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FDSN 507 Midterm 2
Protein Analysis
Give two reasons why protein analysis is important in nutrition labeling. | Can give total protein content, gives protein correction in total dietary fiber determination. |
What are the five characteristics of proteins which allow for analysis methods? | Nitrogen content, peptide bonds, aromatic amino acids, dye binding capacity, ultraviolet absorptivity |
______ method measures the presence of N and utilizes digestion, neutralization, distillation, and titration. | Kjeldahl |
T or F: The Kjeldahl methods measures inorganic nitrogen. | False |
____ methods utilizes combustion and gas chromatography to measure nitrogen. | Dumas |
T or F: The dumas method measures inorganic nitrogen. | True |
_____ method measures the absorbance of radiation by specific functional groups. | IR |
In the BCA method, proteins reduce _____ ions to _____ ions under alkaline conditions and these ions react with BCA to give a purple color. | cupric, cuprous |
What four amino acids does BCA depend on? | Cysteine, cystine, tryptophan, tyrosine |
What three amino acids absorb UV light at 280 nm? | Tyrosine, tryptophan, phenylalanine |
T or F: Heating of a food can reduce its extractabilty of protein. | True |
T or F: Methods other than Kjeldahl, Dumas, and UV require the use of a standard solution. | True |
How is non-protein nitrogen not included in protein analysis? | Extraction under alkaline conditions and then precipitation. |
T or F: Nonprotein nitrogen can cause overestimation of protein efficiency ratio. | False |
What three ways do proteins differ? | Biochemical characteristics, nutritional quality, functional properties |
Name three biochemical differences among proteins. | Solubility, thermal stability, adsorption characteristics, charge, size |
Describe salting out. | Proteins are precipitates from solution as the ionic strength increases, as salt competes with the protein for water. |
Describe isoelectric precipitation. | Proteins precipitate at their isoelectric point because there is no electrostatic repulsion. |
Describe solvent fractionation. | Addition of ethanol or acetone decreases the solubility of proteins. |
Describe protein denaturation. | Heat or extremes in pH cause precipitation. |
What are the four methods of differential solubility? | Salting out, isoelectric precipitation, solvent fractionation, protein denaturation |
In ____ chromatography, adsorption occurs between charged solid molecules in solution and the charged matrix. | ion-exchange |
In ____ chromatography, ligands bind to the support so that proteins can bind to the ligands; changing the pH allows for elution. | affinity |
_____ involves a semipermeable membrane which concentrates the sample with PEG. | Dialysis |
Name three types of membrane processes which separate based on size under pressure. | Microfiltration, ultrafiltration, nanofiltration, reverse osmosis |
T or F: In size-exclusion chromatography, larger molecules travel through the column fastest. | True |
Name three methods of electrophoresis. | SDS-PAGE, isoelectric focusing, 2D electrophoresis |
Describe SDS-PAGE. | SDS is used to denature proteins and then is run through a polyacrylamide gel; separates charged molecules by their molar mass. |
_____ can be used to determine the molecular weight of a protein. | SDS-PAGE |
Describe isoelectric focusing. | A pH gradient is established and a sample is loaded and voltage is applied; proteins migrate to pI, where they have no net charge. |
Describe 2D electrophoresis. | Proteins are separated first by pI and second by molecular weight. |
_____ are used to determine nutritional quality, identify a new protein, or determine molecular weight. | Amino acid analysis |
Describe amino acid analysis. | A protein sample is acid hydrolyzed and then separated using chromatography. |
T or F: Chromatography does not require the use of an internal standard or standard curves. | False |
What three items determine protein nutritional quality? | Amino acid composition, digestability, anti-nutritional factors (eg. enzyme inhibitors/tannins) |
How is protein efficiency ratio determined? | Use a rat feeding test where PER = total weight gain of group/total protein consumed; compared to a casein control. |
Name three tests which can be used to assess nutritional quality of protein. | Essential amino acid index, in vitro protein digestability, lysine availability |
Name four protein functional properties. | Solubility, emulsification, foaming, gelation, dough formation |