Extraction of peroxidase from bitter gourd (Momordica charantia) by three phase partitioning with dimethyl carbonate (DMC) as organic phase

Abstract Three phase partitioning (TPP) is most renowned technique used for extraction and purification of natural products. In previous studies of TPP, t -butanol is mainly used as an organic phase. This is the first report that explores ability of dimethyl carbonate (DMC) in the field of TPP as an alternate solvent for t -butanol. In the present study TPP process with t -butanol and DMC as organic phase along with different salts was applied to waste bitter gourd powder to obtained peroxidase enzyme. DMC was found to be compatible with most of salts such as ammonium sulphate and sodium citrate and explored as more efficient solvent than t -butanol. This TPP system provides 4.84 fold purity of peroxidase enzyme at optimum source concentration of 0.15 g/mL, with a system comprising DMC as organic phase, sodium citrate (20%) as salt, agitation speed 120 rpm, pH 7, temperature 30 °C and extraction time of 3 h. Present study has aimed for extraction and separation of peroxidase from bitter gourd waste with TPP technique and ensures the scope of carbonated solvents in extraction and purification of proteins.

[1]  V. Mulimani,et al.  Three-phase partitioning of α-galactosidase from fermented media of Aspergillus oryzae and comparison with conventional purification techniques , 2008, Journal of Industrial Microbiology & Biotechnology.

[2]  R. Singhal,et al.  Extraction of forskolin from Coleus forskohlii roots using three phase partitioning , 2012 .

[3]  V. Rathod,et al.  Ultrasound‐assisted three‐phase partitioning of polyphenol oxidase from potato peel (Solanum tuberosum) , 2015, Biotechnology progress.

[4]  Eun Yeol Lee,et al.  Dimethyl carbonate-mediated lipid extraction and lipase-catalyzed in situ transesterification for simultaneous preparation of fatty acid methyl esters and glycerol carbonate from Chlorella sp. KR-1 biomass. , 2014, Bioresource technology.

[5]  R. Lovrien,et al.  Three phase partitioning: concentration and purification of proteins. , 1997, Protein expression and purification.

[6]  Q. Husain,et al.  Direct immobilization of peroxidase on DEAE cellulose from ammonium sulphate fractionated proteins of bitter gourd (Momordica charantia) , 2006 .

[7]  Q. Husain,et al.  Catalyzed degradation of disperse dyes by calcium alginate-pectin entrapped bitter gourd (Momordica charantia) peroxidase. , 2011, Journal of environmental sciences.

[8]  Á. Irabien,et al.  Ionic liquid-based three phase partitioning (ILTPP) systems: Ionic liquid recovery and recycling , 2014 .

[9]  Fen-fang Li,et al.  Three phase partitioning for simultaneous purification of aloe polysaccharide and protein using a single-step extraction , 2015 .

[10]  S. Rudloff,et al.  Quantification of antidiabetic extracts and compounds in bitter gourd varieties , 2011 .

[11]  V. Rathod,et al.  Recent advances in enzyme extraction strategies: A comprehensive review. , 2017, International journal of biological macromolecules.

[12]  N. C. Veitch,et al.  Horseradish peroxidase: a modern view of a classic enzyme. , 2004, Phytochemistry.

[13]  R. H. Khan,et al.  A peroxidase from bitter gourd (Momordica charantia) with enhanced stability against organic solvent and detergent: A comparison with horseradish peroxidase , 2007 .

[14]  V. Rathod,et al.  A novel method to augment extraction of mangiferin by application of microwave on three phase partitioning , 2014, Biotechnology reports.

[15]  V. Rathod,et al.  Ultrasound assisted three phase partitioning of a fibrinolytic enzyme. , 2014, Ultrasonics sonochemistry.

[16]  Seçil Önal,et al.  Concentration and purification of α-galactosidase from watermelon (Citrullus vulgaris) by three phase partitioning , 2013 .

[17]  R. Singhal,et al.  Extraction of cocoa butter alternative from kokum (Garcinia indica) kernel by three phase partitioning , 2013 .

[18]  V. Rathod,et al.  Utilization of glycerol for the production of glycerol carbonate through greener route , 2013 .

[19]  S. Rawdkuen,et al.  Three-phase partitioning and proteins hydrolysis patterns of alkaline proteases derived from fish viscera , 2014 .

[20]  V. Rathod,et al.  Three phase partitioning coupled with ultrasound for the extraction of ursolic acid and oleanolic acid from Ocimum sanctum , 2014 .

[21]  L. Ananthanarayan,et al.  Three phase partitioning as a novel method for purification of ragi (Eleusine coracana) bifunctional amylase/protease inhibitor , 2007 .

[22]  Cristian Torri,et al.  Dimethyl carbonate and switchable anionic surfactants: two effective tools for the extraction of polyhydroxyalkanoates from microbial biomass , 2015 .

[23]  M. N. Gupta,et al.  A novel process for extraction of edible oils: Enzyme assisted three phase partitioning (EATPP). , 2007, Bioresource technology.

[24]  V. Rathod,et al.  Three phase partitioning a novel technique for purification of peroxidase from orange peels (Citrus sinenses) , 2015 .

[25]  Mahesh Bule,et al.  Enzyme-assisted three phase partitioning: A novel approach for extraction of turmeric oleoresin , 2011 .