丝瓜络固定化米根霉催化光皮树油制备生物柴油

doi:10.16085/j.issn.1000-6613.2021-2549

摘要/Abstract

摘要：

近年来，全细胞生物催化剂应用于生物柴油生产的研究引起了人们的极大关注，全细胞催化剂的制备过程可省去烦琐的酶纯化过程和降低催化剂的制备成本，且全细胞催化剂使用寿命较长。本研究对一株高产脂肪酶的米根霉进行了固定化研究，并将其应用于生物柴油的制备。首先筛选不同的生物基固定化材料，探索其适宜的固定化条件，以获得的固定化米根霉作为全细胞催化剂催化光皮树油制备生物柴油，探讨了转酯化条件对生物柴油得率的影响。研究结果表明，丝瓜络作为固定化材料的固载率最高，以橄榄油作为碳源，多聚蛋白胨和NaNO₃作为复合氮源，获得的固定化细胞的固载率及脂肪酶活性最强。以丝瓜络固定化米根霉催化光皮树油，在含水量为10%、催化剂用量为12%的反应体系中，总醇油摩尔比为4∶1，甲醇分别在0h、10h、24h、40h以1∶1添加，甲酯得率可达94%以上。固定化全细胞重复使用6次后，转酯化效果依然保持在80%以上。

关键词: 全细胞催化剂, 米根霉, 丝瓜络, 生物柴油, 光皮树油

Abstract:

In recent years, the application of whole-cell biocatalyst in biodiesel production has attracted great attention. Whole-cell catalyst avoids the cumbersome enzyme purification process and saves the preparation cost of the catalyst. Moreover, whole-cell biocatalyst has a long service life. In this study, a strain of Rhizopus oryzae with high lipase production was immobilized and applied. Different immobilization materials were selected and the suitable immobilization conditions were explored. The immobilized Rhizopus oryzae as the whole-cell catalyst was used to catalyze the preparation of biodiesel from Comus wilsoniana fruit oil. The effect of esterification conditions on the yield of biodiesel was discussed. The results showed that the immobilization rate and activity of the immobilized cells were the highest with the loofah sponge as the immobilization material, olive oil as carbon source,and peptone and NaNO₃ as compound nitrogen source. In the reaction system of 10% water content and 12% catalyst dosage, the molar ratio of total alcohol to oil was 4∶1, methanol was added 1∶1 at 0h, 10h, 24h, 40h, respectively, and the yield of methyl ester was more than 94%. After the immobilized whole-cells were reused for 6 times, the transesterification acticity of immobilized cells remained above 80%.

Key words: whole-cell catalyst, Rhizopus oryzae, loofah sponge, biodiesel, Comus wilsoniana fruit oil

中图分类号:

TQ426.97

纪淑兰, 李迅, 王飞. 丝瓜络固定化米根霉催化光皮树油制备生物柴油[J]. 化工进展, 2022, 41(10): 5381-5389.

JI Shulan, LI Xun, WANG Fei. Immobilization of Rhizopus oryzae onto loofah sponge as a whole-cell biocatalyst to preparation of biodiesel from Comus wilsoniana fruit oil[J]. Chemical Industry and Engineering Progress, 2022, 41(10): 5381-5389.

图/表 13

图1 固定化米根霉催化光皮树油的产物气相色谱图1—棕榈酸甲酯；2—硬脂酸甲酯；3—油酸甲酯；4—亚油酸甲酯；5—亚麻酸甲酯

图2 不同固定化材料对米根霉固定化的影响[固定化培养条件：28℃，180r/min，基础培养基。甲酯化反应条件：35℃，180r/min，pH6.8，质量分数10%的缓冲溶液（占油重），质量分数10%的全细胞催化剂（占油重），醇油比为4∶1]

图3 丝瓜络添加时间对细胞固定化的影响[固定化培养条件：28℃，180r/min，基础培养基。甲酯化反应条件：35℃，180r/min，pH6.8，质量分数10%的缓冲溶液（占油重），质量分数10%的全细胞催化剂（占油重），醇油比为4∶1]

图4 丝瓜络添加时间对细胞固定化的影响[固定化培养条件：28℃，180r/min，基础培养基。甲酯化反应条件：35℃，180r/min，pH6.8，质量分数10%的缓冲溶液（占油重），质量分数10%的全细胞催化剂（占油重），醇油比为4∶1]

图5 碳源对米根霉固载率的影响

图6 碳源对米根霉全细胞催化剂催化光皮树油制备生物柴油的影响[反应条件：35℃，180r/min，pH6.8，质量分数10%的缓冲溶液（占油重），质量分数10%的全细胞催化剂（占油重），醇油比为4∶1]

图7 氮源对米根霉固载率的影响（有机氮源为有机氮与0.1％ NaNO3复合）

图8 氮源对米根霉全细胞催化剂催化光皮树油制备生物柴油的影响[反应条件：35℃，180r/min，pH6.8，质量分数10%的缓冲溶液（占油重），质量分数10%的全细胞催化剂（占油重），醇油比为4∶1]

图9 （未）经不同交联剂交联处理的全细胞催化剂的重复使用次数[单批反应条件：35℃，180r/min，pH6.8，质量分数10%的缓冲溶液（占油重），质量分数10%的全细胞催化剂（占油重），醇油摩尔比为4∶1]

图10 温度对全细胞催化光皮树油甲醇分解的影响[反应条件：不同温度，180r/min，质量分数10%的缓冲溶液（占油重），质量分数10%的全细胞催化剂（占油重），醇油摩尔比为4∶1]

图11 含水量对全细胞催化光皮树油甲醇分解的影响[反应条件：35℃，180r/min，质量分数10%的缓冲溶液（占油重），醇油摩尔比为4∶1]

图12 催化剂量对全细胞催化光皮树油甲醇分解的影响[反应条件：35℃，180r/min，质量分数10%的缓冲溶液（占油重），醇油摩尔比为4∶1]

图13 甲醇添加时间对全细胞催化剂催化光皮树油甲酯得率的影响[反应条件：35℃，180r/min，质量分数10%的缓冲溶液（占油重），质量分数12%的全细胞催化剂（占油重），醇油摩尔比为4∶1]

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