g, time: 30 min, stirring rate: 250 r/min.

Fig.3 Effect of time on total lipid yield(A) and FAME yield(B) from Chlorella protothecoides Mass fraction of formic acid: 88%, ratio of liquid to solid: 10 mL/g, stirring rate: 250 r/min, temperature/℃: a. 75; b. 90; c. 100.

Fig.4 Effect of liquid/solid ratio on total lipid yield(A) and FAME yield(B) from Chlorella protothecoides Mass fraction of formic acid: 88%, ratio of liquid to solid: 100 ℃, time: 30 min, stirring rate: 250 r/min.

Table 2 Fatty acid composition of Chlorella protothecoides*

Fatty acid	Amount of fatty acid(%)		Fatty acid	Amount of fatty acid(%)
Fatty acid	Formic acid		HCl	Formic acid	HCl
C16:0	7.6±0.0	7.6±0.1	C18:2	15.9±0.1	16.0±0.0
C18:0	4.2±0.1	4.0±0.0	C20:0	1.7±0.0	1.6±0.0
C18:1	70.6±0.0	70.8±0.1

Table 3 Lipid extraction efficiency according to species with formic acid treatment method*

Series	Chlorella protothecoides		Nannochlorum sp		Nannochloropsis oceanic
Series	Formic acid	HCl	Formic acid	HCl	Formic acid	HCl
Total lipid yield(mass fraction, %)	49.2±0.0	47.2±2.6	21.3±0.7	14.7±0.4	26.0±0.0	21.7±0.1
FAME yield(mass fraction, %)	92.1±0.93	91.0±5.1	92.9±3.1	89.5±2.6	96.2±0.1	97.8±0.7

参考文献 24

[1]	Aransiola E. F., Ojumu T. V., Oyekola O. O., Madzimbamuto T. F., Ikhu-Omoregbe D. I. O., Biomass and Bioenergy,2014, 61, 276—297
[2]	Li S. H., Fang Y. M., Wang F., Li P., Li X. Y., Chem. J. Chinese Universities,2013, 34(7), 1714—1722
	(李树豪, 方亚梅, 王繁, 李萍, 李象远. 高等学校化学学报, 2013, 34(7), 1714—1722)
[3]	Chisti Y., Biotechnol. Advances, 2007, 25(3), 294—306
[4]	Rawat I., Ranjith R., Mutanda T., Bux F., Appl. Energy,2013, 103, 444—467
[5]	Tong M., Zhou Z.G., Agric. Eng. Technol.:New Energy Ind., 2009, (5), 19—26
	(童牧, 周志刚. 农业工程技术: 新能源产业, 2009, (5), 19—26)
[6]	Jiang X. M., Zheng Y. Z., Acta. Hydrob. Sin., 2003, 27(3), 243—247
	(蒋霞敏, 郑亦周. 水生生物学报, 2003, 27(3), 243—247)
[7]	Halim R., Harun R., Danquah M. K., Webley P. A., Appl. Energy,2012, 91(1), 116—121
[8]	Lee A. K., Lewis D. M., Ashman P. J., Biomass and Bioenergy,2012, 46, 89—101
[9]	Halim R., Danquah M. K., Webley P. A., Biotechnol. Adv., 2012, 30(3), 709—732
[10]	Lee J. Y., Yoo C., Jun S. Y., Ahn C. Y., Oh H. M., Bioresour. Technol., 2010, 101(1), 75—77
[11]	Zheng H. L., Yin J. L., Gao Zh., Huang H., Ji X. J., Dou C., Appl. Biochem. Biotechnol., 2011, 164(7), 1215—1224
[12]	Prabakaran P., Ravindran A. D., Lett. Appl. Microbiol., 2011, 53(2), 150—154
[13]	Neto A. M. P., Souza R. A. S., Leon-Nino A. D., Costa J. D. A., Tiburcio R. S., Nunes T. A., Mello T. C. S., Kanemoto F. T., Saldanha F. M. P., Gianesella S. M. F., Renew. Energy,2013, 55, 525—531
[14]	Kong F. M., Zhao X. Y., Tian Y. J., Guo T. Q., Liu J. J., China Brewing,2010, 29(5), 143—145
	(孔凡敏, 赵祥颖, 田延军, 国天庆, 刘建军. 中国酿造, 2010, 29(5), 143—145)
[15]	Xu J.Y., Microbial Conversion of Crude Glycerol to Lipids by Oleaginous Yeast Rhodosporidium. Toruloides, Tsinghua University, Beijing, 2013
	(徐静阳. 圆红冬孢酵母转化粗甘油合成油脂及其调控, 北京: 清华大学, 2013)
[16]	Zhang H., Tang B. K., Row K., Chem. Res. Chinese Universities,2014, 30(1), 37—41
[17]	Steriti A., Rossi R., Concas A., Cao G., Bioresour. Technol., 2014, 164, 70—77
[18]	Lam H. Q., Bigot Y. L., Delmas M., Avignon G., Ind. Crops Prod., 2001, 14, 65—71
[19]	Ligero P., Vega A., Bao M., Ind. Crops Prod., 2005, 21(2), 235—240
[20]	Zhao X. B., Wang L., Liu D. H., J. Chem. Technol. Biotechnol., 2007, 82(12), 1115—1121
[21]	Zhao X.B., Pulping and Pretreatment of Lignocellulosic Biomass with Organic Acids, Tsinghua University, Beijing, 2009
	(赵雪冰. 木质纤维原料的有机酸制浆及生物转化的预处理, 北京: 清华大学, 2009)
[22]	Bligh E. G., Dyer W. J., Can. J. Biochem. Physiol., 1959, 37, 911—917
[23]	Laurens L. M. L., Quinn M., Van W. S., David W. T., Edward J. W., Anal. Bioanal. Chem., 2012, 403(1), 167—178
[24]	Griffiths M. J., Van Hille R. P., Harrison S. T. L., Lipids,2010, 45(11), 1053—1060