复合脂肪酶催化制备生物柴油的工艺研究
|
摘要
生物柴油作为一种对环境友好、无毒、可再生的替代燃料,已受到越来越多的关注。本文研究了复合脂肪酶催化制备生物柴油的新工艺,主要结果如下:
从游离脂肪酶催化制备生物柴油的实验中发现多种酶混合催化制备生物柴油过程中存在协同现象,并利用多种游离脂肪酶对这一现象进行了验证,确认复合脂肪酶比单一脂肪酶对催化酯交换反应有更为显著的促进作用,复合脂肪酶能有效地克服单一脂肪酶催化的位置专一性,提高生物柴油的转酯效率,特别是在转酯能力相当而位置特异性不同的脂肪酶之间表现出更强的协同效应。
进行了复合酶催化制备生物柴油工艺的初步研究,分别进行了无溶剂体系与叔丁醇体系中复合固定化脂肪酶与单一酶转酯效率的对比试验。在无溶剂体系中,Novozym435分别与Lipozyme TLIM和Lipozyme RMIM以70/30比例混合时,甲酯得率分别达到94.52%和96.25%,比Novozym435单独催化时的甲酯得率分别提高了9.52%和9.99%;在叔丁醇体系中,当Novozym435与Lipozyme TLIM和Lipozyme RMIM分别以60/40和80/20的比例混合时,其甲酯得率分别为85.06%和81.5%,比Novozym435单独催化的效率分别提高了9.89%和7.48%。两种体系均表现出较强的协同效果。
通过对叔丁醇体系中Novozym435与Lipozyme TLIM复合酶催化条件进行优化表明:在溶剂添加量为40%、醇油摩尔比4:1、复合酶添加量为Novozym435 1.35%+Lipozyme TLIM 1.80%,温度为40℃,摇床转速为200rpm,反应24小时,甲酯得率达到92%,复合酶在该工艺下连续使用八次时甲酯得率仍保持在90%以上。
进行了复合酶工艺扩大油源底物的研究,复合酶在叔丁醇体系中催化大豆油、菜籽油、棉籽油、蓖麻油、橄榄油、茶油、桐油的甲酯得率均在90%以上,均高于单一脂肪酶催化的甲酯得率。这表明复合酶工艺具有对广泛油源底物的适应性。因此,复合酶催化制备生物柴油工艺具有很大的潜在实用价值。
Increasing attention is being paid to biodiesel as a friendly environmental, non-toxic and renewable diesel fuel. In this study, the biodiesel production with compound lipases has been investigated.
Synergistic effect was found in biodiesel production catalyzed by free compound lipase, and was validated by a series of free lipases. The results showed that the transesterification efficiency of compound lipases in biodiesel production was higher than that of single lipase with high region-selectivity. This synergistic effect is especially obvious among lipases with the similiar transesterification efficiency yet different region-selectivity.
The biodiesel production with compound immobilized lipases was investigated. The transesterification efficiencies between compound lipases and single lipase in solvent-free system and tert-butanol system were compared, and the compound lipases were found to have higher efficiency. And the ratios of lipases in the compound lipases were optimized.
In solvent-free system, when Novozym435 was mixed with Lipozyme TLIM and Lipozyme RMIM both at the ratio of 70/30, the methyl ester yields were higher than that of only with Novozym435 by 9.52% and 9.99% respectively; while in tert-butanol system, when Novozym435 was mixed with Lipozyme TLIM and Lipozyme RMIM at the ratio of 60/40 and 80/20, the methyl ester yields were higher than that of only with Novozym435 by 9.89% and 7.48% respectively. Compound lipases exhibited synergistic effect in isolvent-free system and tert-butanol system.
The reaction condition in tert-butanol system was optimized. The results showed when the optimal conditions for transesterification were 40% solvent quantity, temperature 40℃, 4:1methanol/oil molar ratio, Novozym435 1.35%+Lipozyme TLIM 1.80% lipase, 200rpm, methyl esters yield of 92% was obtained after 24h reaction, and the compound lipases were continuously used for 8 cycles, methyl esters yield were always over 90%, which suggested there was no obvious loss of their activity.
Compound lipases were used to catalyze more oil substrates. The methyl ester yields was all higher than 90%, much higher than that of single lipase in soybean oil, rapseed oil, cottonseed oil, castor oil, olive oil, tea oil and tung oil. The results showed that compound lipases have great potentiality to apply for various oils to make biodiesel.
从游离脂肪酶催化制备生物柴油的实验中发现多种酶混合催化制备生物柴油过程中存在协同现象,并利用多种游离脂肪酶对这一现象进行了验证,确认复合脂肪酶比单一脂肪酶对催化酯交换反应有更为显著的促进作用,复合脂肪酶能有效地克服单一脂肪酶催化的位置专一性,提高生物柴油的转酯效率,特别是在转酯能力相当而位置特异性不同的脂肪酶之间表现出更强的协同效应。
进行了复合酶催化制备生物柴油工艺的初步研究,分别进行了无溶剂体系与叔丁醇体系中复合固定化脂肪酶与单一酶转酯效率的对比试验。在无溶剂体系中,Novozym435分别与Lipozyme TLIM和Lipozyme RMIM以70/30比例混合时,甲酯得率分别达到94.52%和96.25%,比Novozym435单独催化时的甲酯得率分别提高了9.52%和9.99%;在叔丁醇体系中,当Novozym435与Lipozyme TLIM和Lipozyme RMIM分别以60/40和80/20的比例混合时,其甲酯得率分别为85.06%和81.5%,比Novozym435单独催化的效率分别提高了9.89%和7.48%。两种体系均表现出较强的协同效果。
通过对叔丁醇体系中Novozym435与Lipozyme TLIM复合酶催化条件进行优化表明:在溶剂添加量为40%、醇油摩尔比4:1、复合酶添加量为Novozym435 1.35%+Lipozyme TLIM 1.80%,温度为40℃,摇床转速为200rpm,反应24小时,甲酯得率达到92%,复合酶在该工艺下连续使用八次时甲酯得率仍保持在90%以上。
进行了复合酶工艺扩大油源底物的研究,复合酶在叔丁醇体系中催化大豆油、菜籽油、棉籽油、蓖麻油、橄榄油、茶油、桐油的甲酯得率均在90%以上,均高于单一脂肪酶催化的甲酯得率。这表明复合酶工艺具有对广泛油源底物的适应性。因此,复合酶催化制备生物柴油工艺具有很大的潜在实用价值。
Increasing attention is being paid to biodiesel as a friendly environmental, non-toxic and renewable diesel fuel. In this study, the biodiesel production with compound lipases has been investigated.
Synergistic effect was found in biodiesel production catalyzed by free compound lipase, and was validated by a series of free lipases. The results showed that the transesterification efficiency of compound lipases in biodiesel production was higher than that of single lipase with high region-selectivity. This synergistic effect is especially obvious among lipases with the similiar transesterification efficiency yet different region-selectivity.
The biodiesel production with compound immobilized lipases was investigated. The transesterification efficiencies between compound lipases and single lipase in solvent-free system and tert-butanol system were compared, and the compound lipases were found to have higher efficiency. And the ratios of lipases in the compound lipases were optimized.
In solvent-free system, when Novozym435 was mixed with Lipozyme TLIM and Lipozyme RMIM both at the ratio of 70/30, the methyl ester yields were higher than that of only with Novozym435 by 9.52% and 9.99% respectively; while in tert-butanol system, when Novozym435 was mixed with Lipozyme TLIM and Lipozyme RMIM at the ratio of 60/40 and 80/20, the methyl ester yields were higher than that of only with Novozym435 by 9.89% and 7.48% respectively. Compound lipases exhibited synergistic effect in isolvent-free system and tert-butanol system.
The reaction condition in tert-butanol system was optimized. The results showed when the optimal conditions for transesterification were 40% solvent quantity, temperature 40℃, 4:1methanol/oil molar ratio, Novozym435 1.35%+Lipozyme TLIM 1.80% lipase, 200rpm, methyl esters yield of 92% was obtained after 24h reaction, and the compound lipases were continuously used for 8 cycles, methyl esters yield were always over 90%, which suggested there was no obvious loss of their activity.
Compound lipases were used to catalyze more oil substrates. The methyl ester yields was all higher than 90%, much higher than that of single lipase in soybean oil, rapseed oil, cottonseed oil, castor oil, olive oil, tea oil and tung oil. The results showed that compound lipases have great potentiality to apply for various oils to make biodiesel.
引文
[1] Jim R.Sustainable production systems for bioenergy: Forest energy in practice[J]. Biomass and Bioenergy, 2006, (30): 999~1000
[2]贾振航.绿色的生物质能[J].节能与环保, 2004, (5): 53~54
[3] Ma F, Milford A.H. Biodiesel production: a review[J]. Bioresource Technology, 1999, (70): 1~15
[4] Krawczyk T. Biodiesel-Alternative fuel makes inroads but hurdles remain[J]. Information, 1996, (7): 801~829
[5] Kevin J.H. Chemical and physical properties of vegetable oil esters and their effect on diesel fuel performance[J]. Biomass, 1986(9): 1~17
[6] Jose M. E, Juan F G, Eduardo S. Preparation and properties of biodiesel from Canary[J].Carbuncles Cheese,1999,(38): 93~95
[7]袁银南,张恬,梅德清等.柴油机燃用生物柴油燃烧与排放[J].江苏大学学报(自然科学版), 2006, 27(3) : 216~219
[8]左青.生物柴油的应用前景[J].粮油食品科技, 2006, 14(5): 22~24
[9]李玉芹,曾虹燕.生物柴油生产应用现状[J].四川化工, 2005, 8(4) : 23~27
[10]梅德清,孙平,袁银南等.柴油机燃用生物柴油的排放特性研究[J].内燃机学报, 2006, 24(4) : 331~335
[11] Heinrich P. High biodiesel quality required by European standards [J]. European Journal of Lipid Science and Technology, 2002, 104: 371~375
[12]吕鹏梅,袁振宏,廖翠萍等.生物柴油标准分析与制定研究[J].现代化工,2006,26(12): 8~14
[13]罗文,袁振宏,廖翠萍.生物柴油标准及质量评价[J].可再生能源,2006,4(128): 33~37
[14]姜南,张正.生物柴油的现状与发展前景[J].世界农业, 2005, 3(311) : 50~52
[15]罗永宗.扩大储备食油部分替代石油储备,一储二用,具有重大经济意义和战略意义[J].中国油脂, 2006, 31(10) : 18~20
[16] Demirbas A. Progress and recent trends in biofuels[J].Progress in Energy and Combustion Science, 2007,(33): 1~18
[17] Schaeffer R, Szklo A S, Cima F M, et al. Indicators for sustainable energy development: Brazil’s case study[J]. Natural Resources Forum, 2005, (29) : 284～297
[18]沈珺珺,迟晓元,杨庆利等.生物柴油的研究进展[J].中国生物工程杂志, 2006, 26(11): 87~90
[19]孙纯,刘金迪.国内外生物柴油的开发应用现状[J].中外能源, 2006, 11(3): 97~102
[20]郭小熹.国际生物柴油发展状况[J].特写, 2006, (1): 96~97
[21]孙永祥.国外可再生能源发展现状评析[J].天然气经济, 2006, (2) : 17~19
[22] Shay E G. Diesel fuel from vegetable oils: status and opportunities[J]. Biomass Bioenergy, 1993(4) : 227~242
[23] Saka S, Kusdiana D. Biodiesel fuel from rapeseed oil as prepared in supercritical methanol[J]. Fuel, 2001, (80): 225~231
[24] Komers K, Stloukal R, Machek J, et al. Biodiesel from rapeseed oil, methanol and KOH: analysis of composition of actual reaction mixture[J]. European Journal of Lipid Science and Technology, 2001, (103): 363~371
[25] Ayhan D. Biodiesel fuels from vegetable oils via catalytic and non-catalytic supercritical alcohol transesterifications and other methods: a survey[J]. Energy Conversion and Management ,2003, (44): 2093~2109
[26] Kranczyk T. Biodiesel[J]. Inform, 1996,7 (8): 801~803
[27]王一平,翟怡,张金利等.生物柴油制备方法研究进展[J].化工进展, 2003, 22(1): 8~22
[28] Recep A, Selim C, Huseyin S Y.The potential of using vegetable oil fuels as fuel for dieselingines[J].Energy Conversion and Management, 2001, 42: 529~538
[29]何灼成,魏小平,朱瑞国.生物柴油的制备方法及研究进展[J].润滑油与燃料,2006, 16(77) : 7~10
[30] Ziejewski M, Kaufman K R, Schwab A W, et a1. Diesel engine evalution of a nonionic sunflower oil-aqueous ethanol microemulsion[J]. Journal of American Oil Chemical Society, 1984, 61: 1620~l626
[31] Schwab A W, Dykstra G J, Selke E, et al. Diesel fuel from thermal decomposition of soybean oil[J]. Journal of American Oil Chemical Society, 1988, (65) : 1781~ 1786
[32] Crabble E, Nolasco C H, Kobayashi G, et al. Biodiesel production from crude palm oil and evaluation of butanol extanol extraction and fuel properties[J]. Process Biochemistry, 2001, 37(1): 65~71
[33]鞠庆华,曾昌凤,郭卫军.酯交换法制备生物柴油的研究进展[J].化工进展, 2004, 23(10): 1053~1057
[34]盛梅,李为民,邬国英.生物柴油研究进展[J].中国油脂, 2003, 28(4) : 66~70
[35]姜绍通,李岩,邵平.酯交换法生产生物柴油的研究进展[J].农产品加工, 2006, 10 (79): 5~8
[36] Jon V G. Biodiesel processing and production[J].Fuel Processing Technology, 2005,(86): 1097~1107
[37]王志华,孙小嫚,孙桂芳等.碱催化法菜籽油制备生物柴油实验研究[J].现代化工, 2006, 26: 124~126
[38] Alcantara R, Amores J, Canoira L, et al. Catalytic production of biodiesel from soy-bean oil, used frying oil and tallow[J].Biomass and Bioenergy, 2000, 18: 515~527
[39] Ma F, Biodiesel fuel: The transesterification of beef tallow. PhD dissertation, University of Nebraska, Lincoln, 1998
[40] Schuchardt U, Vargas R M, Gelbard G. Alkylguanidines as catalysts for the transesterification of rapeseed oil[J]. Journal of Molecular Catalysis A: Chemical, 1995, 99(2): 65~70
[41] Schuchardt U, Sercheli R, Vargas R M[J]. Journal of Brazil Chemical Society, 1998,9(l): 199~210
[42] Iso M, Chen B, Eguchi M, et al. Production of biodiesel fuel from triglycerides and alcohol using immobilized lipase[J]. Journal of Molecular Catalysis B: Enzymatic, 2001, 16(1): 53~58
[43] Hsu A F, Kerby C J, Thomas A F, et al. Transesterification activity of lipases immobilized in a phyllosilicate sol-gel matrix[J]. Biotechnology Letters, 2004, (26): 917~921
[44] Noureddini H, Gao X, Philkana R.S. Immobilized Pseudomonas cepacia lipase for biodiesel fuel production from soybean oil[J].Bioresource Technology, 2005, (96): 769~777
[45]聂开立,王芳,谭天伟.固定酶法生产生物柴油[J].现代化工, 2005, (9) : 35~38
[46] Yuan Y X, Wei D, Jing Z, et al. Conversion of Soybean Oil to Biodiesel Fuel Using Lipozyme TLIM in a Solvent-free Medium[J]. Biocatalysis and Biotransformation, 2004, 22(1): 45~48
[47] Li L L, Wei D, De H L, et al. Lipase-catalyzed transesterification of rapeseed oils for biodiesel production with a novel organic solvent as the reaction medium[J]. Journal of Molecular Catalysis B: Enzymatic: 2006, (43): 58~62
[48] Matsumoto T, Takahashi S, Kaieda M, et al. Yeast whole-cell biocatalyst constructed by intracellular overproduction of Rhizopus oryzae lipase is applicable to biodiesel fuel production[J]. Microbiology and Biotechnology, 2001, (57): 515~520
[49] Ban K, Kaieda M, Matsumoto T, et al. Whole cell biocatalyst for biodiesel fuel production utilizing Rhizopus oryzae cells immobilized within biomass support particles[J]. Biochemical Engineering Journal, 2001, (8): 39~43
[50] Ban K, Hama S, Nishizuka K, et al. Repeated use of whole-cell biocatalysts immobilized within biomass support particles for biodiesel fuel production[J]. Journal of Molecular Catalysis B: Enzymatic, 2002 (17): 157~165
[51] Hama S, Yamaji H, Kaieda M, et al. Effect of fatty acid membrane composition onwhole-cell biocatalysts for biodiesel-fuel production[J]. Journal of Biochemical Engineering, 2004, (21): 155~160
[52] Hama S, Yamaji H, Fukumizu T, et al. Biodiesel-fuel production in a packed-bed reactor using lipase-producing Rhizopus oryzae cells immobilized within biomass support particles[J]. Biochemical Engineering Journal, 2007, in press
[53] Wei L, Wei D, Liu D H. Optimization of whole cellcatalyzed methanolysis of soybean oil for biodiesel production using response surface methodology[J]. Journal of Molecular Catalysis B: Enzymatic, 2007, in press
[54] Jing Z, Wei D, Xin Y L, et al. Study on the effect of cultivation parameters and pretreatment on Rhizopus oryzae cell-catalyzed transesterification of vegetable oils for biodiesel production[J]. Journal of Molecular Catalysis B: Enzymatic, 2006, (43): 15~18
[55]曾静,杜伟,徐圆圆等.霉菌R.oryzae IFO催化大豆油脂合成生物柴油的研究[J].现代化工, 2005(7) : 228~230
[56] Saka S, Kusdiana D. Biodiesel fuel from rapeseed oil as prepared in supercritical methanol[J].Fuel, 2001, 80(2): 225~231
[57] Warabi Y, Kusdiana D, Saka S. Reactivity of triglycerides and fatty acids of rapeseed oil in supercritical alcohols[J]. Bioresource Technology, 2004, 91(3): 283~287
[58] Kusdiana D, Saka S. Effects of water on biodiesel fuel production by supercritical methanol treatment[J]. Bioresource Technology, 2004, 91 (3): 289~295
[59]吕亮,吾国强,汪金良.新型固体碱催化剂在油脂酯交换反应中的应用[J].皮革化工, 2001, 18(3): 37~40
[60] Gryglewicz S. Rapeseed oil methyl esters preparation using heterogeneous catalysts[J]. Bioresource Technology, 1999, 70 (3): 249~253
[61] Suppes G J, Dasari M A, Doskocil E J. Transesterification of soybean oil with Zeolite and metal catalysts[J]. Applied Catalysis A: General, 2004,(257): 213-223
[62]韦德纳,甘斯温特.用醇解方法使生物来源的脂肪酸和油进行酯交换的工艺[P].CN 1327472, 2001
[63] Schuchardt U, Vargas R M, Gelbard G. Transesterification of soybean oil catalyzed alkyguanidines heterogenized on different substituted polystyrenes [J]. Journal of Molecular Catalysis A: Chemical, 1996, 109(1): 37~44
[64]武海棠,周玉杰,张建安等.动植物油脂制备生物柴油的研究[J].科技导报, 2006, 24(9) : 54~58
[65] Stavarache C, Vinatoru M, Nishimura R, et al. Fatty acids methyl esters from vegetable oil by means of ultrasonic energy[J]. Ultrasonics Sonochemistry, 2005, (12): 367~372
[66] Hernando J, Leton P, Matia M P, et al. Biodiesel and fame synthesis assisted by microwaves[J]. Fuel, 2006, in press
[67]李香春,甄宗园.脂肪酶特性及其应用[J].粮食与油脂, 2003, (3) : 19~20
[68]唐平,雕鸿荪.用于油脂改性的1, 3-定向脂肪酶[J].中国油脂, 1996, 21(4) : 4~9
[69]王萍.脂肪酶专一性在脂质生物转化中应用前景[J].粮食与油脂, 1998, (2) : 21~23
[70] Wang L, Du W, Liu D H, et al. Lipase-catalyzed biodiesel production from soybean oil deodorizer distillate with absorbent present in tert-butanol system[J]. Journal of Molecular Catalysis B: Enzymatic, 2006, in press
[71]邓利,谭天伟,王芳.脂肪酶催化合成生物柴油的研究[J].生物工程学报, 2003, 19(1): 97~101
[72]徐圆圆,杜伟,刘德华.非水相脂肪酶催化大豆油脂合成生物柴油的研究[J].现代化工, 2003, 23: 167~169
[73] Kaieda M, Samukaw T, Matsumoto T, et al. Biodiesel fuel production from plant oil catalyzed by Rhizopus oryzae lipase in a water-containing system without organic solvent[J]. Journal of Bioscience and Bioengineering, 1999, 88: 627~631
[74] Wei D, Yuan Y X, De H L, et al. Study on acyl migration in immobilized lipozyme TL-catalyzed transesterification of soybean oil for biodiesel production[J]. Journal of Molecular Catalysis B: Enzymatic, 2005, 37: 68~71
[75]赵宗保,华艳艳,刘波.中国如何突破生物柴油产业的原料瓶颈[J].中国生物工程杂志, 2005, 25(11) : 1~6
[76]徐宝财,郑福平.日用化学品与原材料分析手册[M].北京:化学工业出版社, 2002
[77]邓张双,王宏勋,杜娟.GLA高产菌株的筛选及菌丝体脂肪酸组成的气相色谱质谱分析[J].河南工业大学学报(自然科学版), 2005, 26(1) : 81~87
[78]鲁明波,余龙江,薛勇等.生物柴油的酶促合成研究[J].中国油脂, 2005, 30(11) : 62~64
[79]寇登民,吕宪禹.气相毛细管色谱柱的进展[J].现代仪器使用与维修, 1998, (2) : 4~8
[80] Ma F, Clements L D, Hanna M A. The effect of catalyst, free fatty acids and water on transesterification of beef tallow[J]. Transactions of the ASAE, 1998, 41: 1261~1264
[81] Y.H.Hui.油脂化学与工艺学第一卷[M].第一版.徐生庚,裘爱泳.中国轻工业出版社: 2001.26~27
[82]王瑞元.中国油脂工业与大豆产业[J].中国食物与营养, 2006, (11) : 4~7
[83]顾琪萍,勇强,叶汉玲等.纤维素酶与木聚糖酶混合用于脱墨的研究[J].林产化学与工业, 2003, 23(3): 19~22
[84] Wei D, XU Y Y, Zeng J, et al. Novozym 435-catalysed transesterification of crude soybean oils for biodiesel production in a solvent-free medium[J]. Applied Biochemistry and Biotechnology, 2004, 40: 187~190
[85] Wei D, XU Y Y, Liu D H. Lipase-catalysed transesterification of soya bean oil for biodiesel production during continuous batch operation[J]. Application of Biochemistry and Biotechnology, 2003, 38: 103~106
[86] Mohamed M. S, Uwe T. B. Improvement in lipase-catalyzed synthesis of fatty acid methyl esters from sunflower oil[J]. Enzyme and Microbial Technology, 2003, (33) : 97~103
[87] Royon D, Daz M, Ellenrieder G, et al. Enzymatic production of biodiesel from cotton seed oil using t-butanol as a solvent[J]. Bioresource Technology, 2007, 9: 648~653
[88] Dossat V, Combes D, Marty A. Continuous enzymatic transesterification of high oleic sunflower oil in a packed bed reactor: influence of the glycerol production[J]. Enzyme and Microbial Technology, 1999, (25): 194~200
[89] Chen J W, Wu W T. Regeneration of immobilized Candida Antarctica lipase for transesterification[J]. Journal of Bioscience and Bioengineering, 2003,95(5) : 466~469
[90] Shimada Y, Watanabe Y, Sugihara A, et al. Enzymatic alcoholysis for biodiesel fuel production and application of the reaction to oil processing[J]. Journal of Molecular Catalysis B: Enzymatic, 2002, (17): 133~142
[91]李俐林,杜伟,刘德华等.新型反应介质中脂肪酶催化多种油脂制备生物柴油[J].过程工程学报, 2006, 6(5) : 799~803
[92]盛梅,郭登峰.固定化酶催化菜籽油合成生物柴油稳定性研究[J].中国油脂, 2005, 30(5) : 68~70
[2]贾振航.绿色的生物质能[J].节能与环保, 2004, (5): 53~54
[3] Ma F, Milford A.H. Biodiesel production: a review[J]. Bioresource Technology, 1999, (70): 1~15
[4] Krawczyk T. Biodiesel-Alternative fuel makes inroads but hurdles remain[J]. Information, 1996, (7): 801~829
[5] Kevin J.H. Chemical and physical properties of vegetable oil esters and their effect on diesel fuel performance[J]. Biomass, 1986(9): 1~17
[6] Jose M. E, Juan F G, Eduardo S. Preparation and properties of biodiesel from Canary[J].Carbuncles Cheese,1999,(38): 93~95
[7]袁银南,张恬,梅德清等.柴油机燃用生物柴油燃烧与排放[J].江苏大学学报(自然科学版), 2006, 27(3) : 216~219
[8]左青.生物柴油的应用前景[J].粮油食品科技, 2006, 14(5): 22~24
[9]李玉芹,曾虹燕.生物柴油生产应用现状[J].四川化工, 2005, 8(4) : 23~27
[10]梅德清,孙平,袁银南等.柴油机燃用生物柴油的排放特性研究[J].内燃机学报, 2006, 24(4) : 331~335
[11] Heinrich P. High biodiesel quality required by European standards [J]. European Journal of Lipid Science and Technology, 2002, 104: 371~375
[12]吕鹏梅,袁振宏,廖翠萍等.生物柴油标准分析与制定研究[J].现代化工,2006,26(12): 8~14
[13]罗文,袁振宏,廖翠萍.生物柴油标准及质量评价[J].可再生能源,2006,4(128): 33~37
[14]姜南,张正.生物柴油的现状与发展前景[J].世界农业, 2005, 3(311) : 50~52
[15]罗永宗.扩大储备食油部分替代石油储备,一储二用,具有重大经济意义和战略意义[J].中国油脂, 2006, 31(10) : 18~20
[16] Demirbas A. Progress and recent trends in biofuels[J].Progress in Energy and Combustion Science, 2007,(33): 1~18
[17] Schaeffer R, Szklo A S, Cima F M, et al. Indicators for sustainable energy development: Brazil’s case study[J]. Natural Resources Forum, 2005, (29) : 284～297
[18]沈珺珺,迟晓元,杨庆利等.生物柴油的研究进展[J].中国生物工程杂志, 2006, 26(11): 87~90
[19]孙纯,刘金迪.国内外生物柴油的开发应用现状[J].中外能源, 2006, 11(3): 97~102
[20]郭小熹.国际生物柴油发展状况[J].特写, 2006, (1): 96~97
[21]孙永祥.国外可再生能源发展现状评析[J].天然气经济, 2006, (2) : 17~19
[22] Shay E G. Diesel fuel from vegetable oils: status and opportunities[J]. Biomass Bioenergy, 1993(4) : 227~242
[23] Saka S, Kusdiana D. Biodiesel fuel from rapeseed oil as prepared in supercritical methanol[J]. Fuel, 2001, (80): 225~231
[24] Komers K, Stloukal R, Machek J, et al. Biodiesel from rapeseed oil, methanol and KOH: analysis of composition of actual reaction mixture[J]. European Journal of Lipid Science and Technology, 2001, (103): 363~371
[25] Ayhan D. Biodiesel fuels from vegetable oils via catalytic and non-catalytic supercritical alcohol transesterifications and other methods: a survey[J]. Energy Conversion and Management ,2003, (44): 2093~2109
[26] Kranczyk T. Biodiesel[J]. Inform, 1996,7 (8): 801~803
[27]王一平,翟怡,张金利等.生物柴油制备方法研究进展[J].化工进展, 2003, 22(1): 8~22
[28] Recep A, Selim C, Huseyin S Y.The potential of using vegetable oil fuels as fuel for dieselingines[J].Energy Conversion and Management, 2001, 42: 529~538
[29]何灼成,魏小平,朱瑞国.生物柴油的制备方法及研究进展[J].润滑油与燃料,2006, 16(77) : 7~10
[30] Ziejewski M, Kaufman K R, Schwab A W, et a1. Diesel engine evalution of a nonionic sunflower oil-aqueous ethanol microemulsion[J]. Journal of American Oil Chemical Society, 1984, 61: 1620~l626
[31] Schwab A W, Dykstra G J, Selke E, et al. Diesel fuel from thermal decomposition of soybean oil[J]. Journal of American Oil Chemical Society, 1988, (65) : 1781~ 1786
[32] Crabble E, Nolasco C H, Kobayashi G, et al. Biodiesel production from crude palm oil and evaluation of butanol extanol extraction and fuel properties[J]. Process Biochemistry, 2001, 37(1): 65~71
[33]鞠庆华,曾昌凤,郭卫军.酯交换法制备生物柴油的研究进展[J].化工进展, 2004, 23(10): 1053~1057
[34]盛梅,李为民,邬国英.生物柴油研究进展[J].中国油脂, 2003, 28(4) : 66~70
[35]姜绍通,李岩,邵平.酯交换法生产生物柴油的研究进展[J].农产品加工, 2006, 10 (79): 5~8
[36] Jon V G. Biodiesel processing and production[J].Fuel Processing Technology, 2005,(86): 1097~1107
[37]王志华,孙小嫚,孙桂芳等.碱催化法菜籽油制备生物柴油实验研究[J].现代化工, 2006, 26: 124~126
[38] Alcantara R, Amores J, Canoira L, et al. Catalytic production of biodiesel from soy-bean oil, used frying oil and tallow[J].Biomass and Bioenergy, 2000, 18: 515~527
[39] Ma F, Biodiesel fuel: The transesterification of beef tallow. PhD dissertation, University of Nebraska, Lincoln, 1998
[40] Schuchardt U, Vargas R M, Gelbard G. Alkylguanidines as catalysts for the transesterification of rapeseed oil[J]. Journal of Molecular Catalysis A: Chemical, 1995, 99(2): 65~70
[41] Schuchardt U, Sercheli R, Vargas R M[J]. Journal of Brazil Chemical Society, 1998,9(l): 199~210
[42] Iso M, Chen B, Eguchi M, et al. Production of biodiesel fuel from triglycerides and alcohol using immobilized lipase[J]. Journal of Molecular Catalysis B: Enzymatic, 2001, 16(1): 53~58
[43] Hsu A F, Kerby C J, Thomas A F, et al. Transesterification activity of lipases immobilized in a phyllosilicate sol-gel matrix[J]. Biotechnology Letters, 2004, (26): 917~921
[44] Noureddini H, Gao X, Philkana R.S. Immobilized Pseudomonas cepacia lipase for biodiesel fuel production from soybean oil[J].Bioresource Technology, 2005, (96): 769~777
[45]聂开立,王芳,谭天伟.固定酶法生产生物柴油[J].现代化工, 2005, (9) : 35~38
[46] Yuan Y X, Wei D, Jing Z, et al. Conversion of Soybean Oil to Biodiesel Fuel Using Lipozyme TLIM in a Solvent-free Medium[J]. Biocatalysis and Biotransformation, 2004, 22(1): 45~48
[47] Li L L, Wei D, De H L, et al. Lipase-catalyzed transesterification of rapeseed oils for biodiesel production with a novel organic solvent as the reaction medium[J]. Journal of Molecular Catalysis B: Enzymatic: 2006, (43): 58~62
[48] Matsumoto T, Takahashi S, Kaieda M, et al. Yeast whole-cell biocatalyst constructed by intracellular overproduction of Rhizopus oryzae lipase is applicable to biodiesel fuel production[J]. Microbiology and Biotechnology, 2001, (57): 515~520
[49] Ban K, Kaieda M, Matsumoto T, et al. Whole cell biocatalyst for biodiesel fuel production utilizing Rhizopus oryzae cells immobilized within biomass support particles[J]. Biochemical Engineering Journal, 2001, (8): 39~43
[50] Ban K, Hama S, Nishizuka K, et al. Repeated use of whole-cell biocatalysts immobilized within biomass support particles for biodiesel fuel production[J]. Journal of Molecular Catalysis B: Enzymatic, 2002 (17): 157~165
[51] Hama S, Yamaji H, Kaieda M, et al. Effect of fatty acid membrane composition onwhole-cell biocatalysts for biodiesel-fuel production[J]. Journal of Biochemical Engineering, 2004, (21): 155~160
[52] Hama S, Yamaji H, Fukumizu T, et al. Biodiesel-fuel production in a packed-bed reactor using lipase-producing Rhizopus oryzae cells immobilized within biomass support particles[J]. Biochemical Engineering Journal, 2007, in press
[53] Wei L, Wei D, Liu D H. Optimization of whole cellcatalyzed methanolysis of soybean oil for biodiesel production using response surface methodology[J]. Journal of Molecular Catalysis B: Enzymatic, 2007, in press
[54] Jing Z, Wei D, Xin Y L, et al. Study on the effect of cultivation parameters and pretreatment on Rhizopus oryzae cell-catalyzed transesterification of vegetable oils for biodiesel production[J]. Journal of Molecular Catalysis B: Enzymatic, 2006, (43): 15~18
[55]曾静,杜伟,徐圆圆等.霉菌R.oryzae IFO催化大豆油脂合成生物柴油的研究[J].现代化工, 2005(7) : 228~230
[56] Saka S, Kusdiana D. Biodiesel fuel from rapeseed oil as prepared in supercritical methanol[J].Fuel, 2001, 80(2): 225~231
[57] Warabi Y, Kusdiana D, Saka S. Reactivity of triglycerides and fatty acids of rapeseed oil in supercritical alcohols[J]. Bioresource Technology, 2004, 91(3): 283~287
[58] Kusdiana D, Saka S. Effects of water on biodiesel fuel production by supercritical methanol treatment[J]. Bioresource Technology, 2004, 91 (3): 289~295
[59]吕亮,吾国强,汪金良.新型固体碱催化剂在油脂酯交换反应中的应用[J].皮革化工, 2001, 18(3): 37~40
[60] Gryglewicz S. Rapeseed oil methyl esters preparation using heterogeneous catalysts[J]. Bioresource Technology, 1999, 70 (3): 249~253
[61] Suppes G J, Dasari M A, Doskocil E J. Transesterification of soybean oil with Zeolite and metal catalysts[J]. Applied Catalysis A: General, 2004,(257): 213-223
[62]韦德纳,甘斯温特.用醇解方法使生物来源的脂肪酸和油进行酯交换的工艺[P].CN 1327472, 2001
[63] Schuchardt U, Vargas R M, Gelbard G. Transesterification of soybean oil catalyzed alkyguanidines heterogenized on different substituted polystyrenes [J]. Journal of Molecular Catalysis A: Chemical, 1996, 109(1): 37~44
[64]武海棠,周玉杰,张建安等.动植物油脂制备生物柴油的研究[J].科技导报, 2006, 24(9) : 54~58
[65] Stavarache C, Vinatoru M, Nishimura R, et al. Fatty acids methyl esters from vegetable oil by means of ultrasonic energy[J]. Ultrasonics Sonochemistry, 2005, (12): 367~372
[66] Hernando J, Leton P, Matia M P, et al. Biodiesel and fame synthesis assisted by microwaves[J]. Fuel, 2006, in press
[67]李香春,甄宗园.脂肪酶特性及其应用[J].粮食与油脂, 2003, (3) : 19~20
[68]唐平,雕鸿荪.用于油脂改性的1, 3-定向脂肪酶[J].中国油脂, 1996, 21(4) : 4~9
[69]王萍.脂肪酶专一性在脂质生物转化中应用前景[J].粮食与油脂, 1998, (2) : 21~23
[70] Wang L, Du W, Liu D H, et al. Lipase-catalyzed biodiesel production from soybean oil deodorizer distillate with absorbent present in tert-butanol system[J]. Journal of Molecular Catalysis B: Enzymatic, 2006, in press
[71]邓利,谭天伟,王芳.脂肪酶催化合成生物柴油的研究[J].生物工程学报, 2003, 19(1): 97~101
[72]徐圆圆,杜伟,刘德华.非水相脂肪酶催化大豆油脂合成生物柴油的研究[J].现代化工, 2003, 23: 167~169
[73] Kaieda M, Samukaw T, Matsumoto T, et al. Biodiesel fuel production from plant oil catalyzed by Rhizopus oryzae lipase in a water-containing system without organic solvent[J]. Journal of Bioscience and Bioengineering, 1999, 88: 627~631
[74] Wei D, Yuan Y X, De H L, et al. Study on acyl migration in immobilized lipozyme TL-catalyzed transesterification of soybean oil for biodiesel production[J]. Journal of Molecular Catalysis B: Enzymatic, 2005, 37: 68~71
[75]赵宗保,华艳艳,刘波.中国如何突破生物柴油产业的原料瓶颈[J].中国生物工程杂志, 2005, 25(11) : 1~6
[76]徐宝财,郑福平.日用化学品与原材料分析手册[M].北京:化学工业出版社, 2002
[77]邓张双,王宏勋,杜娟.GLA高产菌株的筛选及菌丝体脂肪酸组成的气相色谱质谱分析[J].河南工业大学学报(自然科学版), 2005, 26(1) : 81~87
[78]鲁明波,余龙江,薛勇等.生物柴油的酶促合成研究[J].中国油脂, 2005, 30(11) : 62~64
[79]寇登民,吕宪禹.气相毛细管色谱柱的进展[J].现代仪器使用与维修, 1998, (2) : 4~8
[80] Ma F, Clements L D, Hanna M A. The effect of catalyst, free fatty acids and water on transesterification of beef tallow[J]. Transactions of the ASAE, 1998, 41: 1261~1264
[81] Y.H.Hui.油脂化学与工艺学第一卷[M].第一版.徐生庚,裘爱泳.中国轻工业出版社: 2001.26~27
[82]王瑞元.中国油脂工业与大豆产业[J].中国食物与营养, 2006, (11) : 4~7
[83]顾琪萍,勇强,叶汉玲等.纤维素酶与木聚糖酶混合用于脱墨的研究[J].林产化学与工业, 2003, 23(3): 19~22
[84] Wei D, XU Y Y, Zeng J, et al. Novozym 435-catalysed transesterification of crude soybean oils for biodiesel production in a solvent-free medium[J]. Applied Biochemistry and Biotechnology, 2004, 40: 187~190
[85] Wei D, XU Y Y, Liu D H. Lipase-catalysed transesterification of soya bean oil for biodiesel production during continuous batch operation[J]. Application of Biochemistry and Biotechnology, 2003, 38: 103~106
[86] Mohamed M. S, Uwe T. B. Improvement in lipase-catalyzed synthesis of fatty acid methyl esters from sunflower oil[J]. Enzyme and Microbial Technology, 2003, (33) : 97~103
[87] Royon D, Daz M, Ellenrieder G, et al. Enzymatic production of biodiesel from cotton seed oil using t-butanol as a solvent[J]. Bioresource Technology, 2007, 9: 648~653
[88] Dossat V, Combes D, Marty A. Continuous enzymatic transesterification of high oleic sunflower oil in a packed bed reactor: influence of the glycerol production[J]. Enzyme and Microbial Technology, 1999, (25): 194~200
[89] Chen J W, Wu W T. Regeneration of immobilized Candida Antarctica lipase for transesterification[J]. Journal of Bioscience and Bioengineering, 2003,95(5) : 466~469
[90] Shimada Y, Watanabe Y, Sugihara A, et al. Enzymatic alcoholysis for biodiesel fuel production and application of the reaction to oil processing[J]. Journal of Molecular Catalysis B: Enzymatic, 2002, (17): 133~142
[91]李俐林,杜伟,刘德华等.新型反应介质中脂肪酶催化多种油脂制备生物柴油[J].过程工程学报, 2006, 6(5) : 799~803
[92]盛梅,郭登峰.固定化酶催化菜籽油合成生物柴油稳定性研究[J].中国油脂, 2005, 30(5) : 68~70