Citation: Kumar R, Kumar R, Chauhan A, Kumar M, Goyal MK, et al. (2015) Characterization and Isolation of Fungi for Removal of Color from Pulp DQG3DSHU0LOO(IÀXHQW0HHUXW ,QGLD -(QYLURQ$QDO7R[LFRO GRL 10.4172/2161-0525.1000324
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supernatant was then adjusted to 7.0 with 2M NaOH. The sample (50 ml) was mixed with 1 ml of CH 3 COOH (10%) and 1 ml of NaNO 2 (10%). After 15 minutes, 2 ml of NH 4 OH was added. It was then left for five minutes and absorbance (OD) was measured at 430 nm. For blank, 1 ml of CH 3 COOH (10%) was added in 50 ml of distilled water and 2 ml of NH 4 OH. After 15 minutes, 1 ml of NaNO 2 (10%) was added. After waiting for 5 minutes optical density (OD) was taken at 430 nm. The absorbance value was transformed into lignin content (ppm) using the following formulate [3-8]. Lignin (ppm)=Absorbance/0.000247 Enzyme analysis Finally the three most potent strains were selected for enzyme analysis. Two enzymes Lignin peroxidase and Manganese peroxidase were studied [3-8]. Lignin peroxidase (Lip): Veratryl alcohol (VA) assy. Lip can be measured photometrically through the oxidation of VA to veratryl aldehyde at 310 nm. The reaction mixture will contain 2 mM VA, 35 mM sodium titrate buffer (pH 3.0) and enzyme. The reaction starts with the addition of 0.36 mM H 2 O 2 . Manganese peroxidase (MnP): MnP activity can be measured directly through the oxidation of Mn (II) to Mn (III) at 270 nm. The reaction mixture consists of 0.5 mM MnSO 4 .H 2 O and enzyme, 45 mM sodium malonate buffer (pH-4.5) and enzyme. The reaction starts with the addition of 0.1 mM H 2 O 2 . Microscopic study and identification of fungal strains In this the isolated fungal strains were grown on PDA plate. A very small amount of mycelium was picked up from plate and kept on clear glass slide having a drop of water. Then observing under microscope (2X) the mycelium was teased and the fungal hyphae were separated from each other. Finally this mycelium was transferred on second glass slide having a drop of water. The cover slip was placed to make a temporary slide. This slide was observed under a microscope, Olympus and Magnus MLX-TR at 40X and 100X having camera attached with it. Fungal mycelium, spores and the spore attachment was observed and photographs were taken for the purpose of identification. Results Isolation of fungal strains Sampling was done from two industries namely as centrally pulp and paper mill, Lalkuan and Anand tissue paper mill, Meerut where sediment samples were collected. Fungal strains were isolated by serial dilution and grown on Potato Dextrose Agar (PDA) pates. Three fungal strains F1, F2 and F3 were isolated from century pulp and paper mill, Lalkuan and three fungal strains 1, 5 and 7 were isolated from Anand tissue paper mill, Meerut. One fungal strain AF3 was available in laboratory. Hence total seven fungal strains were used in this study. Figures 1 and 2 shows the isolated and purified fungal strains. The characteristics of these strains are given in the Tables 2 and 3 [1-4]. Microscopic study of fungal strains 1, F1 and AF3 The fungal strains were inoculated on the PDA plates. They were allowed to mature and form spores. Then temporary slides were made and observed under microscope and photographs were taken for the purpose of identification. The morphological characteristics of the three fungal strains are in the Table 3, Figures 3-5.
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The three fungi on the basis of their morphology have been identified as 1: Aspergillus sp.
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