Synthetic Gene-Based Heterologous Expression, Proteolytic, and Structural Characterization of Caseinolytic Protease of Lactobacillus plantarum IIA-1A5

Tropical Animal Science Journal

View Publication Info
 
 
Field Value
 
Title Synthetic Gene-Based Heterologous Expression, Proteolytic, and Structural Characterization of Caseinolytic Protease of Lactobacillus plantarum IIA-1A5
 
Creator Yusuf, M.
Budiman, C.
Arief, I. I.
Sumantri, C.
 
Subject Clp-Protease
heterologous expression
Lactobacillus plantarum IIA-1A5
structural homology modelling
synthetic gene
 
Description Genome sequence of Indonesian probiotic of Lactobacillus plantarum II1A5 contains a gene encoding a proteolytic subunit of caseinolytic protease, designated as ClpP_LP. This study aims to express the Clp gene heterological and apply its proteolytic activity to some livestock products. To address this, the gene encoding ClpP_LP was optimized in silico by improving its Codon Adaptation Index and GC content to 0.94 and 53.62%, respectively. The optimized gene was then inserted into pET28a, transformed into Escherichia coli BL21(DE3), and over-expressed by induction of 1 mM Isopropyl β-D-1-thiogalactopyranoside at 37°C. The result showed that ClpP_LP was successfully over-expressed in a fully soluble form with the specific activity towards milk casein was 7739.89 AU mg-1. This activity was significantly greater than that of chymotrypsin. Further, the three-dimensional model of ClpP_LP was built using SWISS MODEL, which showed that this protein formed a homo-tetradecameric (14-mer) structure with each monomer consisting of 7 α-helix and 10 β-sheets. The identification of the active side showed that the active side of ClpP_LP is Ser-97, His-122, Asp-171, and forms a substrate-binding cavity with a size of about 29.5 Ǻ. Overall, our approach can serve as an appropriate platform for the production of ClpP_LP in a large-scale production for various applications in dairy products and derivatives.
 
Publisher Faculty of Animal Science, Bogor Agricultural University
 
Date 2021-11-12
 
Type info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Peer-reviewed Article
 
Format application/pdf
 
Identifier http://journal.ipb.ac.id/index.php/tasj/article/view/34024
10.5398/tasj.2021.44.4.520
 
Source Tropical Animal Science Journal; Vol. 44 No. 4 (2021): Tropical Animal Science Journal; 520-530
2615-790X
2615-787X
10.5398/tasj.2021.44.4
 
Language eng
 
Relation http://journal.ipb.ac.id/index.php/tasj/article/view/34024/22500
 
Rights Copyright (c) 2021 Tropical Animal Science Journal
http://creativecommons.org/licenses/by-sa/4.0
 

Contact Us

The PKP Index is an initiative of the Public Knowledge Project.

For PKP Publishing Services please use the PKP|PS contact form.

For support with PKP software we encourage users to consult our wiki for documentation and search our support forums.

For any other correspondence feel free to contact us using the PKP contact form.

Find Us

Twitter

Copyright © 2015-2018 Simon Fraser University Library