Explore a groundbreaking method for simultaneous peptide parameter measurement, enhancing peptide production efficiency and quality with innovative decolorization and hydrolysis strategies.

Key words: Rice protein, Rice peptide, Decolorization, Enzymatic hydrolysis, Debittering

Abstract

In the enzymatic production of peptides, controlling the degree of hydrolysis is crucial. Traditionally, parameters such as peptide content, free amino acid content, degree of hydrolysis, and molecular weight distribution are measured separately, which is time-consuming and inefficient. This study introduces a new method for simultaneously characterizing these parameters using gel filtration chromatography. Additionally, the research explores the decolorization of alkaline protease solutions for peptide production and the enzymatic preparation kinetics of rice peptides, addressing the challenge of improving peptide yield and reducing bitterness.

Introduction

The production of peptides requires the measurement of various parameters to control the hydrolysis process effectively. Current methods measure these parameters separately, leading to inefficiencies. There is a need for a unified method that can simultaneously measure multiple peptide parameters, enhancing the efficiency and convenience of the peptide production process.

Materials and Methods

The study employed gel filtration chromatography to simultaneously determine the molecular weight distribution, peptide content, degree of hydrolysis, and free amino acid content of peptide products. The method’s feasibility and accuracy were evaluated using bovine serum albumin and rice protein as substrates for enzymatic hydrolysis.

Peptide Molecular Weight Distribution Determination

Gel filtration chromatography was utilized to determine the molecular weight distribution of peptides at different enzymatic hydrolysis times. The method proved comparable to traditional techniques, with a good correlation (r=0.98) between the methods.

Degree of Hydrolysis Determination

A formula was derived to estimate the degree of hydrolysis based on the peptide molecular weight distribution obtained from gel filtration chromatography. The method showed a high correlation with conventional colorimetric methods, indicating its accuracy and feasibility.

Results and Discussion

The gel filtration chromatography method demonstrated reliable performance in simultaneously measuring the targeted peptide parameters. Additionally, the study explored the decolorization of alkaline protease solutions, a crucial step for producing high-quality peptides. Among the decolorization methods tested, a combination of salt precipitation and ultrafiltration yielded the best results, significantly improving the enzyme’s color and purity without compromising its activity.

Peptide Yield and Bitterness Reduction

The enzymatic preparation kinetics of rice peptides revealed that the peptide yield is initially high but plateaus over time, suggesting that pH changes and product inhibition might limit yield improvement. Strategies to overcome these challenges, including pH adjustment and double-enzyme hydrolysis, were explored, showing potential for increasing peptide yield and reducing bitterness.

Conclusion

This study successfully established a method for the simultaneous measurement of multiple peptide parameters, significantly improving the efficiency of peptide production. Additionally, the research provided insights into improving peptide yield and taste profile, which are critical for the commercial viability of peptide products.

Acknowledgments

The study acknowledges the support from Tianjin University of Science and Technology and collaboration with NovaTech Technologies Ltd. for providing the alkaline protease solutions.

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Original authors: Lu Zhijia, Lu Fuping (Tianjin University of Science and Technology, Tianjin, 300457)

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