Proteolytic and Lipolytic Changes in Beef Inoculated with Spoilage Microorganisms and Bioprotective Lactic Acid Bacteria

Abstract

The use of a controlled lactic fermentation has been studied by several authors as a means to extend meat shelf‐life without notably altering its sensory quality. Growth and metabolite production by spoilage microorganisms as well as by bioprotective strains are influenced by storage conditions. The objective of this work was to study the effect of temperature and storage time upon the growth and metabolite production of four spoilage bacteria and three bioprotective lactic acid strains in vacuum packaged, finely cut beef. Lipolysis was mainly due to the presence of Brochothrix thermosphacta at 4°C and at 20°C, whereas proteolysis was mainly caused by pseudomonads. Presence of Lactobacillus minor, a heterofermentative strain, did not promote proteolysis nor lipolysis at either temperature. No considerable lipolysis was observed until day 8 of storage in samples inoculated with bioprotective strains, but increased at day 12 when stored at 20°C; no proteolysis was observed at any storage temperature.     

Biochemical changes during processing of traditional Jinhua ham

Jinhua ham is the most famous traditional meat product of China and one of the most famed dry-cured hams in the world. Its processing consists of six stages: green ham preparation, salting, washing and sun-drying and shaping, ripening, and post-ripening. Intense proteolysis and lipolysis occur during processing period. As a result, the content of free amino acids in final ham products is 14-16 times that of green ham, and 191 volatile compounds have been identified during processing, which make a major contribution to the flavor of Jinhua ham.     

Plasmin activity, β-lactoglobulin denaturation and proteolysis in high pressure treated milk

The effects of high pressure (HP) on plasmin activity, β-lactoglobulin denaturation and proteolysis during subsequent storage of HP treated milk, were studied. Fresh raw milk samples were exposed to a range of pressures from 50 to 800 MPa, for times of 1, 10 or 30 min, at 20°C. Residual plasmin activity and whey protein denaturation were measured immediately post HP-treatment. Indices of proteolysis were measured during post-HP storage. Treatment at pressures >300 MPa resulted in extensive β-lactoglobulin denaturation. Plasmin activity decreased in milk treated at pressures 400 MPa; the loss of activity was not well correlated with β-lactoglobulin denaturation. Compared to raw milk, treatment at 50 MPa had little effect on proteolysis during storage of treated milk measured as increases in pH 4.6-soluble N and liberation of proteose peptones, but at pressures of 300–400 MPa, proteolysis was increased relative to raw milk. After pressurisation >500 MPa, proteolysis during storage of milk was less than that observed in raw milk. Overall, HP influenced proteolysis in milk in a way which is different from that produced by heat, in terms of subsequent susceptibility of casein to proteolysis during storage or incubation. In particular, HP treatment at pressures of 300–500 MPa can increase proteolysis in milk, possibly through changes in micelle structure facilitating increased availability of substrate bonds to plasmin, which has implications for products prepared from milk thus treated.     

Proteolysis of ultra-high pressure homogenised treated milk during refrigerated storage

Plasmin residual activity and its relation to proteolysis of milk subjected to ultra-high pressure homogenisation (UHPH; 200–300 MPa, inlet temperature = 30 °C and 40 °C) and to a high-pasteurisation treatment (90 °C, 15 s) were studied during refrigerated storage. Proteolysis was examined by capillary electrophoresis, HPLC peptide profiles, pH 4.6-soluble nitrogen and free amino acids. Inactivation of plasmin increased as homogenisation pressure did. Extensive proteolysis, was observed in 200 MPa 40 °C milk, due to its higher native and microbial enzyme contents, compared with the other samples. In general, hydrolysis of β-casein, hydrophobic peptide and pH 4.6-soluble nitrogen levels increased with higher residual plasmin activity, while hydrophilic peptides were not affected by the different treatments applied. β-Lactoglobulin was denatured to a greater extent by thermal treatment than by UHPH treatments. This study provides further insight into how UHPH treatments influence milk properties.     

泌乳月份及部分乳成分与牛乳中体细胞数关系的相关性研究

本研究利用陕西省西安市草滩牛场第五奶牛场62头泌乳牛2003年8个月的DHI记录资料,对泌乳月份及部分乳成分与牛乳中体细胞数的关系进行了研究.结果表明,体细胞数与日产奶量,日产奶量与乳蛋白率、乳脂率呈极显著的负相关(P＜0.01),体细胞数与乳蛋白率与乳脂率之间无显著相关关系(P＞0.05),乳蛋白率与乳脂率呈极显著的正相关(P＜0.01).5、7和10月份样品平均体细胞数较高,分别为789×103mL-1,696×103mL-1、1031×103mL-1,3月份样品平均体细胞数最低为390×103mL-1.     

Effect of selected yeast strains on the sensory properties of dry fermented sausages

This study examines the role of yeasts in the sensorial characteristics of dry fermented sausages. A total of 172 yeast strains were isolated from commercially obtained dry fermented sausages and their enzymatic activities studied. Those most active against meat proteins and pork fat were selected. Experimental sausages were manufactured and the yeasts inoculated onto the surface or into the mince. Control batches were manufactured similarly, but without inoculation. After ripening, the composition of the protein and lipid fractions were studied. The sensory properties of the products were evaluated by trained judges. Despite the enzymatic activity of the yeasts, the organoleptic characteristics of the sausages—irrespective of inoculation site—did not differ significantly from those of the controls. The influence of yeasts on the ripening of dry fermented sausages is, therefore, doubtful.     

Proteolytic and lipolytic starter cultures and their effect on traditional fermented sausages ripening and sensory traits

In this study, three starter formulations including Lactobacillus curvatus and Staphylococcus xylosus strains selected in vitro on the basis of their lipolytic and proteolytic activities were employed for the manufacture of traditional fermented sausages of southern Italy. Microbial population, proteolysis, lipolysis, changes in free amino acids (FAA) and free fatty acids (FFA) and development of characteristic taste and flavor of the final product were investigated. Proteolysis and lipolysis were observed in sausages inoculated with proteolytic and lipolytic S. xylosus coupled with L. curvatus, while the sausage started with only S. xylosus without lactobacilli was identical to the non-inoculated control, indicating that the proteolysis could be due to both microbial activity and endogenous proteases activated by the decrease in pH. The statistical analysis applied to the instrumental and sensory data showed that there was an effect of the starter used on the characteristics of the sausage obtained. In particular, the control samples showed very close features different from the sausages obtained by adding starter cultures. Finally, analyzing the sensory parameters the sausages ripened without starter addition and those started without the L. curvatus AVL3 showed similar features indicating an influence of the presence of the lactobacilli on the final organoleptic quality of the sausages. An appropriate choice of a combination of strains in a starter formulation is fundamental to obtain products of the expected quality.     

Cysteine protease activity in bovine milk

Proteolytic activity of native cysteine proteases was studied in bovine milk. Five fractions (fI–fV) with cysteine protease activity were separated from acid whey prepared from raw bovine milk by ion-exchange chromatography on Q-Sepharose. The hydrolytic action of the most active fractions (fIII and fV), after further purification using gel permeation chromatography on Superdex S75, was studied against individual caseins. The two fractions contained different cysteine protease activities capable of hydrolyzing both α_s1- and β-casein. Studies of the effects of different reagents on the activity of partially purified fIII showed that the activity in this fraction was unaffected by aprotinin, slightly inhibited by p-chloromercuribenzoate and o-phenanthroline and completely inhibited by -trans-epoxysuccinyl- -leucylamido (4-guanidino) butane, consistent with identification as a cysteine protease. Phenylmethylsulphonyl fluoride and pepstatin A reduced activity of fIII by 40% and 50%, respectively. The partially purified fIII retained 20% of its cysteine protease activity after heating at 55°C, 60°C, 65°C and 72°C for 40 min, 20 min, 10 min and 30 s, respectively. Immunoblotting of fIII with antibodies to the bovine lysosomal cysteine protease, cathepsin B, clearly indicated the presence of immunoreactive cathepsin B in this fraction. This study presents strong evidence for the presence of a heterogeneous group of cysteine proteases in bovine milk, with one of these enzymes probably being cathepsin B.     

Study of the counts,species and characteristics of the yeast population during the manufacture of dry-cured “lacón”. Effect of salt level

The aim of this work was to study the yeast population during the manufacture of dry-cured “lacón” (a Spanish traditional meat product) and the effect of the salting time. For this study, six batches of “lacón” were manufactured with three different salting times (LS (3 days of salting), MS (4 days of salting) and HS (5 days of salting)). Yeast counts increased significantly (P < 0.001) during the whole process from 2.60 to 6.37 log cfu/g. An increased length of salting time did not affect yeast counts throughout the manufacture of dry-cured “lacón”, although the highest yeast counts were obtained from LS batches. A total of 226 isolates were obtained from dry-cured “lacón” during drying-ripening stage, of which 151 were yeasts and were identified at the species level using molecular techniques. The total of 151 identified yeasts belonged to 4 different genera: Debaryomyces, Candida, Cryptococcus and Rhodotorula. Debaryomyces hansenii was the most abundant species isolated throughout the whole process as much in the interior as in the exterior of the pieces of three salt levels of “lacón” studied, while Candida zeylanoides was only isolated from the interior of MS and HS batches and from the exterior of LS and HS groups, but at lesser proportion than D. hansenii.     

Modeling of proteolysis and lipolysis in Iranian white brine cheese

The simultaneous effects of processing variables such as ripening time (20–60 days), ripening temperature (6–10 °C), level of rennet added (1–2 g/100 kg milk) and brine concentration (8–14%, w/v) on the proteolysis, lipolysis and sensory score of Iranian white brined cheese (Feta type) were explored by the means of response surface methodology. The most important effect in proteolytical terms was produced by ripening temperature and ripening time in linear form, but level of rennet added and brine concentration were also significant at the 5% level. In terms of lipolysis, ripening time was dominant factor in both linear and quadratic forms; quadratic effect of ripening temperature was greater than its linear effect.