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CLC number: Q556.2; R284.1

On-line Access: 2019-03-01

Received: 2018-08-06

Revision Accepted: 2018-10-11

Crosschecked: 2019-01-10

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Citations:  Bibtex RefMan EndNote GB/T7714

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Journal of Zhejiang University SCIENCE B 2019 Vol.20 No.3 P.264-272


A simplified and miniaturized glucometer-based assay for the detection of β-glucosidase activity

Author(s):  Min-Yi Jin, Tong Zhang, Yi-Shun Yang, Yue Ding, Jun-Song Li, Gao-Ren Zhong

Affiliation(s):  Experiment Center for Teaching and Learning, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; more

Corresponding email(s):   zhangtdmj@hotmail.com

Key Words:  Glucometer-based assay, β, -Glucosidase, Activity detection

Min-Yi Jin, Tong Zhang, Yi-Shun Yang, Yue Ding, Jun-Song Li, Gao-Ren Zhong. A simplified and miniaturized glucometer-based assay for the detection of β-glucosidase activity[J]. Journal of Zhejiang University Science B, 2019, 20(3): 264-272.

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author="Min-Yi Jin, Tong Zhang, Yi-Shun Yang, Yue Ding, Jun-Song Li, Gao-Ren Zhong",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T A simplified and miniaturized glucometer-based assay for the detection of β-glucosidase activity
%A Min-Yi Jin
%A Tong Zhang
%A Yi-Shun Yang
%A Yue Ding
%A Jun-Song Li
%A Gao-Ren Zhong
%J Journal of Zhejiang University SCIENCE B
%V 20
%N 3
%P 264-272
%@ 1673-1581
%D 2019
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1800416

T1 - A simplified and miniaturized glucometer-based assay for the detection of β-glucosidase activity
A1 - Min-Yi Jin
A1 - Tong Zhang
A1 - Yi-Shun Yang
A1 - Yue Ding
A1 - Jun-Song Li
A1 - Gao-Ren Zhong
J0 - Journal of Zhejiang University Science B
VL - 20
IS - 3
SP - 264
EP - 272
%@ 1673-1581
Y1 - 2019
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1800416

β;-Glucosidase activity assays constitute an important indicator for the early diagnosis of neonatal necrotizing enterocolitis and qualitative changes in medicinal plants. The drawbacks of the existing methods are high consumption of both time and reagents, complexity in operation, and requirement of expensive instruments and highly trained personnel. The present study provides a simplified, highly selective, and miniaturized glucometer-based strategy for the detection of β;-Glucosidase activity. Single-factor experiments showed that optimum β;-Glucosidase activity was exhibited at 50 °C and pH 5.0 in a citric acid-sodium citrate buffer when reacting with 0.03 g/mL salicin for 30 min. The procedure for detection was simplified without the need of a chromogenic reaction. Validation of the analytical method demonstrated that the accuracy, precision, repeatability, stability, and durability were good. The linear ranges of β;-Glucosidase in a buffer solution and rat serum were 0.0873–1.5498 U/mL and 0.4076–2.9019 U/mL, respectively. The proposed method was free from interference from β;-dextranase, snailase, β;-galactosidase, hemicellulase, and glucuronic acid released by baicalin. This demonstrated that the proposed assay had a higher selectivity than the conventional dinitrosalicylic acid (DNS) assay because of the specificity for salicin and unique recognition of glucose by a personal glucose meter. Miniaturization of the method resulted in a microassay for β;-Glucosidase activity. The easy-to-operate method was successfully used to detect a series of β;-Glucosidases extracted from bitter almonds and cultured by Aspergillus niger. In addition, the simplified and miniaturized glucometer-based assay has potential application in the point-of-care testing of β;-Glucosidase in many fields, including medical diagnostics, food safety, and environmental monitoring.


结论:酶活性测定最优条件为以柠檬酸-柠檬酸钠缓冲液(0.05 mol/L,pH 5.0)为反应溶液,0.03 g/mL水杨苷为底物,在50 °C条件下,反应30 min.新方法具有良好的精密度、准确性、重复性、稳定性和耐用性,且不受β-葡聚糖酶、蜗牛酶、β-半乳糖苷酶、半纤维素酶和葡萄糖醛酸的影响, 专一性较强.β-葡萄糖苷酶在缓冲液和大鼠血清中线性关系良好,线性范围分别为0.0873~ 1.5498 U/mL和0.4076~2.9019 U/mL.该方法已成功应用于苦杏仁、燀苦杏仁、酶制剂以及黑曲霉培养的一系列β-葡萄糖苷酶活性检测中,结果与现有方法所测的基本相符.


Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


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[26]List of electronic supplementary materials

[27]Data S1 Preparation of the DNS reagent

[28]Data S2 Preparation of the glucose calibration standards

[29]Fig. S1 Experimental protocol design

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