Citation: LI Shuanghua, FENG Si, CHEN Yinjia, LI Tong. Determination of activity of transglycosidase in diastatic enzyme by high performance liquid chromatography[J]. Chinese Journal of Chromatography, ;2014, 32(5): 539-542. doi: 10.3724/SP.J.1123.2013.12031 shu

Determination of activity of transglycosidase in diastatic enzyme by high performance liquid chromatography

LI Shuanghua ¹ ,
FENG Si ¹ ,
CHEN Yinjia ¹ ,
LI Tong ^1,2,,

1.
1. Dalian Elite Analytical Instruments Co., Ltd., Dalian 116023, China;

Corresponding author: LI Tong,
Received Date: 25 December 2013
Available Online: 27 February 2014

An analytical method for the determination of the activity of transglycosidase in diastatic enzyme by high performance liquid chromatography (HPLC) was established. Taken as the substrate, maltose was transformed into trisaccharide by transglycosidase in a 37 ℃ water bath and acetic acid buffer solution (pH=4.8) with acarbose as transglycosidase inhibitor. The transformation of the trisaccharide was detected on a SUGAR SH1011 column (300 mm×8.0 mm, 6 μm) with 0.01 mol/L sulfuric acid solution as mobile phase at a flow rate of 0.6 mL/min and a differential refractive index detector (RID), in order that the activity of transglycosidase can be measured indirectly. The conditions such as the chromatographic conditions, the concentration of substrate, the usage of inhibitor, and the incubation time were investigated. Under the optimized separation conditions, the calibration curve of the trisaccharide showed good linearity within the mass concentrations of 0.1-10 g/L (r=0.9998). The limit of detection and the limit of quantitation for transglycosidase activity were 0.013 U and 0.043 U, respectively. The relative standard deviation was 0.63% for six parallel tests. The activities of transglycosidase from different batches of diastatic enzyme were also determined with good result. The method can be applied to determine the activity of transglycosidase in the diastatic enzyme of the producers’ raw materials with the advantages of convenience, simplicity and stability.
- high performance liquid chromatography (HPLC),
- transglycosidase,
- activity,
- trisaccharide,
- diastatic enzyme
1. [1]
  [1] Zhang S M, Wang H J, Lü Y, et al. Journal of Henan Agricultural University (张世敏, 王会娟, 吕洋, 等. 河南农业大学学报), 2006, 40(5): 520
2. [2]
  [2] Wang L D. Grain Processing (王良东. 粮食加工), 2008, 33(6): 65
3. [3]
  [3] QB 2521-2001
4. [4]
  [4] Zhang Q. [MS Dissertation]. Zhengzhou: Zhengzhou University (张茜. [硕士学位论文]. 郑州: 郑州大学), 2011
5. [5]
  [5] Zhao K, Lou Z H. Light Industry Science and Technology (赵柯, 楼志华. 轻工科技), 2013(12): 19
6. [6]
  [6] Chen Y P, Wang Y, Hu X Z. Journal of Wuxi University of Light Industry (陈烨璞, 汪云, 胡学铮. 无锡轻工大学学报), 1998, 17(3): 86
7. [7]
  [7] Lin W X, Yun X, Liu Y X, et al. Journal of Dalian Institute and Light Industry (林维宣, 云霞, 刘英新, 等. 大连轻工业学院学报), 1996, 15(1): 73
8. [8]
  [8] Fan H M, Yu S, Tang Y F. Journal of Food Safety and Quality (樊惠民, 余实, 谭远方. 食品安全质量检测学报), 2011, 2(3): 174
9. [9]
  [9] Dai J. Chinese Journal of Chromatography (戴军. 色谱), 2012, 30(2): 113
10. [10]
  [10] Zou Y H, Ju H F, Chen A Y. Journal of Instrumental Analysis (邹耀洪, 居红芳, 陈爱英. 分析测试学报), 2005, 21(3): 34
11. [11]
  [11] Bi J F, Huang Y. Science and Technology of Food Industry (毕金峰, 黄颖. 食品工业科技), 2004, 25(2): 119
1. [1]
  Fan Wu , Wenchang Tian , Jin Liu , Qiuting Zhang , YanHui Zhong , Zian Lin . Core-Shell Structured Covalent Organic Framework-Coated Silica Microspheres as Mixed-Mode Stationary Phase for High Performance Liquid Chromatography. University Chemistry, 2024, 39(11): 319-326. doi: 10.12461/PKU.DXHX202403031
2. [2]
  Siming Bian , Sijie Luo , Junjie Ou . Application of van Deemter Equation in Instrumental Analysis Teaching: A New Type of Core-Shell Stationary Phase. University Chemistry, 2025, 40(3): 381-386. doi: 10.12461/PKU.DXHX202406087
3. [3]
  Quanliang Chen , Zhaohui Zhou . Research on the Active Site of Nitrogenase over Fifty Years. University Chemistry, 2024, 39(7): 287-293. doi: 10.3866/PKU.DXHX202310133
4. [4]
  Yanhui Zhong , Ran Wang , Zian Lin . Analysis of Halogenated Quinone Compounds in Environmental Water by Dispersive Solid-Phase Extraction with Liquid Chromatography-Triple Quadrupole Mass Spectrometry. University Chemistry, 2024, 39(11): 296-303. doi: 10.12461/PKU.DXHX202402017
5. [5]
  Chunmei GUO , Weihan YIN , Jingyi SHI , Jianhang ZHAO , Ying CHEN , Quli FAN . Facile construction and peroxidase-like activity of single-atom platinum nanozyme. Chinese Journal of Inorganic Chemistry, 2024, 40(9): 1633-1639. doi: 10.11862/CJIC.20240162
6. [6]
  Lihua HUANG , Jian HUA . Denitration performance of HoCeMn/TiO₂ catalysts prepared by co-precipitation and impregnation methods. Chinese Journal of Inorganic Chemistry, 2024, 40(3): 629-645. doi: 10.11862/CJIC.20230315
7. [7]
  Changqing MIAO , Fengjiao CHEN , Wenyu LI , Shujie WEI , Yuqing YAO , Keyi WANG , Ni WANG , Xiaoyan XIN , Ming FANG . Crystal structures, DNA action, and antibacterial activities of three tetranuclear lanthanide-based complexes. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2455-2465. doi: 10.11862/CJIC.20240192
8. [8]
  Heng Zhang . Determination of All Rate Constants in the Enzyme Catalyzed Reactions Based on Michaelis-Menten Mechanism. University Chemistry, 2024, 39(4): 395-400. doi: 10.3866/PKU.DXHX202310047
9. [9]
  Ruiqing LIU , Wenxiu LIU , Kun XIE , Yiran LIU , Hui CHENG , Xiaoyu WANG , Chenxu TIAN , Xiujing LIN , Xiaomiao FENG . Three-dimensional porous titanium nitride as a highly efficient sulfur host. Chinese Journal of Inorganic Chemistry, 2024, 40(5): 867-876. doi: 10.11862/CJIC.20230441
10. [10]
  Yiling Wu , Peiyao Jin , Shenyue Tian , Ji Zhang . The Star of Sugar Substitutes: An Interview of Erythritol. University Chemistry, 2024, 39(9): 22-27. doi: 10.12461/PKU.DXHX202404034
11. [11]
  Liwei Wang , Guangran Ma , Li Wang , Fugang Xu . A Comprehensive Analytical Chemistry Experiment: Colorimetric Detection of Vitamin C Using Nanozyme and Smartphone. University Chemistry, 2024, 39(8): 255-262. doi: 10.3866/PKU.DXHX202312094
12. [12]
  Zhuoya WANG , Le HE , Zhiquan LIN , Yingxi WANG , Ling LI . Multifunctional nanozyme Prussian blue modified copper peroxide: Synthesis and photothermal enhanced catalytic therapy of self-provided hydrogen peroxide. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2445-2454. doi: 10.11862/CJIC.20240194
13. [13]
  Zunxiang Zeng , Yuling Hu , Yufei Hu , Hua Xiao . Analysis of Plant Essential Oils by Supercritical CO₂Extraction with Gas Chromatography-Mass Spectrometry: An Instrumental Analysis Comprehensive Experiment Teaching Reform. University Chemistry, 2024, 39(3): 274-282. doi: 10.3866/PKU.DXHX202309069
14. [14]
  Mingyang Men , Jinghua Wu , Gaozhan Liu , Jing Zhang , Nini Zhang , Xiayin Yao . 液相法制备硫化物固体电解质及其在全固态锂电池中的应用. Acta Physico-Chimica Sinica, 2025, 41(1): 2309019-. doi: 10.3866/PKU.WHXB202309019
15. [15]
  Haitang WANG , Yanni LING , Xiaqing MA , Yuxin CHEN , Rui ZHANG , Keyi WANG , Ying ZHANG , Wenmin WANG . Construction, crystal structures, and biological activities of two Ln^Ⅲ₃ complexes. Chinese Journal of Inorganic Chemistry, 2024, 40(8): 1474-1482. doi: 10.11862/CJIC.20240188
16. [16]
  Yukai Jiang , Yihan Wang , Yunkai Zhang , Yunping Wei , Ying Ma , Na Du . Characterization and Phase Diagram of Surfactant Lyotropic Liquid Crystal. University Chemistry, 2024, 39(4): 114-118. doi: 10.3866/PKU.DXHX202309033
17. [17]
  Yuhang Jiang , Weijie Liu , Jiaqi Cai , Jiayue Chen , Yanping Ren , Pingping Wu , Liulin Yang . A Journey into the Science and Art of Sugar: “Dispersion of Light and Optical Rotation of Matter” Science Popularization Experiment. University Chemistry, 2024, 39(9): 288-294. doi: 10.12461/PKU.DXHX202401054
18. [18]
  Jingjing QING , Fan HE , Zhihui LIU , Shuaipeng HOU , Ya LIU , Yifan JIANG , Mengting TAN , Lifang HE , Fuxing ZHANG , Xiaoming ZHU . Synthesis, structure, and anticancer activity of two complexes of dimethylglyoxime organotin. Chinese Journal of Inorganic Chemistry, 2024, 40(7): 1301-1308. doi: 10.11862/CJIC.20240003
19. [19]
  Congying Lu , Fei Zhong , Zhenyu Yuan , Shuaibing Li , Jiayao Li , Jiewen Liu , Xianyang Hu , Liqun Sun , Rui Li , Meijuan Hu . Experimental Improvement of Surfactant Interface Chemistry: An Integrated Design for the Fusion of Experiment and Simulation. University Chemistry, 2024, 39(3): 283-293. doi: 10.3866/PKU.DXHX202308097
20. [20]
  Ping Song , Nan Zhang , Jie Wang , Rui Yan , Zhiqiang Wang , Yingxue Jin . Experimental Teaching Design on Synthesis and Antitumor Activity Study of Cu-Pyropheophorbide-a Methyl Ester. University Chemistry, 2024, 39(6): 278-286. doi: 10.3866/PKU.DXHX202310087

Get Citation

PDF

XML

Metrics

PDF Downloads(0)
Abstract views(419)
HTML views(22)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142