Abstract

This study was intended to understand suitability of Wrightia tinctoria calli proteases (CCE) as vegetable coagulant through delineating their milk clotting potential and casein (whole and κ-casein) hydrolysis pattern in comparison to their plant stem crude enzyme (PSCE), rennin and Enzeco^® (positive controls). Stem calli was induced using two concentrations of naphthalene acetic acid (NAA) [0.5 mg/L (M1), 1 mg/L (M2)] with fixed concentration (1 mg/L) of 6-benzylaminopurine (BAP). Healthy calli, with significant growth at the end of 14th, 28th, and 48th days, were chosen for enzyme extraction. Overall activity calculated as per gram of fresh weight of callus revealed 3-fold high milk clotting index (MCI) by 28th day CCE in comparison to PSCE (p < 0.5). 14th day CCE mimicked rennin, 28th day CCE partially mimicked rennin and Enzeco^® in their casein hydrolytic pattern. 28th day CCE however exhibited a closer similarity to rennin in its κ- casein hydrolytic pattern. 48th day CCE (M1) presented high MCA, however, with low MCI in comparison to other CCEs. High affinity toward caseins similar to that of Enzeco^® was observed from 48th day CCE. Results from this study indicate 28th day callus protease may serve as a prospective vegetable milk coagulant alternative.

摘要

本研究旨在通过描绘Wrightia tinctoria愈伤组织蛋白酶 (CCE) 作为植物凝固剂的适用性，与它们的植物茎粗酶 (PSCE)、凝乳酶和 Enzeco 相比，它们的牛奶凝固潜力和酪蛋白（全酪蛋白和 κ-酪蛋白）水解模式^®（阳性对照）。使用两种浓度的萘乙酸 (NAA) [0.5 mg/L (M1)、1 mg/L (M2)] 和固定浓度 (1 mg/L) 的 6-苄基氨基嘌呤 (BAP) 诱导茎愈伤组织。选择在第 14 天、第 28 天和第 48 天时有显着生长的健康愈伤组织进行酶提取。按每克愈伤组织鲜重计算的总活性显示，与 PSCE 相比，到第 28 天 CCE 时，牛奶凝固指数 (MCI) 高 3 倍（p < 0.5)。第 14 天 CCE 模拟了凝乳酶，第 28 天 CCE 部分模拟了凝乳酶和 Enzeco ^®的酪蛋白水解模式。然而，第 28 天的 CCE 在其 κ-酪蛋白水解模式中表现出与凝乳酶更接近的相似性。然而，与其他 CCE 相比，第 48 天 CCE (M1) 呈现高 MCA，但 MCI 较低。从第 48 天 CCE 开始观察到与 Enzeco ^{®相似的酪蛋白的高亲和力。}这项研究的结果表明，第 28 天愈伤组织蛋白酶可以作为一种潜在的植物奶凝固剂替代品。