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Critical Reviews™ in Eukaryotic Gene Expression
Factor de Impacto: 2.156 Factor de Impacto de 5 años: 2.255 SJR: 0.649 SNIP: 0.599 CiteScore™: 3

ISSN Imprimir: 1045-4403
ISSN En Línea: 2162-6502

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Critical Reviews™ in Eukaryotic Gene Expression

DOI: 10.1615/CritRevEukaryotGeneExpr.2020034410
pages 411-425

Molecular Cloning and Characterization of an Acidic Polygalacturonase from Grapes and Its Potential in Industry

Zahra Nazir
Institute of Biochemistry and Biotechnology, University of the Punjab, Lahore, Pakistan
Riffat Chohan
Institute of Biochemistry and Biotechnology, University of the Punjab, Lahore, Pakistan
Malik Siddique Mahmood
Institute of Biochemistry and Biotechnology, University of the Punjab, Lahore, Pakistan
Hafiz Muzzammel Rehman
Institute of Biochemistry and Biotechnology, University of the Punjab, Lahore, Pakistan
Roquyya Gul
Institute of Biological Sciences, Gulab Devi Educational Complex, Lahore, Pakistan
Mahjabeen Saleem
Institute of Biochemistry and Biotechnology, University of the Punjab, Lahore, Pakistan

SINOPSIS

Pectinase enzymes from different plants have many possible biotechnological applications in various industries. Considering industrial significance of pectinolytic enzymes, the polygalacturonase (PG) gene from grapes was cloned into Escherichia coli DH5α using pTZ57R/T vector. Homologous sequences established a close link to Vitis vinifera. Conserve domain analysis predicted PLN02218 domain belongs to the cl31843 superfamily, showing its function as polygalacturonase. After confirmation by PCR and restriction analysis, the PG gene was expressed in E. coli BL21 and induced by IPTG. Expression level was assessed by 12% SDS-PAGE, which showed a 47 kDa protein. High expression level in the soluble fraction indicated that the protein is intracellular or transmembrane. Maximum expression was obtained with 1 mM IPTG and 6 hour induction time, and autoinduction with lactose increased production of the recombinant enzyme. Zymogram analysis revealed that the induced protein was an active enzyme. Expressed PG showed pectinolytic effect on the physiochemical properties of lemon juice. Natural biopolymers are greatly needed because synthetic fibers can negatively affect health. Pectin hydrolysis of banana pseudostem by the PG enzyme produced better quality fiber. Morphological studies by scanning electron microscopy revealed pectin degradation within the fiber cell architecture, showing the effectiveness of PG treatment on banana pseudostem.

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