DECOLORISATION OF INDUSTRIAL YE BY MOMORDICA CHARANTIA PEROXIDASES AND ASCORBATE PEROXIDASES - PHYTOREMEDIATION

DR.S.SIVA KUMAR*M.SIVA ARUMUGAM

DEPARTMENT OF BIOCHEMISTRYSRI SANKARA ARTS AND SCIENCE COLLEGE

ENATHURKANCHIPURAM- 631561

INTRODUCTIONThe discharge of waste water from textile industry

contains high concentrations of reactive dyesReactive dyes contain chromophoric groups such

as azo, anthraquinone, triaryl methane, etc..Reactive groups such as vinyl sulfonate,

chlorotrizaone, tricholoropyrimidine, etc.,Biodegradation is a promising technology.Microorganisms also affected by the

contamination and could not properly metabolizing the pollutants.

MOMORDICA CHARANTIA

NEED FOR ENZYME STUDIESOxido-reductive enzymes such as peroxidases,

and polyphenol oxidases are involved in degradation process.

These enzymes can react with low concentration of contaminated sites.

The treatment of a textile waste water performed on the laboratory scale using fruit pulp extract of Momordica charantia and the main constituents of phytoremediation are Momordica charantia peroxidases (MCP) and ascorbate peroxidases (APx).

AIM OF THE STUDYThe primary objective of the study is to evaluate

the efficiency of Momordica charantia peroxidase and ascorbate peroxidase in decolorization of textile waste water. The present study will conclusively compares MCP and APx activity in dye decolorization studies and find out the effective role of both enzymes activity.

AIM OF THE STUDYThe following steps were planned in the present study:Qualitative analysis of aqueous and ethanolic

extractsPreparation of industrial dye for decolorizationPartial purification of enzymesDetermination of enzyme activity in both crude

and purified extractsDye decolrization studies of both enzymesComparison of dye decolrization results. 

MATERIAL AND METHODSMATERIALSMomordica charantia fruits collected from the environment

of enathur village, kanchipuram, Tamilnadu, India. Textile waste water was collected from a textile plant located at Erode,

Tamilnadu.These units were used to colour of the textile fibers of making lungies.

METHODS

. Aqueous extraction (Ullah et al., 2011)Ethanol extraction(Pitipanapong et al., 2007).Momordica charantia peroxidase Activity was carried out

colorimetric ally using Guaiacol as electron donor substrate. (Li Ou et al., 2003 and Iolandaet al., 1998).

Ascorbate peroxidase assay was performed by using ascorbate as a substrate. (Khan and Husain, 2006).

Partial purification of MCP enzyme was done by (Khan and Husain, 2006).

Partial Purification Of APX enzyme was estimated byEffulent Treatment by MCP was followed by((Akhtar et al.,

2005).Effulent Treatment by APX was estimated by (Akhtar et al.,

2005).

CALCULATION OF PERCENT DYE DECOLORIZATION

To compare various experiments, the decolorization was calculated for each dye or mixtures of dyes. Parameter percent decolorization was defined as given bellow (Khan and Husain, 2006).

STATISTICAL ANALYSISStatistical analysis was carried out on a personal computer with the use of SPSS software evaluation version 17.0 for windows (SPSS Inc. Chicago, IL, USA). Data were expressed as Mean and Standard Deviation (SD). Data were analyzed using one way ANOVA by applying mean comparison method. Significant difference were defined as two tailed p<0.05. Data from all experiments were processed in the program Microsoft Excel to represent tables and graphs (Satar and Husain, 2009).

QUALITATIVE PHYTOCHEMICAL ANALYSIS OF MOMORDICA CHARANTIA

Phytochemical

compounds

Aqueous extract

Fruit pulp

Ethanolic extract

Fruit pulp

Flavonoids ++ ++

Terpenoids ++ ++

Tannins - -

Polyphenol ++ ++

Carbohydrate ++++ +++

Alkaloids ++ +++

Saponins +++ +

Cardiacglycoside - +

Protein ++ -

Amino acid ++++ -

Anthraquinones - -

Soluble Momordica charantia peroxidase peroxidase enzyme activity$

S. No. Sample Nature Momordica charantia activitiy (units/mg)

1. Activity in crude extract 134.40 ± 15.52

2. Activity in fractionated extract

333.33 ± 34.02

$

 Soluble Ascorbate peroxidase enzyme activity 

S. No.

Sample Nature Ascorbate activitiy (units/mg)

1. Activity in crude extract 27.32 ± 1.48

2. Activity in fractionated extract

86.74 ± 2.21

Absorbance of Textile Dye

S.No 1 2 3 4 5 6 7 8

Wavelength (nm) 430 470 490 520 550 580 610 700

Absorbance Units 0.64 0.52 0.04 0.07 0.11 0.71 0.16 0.24

Absorbance of Textile Dye

Groups \ Time in hour

1 2 3 4 5

Group – I 26.269 ± 1.11

49.24 ± 2.08

65.66 ± 2.78

68.94 ± 2.92

72.22 ± 3.06

Group – II 20.31 ± 0.76

37.72 ± 1.41

60.93 ± 2.28

66.73 ± 2.49

69.63 ± 2.60

Group –III 35.73 ± 1.04

60.75 ± 1.77

82.19 ± 2.39

88.48 ± 4.12

92.91 ± 2.71

Group –IV 29.28 ± 0.90

55.32 ± 1.70

74.85 ± 2.30

78.10 ± 2.40

81.35 ± 2.50

Soluble Momordica Charantia Peroxidase for Dye Decolorization at 470nm

Dye Decolorization by Momardica charantia peroxidases

  Soluble Ascorbate Peroxidase for Dye Decolorization at 580nm

Groups \ Time in hour 1 2 3 4 5

Group – I 19.05 ± 0.49

46.28 ± 1.19

51.72 ± 1.33

54.44 ± 1.40

57.17 ± 1.47

Group – II 17.07 ± 0.34

39.03 ± 0.77

46.35 ± 0.92

56.11 ± 1.11

56.11 ± 1.11

Group –III 23.23 ± 0.56

54.21 ± 1.32

64.54 ± 1.57

67.12 ± 1.64

69.70 ± 1.70

Group –IV 21.56 ± 0.48

45.52 ± 1.03

55.11 ± 1.25

62.29 ± 1.41

64.69 ± 1.46

Dye Decolorization by Ascorbate Peroxidase

SUMMARYFresh Momordica charantia fruits were collected in Enathur. Textile waste water was collected from a textile plant located at Erode,

TamilnaduQualitative analysis was carried out in two extracts.Dye decolorization studies were studied by treating the dye MCP in crude

and pure form, and APX in crude and pure forms.Using MCP and APX dye decolorization was achieved.

CONCLUSIONFrom the present study, it was concluded that MCP and APX has an ability

to decolorize the textile dye.Optimal conditions for the action of MCP was identified as incubation time

3 hours, pH is 5.6 and 1.0ml of enzyme.The absorbance is read at 470nmOptimal conditions for the action of APX was identified as incubation time

2 hours ,pH is 7.0 and 1.0ml of enzyme.The absorbance is read at 540nm.Thus, by using MCP, and APX toxic dyes in the textile waste water might

be detoxified.

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