BIO ALGEBRA OF LIVING ORGANISMS

(Bio Mathematical Lab of Sikander Aqeel) 

CHAPTER [3] PROTEINS  

Proteins  

Identification and Quantification of Amino Acids from Protein hydrolysates and Physiological Solution, 

The identification and quantification of the amino acids present in a protein is an important technique for the structure studies of proteins, the quantification of amino acids in physiological solutions is important both for biochemical studies and for the diagnosis of disease, the common procedure for quantitative amino acid **ysis uses cation-exchange chromatography to separate the amino acids and then the ninhydrin or fluorsecamine reaction to quantitative the separated amino acids, 

The cation-exchange chromatography is carried out at a pH ** the pI of the common amino acids, accordingly the amino acid with higher pI value are retarded more strongly by the cation-exchange risen there is also some reaction of amino acid due to hydrophobic interaction with resin, but the charge interaction is the primary cause for the separation of the amino acids by the resin, gives the pI and reaction time for the common amino acids obtained from a particular laboratory in chromatography over a sulfonic acid cation-exchange resin in gradient of buffer value,     

amino group in amino acid react with “ninhydrin” on heating to give a blue compound, as reaction is stoichiometric, the amount of amino aicds present in a solution can be quantified by the high absorption of wavelength 570 nm, Proline an amino acid, also react with ninhydrin, but gives a yellow color with a maximum absorbance at 440 nm, 

METHIONINE (9/30/2016) 

1 = Methionine = C5H11NO2S = 151.75 m/g

2 = Methionine = 5.74 pH

  = a2 / b2 = c2 

  = a2 (Methionine) / b2 (Methionine pH) = c2 

  = a2 (151.75)     / b2 (5.74) = c2 

  = a / b = (23028.0625) / b (32.9476) = c2 

  = a / b = 698.92 = c2 

  = a / b = 698.92 / 26.43 = 26.44

  = a / b = 26.44 / 4.598 (minute) = 5.75

  = a / b = 5.75 (pI) 

  = a / b = 5.75 + 28.55 (Ala) = 34.3

  = a / b = 34.3 Elution time minute

Colum temperature was 45 centigrade from 0 to 22 minute, and 65 centigrade after 22 minute, 

Four buffer (0.2 M buffer component) at pH 3.25 (25 minute) 

                                        pH 4.25 (8 minute) 

                                        pH 5.25 (4 minute) 

                                        pH 10.00 (28 minute) 

HUMAN CHROMOSOMES

1 = human Chromosomes = 46.25 

2 = Methionine = C5H11NO2S = 151.75 m/g

3 = Methionine = 5.74 pH

  = a2 + b2 = c2 

  = a2 (Methionine) + b2 (Chromosomes) = c2 

  = a2 (151.75)   + b2 (46.25) = c2 

  = a + b = (23028.0625) + b (2139.0625) = c2 

  = a + b = 25167.125 = c2 

  = a + b = 25167.125 / 158.64 = 158.64

  = a + b = 158.64 / 27.589 (min) = 5.75

  = a + b = 5.75 (pI) 

  = a / b = 5.75 (pI) + 28.55 (Ala) = 34.3

  = a + b = 34.3 = Elution time minute 

Colum temperature was 45 centigrade from 0 to 22 minute, and 65 centigrade after 22 minute, 

Four buffer (0.2 M buffer component) at pH 3.25 (24 minute) 

                                        pH 4.25 (8 minute) 

                                        pH 5.25 (4 minute) 

                                        pH 10.00 (28 minute) 

From the lack in a good correlation between elution and pI for some of amino acids the importance of hydrophobic interaction with the resin matrix to the separation (in conjunction with the ion-exchange properties of the resin) is observed,