Figure 1: SDS-PAGE of the partially purified extracellular enzyme from five isolated strains. Lane 1, marker proteins (from top to bottom) phosphorylase b (Mr, 97,400), bovine serum albumin (Mr, 67,000), ovalbumin (Mr, 43,000), carbonic anhydrase (Mr, 30,000) and Soyabean Trypsin Inhibitor (Mr, 20,100), respectively; lane 2, purified enzyme G-100, lane 3, low molecular weight protein DEAE CL6B, lane 4, high molecular weight protein DEAE CL6B. The gel was stained for protein with Coomassie brilliant blue R-250 and destained in methanol-acetic acid-water (7:2:1).

Figure 2: Optimization of hydrolase activity at different protein concentrations 50, 100, 150, 200, 250, 300 µgml-1 of protein respectively for different subunits of purified M1 viz. 33, 67 and both combined. Error bars indicates standard deviation.

Figure 3: Residual MCP concentration (µgml-1) by chemical, fungal and enzymatic method at various incubation periods in phosphorus free liquid culture medium. Error bars indicates standard deviation.

Figure 4: HPTLC chromatograms of residual MCP concentration in CZM medium by (a) Control, (b) Chemical, (c) Microbial and (d) Enzymatic after10 days of incubation.

Figure 5: FTIR spectrum of MCP degradation by (a) Control, (b) Chemical, (c) Microbial and (d) Enzymatic methods in CZM medium after 10 days of incubation.

Figure 6: Degradation kinetics of MCP by (a) Control, (b) Chemical, (c) Microbial and (d) Enzymatic methods in CZM medium at different incubation periods. Straight line equation shows that degradation of MCP follows first order kinetics.

Total activity (U)
Total protein (mg)
Sp. Activity (U mg-1)
Purification fold
Yield %
Crude
812.63± 2.81
351.33±5.97
2.31±.034
1
100
AmSO4 ppt
Intracellular
152.13±3.36
112.21±3.79
1.35±0.01
0.58±0.04
18.72±0.2
Extracellular
553.73±3.52
221.9±8.45
2.49±.11
1.07±.03
68.14±.67
G-100
OPH 33
320.12±4.46
5.26±.35
60.85±3.46
26.34±2.34
39.39±1.16
OPH 67
450.02±2.64
5.26±.35
85.69±6.29
37.04±3.07
55.37±.16
DEAE CL6B
OPH 33
297.14±5.52
2.15±0.01
138.2±2.42
59.82±1.14
40.32±1.34
OPH 67
425.11±4.46
2.26±0.04
188.1±3.36
81.42±2.34
52.31±1.13

Table 1: Purification profile of extracellular fungal hydrolases from Aspergillus niger