Browsing by Subject "proteolysis"

Plasmin is an enzyme which is an important factor affecting both flavor and shelf life of UHT milk. Plasmin hydrolyzes mainly β, αs1- and αs2-caseins into γ-caseins and proteose peptones. During storage, proteolysis by plasmin causes bitterness and gelation. The objective of present study was to investigate time-temperature combinations for preheat treatment of UHT milk to decrease plasmin activity thus improving the shelf life and quality of existing product. In experimental research, 14 different UHT milk samples were manufactured with 7 different preheat processes and two different types of milk: prehydrolyzed (A) and posthydrolyzed (B). Samples were analyzed for their extent of proteolysis by two different methods: SDS-PAGE and RP-HPLC. SDS-PAGE gave qualitative information on the casein degradation and RP-HPLC gave quantitative information. Also, sensory analyses were made by eight trained employees at Arla Ltd (Sipoo, Finland). There were significant differences in casein degradation of samples at 2 and 4 months. All samples had some proteolysis after 4 months of storage. The largest amount of casein degradation was found in samples 5, 7 and 11, whereas the least proteolysis found in samples 1, 2, 13 and 14. Same results came from both SDS-PAGE and RP-HPLC. The sensory evaluation showed differences between the samples regarding bitterness at 2 and 3 months. The samples which were evaluated most bitter in sensory analysis, showed also most proteolysis in other analyses (samples 5, 7 and 11). The rest of the samples were quite equal in all evaluated features. The impact of hydrolysis point of lactose was unclear in this study. The aim of this study was accomplished since optimal processing conditions were found: 1, 2, 13 and 14 and the optimization of the process was possible based on the results from this study.

In atherosclerosis, cholesterol accumulates in cholesterol-loaded macrophages (foam cells) forming cholesterol plaques in the arterial intima. Reverse cholesterol transport (RCT) is a mechanism in which HDL and its major structural protein apolipoprotein-A-1 (apoA-1) remove cholesterol from the foam cells and take it to the liver for its final excretion from the body in the faeces. An impaired removal of cholesterol from the foam cells is a potential contributor to a reduced RCT, which is related to a higher incidence of coronary heart disease. Chymase, a neutral protease of mast cells (MCs), is widely distributed in the connective tissue of most vertebrates and able to degrade apoA-1. After the degradation, HDL-particles are unable to interact with the ABCA-1 transporter protein on the surface of macrophages, which mediates efflux of cholesterol from the macrophage foam cells to HDL particles. It has been shown that chymase derived from rat peritoneal MCs is able to degrade apoA-1 even in the presence of blood plasma which contains natural inhibitors for chymase (α-2-macroglobulin and α-1-antichymotrypsin). In the present study we wanted to find out if mouse mast cell protease 4 (mMCP-4) isolated from peritoneal mast cells is able to maintain its enzymatic activity even in the presence of mouse serum and intraperitoneal fluid. A small molecular weight compound (S-2586) was used as a substrate. In the in vitro experiments a sonicated MC preparation that contains active chymase was used and the activity of chymase was measured in the presence of varying concentrations of plasma and intraperitoneal fluid. In the in vivo experiments we evaluate whether mast cell-dependent proteolysis of HDL particles does occur, and whether such modification inhibits their efficiency in inducing cellular cholesterol efflux in vitro. We found that both serum and intraperitoneal fluid inhibited chymase activity, serum to a higher extent. Systemic activation of MCs in mast cell-competent mice, but not in mast cell-deficient mice, in vivo led to a decreased ability of plasma and intraperitoneal fluid to act as cholesterol acceptors from cultured cholesterol-loaded macrophages. Local activation of peritoneal mast cells also blocked the cholesterol efflux-inducing effect of intraperitoneally injected human apoA-1. This work was performed at the Wihuri Research Institute. Licenses for animal work were approved by the Finnish Laboratory Animal Experiment Committee (Suomen eläinkoelautakunta, ELLA). Laboratory animals (female NMRI mice) were from the Viikki Laboratory Animal Centre of the University of Helsinki and the mast cell deficient strain of mice W-sash c-kit mutant KitW-sh/W-sh were from the Jackson Laboratory (BarHarbor, Maine). The work was supervised by the director of the Research Institute Petri Kovanen MD PhD and Miriam Lee-Rueckert PhD. Laboratory assistance was perceived from the technicians of the Wihuri Research Institute.

The use of sourdough has numerous benefits, including improvement of the sensory attributes of baked bread in terms of flavour, texture, volume, enhanced nutritional value and extended shelf life of bread. To achieve the desired sourdough performance and bread with optimal quality and improved flavour, it is essential to understand how the starter cultures behave in specific conditions. In a previous part of this research, the metabolic traits of lactic acid bacteria starters and yeast from the food company S.P.C. (S. Korea) were studied. This thesis aimed to explore the pro-technological properties of the selected starter associations of bacteria/yeast, i.e., Lactiplantibacillus plantarum + Fructilactibacillus sanfransciscensis + Saccharomyces cerevisiae (PSY) and Latilactobacillus curvatus + Levilactobacillus brevis + Saccharomyces cerevisiae (CBY). Consequently, analysis of acidification, proteolysis analysis (including free amino acids), and volatile compound profile were done. PSY and CBY grew at the expected cell density. pH of the sourdoughs fermented by PSY decreased along the same line and slower than that of CBY over the course of 24 h. PSY sourdough had the highest TTA value (11.12 ± 0.03 ml) and organic acid production (148.6 ± 2.4 mmol/kg and 25.1 ± 1.5 mmol/kg) than CBY sourdough TTA value (9.01 ± 0.11 ml) and organic acid production (110.6 ± 1.6 mmol/kg and 20.2 ± 0.9 mmol/kg). This shows PSY as having a relatively high capacity for producing acids during sourdough fermentation among the two associations. After assessing their proteolysis capabilities, PSY sourdough had a presumptively higher peptide content while CBY produced the highest free amino acid content (i.e., Orn having a potential repercussion on bread flavour). Several volatile compounds belonging to different chemical classes, such as acids, aldehydes, ketones, alcohols, esters, and other compounds, were produced by PSY and CBY. In PCA, the control sourdough had a distinctive volatile profile from PSY and CBY. Both PSY and CBY show much correlation with about 4% variation. Ethanol, acetic acid, benzene ethanol, 2(3H)-furanone, dihydro-5-pentyl showed their strongest influence on both sourdoughs as they are found in high amount. Finally, during sourdough fermentation, the associations performed in a desired way, and they showed differences in acidity and content of free amino acids that might have a strong influence on bread flavour. Less differences were observed in the volatile profile compounds of the two associations. Proper sensory analysis and consumer test (by the company) will be the most revealing of the differences observed in this experimental study.