Prediction of the percentages of cows'', goats'' and ewes'' milk in “Iberico” cheese by electrophoretic analysis of whey proteins

Stepwise multiple linear regression (SMLR) and principal components regression (PCR) have been used to predict the percentages of cows', goats' and ewes' milk in Iberico cheese, using the results obtained by electrophoretic analysis (PAGE and IEF) of whey proteins, using standard cheeses. Similar predictions of the percentages of milks from the three species were obtained when either SMLR or PCR were applied to the electrophoretic data, i.e. the optical intensity of the electrophoretic bands (PAGE or IEF) of the whey proteins. The root mean square error of prediction in cross-validation (RMSEPCV) was lower than 4% in all cases.     

Enzymatic production of human milk fat substitutes containing palmitic and docosahexaenoic acids at sn-2 position and oleic acid at sn-1,3 positions

This paper studies the synthesis of structured triacylglycerols (STAGs) rich in palmitic and docosahexaenoic acids (PA and DHA) at sn-2 position and oleic acid (OA) at sn-1,3 positions by a four step process. First, triacylglycerols (TAGs) were obtained with 63–66 mol PA/100 mol total fatty acids and 10 mol DHA/100 mol by acidolysis of tuna oil and commercial PA, catalyzed by the non-positionally specific lipase Novozym 435. Then these TAGs were purified neutralizing the free fatty acids (FFAs) by KOH hydroethanolic solutions and extracting TAGs with hexane; these TAGs were completely recovered as pure TAGs (without FFAs). The third step involved the displacement of fatty acids located at sn-1,3 positions by acidolysis of PA and DHA enriched TAGs with OA rich FFAs, catalyzed by the sn-1,3 specific lipase DF from Rhizopus oryzae, immobilized on Accurel MP-1000; TAGs with 67 mol OA/100 mol at sn-1,3 positions and 52.1 and 15.4 mol PA and DHA, respectively, per 100 mol at sn-2 position were obtained. Both acidolysis reactions were carried out in stirred tank reactors (STRs) with lipase both dispersed in the reaction medium and contained in a cartridge filter attached to the stirrer rod. Finally STAGs were purified and obtained with yields of over 80 mol STAGs/100 mol STAGs in the reaction product (no FFAs were detected).     

Functional analysis of the cysteine residues and the repetitive sequence of Saccharomyces cerevisiae Pir4/Cis3: the repetitive sequence is needed for binding to the cell wall beta-1,3-glucan

Identification of PIR/CIS3 gene was carried out by amino-terminal sequencing of a protein band released by beta-mercaptoethanol (beta-ME) from S. cerevisiae mnn9 cell walls. The protein was released also by digestion with beta-1,3-glucanases (laminarinase or zymolyase) or by mild alkaline solutions. Deletion of the two carboxyterminal Cys residues (Cys(214)-12aa-Cys(227)-COOH), reduced but did not eliminate incorporation of Pir4 (protein with internal repeats) by disulphide bridges. Similarly, site-directed mutation of two other cysteine amino acids (Cys(130)Ser or Cys(197)Ser) failed to block incorporation of Pir4; the second mutation produced the appearance of Kex2-unprocessed Pir4. Therefore, it seems that deletion or mutation of individual cysteine molecules does not seem enough to inhibit incorporation of Pir4 by disulphide bridges. In fks1Delta and gsc2/fks2Delta cells, defective in beta-1,3-glucan synthesis, modification of the protein pattern found in the supernatant of the growth medium, as well as the material released by beta-ME or laminarinase, was evident. However, incorporation of Pir4 by both disulphide bridges and to the beta-1,3-glucan of the cell wall continued. Deletion of the repetitive sequence (QIGDGQVQA) resulted in the secretion and incorporation by disulphide bridges of Pir4 in reduced amounts together with substantial quantities of the Kex2-unprocessed Pir4 form. Pir4 failed to be incorporated in alkali-sensitive linkages involving beta-1,3-glucan when the first repetitive sequence was deleted. Therefore, this suggests that this sequence is needed in binding Pir4 to the beta-1,3-glucan.     

Short communication: Effect of subclinical mastitis on proteolysis in ovine milk

The aim of this work was to evaluate the effect of intramammary infection (IMI) on the endogenous proteolysis of milk. Four control checks were carried out in the half-udder milk of 10 ewes that acquired unilateral subclinical mastitis. Two of these checks were conducted before the infection was established and 2 after. Ten healthy ewes were tested as a control group. The presence of a subclinical IMI involved an increase of the products of casein hydrolysis, the proteose-peptone (p-p) fraction and minor (m) caseins, and a decrease of β-casein. As a result, a significant increase in the proteolysis index (PI), calculated as the ratio of m-casein to the sum of caseins (α + β + κ), took place. α-Casein and κ-casein were not significantly affected by IMI. Correlations confirmed the scenario: log₁₀ of somatic cell count (SCC) was positively correlated with p-p content and negatively with β-casein, whereas log₁₀ SCC was not correlated with α-casein or κ-casein. On the other hand, p-p content was positively correlated with m-casein and PI and negatively with β-casein, but no correlation was detected between p-p content and α- or κ-casein. Furthermore, between casein fractions, m-casein was only significantly correlated with β-casein. These results suggest that use of indices of proteolysis of caseins such as p-p, m-casein, and PI, could be applied together with SCC to evaluate the cheese-making quality of milk.     

Detection and quantification of Escherichia coli and Pseudomonas aeruginosa in cow milk by near‐infrared spectroscopy

This work investigated the potential of NIR technology to be applied in the dairy industry for the detection of micro‐organisms. To this end, two types of cow milk samples were studied, one in which only bacterial biomass was considered and the other in which bacteria were cultured and grown in milk for 24 h. The study was carried out using two micro‐organisms Escherichia coli and Pseudomonas aeruginosa. Both types of samples with different counts of both micro‐organisms were analysed by a NIR analyser in the range 10 000–4000 cm^?1 based on transmittance measurements. Multivariate models indicated that a better discrimination between micro‐organisms was attained in those milk samples in which micro‐organisms have been grown.     

The Effect of Food Type (Fish Nuggets or French Fries) on Oil Blend Degradation during Repeated Frying

Abstract: Oil that is reused multiple times for deep frying goes through changes in chemical composition and physical characteristics, affecting the quality of the fried foods. In this study, the effect of the food type (fish nuggets or French fries) on the degradation of an oil blend during the deep‐fat frying of each food at 180°C during 12 days was determined, and the characteristics of the fried products were evaluated. The degradation of oil during repeated use was relatively faster when fish nuggets were fried than when French fries were fried, as higher values of total polar compounds were obtained. Practical Application: The results are useful for producers of French fries and fish nuggets, such as restaurants or fast foods sellers, providing them with practical guidelines within the permitted values established by the regulatory authorities. The studied foods have high economic importance and are different in their composition. Under the studied conditions, the tested oil blend may be used during 4 d (4 h per day) with a daily replenishment, without discarding the oil when frying fish nuggets, and must be discarded after 8 d when French fries are processed. This suggestion allows preparing safe fried foods for consumers.     

Evaluation of the chemical composition and antimicrobial activity of Mentha pulegium,Juniperus phoenicea,and Cyperus longus essential oils from Morocco

The present study evaluates the chemical composition and antimicrobial activity of the essential oils (EOs) of Mentha pulegium L., Juniperus phoenicea L. and Cyperus longus L. from Morocco. The composition of these species was analyzed by GC/MS and 84 components were identified. M. pulegium EO showed a great similarity with EOs coming from other regions, as pulegone, the major component, accounted for about 70% of the EO. The EO of J. phoenicea had as main components α-pinene (24.9%), β-phellandrene (24.4%) and α-terpinyl acetate (12.9%). The EO extracted from C. longus was remarkably rich in sesquiterpene hydrocarbons (83.2%), which included β-himachalene (46.6%), α-humulene (16.9%), and γ-himachalene (10.1%). The antimicrobial activity of these EOs has been evaluated against seven bacteria of significant importance for food hygiene. According to the results, M. pulegium showed the best bacteriostatic and bactericidal effect, followed by J. phoenicea and C. longus. So far as we know, this is the first report on the quantitative composition and biological activity of the essential oil from C. longus. The tested EOs showed a variable degree of antimicrobial activity being M. pulegium the most effective one.     

Cloning and sequence analysis of the Pichia pastoris TRP1, IPP1 and HIS3 genes

The Pichia pastoris TRP1 and HIS3 genes were cloned by complementation of the Saccharomyces cerevisiae trp1 and his3 mutants, respectively, and their nucleotide sequence was determined. The P. pastoris TRP1 gene includes an open reading frame (ORF) of 714 nucleotides corresponding to a polypeptide of 237 amino acids whose sequence shares about 40% identity with that of TRP1 encoding proteins in other yeast species. DNA sequencing showed that an ORF of 858 nucleotides, encoding a protein of 285 amino acids with high homology to inorganic pyrophosphatases (IPP1), is located downstream of the P. pastoris TRP1 gene. Both genes converge in this chromosomal region, showing a genetic organization analogous to that found in the Kluyveromyces lactis genome. The P. pastoris HIS3 gene possesses an ORF of 675 nucleotides, encoding a polypeptide of 224 amino acids which shows 74·1% identity to the homologous S. cerevisiae protein. The hexameric consensus GCN4 binding sequence (TGACTC), characteristic of many amino acid biosynthetic genes, is present in the promoter region. The TRP1 and IPP1 sequences were deposited in the EMBL databank under Accession Number AJ001000. The Accession Number of the HIS3 gene is U69170. © 1998 John Wiley & Sons, Ltd.     

A spheroplast rate assay for determination of cell wall integrity in yeast

The rate of formation of spheroplasts of yeast can be used as an assay to study the structural integrity of cell walls. Lysis can be measured spectrophotometrically in hypotonic solution in the presence of Zymolyase, a mixture of cell wall-digesting enzymes. The optical density of the cell suspension decreases as the cells lyse. We optimized this assay with respect to enzyme concentration, temperature, pH, and growth conditions for several strains of Saccharomyces cerevisiae. The level of variability (standard deviation) was 1–5% between trials where the replications were performed on the same culture using enzyme prepared from the same lot, and 5–15% for different cultures of the same strain. This assay can quantitate differences in cell wall structure (1) between exponentially growing and stationary phase cells, (2) among different S. cerevisiae strains, (3) between S. cerevisiae and Candida albicans, (4) between parental and mutated lines, and (5) between drug- or chemically-treated cells and controls. © 1998 John Wiley & Sons, Ltd.