JOURNAL OF LIGHT INDUSTRY

CN 41-1437/TS  ISSN 2096-1553

固定化卤醇脱卤酶的制备及催化合成ATS-5的稳定性考察

梁国斌 ,  翟良创 ,  汪斌 ,  姚佳 ,  周全法

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梁国斌, 翟良创, 汪斌, 等. 固定化卤醇脱卤酶的制备及催化合成ATS-5的稳定性考察[J]. 轻工学报, 2017, 32(2): 26-32. doi: 10.3969/j.issn.2096-1553.2017.2.005
引用本文: 梁国斌, 翟良创, 汪斌, 等. 固定化卤醇脱卤酶的制备及催化合成ATS-5的稳定性考察[J]. 轻工学报, 2017, 32(2): 26-32. doi: 10.3969/j.issn.2096-1553.2017.2.005
shu
LIANG Guo-bin, ZHAI Liang-chuang, WANG Bin, et al. Preparation of immobilized halohydrin dehalogenase and the stability testing of catalytic systhesis ATS-5[J]. Journal of Light Industry, 2017, 32(2): 26-32. doi: 10.3969/j.issn.2096-1553.2017.2.005
Citation: LIANG Guo-bin, ZHAI Liang-chuang, WANG Bin, et al. Preparation of immobilized halohydrin dehalogenase and the stability testing of catalytic systhesis ATS-5[J]. Journal of Light Industry, 2017, 32(2): 26-32. doi: 10.3969/j.issn.2096-1553.2017.2.005
shu

固定化卤醇脱卤酶的制备及催化合成ATS-5的稳定性考察

  • 江苏理工学院 化学与环境工程学院, 江苏 常州 213001

  • 基金项目: 江苏省产学研联合创新基金项目（BY2015028-04）

  • 中图分类号: Q814.2

Preparation of immobilized halohydrin dehalogenase and the stability testing of catalytic systhesis ATS-5

  • School of Chemical and Environmental Engineering, Jiangsu University of Technology, Changzhou 213001, China

  • Received Date: 2016-06-07
    Available Online: 2017-03-15

    CLC number: Q814.2

  • 摘要: 研究了海藻酸钠固定化卤醇脱卤酶（HHDH）的制备条件，确定其最佳工艺条件为：海藻酸钠浓度3.5%，HHDH给酶量2.4×104 U/mL，戊二醛浓度1.00%，氯化钙浓度1.5%和固化时间2 h.在此条件下，HHDH包埋率为52.1%，活力回收率为75.2%.固定化HHDH间歇催化合成（R）-4-氰基-3-羟基丁酸乙酯（ATS-5），经5批次反应后，相对活力为51.2%；固定化HHDH连续催化合成ATS-5，在10 d后底物转化率为90.6%，ATS-5精馏提纯收率为98.2%，纯度达到99.3%，光学纯度达到99.1%.固定化HHDH在4℃下储存80 d后无明显酶活力损失.
    Abstract: The immobilization conditions of halohydrin dehalogenase (HHDH) by the sodium alginate were studied.The optimum conditions of immobilization process were as follows:sodium alginate concentration 3.5%, HHDH enzymatic amount 2.4×104 U/mL, glutaraldehyde concentration 1.00%, calcium chloride solution concentration 1.5% and immobilization time 2 h.Under the optimum immobilization conditions, the encapsulation efficiency of HHDH was 52.1%, the activity recovery of HHDH was 75.2%.After five batches of intermittent catalytic synthesis of (R)-4-cyano-3-hydroxybutyric acid ethyl (ATS-5), the residual activity of immobilized HHDH was 51.2%. After the continuous catalytic synthesis of ATS-5 for 10 d,the substrate conversion efficiency was 90.6%, the distillation purification yield of ATS-5 was 98.2%, the purity and optical purity of ATS-5 were 99.3% and 99.1%, respectively.No obvious activity loss was obtained for immobilized HHDH after 80 d storage at 4℃.
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  • 收稿日期:  2016-06-07
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梁国斌, 翟良创, 汪斌, 等. 固定化卤醇脱卤酶的制备及催化合成ATS-5的稳定性考察[J]. 轻工学报, 2017, 32(2): 26-32. doi: 10.3969/j.issn.2096-1553.2017.2.005
引用本文: 梁国斌, 翟良创, 汪斌, 等. 固定化卤醇脱卤酶的制备及催化合成ATS-5的稳定性考察[J]. 轻工学报, 2017, 32(2): 26-32. doi: 10.3969/j.issn.2096-1553.2017.2.005
shu
LIANG Guo-bin, ZHAI Liang-chuang, WANG Bin, et al. Preparation of immobilized halohydrin dehalogenase and the stability testing of catalytic systhesis ATS-5[J]. Journal of Light Industry, 2017, 32(2): 26-32. doi: 10.3969/j.issn.2096-1553.2017.2.005
Citation: LIANG Guo-bin, ZHAI Liang-chuang, WANG Bin, et al. Preparation of immobilized halohydrin dehalogenase and the stability testing of catalytic systhesis ATS-5[J]. Journal of Light Industry, 2017, 32(2): 26-32. doi: 10.3969/j.issn.2096-1553.2017.2.005
shu

固定化卤醇脱卤酶的制备及催化合成ATS-5的稳定性考察

  • 江苏理工学院 化学与环境工程学院, 江苏 常州 213001
  • 收稿日期: 2016-06-07
  • 网络出版日期: 2017-03-15
基金项目:  江苏省产学研联合创新基金项目（BY2015028-04）

摘要: 研究了海藻酸钠固定化卤醇脱卤酶（HHDH）的制备条件，确定其最佳工艺条件为：海藻酸钠浓度3.5%，HHDH给酶量2.4×104 U/mL，戊二醛浓度1.00%，氯化钙浓度1.5%和固化时间2 h.在此条件下，HHDH包埋率为52.1%，活力回收率为75.2%.固定化HHDH间歇催化合成（R）-4-氰基-3-羟基丁酸乙酯（ATS-5），经5批次反应后，相对活力为51.2%；固定化HHDH连续催化合成ATS-5，在10 d后底物转化率为90.6%，ATS-5精馏提纯收率为98.2%，纯度达到99.3%，光学纯度达到99.1%.固定化HHDH在4℃下储存80 d后无明显酶活力损失.

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