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]
  Yifan Xie , Liyun Yao , Ruolin Yang , Yuxing Cai , Yujie Jin , Ning Li . Exploration and Practice of Online and Offline Hybrid Teaching Mode in High-Performance Liquid Chromatography Experiment. University Chemistry, 2025, 40(11): 100-107. doi: 10.12461/PKU.DXHX202412133
2. [2]
  Haiyuan Wang , Xiaoning Jin , Yajing Sun , Zhen Zhang , Wentao Zhao , Yi Li . Practical Exploration of High-Performance Liquid Chromatography Experiment Teaching Reform Empowered by Artificial Intelligence. University Chemistry, 2026, 41(4): 45-51. doi: 10.12461/PKU.DXHX202505076
3. [3]
  Tong Wang , Liangyu Hu , Shiqi Chen , Xinqiang Fu , Rui Wang , Kun Li , Shuangyan Huan . Determination of Benzenediol Isomers in Cosmetics Using High-Performance Liquid Chromatography Empowered by “Mathematical Separation”. University Chemistry, 2026, 41(1): 9-19. doi: 10.12461/PKU.DXHX202503128
4. [4]
  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
5. [5]
  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
6. [6]
  Gengjia Chen , Junjie Ou . Application of the van Deemter Equation in Instrumental Analysis Teaching: A Case of Organic Polymer Monolithic Columns. University Chemistry, 2025, 40(11): 362-368. doi: 10.12461/PKU.DXHX202502003
7. [7]
  Pengli GUAN , Renhu BAI , Xiuling SUN , Bin LIU . Trianiline-derived aggregation-induced emission luminogen probe for lipase detection and cell imaging. Chinese Journal of Inorganic Chemistry, 2025, 41(9): 1817-1826. doi: 10.11862/CJIC.20250058
8. [8]
  Bin SUN , Heyan JIANG . Glucose-modified bis-Schiff bases: Synthesis and bio-activities in Alzheimer′s disease therapy. Chinese Journal of Inorganic Chemistry, 2025, 41(7): 1338-1350. doi: 10.11862/CJIC.20240428
9. [9]
  Zhuoyue Guo , Jinxin Guo . The Amazing Journey of Glucose. University Chemistry, 2026, 41(2): 279-285. doi: 10.12461/PKU.DXHX202502082
10. [10]
  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
11. [11]
  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
12. [12]
  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
13. [13]
  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
14. [14]
  Yihui Song , Shangshang Qin , Kai Wu , Chengyun Jin , Bin Yu . 生物化学在高水平创新型药学人才培养中的交叉融合应用——以去甲基化酶LSD1抑制剂的活性评价为例. University Chemistry, 2025, 40(6): 341-352. doi: 10.12461/PKU.DXHX202406018
15. [15]
  Yajuan Xing , Hui Xue , Jing Sun , Niankun Guo , Tianshan Song , Jiawen Sun , Yi-Ru Hao , Qin Wang . Cu₃P-Induced Charge-Oriented Transfer and Surface Reconstruction of Ni₂P to Achieve Efficient Oxygen Evolution Activity. Acta Physico-Chimica Sinica, 2024, 40(3): 2304046-0. doi: 10.3866/PKU.WHXB202304046
16. [16]
  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
17. [17]
  Zhiyang Li , Hui Deng , Xinqi Cai , Zhuo Chen . Magnetic Core/Shell-Capsules Locally Neutralize Gastric Acid for Efficient Delivery of Active Probiotics. Acta Physico-Chimica Sinica, 2024, 40(7): 2306051-0. doi: 10.3866/PKU.WHXB202306051
18. [18]
  Chunhui Gao , Lurong Li , Guanwei Peng , Jinni Shen , Wenxin Dai , Zizhong Zhang . Efficient photocatalytic NADH regeneration and enzymatic CO₂ reduction over[Cp^*Rh(bpy)H₂O]²⁺ self-assembled CdIn₂S₄ flower-like microspheres. Acta Physico-Chimica Sinica, 2026, 42(3): 100165-0. doi: 10.1016/j.actphy.2025.100165
19. [19]
  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
20. [20]
  Yerong Chen , Bingbin Yang , Xinglei He , Yuqi Lin , Keyin Ye . Enzyme-Directed Evolution Enables Bioconversion of Organosilicon Compounds. University Chemistry, 2025, 40(10): 121-129. doi: 10.12461/PKU.DXHX202411054

Get Citation

PDF

XML

Metrics

PDF Downloads(0)
Abstract views(860)
HTML views(48)

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