Explore the optimal enzymatic hydrolysis conditions for producing rice peptides with enhanced yield and functionality using alkaline and neutral proteases.

Keywords: rice dregs, rice peptide, enzymolysis , physicochemical property, antioxidant activity

Introduction

Proteins, when hydrolyzed, yield water-soluble, small molecule active peptides and free amino acids. This not only enhances and facilitates their digestion and absorption by the human body but also endows them with various biological activities such as antibacterial, immune promotion, antioxidation, antiviral, hormone regulation, blood pressure reduction, and lipid-lowering functions. Utilizing various proteins and selecting appropriate proteases for hydrolysis to obtain peptides with nutritional and functional properties has become a key direction in protein development and utilization. Given the specificity of protease hydrolysis sites, the same type of protein can yield different peptide fragments under the action of different proteases. This study compares the hydrolysis of rice peptides using alkaline protease, neutral protease, pepsin, and trypsin, selecting the two with the best hydrolysis effects for dual enzyme hydrolysis. It analyzes the optimal process conditions for single enzyme hydrolysis and combines them to improve the hydrolysis degree of rice dregs further.

Materials and Equipment

Experimental Materials and Main Reagents

The study utilized rice dregs and various enzymes including alkaline protease, neutral protease, trypsin, and pepsin, alongside analytical grade compounds such as copper sulfate, potassium sulfate, boric acid, hydrochloric acid, and formaldehyde.

Instrumentation

Instruments used included a LXJ-II type centrifuge, an HH-SB digital electric heating constant temperature water bath, and a precision force-increasing electric stirrer.

Research Methods

Enzymatic Hydrolysis Process of Rice Dregs Protein

The study outlined the processes for single enzyme hydrolysis reactions and dual enzyme hydrolysis reactions, detailing the conditions under which each enzyme was added and the subsequent steps taken to measure peptide yield and hydrolysis degree.

Analytical Detection Methods

Methods for determining hydrolysis degree and the determination of amino nitrogen were included, using titration and established standards.

Results and Discussion

Hydrolysis by Different Proteases

The study compared the hydrolysis effects of four different proteases on rice dregs protein, revealing that alkaline protease showed the best hydrolysis effect, followed by neutral protease and trypsin, with pepsin performing the least effectively. The paper discusses the implications of these results in detail.

Analysis of Single Enzyme Hydrolysis Conditions

The effects of temperature, pH, reaction time, and enzyme dosage on the hydrolysis of rice dregs protein were analyzed. Optimal conditions were determined for both alkaline and neutral proteases.

Orthogonal Experimental Design

The study utilized an orthogonal design to refine the optimal conditions for enzymatic hydrolysis, identifying the most significant factors influencing the process.

Verification Experiments

Experiments were conducted under optimal conditions to verify the results of the orthogonal experiments, yielding specific hydrolysis degrees for both alkaline and neutral protease treatments.

Conclusion

The study successfully identified the optimal conditions for the enzymatic hydrolysis of rice dregs using alkaline and neutral proteases, both individually and in combination. It was found that using a combination of these enzymes significantly improved the hydrolysis degree, offering a promising method for producing rice peptides with enhanced yield and functionality.

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Original research by Xiong Weijie, Wu Weiguo(Hunan Agricultural University, Changsha 410128)

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