响应面优化产油沙漠链带藻(Desmodesmus sp.XJ842)的培养基组成
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摘要
以一株从新疆荒漠地区筛选出来的沙漠产油微藻为实验对象,经过形态学和分子生物学手段(18s rRNA及ITS测序)鉴定为链带藻属(Desmodesmus sp.),并命名为XJ842。为提高其油脂产率,采用响应面对其培养基组成(碳源、氮源和磷源)进行优化。单因素实验分析结果显示,其最适产油的碳源为Na2CO3,最适氮源为Na NO_3。运用Box-Behnken中心组合实验以及Design-Expert分析软件建立了以总脂含量和油脂产率为响应值的响应面方程,得到最适合产油的Na2CO3浓度为1.98 mmol/L,NaNO_3浓度为48.00 mmol/L,K_2HPO_4浓度为0.46 mmol/L。在最优培养基条件下,Desmodesmus sp.XJ842总脂含量达到67.73%,油脂产率达到29.27 mg/(L·d)。优化后的总脂含量、油脂产率比优化前的分别提高了2.08倍和2.61倍。优化后Desmodesmus sp.XJ842油中饱和脂肪酸和单不饱和脂肪酸占总脂肪酸的88.41%,且亚麻酸的含量小于12%,符合欧洲生物柴油标准EN14214,具有生产生物柴油的潜力。
An oil- rich desert microalgae isolated from desert regions in Xinjiang province was identified as Desmodesmus sp. by morphology and molecular biology( 18 s rRNA and ITS sequencing) and named XJ842. Its composition of culture medium( carbon source,nitrogen source and phosphorus source) was optimized to enhance its oil yield. The analysis results of the single factor experiment showed that the optimal carbon source and nitrogen source were Na_2CO_3 and NaNO_3,respectively. With the total oil content and oil yield as response values,the response surface equation was established by Box- Behnken center combination experiment combined with Design- Expert software. And the optimal concentrations of Na_2CO_3,Na NO_3 and K_2HPO_4were 1. 98,48. 00 mmol / L and 0. 46 mmol / L,respectively. Under the optimal condition of culture medium,the total oil content and oil yield of Desmodesmus sp. XJ842 reached67. 73% and 29. 27 mg /( L·d),respectively. After optimization,the total oil content and oil yield respectively increased by 2. 08 times and 2. 61 times as many as those before optimization. Meanwhile,the contents of saturated fatty acid and monounsaturated fatty acid in Desmodesmus sp. XJ842 oil accounted for 88. 41% of the total fatty acid,and the content of linolenic acid was less than 12%,which complied the European Biodiesel Standards( EN14214)and had potential in biodiesel production.
An oil- rich desert microalgae isolated from desert regions in Xinjiang province was identified as Desmodesmus sp. by morphology and molecular biology( 18 s rRNA and ITS sequencing) and named XJ842. Its composition of culture medium( carbon source,nitrogen source and phosphorus source) was optimized to enhance its oil yield. The analysis results of the single factor experiment showed that the optimal carbon source and nitrogen source were Na_2CO_3 and NaNO_3,respectively. With the total oil content and oil yield as response values,the response surface equation was established by Box- Behnken center combination experiment combined with Design- Expert software. And the optimal concentrations of Na_2CO_3,Na NO_3 and K_2HPO_4were 1. 98,48. 00 mmol / L and 0. 46 mmol / L,respectively. Under the optimal condition of culture medium,the total oil content and oil yield of Desmodesmus sp. XJ842 reached67. 73% and 29. 27 mg /( L·d),respectively. After optimization,the total oil content and oil yield respectively increased by 2. 08 times and 2. 61 times as many as those before optimization. Meanwhile,the contents of saturated fatty acid and monounsaturated fatty acid in Desmodesmus sp. XJ842 oil accounted for 88. 41% of the total fatty acid,and the content of linolenic acid was less than 12%,which complied the European Biodiesel Standards( EN14214)and had potential in biodiesel production.
引文
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[2]HSIEH C H,WU W T.Cultivation of microalgae for oil production with a cultivation strategy of urea limitation[J].Bioresour Technol,2009,100:3921-3926.
[3]YONGMANITCHAI W C,WARD O P.Omega-3 fatty acids-alternative sources of production[J].Process Biochem,1989,24(4):117-125.
[4]赵涛,苏青福,刘朝.中国荒漠化地区发展生物柴油的最优选择——微藻[J].资源科学,2011,33(8):1529-1536.
[5]HUANG Y T,SU C P.High lipid content and productivity of microalgae cultivating under elevated carbon dioxide[J].Int J Environ Sci Technol,2014,11(3):703-710.
[6]GAO Y Y,YANG M C,WANG C H.Nutrient deprivation enhances lipid content in marine microalgae[J].Bioresour Technol,2013,147:484-491.
[7]梁海斌.气相色谱法测定生物柴油中脂肪酸甲酯的含量[J].化学工程与装备,2010(8):178-179.
[8]YE C L,JIANG C J.Optimization of extraction process of crude polysaccharides from Plantago asiatica L.by response surface methodology[J].Carbohydr Polym,2011,84(1):495-502.
[9]FENG C,JOHNS M R.Effect of C/N ratio and aeration on the fatty acid composition of heterotrophic Chlorella sorokiniana[J].J Appl Phycol,1991,3(3):203-209.
[10]孔倩,杨柳燕,肖琳,等.黑暗条件下不同氮源对铜绿微囊藻(Microcystis aeruginosa)生长和pH的影响[J].生态学报,2008,28(5):2060-2064.
[11]FENG D N,CHEN Z A,XUE S,et al.Increased lipid production of the marine oleaginous microalgae Isochrysis zhangjiangensis(Chrysophyta)by nitrogen supplement[J].Bioresour Technol,2011,102:6710-6716.
[12]JIANG H M,GAO K S.Effects of nitrogen sources and concentrations on the growth and fatty acid composition of Phaeoda-ctylum tricornutum[J].Acta Pharmacol Sin,2004,28(5):545-551.
[13]肖华山,谢志强,郭建生,等.5种不同形式氮源对紫球藻生长影响的研究[J].福建师范大学学报,2001,17(2):78-81.
[14]BOHUTSKYI P,KULA T,KESSLER B,et al.Mixed trophic state production process for microalgal biomass with high lipid content for generating biodiesel and biogas[J].Bioenerg Res,2014,7(4):1174-1185.
[15]徐檬,徐小琳,李春.氮、磷对富油荒漠微藻混养生长及总脂的影响[J].石河子大学学报,2013,30(4):401-406.
[16]陈林,王忠,李铭迪,等.生物柴油组分与十六烷值关系研究[J].中国油脂,2012,37(1):53-56.