Studies on effect of additives on protein profile, microstructure and quality characteristics of pasta.

Wheat storage proteins play a vital role in pasta making quality. In the present study, SDS-PAGE, Gel filtration chromatography and Scanning electron microscopy techniques were employed to understand the changes in the wheat protein fractions and their interactions with additives namely Sodium Steroyl Lactate (SSL), Glycerol Monostearate (GMS) and Hydroxy Propyl Methyl Cellulose (HPMC) during processing of pasta. SDS-PAGE studies indicated changes in High Molecular Weight Glutenin (HMW) fractions during drying stages of pasta preparation and in cooked pasta samples. In uncooked pasta, gel filtration patterns showed four peaks corresponding to different storage proteins whereas in the case of cooked pasta, these peaks were merged into three peaks. Pasta quality characteristics studies indicated that pasta with HPMC was found to have minimum percentage of cooking loss (5.6%), increased cooked weight (82 g), firmness (2.97 N) and high overall quality score (27) than GMS, SSL and control. Microstructure studies confirm the beneficial effect of HPMC. The present study indicated that HPMC is better additive for pasta manufacture followed by GMS. This could be due to interaction of HPMC with starch and protein matrix is different from that of GMS and SSL.

Wheat bran stabilization and its use in the preparation of high-fiber pasta.

Wheat bran was explored as a source of fiber in the preparation of high-fiber pasta. Ground raw wheat bran having an ash content 5.99%, crude protein 15.1% and fat content 5.83% was subjected to moist heat treatment (steam heat-treated bran) and dry heat treatment (dry heat-treated bran), wherein the lipase activity was reduced by 50%. Treated bran samples were stable for 3 months without developing any rancid flavor and bitterness. Pasta samples were prepared by substituting semolina with 40% and 50% of bran samples. There was no further significant inactivation of lipase activity upon extrusion followed by drying of pasta, irrespective of the type and the amount of bran sample used. The cooked weights of the pasta were in the range 257-268 g/100 g, whereas the cooking loss decreased from 12.8% to 9.3% for treated bran-incorporated pasta. Sensory scores for pasta containing treated bran samples were higher. The total dietary fiber increased by 5.2 times upon replacement of semolina by 40% of treated wheat bran. Sodium dodecyl sulfate polyacrylamide gel electrophoresis studies showed faint bands in treated bran samples as well as treated bran-incorporated pasta samples.

Role of disulfide linkages in structure and activity of proteinase inhibitor from horsegram (Dolichos biflorus).

Proteinase inhibitor isolated from horsegram (Dolichos biflorus or Macrotyloma uniflorum) inhibited specifically the enzymes trypsin and chymotrypsin. The inhibitor contained seven disulfide linkages and was free from thiol groups. The inhibitor is resistant to denaturation by urea, guanidine hydrochloride or sodium dodecyl sulfate. Reduction of the inhibitor with dithiothreitol abolished both trypsin and chymotrypsin inhibitory activities. The kinetic plots of the reduction as followed by activity and loss in structure as reflected in the 257 nm CD band could be superposed; loss in the activity paralleled the loss in structure. The kinetics of the reduction process was complex; reduction of the inhibitor was slow and depended on the concentration of DTT. Reduction of the disulfide linkages with DTT affected the tertiary structure significantly and secondary structure was not affected considerably. Fluorescence quenching by acrylamide and potassium iodide suggested the unfolding of the molecule due to reduction. Thus, disulfide linkages play a predominant role in maintaining the three-dimensional structure of the inhibitor.

Detection of lectin activity on western blots using erythrocytes.

A method is described for the direct detection of lectins, agglutinating erythrocytes, on nitrocellulose membranes after Western blotting, thus avoiding protein extraction from specific bands in the gel, followed by agglutination assays. The methodology essentially involves exposing the lectin band on a nitrocellulose strip to trypsinized rabbit erythrocytes (2%, in 0.15 M NaCl) for 30 min at 37 degrees C and then carefully transferring the membrane to saline (4 degrees C) for a few gentle washes and then fixing it in a solution (0.2% glutaraldehyde in 0.15 M NaCl) for 30 min. Later, the membrane is gently washed several times in 0.15 M NaCl containing 10 mM beta-alanine. The lectin band is visualized as a red agglutinated patch. The method is specific for lectins that can agglutinate red blood cells and virtually has no cross reactivity with the various nonlectin proteins tested. Binding of erythrocytes to the lectin band on the nitrocellulose strip can be prevented by specific competing sugars. The method can be applied to screen for the presence of lectins in natural materials and to monitor lectin fractions during purification.

Binding of Cu(2+)-beta-cyclodextrin complex to calf thymus DNA.

Cu(2+)-beta-cyclodextrin (1:1) complex has been found by UV, fluorescence and CD spectroscopy, polarimetry and gel electrophoresis to bind reversibly to calf thymus DNA. Using UV the binding constant was found to be 45280 +/- 7100 M-1. The binding of the complex Cu(2+)-BCD with DNA was stronger than that of free Cu2+. However the ternary complex formed thus was destroyed by EDTA.