Characterization and Industrial Application of the Enzyme Papain Through the Use of a Biosensor.

IF 2 Q3 CELL BIOLOGY

Cellular Physiology and Biochemistry Pub Date : 2025-09-13 DOI:10.33594/000000813

Guilber Vergara Velez, Lorena Daniela Domínguez Brito, Melissa Lisbeth Cerón Erazo, Rodolfo Andrés Rivadeneira Z, Frank Guillermo Intriago Flor, Andrés Miguel Anchundia Loor, Yanelis Ramos Alfonso, Vladimir Álvarez Ojeda, José Francisco Díaz Izurieta, Jorge Tumbaco, Felipe Jadán Piedra

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引用次数: 0

Abstract

Background/aims: Casein influences coagulation, yield, and product quality. This study aimed to develop and validate a papain (EC 3.4.22.2)-based biosensor to quantify casein in whole milk, evaluating its kinetic behavior and comparing it with HPLC.

Methods: Papain was immobilized on nylon membranes and a cassava starch biopolymer. Enzyme kinetics (KM), stability, sensitivity, precision, and linearity of the system were evaluated. A t-test (α = 0.05) was applied to compare the casein concentration obtained by the biosensor versus HPLC (n = 20). Interference from milk proteins and calcium was also analyzed.

Results: The biosensor showed high affinity for casein (KM = 0.037 mM), with a linear range of 0.001-0.03 mM (R² = 0.9974) and a response time <5 s. No significant differences were found with HPLC (p = 0.0665). Stability reached 70 days at 4°C and was reusable up to 15 times.

Conclusion: The developed biosensor proved to be an accurate, fast, and sustainable analytical alternative for the determination of casein in complex milk matrices.

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利用生物传感器表征木瓜蛋白酶及其工业应用。

背景/目的：酪蛋白影响凝血、产率和产品质量。本研究旨在建立并验证一种基于木瓜蛋白酶（EC 3.4.22.2）的生物传感器，用于定量全脂牛奶中的酪蛋白，评估其动力学行为并与HPLC进行比较。方法：将木瓜蛋白酶固定在尼龙膜和木薯淀粉生物聚合物上。对酶动力学（KM）、稳定性、灵敏度、精密度和线性度进行了评价。采用t检验（α = 0.05）比较生物传感器与高效液相色谱法测定的酪蛋白浓度（n = 20）。还分析了乳蛋白和钙的干扰。结果：该传感器对酪蛋白具有较高的亲和力（KM = 0.037 mM），在0.001 ~ 0.03 mM的线性范围内（R²= 0.9974），响应时间长。结论：该传感器可用于复杂乳基质中酪蛋白的准确、快速、可持续的分析方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Cellular Physiology and Biochemistry 生物-生理学

CiteScore

5.80

自引率

0.00%

发文量

审稿时长

1 months

期刊介绍： Cellular Physiology and Biochemistry is a multidisciplinary scientific forum dedicated to advancing the frontiers of basic cellular research. It addresses scientists from both the physiological and biochemical disciplines as well as related fields such as genetics, molecular biology, pathophysiology, pathobiochemistry and cellular toxicology & pharmacology. Original papers and reviews on the mechanisms of intracellular transmission, cellular metabolism, cell growth, differentiation and death, ion channels and carriers, and the maintenance, regulation and disturbances of cell volume are presented. Appearing monthly under peer review, Cellular Physiology and Biochemistry takes an active role in the concerted international effort to unravel the mechanisms of cellular function.