Cloning, expression and purification of collagen-like protein from Methylobacteria sp. 4-46 as alternative substance for collagen-based industries

Salleh, Faqihah and Amid, Azura and Nordin, Noor Faizul Hadry (2021) Cloning, expression and purification of collagen-like protein from Methylobacteria sp. 4-46 as alternative substance for collagen-based industries. In: Virtual International Halal Science Conference 21: "Enhancing and Strengthening Halal Industry through Science", 13th-14 July 2021, Kuala Lumpur.

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Abstract

Nowadays, collagen is widely introduced into oral and topical relief cosmeceutical products and is applied in the pharmaceutical industry in the form of hydrogels and collagenated scaffolds. However, due to the source of collagen in the market, it is a big concern for the Muslim community in terms of Halalan and Toyyiban since it is mainly extracted from mammals, especially porcine (Haram) and it raises health issues of animal-to-human disease transmission such as mad cow disease, swine, and avian flu. Collagen from bacteria would be the best alternative for halal and non-animal collagen. In this study, the bacterial collagen that produces collagen-like protein (CLP) of Methylobacteria sp. 4-46 was synthetically synthesized by Genscript. The CLPM4-46 in pUC57 was transformed into Escherichia coli DH5α and digested with BamHI/NdeI. The digested product (CLPM4-46, 1.04kbp) was successfully ligated with pCold II plasmid (4.39kbp) and transformed into E. coli BL21 (DE3). The ligated CLP gene was expressed using M9 minimal media. The protein expressed by the recombinant E. coli was measured with the Bradford Assay and SDS-PAGE where the size of the protein was 34.9k Da, and the CLP recombinant protein was purified using Fast Protein Liquid Chromatography and the collagen was measured using Enzyfluo™ Collagen Assay

Item Type:	Conference or Workshop Item (Plenary Papers)
Uncontrolled Keywords:	Halalan and Toyyiban, non-animal collagen, Methylobacteria sp. 4-46, Collagen-Like Protein
Subjects:	Q Science > Q Science (General)
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button):	International Institute for Halal Research and Training (INHART)
Depositing User:	Prof Ts Dr Azura Amid
Date Deposited:	09 Sep 2021 15:54
Last Modified:	09 Sep 2021 15:55
URI:	http://irep.iium.edu.my/id/eprint/92122

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