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SBA-15 吸附固定甲羟戊酸途径下游五种酶体外合成异戊二烯

作者:时间:2025-07-04点击数:



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文章编号:1672-6987(2025)03-0027-08;DOI:10. 16351/j. 1672-6987. 2025. 03. 004


何明岩,苏忠亮* ,徐 梅,姬 越,韦 敏 (青岛科技大学 化工学院,山东 青岛 266042)


要:甲羟戊酸激酶(MVK)、磷酸甲羟戊酸激酶(PMK)、二磷酸甲羟戊酸脱羧酶(MVD)、

异戊烯基二磷酸异构酶(IDI)和异戊二烯合成酶(ISPS)为甲羟戊酸(MVA)下游反应途径的 5 种酶。本研究以 SBA-15 作为固定化载体,运用物理吸附方法,分别制备 MVK、PMK、MVD、 IDI 和 ISPS 的固定化单酶;以双酶固定的方法分别制备 MVK 和 PMK(浓度比 1∶1),MVD 和 IDI(浓度比 1∶2)的固定化双酶;将 MVK、PMK、MVD 和 IDI(浓度比 1∶1∶1∶2)固定在 SBA-15 上制备固定化四酶;多酶固定制备 MVK、PMK、MVD、IDI 和 ISPS(浓度比 1∶1∶1∶2∶6)的固定 化五酶。利用游离酶、固定化单酶、固定化双酶、固定化四酶和固定化五酶在相同反应条件下 (40 ℃,pH7. 0)分别反应 48 h 体外催化甲羟戊酸合成异戊二烯,其中固定化五酶异戊二烯产量 为 401 mg · L−1 ,产量最高。用单因素变量法考察游离酶和固定化五酶在不同温度、pH 值下 的相对活性,研究固定化五酶的酶催化性能。结果表明,固定化五酶催化活性高于游离酶,重 复利用 5 次后的相对酶活为 46. 34%,具有较高工业应用价值。


关键词:异戊二烯;甲羟戊酸; SBA-15;酶法

中图分类号:O 180 文献标志码:A


引用格式:何明岩,苏忠亮,徐梅,等 . SBA-15 吸附固定甲羟戊酸途径下游五种酶体外合成 异戊二烯[J]. 青岛科技大学学报(自然科学版),2025,46(3):27-34.


HE Mingyan, SU Zhongliang, XU Mei, et al. In vitro synthesis of isoprene by SBA-15

adsorption and immobilization of five enzymes downstream of MVA[J]. Journal of Qingdao University of Science and Technology(Natural Science Edition),2025,46(3):27-34.


In Vitro Synthesis of Isoprene by SBA-15 Adsorption and Immobilization of Five Enzymes Downstream of MVA


HE Mingyan,SU Zhongliang,XU Mei,JI Yue,WEI Min (College of Chemical Engineering, Qingdao University of Science and Technology, Qingdao 266042, China)


Abstract:Mevalonate kinase (MVK), phosphate mevalonate kinase (PMK), mevalonate

diphosphate decarboxylase (MVD), isopentenyl diphosphate isomerase (IDI) and isoprene synthase (ISPS) are the five enzymes in the downstream pathway of mevalonate (MVA). In this paper, the immobilized single enzyme of MVK, PMK, MVD, IDI and ISPS was prepared by physical adsorption method using SBA-15 as immobilized carrier. The immobilized double enzymes of MVK and PMK (concentration ratio 1∶1), MVD and IDI (concentration ratio 1∶2) were prepared by double enzyme immobilization method. MVK, PMK, MVD and IDI (con centration ratio 1∶1) were immobilized on SBA-15 to prepare immobilized quadruenzyme. The immobilized five enzymes of MVK, PMK, MVD, IDI and ISPS (1∶1∶1∶2∶6) were prepared by multi-enzyme immobilization. Free enzyme, immobilized single enzyme, immobilized double enzyme, immobilized four enzyme and immobilized five enzyme were used to catalyze MVA to synthesize isoprene in vitro under the same reaction conditions (40 ℃ , pH7. 0).

Among them, the yield of immobilized five enzyme isoprene was 401 mg· L−1 , which was the highest. The relative activities of free enzyme and immobilized five enzymes at different tem peratures and pH values were investigated by single factor variable method, and the enzyme catalytic performance of immobilized five enzymes was studied. The results showed that the catalytic activity of immobilized five enzymes was higher than that of free enzymes, and the relative enzyme activity was 46. 34% after 5 times of reuse, which had high industrial applica tion value.


Key words:isoprene; MVA; SBA-15; enzymatic method


收稿日期:2024-07-22

基金项目:国家科技攻关计划项目(2022YFC2104701).

作者简介:何明岩(1997—),男,硕士研究生 . 通信联系人 .


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