Nutcracker

Dhara Patel
Results
Protein Purification of Fumarase
The total Fumarase activity in the sample was unconquerable by the absorbance of Fumarate at 250nm which was measured as a run of time using a UV spectrometer. A chromatography column fraction table was used to summarize the purification result. The tabularise 1 contains the information about each fraction note A, B, and C and it shows that the altitude B is the most efficient mince at removing unwanted proteins hence the % yield is less than deoxycytidine monophosphate%. It also shows the starting and the purest final Fumarase activity.

Table 1: Ni - NTA Fraction Table

Fraction| Volume (ml)| Fumrase Activity (Units/ml)| inwardness Fumarse (Units)| Protein Conc. (mg/ml)| Total Protein (mg)|
CFE| 1.28| 248| 317.44| 13.383| 17.13024|
Peak A| 21| 0.389| 8.169| 0.5392| 11.3232|
Peak B| 15| 1.348| 20.22| 0.2551| 3.8265|
Peak C| 15| 23.65| 354.75| 0.4081| 6.1215|

Fumarase Specific activity (Units/mg)| % behave| Fold Purification|
18.53097213| 0| 0|
0.721439169| 2.5734| 0.038931534|
5.284202274| 6.369708| 0.285155157|
57.95148248| 111.7534| 3.127276976|

Fractions were eluted from Ni-NTA column with next buffers: Fraction A (50 mM NaPhosphate, pH 7.8, 300 mM NaCl), Fraction B (50 mM NaPhosphate, pH 7.



8, 300mM NaCl, 60mM Imidazole) and Fraction C (50 mM NaPhosphate, pH 7.8, 300mM NaCl, 400mM Imidazole). One unit of Fumarase activity is the pith needed to produce 1µmol of fumarase/minute.

Ni-NTA chromatography

Peak A
Peak A

Peak C
Peak C

Peak B
Peak B

Figure 1. electric stall extract was thawed and centrifuged to separate supernatant from the pellet , supernatant was used for the column. Base line was equilibrated and established at 1.5ml/min using 1ml of sluggish green solution and Buffer A (50 mM NaPhosphate, pH 7.8, 300 mM NaCl). 1280µl of cell free extract was added and the final total fraction stack of 21 ml was collected for beak A. Similarly for peak B and peak C, Buffer...If you want to get a to the full essay, order it on our website: Orderessay



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