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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  &lt; 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.

Potential utility of callus proteases as a milk clotting alternative to naturally propagated Wrightia tinctoria proteases