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Kinetic studies on recombinant stem bromelain

Bala, Muntari and Mel, Maizirwan and Jami, Mohammed Saedi and Amid, Azura and Salleh, Hamzah Mohd. (2013) Kinetic studies on recombinant stem bromelain. Advances in Enzyme Research, 1 (3). pp. 52-60. ISSN 2328-4846

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Abstract

Stem bromelain is a plant thiol protease with several industrial and therapeutic applications. This current work presents kinetic studies of recombinant bromelain (recBM) expressed in Escherichia coli BL21-AI on foursynthetic substrates, N-α-carbobenzoxy-L-alanyl-p-nitrophenylester (ZANPE), N-α-carbobenzoxy-L-arginyl-L-ar-ginine-p-nitroanilide (ZAANA), N-α-carbobenzo-xy-L-phenylalanyl-L-valyl-L-arginine-p-nitroanili-de (ZPVANA) and L-pyroglutamyl-L-phenylalanyl-L-leucine-p-nitroanilide (PFLNA). Hydrolytic activities of recBM at various pH and temperature conditions were compared to that of commercial bromelain (cBM). Both enzymes demonstrated high activities at 45o C and pH 5 - 8 for recBM and pH 6 - 8 for cBM. recBM showed marginally lower Kmand slightly higher kcat/Kmfor ZAANA, ZANPE and ZPVANA in comparison to cBM.trans Epoxysuccinyl-L-leucylamido {4- guanidino}butane (E-64) severely affected recBM and cBM hydrolysis of the synthetic substrates by competitive inhibition with Kivalues of 3.6 - 5.1 μM and 5.5 - 6.9 μM for recBM and cBM, respectively. The evaluated properties of recBM including temperature and pH optima, substrate specificity and sensitivity to inhibitors or activators, satisfy the requisites required for food industries.

Item Type: Article (Journal)
Additional Information: 4085/32114
Uncontrolled Keywords: Protease; Recombinant Bromelain; Substrates; Inhibitors; Activators
Subjects: T Technology > TP Chemical technology > TP248.13 Biotechnology
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button): Kulliyyah of Engineering > Department of Biotechnology Engineering
Depositing User: Prof Ts Dr Azura Amid
Date Deposited: 02 Oct 2013 12:41
Last Modified: 14 Jul 2021 17:39
URI: http://irep.iium.edu.my/id/eprint/32114

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