Caseinolytic Mechanism of Serine Protease MCP746 from Tamarillo (Solanum betacea Cav.) and Potential Application in Fresh Cheese.

IF 6.2 1区农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY

Journal of Agricultural and Food Chemistry Pub Date : 2025-07-24 DOI:10.1021/acs.jafc.5c04993

Tong Zhang,Xinxin Bao,Siying Yang,Chunli Lu,Qiong Zhao,Ying Gu,Shuo Wang

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Abstract

The serine protease MCP746 was previously found to have potential for cheese making in our previous research, but its caseinolytic mechanism is still unclear. This study aims to identify the cleavage sites of MCP746 on κ-casein (κ-CN), elucidate its caseinolytic mechanism, and evaluate the quality of the cheese. The Michaelis constant (Km) and maximum hydrolysis rate (Vmax) of MCP746 were 0.92 mg/mL and 833.33 U/min, respectively. MCP746 catalytic triad consists of Asp145/His202/Ser531, enabling specific cleavage at Lys133-Asn134 of κ-CN. Additionally, compared to calf rennet, cheese produced with MCP746 demonstrated higher adhesiveness, springiness, and cohesiveness (p < 0.05). Cryogenic scanning electron microscopy (Cryo-SEM) revealed that cheese made with MCP746 exhibited smaller and denser pore structures. The content of bitter amino acids in cheese made with MCP746 (24.52%) was lower than that in the control (35.61%). These results illustrate that the plant protease MCP746 has promising applications in the production of fresh cheese.

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柽柳丝氨酸蛋白酶MCP746的酪蛋白溶解机制及其在新鲜奶酪中的潜在应用

我们之前的研究发现丝氨酸蛋白酶MCP746在奶酪制作中具有潜力，但其酪蛋白溶解机制尚不清楚。本研究旨在鉴定MCP746在κ-酪蛋白（κ-CN）上的裂解位点，阐明其溶酪蛋白机制，并对奶酪的质量进行评价。MCP746的米切里斯常数（Km）和最大水解速率（Vmax）分别为0.92 mg/mL和833.33 U/min。MCP746催化三元组由Asp145/His202/Ser531组成，能够特异性裂解κ-CN的Lys133-Asn134。此外，与小牛凝乳酶相比，MCP746生产的奶酪具有更高的粘连性、弹性和内聚性（p < 0.05）。低温扫描电镜（cro - sem）显示，用MCP746制成的奶酪具有更小、更致密的孔隙结构。用MCP746制作的奶酪苦味氨基酸含量（24.52%）低于对照（35.61%）。这些结果表明，植物蛋白酶MCP746在新鲜奶酪的生产中具有广阔的应用前景。

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来源期刊

Journal of Agricultural and Food Chemistry 农林科学-农业综合

CiteScore

9.90

自引率

8.20%

发文量

1375

审稿时长

2.3 months

期刊介绍： The Journal of Agricultural and Food Chemistry publishes high-quality, cutting edge original research representing complete studies and research advances dealing with the chemistry and biochemistry of agriculture and food. The Journal also encourages papers with chemistry and/or biochemistry as a major component combined with biological/sensory/nutritional/toxicological evaluation related to agriculture and/or food.