Evaluation of Anticancer and Anti-hemolytic Activity of Azurin, a Novel Bacterial Protein from Pseudomonas aeruginosa SSj

  • M. C. Sereena
  • Denoj SebastianEmail author


Cancer is the second leading cause of death in the world. About 18 milllion people were affected with cancer in 2018. Research in novel anti-cancer agents of bacterial origin thus gain importance. Azurin is a novel bacterial protein that has unique ability to specifically target cancer cells without any harmful effect to normal cells. In this study, cytotoxic effect of azurin from Pseudomonas aeruginosa SSj expressed in pET22b (+) vector and standard azurin (Sigma) were studied on (MCF7), (HCT15), and (VERO) cell lines by MTT assay. Apoptosis assay was performed using Hoechst staining and confirmed by FRET based assay on colon cancer cell lines (SW480). In vivo ascites antitumor model assay was also done to detect the in vivo cytotoxicity of azurin. Anti-hemolytic potential of purified azurin on human erythrocytes was also evaluated. Azurin induced cytotoxicity in both HCT15 and MCF7 cell lines, without having any adverse effect on the normal cells. Cell cycle analysis by Hoechst staining and FRET analysis, both showed evidence of apoptosis induced by azurin. In vivo ascites antitumor model proved that azurin treatment can increase the lifespan of tumor-bearing mice. Anti-hemolytic assay showed a significant increase of inhibition of hemolysis.


Azurin MTT assay FRET Hemolytic assay SW 480 



Michigan Cancer Foundation-7

HCT 15

Human colorectal carcinoma


Spectral karyotyping of the human colon cancer cell line


3-(4,5-Dimethylthiazol-2-Yl)-2,5-diphenyl tetrazolium bromide


Dulbecco’s modified Eagle medium


Fetal bovine serum


Phosphate-buffered saline


Daltons lymphoma ascites


Enhanced yellow fluorescent protein


Enhanced cyan fluorescent protein




Forward scatter


Side scatter


Fluorescence resonance energy transfer


Aspartic acid–glutamic acid–valine–aspartic acid


Compliance with Ethical Standards

Conflict of interest

Authors have no conflict of interest regarding the publication of paper.

Research Involving Animal Rights

All applicable institutional guidelines for the care and use of animals were followed.


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© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Life SciencesUniversity of CalicutThenjipalamIndia

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