Production of biodiesel fuel from soybean oil catalyzed by fungus whole-cell biocatalysts in ionic liquids
Production of biodiesel fuel from soybean oil catalyzed by fungus whole-cell biocatalysts in ionic liquids
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DOI：
10.1016/j.enzmictec.2009.08.008
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发表时间：
2010-01-07
影响因子：
3.4
通讯作者：
Fukuda, Hideki
中科院分区：
文献类型：
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作者：
Arai, Shogo;Nakashima, Kazunori;Fukuda, Hideki
在离子液体存在下，固定在生物质载体颗粒（BSP）上的产脂肪酶丝状真菌作为全细胞生物催化剂，催化大豆油的甲醇分解生产脂肪酸甲酯（ME，即生物柴油燃料）。我们使用了四种类型的全细胞生物催化剂：产生三酰甘油脂肪酶的野生型米根霉（w-ROL）、表达异孢镰孢脂肪酶的重组米曲霉（r-FHL）、南极假丝酵母脂肪酶B（r-CALB）以及来自米曲霉的单酰基甘油和二酰基甘油脂肪酶（r-mdlB）。反应 24 小时后，w-ROL 在离子液体 [Emim][BF4] 或 [Bmim][BF4] 双相体系中产生高产率的脂肪酸甲酯 (ME)。虽然已知在传统系统中过量的甲醇（例如，1.5当量的甲醇对油）会使脂肪酶严重失活，但在离子液体双相系统中，即使甲醇/油的比例为4，甲醇分解也能成功进行，其中离子液体将作为甲醇的储存库以抑制酶失活。当仅使用w-ROL作为甲醇分解的生物催化剂时，由于米根霉脂肪酶的1,3位特异性，未反应的单酸甘油酯仍然存在。通过联合使用两种类型的全细胞生物催化剂 w-ROL 和 r-mdlB，实现了高 ME 转化率。在稳定性测试中，在[Bmim][BF4]中孵育72小时后，w-ROL的活性降低至原始值的三分之一。通过将生物催化剂与戊二醛交联，大大增强了[Bmim][BF4]中w-ROL的稳定性。本研究表明，离子液体是全细胞生物催化剂生产生物柴油燃料中用作第二种溶剂的有前途的候选者。 (C) 2009 Elsevier Inc. 保留所有权利。
The methanolysis of soybean oil to produce a fatty acid methyl ester (ME, i.e., biodiesel fuel) was catalyzed by lipase-producing filamentous fungi immobilized on biomass support particles (BSPs) as a whole-cell biocatalyst in the presence of ionic liquids. We used four types of whole-cell biocatalysts: wildtype Rhizopus oryzae producing triacylglycerol lipase (w-ROL), recombinant Aspergillus oryzae expressing Fusarium heterosporum lipase (r-FHL), Candida antarctica lipase B (r-CALB), and mono- and diacylglycerol lipase from A. oryzae (r-mdlB). w-ROL gave the high yield of fatty acid methyl ester (ME) in ionic liquid [Emim][BF4] or [Bmim][BF4] biphasic systems following a 24h reaction. While lipases are known to be severely deactivated by an excess amount of methanol (e.g. 1.5 Mequiv. of methanol against oil) in a conventional system, methanolysis successfully proceeded even with a methanol/oil ratio of 4 in the ionic liquid biphasic system, where the ionic liquids would work as a reservoir of methanol to suppress the enzyme deactivation. When only w-ROL was used as a biocatalyst for methanolysis, unreacted monoglyceride remained due to the 1,3-positional specificity of R. oryzae lipase. High ME conversion was attained by the combined use of two types of whole-cell biocatalysts, w-ROL and r-mdlB. In a stability test, the activity of w-ROL was reduced to one-third of its original value after incubation in [Bmim][BF4] for 72 h. The stability of w-ROL in [Bmim][BF4] was greatly enhanced by cross-linking the biocatalyst with glutaraldehyde. The present Study demonstrated that ionic liquids are promising candidates for use as the second solvent in biodiesel fuel production by whole-cell biocatalysts. (C) 2009 Elsevier Inc. All rights reserved.