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Functional Characterization of Recombinant Bromelain of Ananas comosus Expressed in a Prokaryotic System

Molecular biotechnology, 2014-02, Vol.56 (2), p.166-174 [Peer Reviewed Journal]

Springer Science+Business Media New York 2013 ;Springer Science+Business Media New York 2014 ;ISSN: 1073-6085 ;EISSN: 1559-0305 ;DOI: 10.1007/s12033-013-9692-2 ;PMID: 23921698

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Title:
Functional Characterization of Recombinant Bromelain of Ananas comosus Expressed in a Prokaryotic System
Author: George, Susan ; Bhasker, Salini ; Madhav, Harish ; Nair, Archana ; Chinnamma, Mohankumar
Subjects: amino acids ; Ananas - enzymology ; Ananas comosus ; Anti-Bacterial Agents - pharmacology ; Biochemistry ; Biodiversity ; Biological Techniques ; Biotechnology ; bromelains ; Bromelains - chemistry ; Bromelains - genetics ; Bromelains - metabolism ; Bromelains - pharmacology ; Cell Biology ; Chemistry ; Chemistry and Materials Science ; Cloning, Molecular ; E coli ; Escherichia coli ; Escherichia coli - drug effects ; Escherichia coli - enzymology ; Escherichia coli - genetics ; genes ; Genetic Variation ; Human Genetics ; immunoglobulin G ; pharmaceutical industry ; Phylogeny ; physicochemical properties ; pineapples ; Polymorphism, Single Nucleotide ; Proteases ; Protein Conformation ; Protein Science ; Protein Structure, Secondary ; Protein Structure, Tertiary ; Proteins ; Recombinant Proteins - pharmacology ; reverse transcriptase polymerase chain reaction ; sequence analysis ; single nucleotide polymorphism ; Streptococcus agalactiae ; Streptococcus agalactiae - drug effects ; Streptococcus infections ; Western blotting
Is Part Of: Molecular biotechnology, 2014-02, Vol.56 (2), p.166-174
Description: Bromelain (BRM) is a defense protein present in the fruit and stem of pineapple (Ananas comosus) and it is grouped as a cysteine protease enzyme with diversified medicinal uses. Based on its therapeutic applications, bromelain has got sufficient attention in pharmaceutical industries. In the present study, the full coding gene of bromelain in pineapple stem (1,093 bp) was amplified by RT-PCR. The PCR product was cloned, sequenced, and characterized. The sequence analysis of the gene revealed the single nucleotide polymorphism and its phylogenetic relatedness. The peptide sequence deduced from the gene showed the amino acid variations, physicochemical properties and secondary and tertiary structural features of the protein. The full BRM gene was transformed to prokaryotic vector pET32b and expressed in Escherichia coli BL21 DE3pLysS host cells successfully. The identity of the recombinant bromelain (rBRM) protein was confirmed by Western blot analysis using anti-BRM-rabbit IgG antibody. The activity of recombinant bromelain compared with purified native bromelain was determined by protease assay. The inhibitory effect of rBRM compared with native BRM in the growth of Gram-positive and Gram-negative strains of Streptococcus agalactiae and Escherichia coli O111 was evident from the antibacterial sensitivity test. To the best of our knowledge, this is the first report showing the bactericidal property of rBRM expressed in a prokaryotic system.
Publisher: Boston: Springer-Verlag
Language: English
Identifier: ISSN: 1073-6085
EISSN: 1559-0305
DOI: 10.1007/s12033-013-9692-2
PMID: 23921698
Source: MEDLINE
Alma/SFX Local Collection
ProQuest Central

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Functional Characterization of Recombinant Bromelain of Ananas comosus Expressed in a Prokaryotic System

Springer Science+Business Media New York 2013 ;Springer Science+Business Media New York 2014 ;ISSN: 1073-6085 ;EISSN: 1559-0305 ;DOI: 10.1007/s12033-013-9692-2 ;PMID: 23921698

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