Page 65 - 201808

第 35 卷第 8 期 精 细 化 工 Vol.35, No.8
201 8 年 8 月 FINE CHEMICALS Aug. 2018

生物工程
金黄节杆菌 CYC705 腈水解酶催化

亚氨基二乙酸的合成

2
1
1
1
杨 猛 ，梅娜娜 ，陈璐瑶 ，蔡雯雯 ，苏二正 1,2*
（1. 南京林业大学 轻工与食品学院，江苏 南京 210037；2. 华东理工大学 生物反应器工程国家重点实
验室，上海 200237）
摘要：将金黄节杆菌 CYC705（Arthrobacter aurescens CYC705）腈水解酶用于生物催化合成亚氨基二乙酸（IDA），
从生物催化剂的形式、生物催化反应过程优化和反应体系放大 3 个方面进行了考察。在氨基载体固定化酶、环
氧基载体固定化酶、海藻酸钠固定化细胞、壳聚糖固定化细胞和游离全细胞几种生物催化剂形式中，壳聚糖固
定化细胞催化效率最高、稳定性最好。通过反应体系、反应温度、金属离子、底物浓度、固定化细胞投量等因
素的优化，确定了最佳的生物催化反应条件：以 50 mmol/L pH=6.6 磷酸氢二钠-柠檬酸缓冲液作为反应体系，底
物亚氨基二乙腈（IDAN）的浓度为 200 mmol/L，添加 CoCl 2 至终浓度为 1 mmol/L，反应温度 37 ℃，固定化细
胞投量为 0.25 g 每 5 mL 反应体积。在该条件下，反应 2 h 可将 IDAN 完全转化为 IDA。进一步将反应体系放大
10 倍，催化 200 mmol/L 的 IDAN 完全转化为 IDA 仅需 1 h。
关键词：金黄节杆菌；腈水解酶；固定化；亚氨基二乙酸；生物催化；优化
中图分类号：TQ033 文献标识码：A 文章编号：1003-5214 (2018) 08-1313-07

Synthesis of Iminodiacetic Acid Catalyzed by a Nitrilase from

Arthrobacter aurescens CYC705

1
1
1
YANG Meng , MEI Na-na , CHEN Lu-yao , CAI Wen-wen, SU Er-zheng 1,2*
（1. College of Light Industry and Food Engineering, Nanjing Forestry University, Nanjing 210037, Jiangsu, China; 2. State
Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China）

Abstract: A nitrilase from Arthrobacter aurescens CYC705 was used to synthesize iminodiacetic acid
(IDA). The biocatalyst form, optimization of biocatalysis process and scale-up of the reaction system were
investigated in detail. Several biocatalyst forms including aminated carrier immobilized nitrilase, epoxy
carrier immobilized nitrilase, sodium alginate immobilized cells, chitosan immobilized cells and free whole
cells were prepared and compared. It was found that the chitosan immobilized cells had the highest catalytic
efficiency and the best stability. After optimizing reaction system, reaction temperature, metal ion
concentration, substrate concentration and additive amount of chitosan immobilized cells, the optimal
reaction conditions were obtained as follows: 50 mmol/L pH 6.6 Na 2HPO 4-citric acid buffer as reaction
medium, iminodiacetonitrile (IDAN) concentration 200 mmol/L, final CoCl 2 concentration in the reaction
medium 1 mmol/L, reaction temperature 37 ℃, addition amount of chitosan immobilized cells 0.25 g/5 mL
reaction volume. Under these conditions, IDAN could be completely hydrolyzed to IDA within 2 h.
However, when the reaction system was amplified 10-fold, the time required to catalyze the full conversion
of 200 mmol/L IDAN to IDA was only 1 h.
Key words: Arthrobacter aurescens; nitrilase; immobilization; iminodiacetic acid; biocatalysis; optimization
Foundation items: Six Talent Peaks Project in Jiangsu Province (2015-JY-016); China Postdoctoral Science

收稿日期：2017-09-12; 定用日期：2017-12-04; DOI: 10.13550/j.jxhg.20170739
基金项目：江苏省“六大人才高峰”高层次人才项目（2015-JY-016）；中国博士后科学基金（2016M600417，2017T100373）；江苏省
第五期“333 工程”培养资金资助项目（BRA2017458）；生物反应器工程国家重点实验室开放基金
作者简介：杨 猛（1992—），男，硕士生。联系人：苏二正（1976—），男，教授，025-85428906，E-mail：ezhsu@njfu.edu.cn。