Combined use of Pronase E and a fungal extract (Penicillium aurantiogriseum) to potentiate the sensory characteristics of dry fermented sausages

A combination of an extract of Penicillium aurantiogriseum and Pronase E was used in an attempt to enhance the sensory characteristics of dry fermented sausages, specifically, a salami-like one named "salchichón". The addition of Pronase E alone (600 units/kg) produced a rise in free amino acids and biogenic amines and also an increase in the ammonia content. Addition of the protease and fungal extract (100.87 mg protein/kg of mixture) brought about a decrease in the level of free amino acids and a larger increase in the ammonia content than the batch added with only Pronase E. There was also an increase in the amount of specific volatile compounds such as 2- and 3-methylbutanal, 2- and 3-methylpropanal and 2- and 3-methyl-1-butanol produced by the breakdown of amino acids in these fermented sausages. Values of pH, a(w) and dry matter were not affected by addition of the protease or fungal extract. In contrast, addition of Pronase E produced a notable change in the textural characteristics, reducing the hardness, cohesiveness, gumminess, chewiness and cutting force. Sensory analysis showed that fermented sausages manufactured with Pronase E and Penicillium aurantiogriseum extract had better odour, flavour, texture and, as a consequence, better general acceptability.     

Impact of pre-selected autochthonous starter cultures on the flavor quality of Iberian dry-fermented "salchichón" sausage with different ripening processes

The purpose of this study was to investigate the impact on flavor quality of 2 autochthonous starter cultures in the manufacture of Iberian dry-fermented "sálchichón." A total of 2 strains of Pediococcus acidilactici (MS200 and MS198) were combined individually with a strain of Staphylococcus vitulus (RS34) to obtain 2 starter cultures: P200S34 and P198S34. The ability of both starter cultures to implant during 2 different manufacturing procedures of "salchichón" was evaluated by 16S rRNA gene sequence analysis. Changes due to starter culture inoculations on volatile compounds were determined by gas chromatography/mass spectrometry, and the impact on flavor quality evaluated by sensorial analysis. The implantation of starter cultures was adequate and did not significantly modify the flavor of the traditional Iberian dry-fermented "salchichón," while modulating the volatile compound profile with respect to that found in sausages with wild microbial populations. The influence of the starter cultures studied was more evident in the shorter sausage processing time. P198S34 was related with the increase in some volatile compounds deriving from lipid degradation. P200S34 contributed to the formation of several volatile compounds arising from lipid degradation, amino acid catabolism, and microbial esterase activity. PRACTICAL APPLICATION: P200S34 and P198S34 are autochthonous starter cultures for specific use in the processing of Iberian dry-fermented sausages. The application of these starters allows obtaining Iberian dry-fermented sausages with sensorial characteristics standardized. It is greatly interesting for processing and marketing industry of these high-quality meat products.     

Improvement of the Sensory Properties of Dry Fermented Sausages by the Superficial Inoculation and/or the Addition of Intracellular Extracts of Mucor racemosus

ABSTRACT: The effect of inoculation of the "salchichón" (dry fermented sausage) surface with an atoxigenic, proteolytic, and lipolytic strain of Mucor racemosus and/or the addition of a intracellular extract of the same mold on the ripening process was studied. Four batches of salchichón were manufactured: control, superficially inoculated, added with intracellular extract, and a combination of both treatments. Superficial mold modified the amino and free fatty acid contents, volatile compounds, and sensory properties. Intracellular extract degraded free amino acids, producing a rise in ammonia and volatile compounds. Both treatments combined improved significantly the sensory properties, which were given scores higher than control sausages at the end of ripening.     

Use of Lactococcus lactis subsp. cremoris NCDO 763 and α-ketoglutarate to improve the sensory quality of dry fermented sausages

The aim of the present work was to enhance the degradation of free amino acids in dry fermented sausages as precursors of volatile compounds responsible for the ripened flavour. For this purpose, Lactococcus lactis subsp. cremoris NCDO 763, its intracellular cell free extract (ICFE) and α-ketoglutarate were added to sausages. Papain was also used to increase the amount of free amino acids. When L. lactis was inoculated in sausages, an increase in the proteolytic phenomena was observed. The addition of α-ketoglutarate increased transamination phenomena in batches where it was added. The enhancement of these phenomena determined a noticeable rise in the content of glutamic acid (the main final product in transamination reactions) and a decrease, among other amino acids, of valine and leucine, with the formation of high amounts of their derivatives 2-methylpropanal and 3-methylbutanal. These aldehydes are responsible for the ripened flavour of dry fermented sausages. Sensory analysis showed an improvement of odour and flavour when L. lactis and α-ketoglutarate were combined. On the other hand, the intracellular cell free extract of L. lactis did not show any important activity in relation to amino acid breakdown even when used together with α-ketoglutarate and/or papain.     

Effects of muscle shortening and proteolysis on Warner-Bratzler shear force in beef longissimus and semitendinosus

The effect of the addition of the fungal protease EPg222 on the sensory characteristics of dry fermented sausage "salchichón" ripened with commercial starter cultures was investigated. Sausages were prepared with purified EPg222 and Staphylococcus carnosus, Staphylococcus xylosus, and Lactobacillus sakei as starter cultures, ripened for 145 days and compared with a control batch only inoculated with the starter cultures. Dry fermented sausages ripened with EPg222 and starter cultures showed higher amount of NPN and volatile compounds derived from amino acid catabolism, than control ripened only with starter cultures. Several branched aldehydes, acids and alcohols such as 2- and 3-methylbutanoic acid and 2-methylpropanol were detected only in enzyme treated samples. Sensory analysis reflected higher values for aroma intensity of sausages treated with EPg222 and lower values of hardness than control. The effect of EPg222 may be of great interest to improve sensory characteristics of dry fermented sausages ripened with starter cultures.     

Lipolysis, proteolysis and sensory characteristics of a Spanish fermented dry-cured meat product (salchichón) with oregano essential oil used as surface mold inhibitor

The superficial antifungal activity of oregano essential oil (OEO) in Spanish fermented dry-cured sausages ("salchichón"), and its effects on lipolysis, proteolysis and sensory characteristics were evaluated. The surface application of OEO reduced mold contamination on the surface, without significantly affecting the drying process. To evaluate the intensity of lipolysis during the ripening process, the profile and content of free fatty acids were determined. The addition of OEO led to a higher amount of unsaturated fatty acids, but lipolysis was not greatly affected. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), which was used to qualitatively assess the proteolytic changes in the sarcoplasmic and myofibrillar proteins during the process, pointed to very similar patterns in all the sausages. OEO did not significantly affect the sensory properties, but increased hardness, resulting in a better texture. Therefore, a shorter ripening time may be necessary for sausages treated with OEO.     

Use of bacterial extracts to enhance amino acid breakdown in dry fermented sausages

The effect of the intracellular cell-free extracts (ICFEs) of two bacterial strains (Lactobacillus sakei GO and Bacillus pumilus) on the amino acid catabolism and the sensory properties of dry fermented sausages, was investigated. Extracts were added to sausages alone or in combination with a protease, papain. Amino acid breakdown was monitored by the changes in free amino acids, ammonia and amine content during the ripening process. A 15% decrease in the content of free amino acids was observed in sausages added with the ICFE from L. sakei GO. Furthermore, the extract of L. sakei GO significantly reduced (54-68%) the content of the amino acids considered as precursors of the typical ripened flavour, i.e., valine, leucine and isoleucine. Chemical changes were not reflected in a significant improvement of the sensory quality of sausages added with the ICFEs. The potential use of the bacterial ICFEs studied in the present work for the manufacture of dry fermented sausages, and its comparison with the use of fungal extracts, are discussed.     

Bactometer system versus traditional methods for monitoring bacteria populations in salchichón during its ripening process

The performance of the Bactometer system (an impedimetric microbial monitoring system) compared with traditional methods (microbial colony counts) for monitoring bacterial populations (lactic acid bacteria [LAB], Enterobacteriaceae, and coliforms) was studied in 90 samples of an experimental salchichón (a type of Spanish ripened dry sausage) during its ripening process. The population quantitations were carried out with fresh sausage, semiripened sausage (14 days of ripening), and finished product (28 days of ripening). The results showed a high correlation between the traditional microbial colony count (in CFU per gram) and the impedance detection time: -0.98, -0.97, and -0.94 for coliforms, Enterobacteriaceae, and LAB, respectively (P < 0.01). Considering the results obtained with regard to the enumeration of populations of Enterobacteriaceae, coliforms, and LAB in salchichón during its ripening process, the advantages of impedance with respect to plate counts for monitoring the microbial dynamics of ripening processes are notable, especially in its time-saving aspects: between 19 and 21 h in the case of Enterobacteriaceae, between 7 and 20 h for coliforms, and between 32 and 46 h for LAB.     

The influence of manufacturing and drying conditions on the survival and toxinogenesis of Staphylococcus aureus in two Spanish dry sausages (chorizo and salchichón)

The effects of formulation, starter culture and fermentation temperature on growth and synthesis of toxin A (SEA) and TNase by Staphylococcus aureus during fermentation and drying of Spanish chorizo were investigated. Inhibitory factors able to inhibit SEA synthesis in culture media were unable to prevent SEA production in chorizo fermented at 20 and 30°C, though a lower temperature and starter culture SP318 (Lactobacillus sake, Pediococcus pentosaceous and Staphylococcus xylosus) decreased staphylococcal growth and SEA formation. Reduction and even disappearance of the SEA during ripening was observed. In most batches, TNase was a reliable indicator of staphylococcal growth and SEA production. Dextrose added to the salchichón formulation repressed S. aureus growth during drying. Lactobacillus curvatus in combination with dextrose was an effective anti-staphylococcal agent during fermentation.     

Changes in selected biochemical and sensory parameters as affected by the superficial inoculation of Penicillium camemberti on dry fermented sausages

An atoxigenic strain of Penicillium camemberti was superficially inoculated on fermented sausages in an attempt to improve their sensory properties. The growth of this mould on the surface of the sausages resulted in an intense proteolysis and lipolysis, which caused an increase in the concentration of free amino acids, free fatty acids (FFA) and volatile compounds. Many of these were derived from amino acid catabolism and were responsible for the "ripened flavour", i.e. branched aldehydes and the corresponding alcohols, acids and esters. The development of the fungal mycelia on the surface of the sausages also protected lipids from oxidation, resulting in both lower 2-thiobarbituric acid (TBARS) values and lipid oxidation-derived compounds, such as aliphatic aldehydes and alcohols. The sensory analysis of superficially inoculated sausages showed clear improvements in odour and flavour and, as a consequence, in the overall quality of the sausages. Therefore, this strain is proposed as a potential starter culture for dry fermented sausage production.     

The contribution of Penicillium aurantiogriseum to the volatile composition and sensory quality of dry fermented sausages

An atoxygenic, proteolytic and lipolytic strain of Penicillium aurantiogriseum was tested for its ability to accelerate the production of volatile compounds and to improve the sensory properties of dry fermented sausages. The following batches of sausages were prepared: control; superficially inoculated with a spore suspension; added with an intracellular cell free extract; and superficially inoculated and added with the intracellular cell free extract. Higher levels of lipid oxidation products were found in the aroma extracts of sausages without a mould cover. In contrast, branched aldehydes and alcohols presented higher concentrations in superficially inoculated and extract added sausages, while esters only showed higher concentration in the first ones. The sensory analysis showed that sausages prepared with both treatments received the highest scores in all the properties evaluated, which demonstrated both the potential of this mould as producer of volatile compounds and the effectiveness of combining both treatments.     

Metabolism of peptides and amino acids during in vitro protein degradation by mixed rumen organisms

In vitro inoculum enriched with particle-associated organisms was prepared using rumen contents from a cow fed a 60% forage, 40% concentrate diet. Treatment of in vitro inoculum with cetyltrimethylammonium bromide was used to release intracellular free amino acids from mixed rumen organisms. Addition of 10 mM tosylarginine methyl ester, a competitive inhibitor of trypsin, decreased degradation rate and intracellular free amino acids in incubations containing either casein or serum albumin. Extracellular peptides increased rapidly to a maximum at 60 min in casein incubations but were not different from zero in albumin incubations. Accumulation of intracellular free amino acids was maximal at 60 min in casein and albumin incubations; the concentration observed with albumin was about one-fourth that with casein. Ammonia production from intact casein was slightly greater than that from acid and enzymatically hydrolyzed casein and about 80% greater than that from albumin. Ammonia production and appearance of extracellular free amino acids lagged behind accumulation of intracellular free amino acids. Results suggest that formation and metabolism of extracellular peptides are important in controlling the rate of protein degradation by mixed rumen organisms.     

Influence of the salting time on volatile compounds during the manufacture of dry-cured pork shoulder "lacón"

The effect of the length of salting time on volatile compounds throughout the manufacture of dry-cured "lacón" was studied using a purge-and-trap extraction and GC/MS analysis. For this study, six treatments of "lacón" were salted with different amounts of salt (LS (3 days of salting), MS (4 days of salting) and HS (5 days of salting)). The total area of volatile compounds increased significantly (P<0.001) as the length of salting time increased. Significant differences were found for the total area of all aldehydes for each salting time at the end of process. Hexanal presented the most relative abundance, showing highest content in HS treatments while LS treatments showed the lowest one at the end of salting, post-salting and drying-ripening stage. The influence of salting time on the content of 3-Octen-2-one was significant on salting stage (P<0.001) and drying-ripening stage (P<0.01). The salting time affected (P<0.001) the relative abundance of esters at the end of salting and at the end of drying-ripening stage. The salting time affected significantly (P<0.001) on alcohols content at the end of drying-ripening process. However, no significant effect of salt level (P>0.05) was detected on total hydrocarbons and total furans at the end of process.     

茶鲜叶萎凋时期乙烯利溶液对游离氨基酸组分的影响

探究萎凋过程中乙烯利溶液对离体茶鲜叶游离氨基酸含量的影响。以贵州省自主培育品种“黔茶1号”为试材,设置对照组、喷施乙烯利组及喷施乙烯抑制剂组。采用邻苯二甲醛（ophthalaldehyde,OPA）法、9-芴甲基氯甲酸酯（9-fluorenylmethyl chloroformate,FMOC）法进行高效液相色谱在线衍生检测游离氨基酸。结果表明,鲜爽味氨基酸含量随萎凋时间延长而减少,甜味、苦味氨基酸含量随时间推移呈上升趋势。3类呈味氨基酸含量在萎凋期间对喷施乙烯处理响应不同。萎凋期间,鲜味氨基酸含量受乙烯诱导上调,差异显著发生在4、8 h;甜味氨基酸含量在各处理间总体差异不显著;各处理组苦味氨基酸含量在4、8 h无显著变化,12、20、24 h含量有所下降,差异不显著。鲜味氨基酸以茶氨酸为主,除谷氨酸含量在4 h时各处理间无显著差异外,茶氨酸、天冬氨酸及谷氨酸含量受乙烯诱导上调。甜味氨基酸中丝氨酸、脯氨酸含量不受乙烯调控,甘氨酸含量受乙烯诱导上调,丙氨酸含量不完全受乙烯调控。苦味氨基酸中组氨酸、酪氨酸、缬氨酸、异亮氨酸及亮氨酸含量受乙烯诱导上调。因此,萎凋期间喷施乙烯利溶液可以显著提高...     

Proteolysis of Mahon cheese as affected by acoustic-assisted brining

 Mahon cheeses were brined in the presence of an ultrasonic field and ripened during 75 days at 12  °C and 85% RH. Secondary proteolysis (water-soluble N, non-protein N, and free amino acids) was measured and compared to that obtained for cheeses conventionally brined. There were no differences in water-soluble and non-protein N attributable to the brining treatment. However, cheeses acoustically brined exhibited higher concentrations of free amino acids. The release of total free amino acids was more pronounced during the first 15 days of ripening for both types of brining treatments. The changes in proteolysis (free amino acids) during cheese ripening caused by acoustic-assisted brining are indicative of a higher extent of proteolysis and may also improve cheese flavor. Received: 13 March 2000     

Effect of glucose and glycine addition to cocoa mass before roasting on Maillard precursor consumption and pyrazine formation

Cocoa mass was treated with glucose and/or glycine before roasting. For glucose‐treated samples the reductions in reducing sugars and free amino acids were 41.1 and 56.9% respectively, whereas for glycine‐treated samples the respective reductions were 16.2 and 37.4%. The combination of glucose plus glycine treatment led to 39.0 and 48.9% reductions in reducing sugars and free amino acids respectively. For free amino acids the reductions in control and glucose‐treated samples were similar, 56.2 and 56.9% respectively, indicating that the added glucose did not significantly (P < 0.05) influence the free amino acid uptake. When glycine was added, the consumption of free amino acids (37.4%) was lower than for the control and glucose treatments. When glucose and glycine were added, the reduction in free amino acids (48.9%) was lower than for the control and glucose treatments but higher than for the glycine treatment. Apparently, glycine addition inhibited the reducing sugar and free amino acid consumption, whereas glucose addition enhanced the reducing sugar and free amino acid consumption. The glucose plus glycine treatment was similar to the glycine treatment in the contents of 2,5‐dimethyl‐ and trimethylpyrazines. For 2‐methylpyrazine the value for glucose‐treated samples was lower than for control and glycine‐treated samples but almost equal to the value for the glucose plus glycine treatment. It is concluded that the addition of glucose and/or glycine does not affect pyrazine production. © 2002 Society of Chemical Industry     

不同包装材料对腐乳质量的影响

该文研究了腐乳生产过程中,采用不同包装材料,对其游离氨基酸态氮、食盐、总酸、游离脂肪酸和可溶性无盐固形物含量的影响。实验结果表明,在保质期中,采用陶瓷和玻璃包装的腐乳,其质量优于采用塑料容器包装的腐乳,贮藏12个月无霉变,塑料制品包装的腐乳在12个月内有8%发生霉变。     

Influence of salt content and processing time on sensory characteristics of cooked "lacón"

The influence of salt content and processing time on the sensory properties of cooked "lacón" were determined. "Lacón" is a traditional dry-cured and ripened meat product made in the north-west of Spain from the fore leg of the pig, following a similar process to that of dry-cured ham. Six batches of "lacón" were salted with different amounts of salt (LS (3 days of salting), MS (4 days of salting) and HS (5 days of salting)) and ripened during two times (56 and 84 days of dry-ripening). Cured odour in all batches studied, red colour and rancid odour in MS and HS batches, flavour intensity in MS batch and fat yellowness, rancid flavour and hardness in the HS batch were significantly different with respect to the time of processing. Appearance, odour, flavour and texture were not significantly affected by the salt content (P>0.05). However, the saltiness score showed significant differences with respect to the salt levels in all studied batches (56 and 84 days of process). The principal component analysis showed that physicochemical traits were the most important ones concerning the quality of dry-cured "lacón" and offered a good separation of the mean samples according to the dry ripening days and salt level.     

Role of amino acids and glucose in development of burnt off-flavours in liver sausage during heat processing

The effect of amino acids and glucose on the development of burnt off-flavours was investigated in meat model systems for liver sausage. At a constant glucose concentration, the burnt flavour was intensified by addition of an amino acid mixture. In a batter without liver, the development of burnt off-flavours, similar to those in liver sausage, could be induced through addition of exogenous amino acids and glucose. The endogenous amino acids from liver reacted more intensely than the exogenous amino acids in the formation of the burnt flavour components. After heating batches with added exogenous amino acids and glucose the free amino acid contents (73–89% of initial content) were higher than in batches to which liver was added (42–53% of initial content). The differences in the free amino acid content of glycine, glutamic acid and threonine were linearly related to the intensity of burnt flavour. Results indicate that burnt flavour is due to the Maillard reaction.     

烟叶烘烤过程中蛋白质的降解及相关酶活性的变化

  目的  明确不同成熟度烤烟烘烤过程中蛋白质降解规律。  方法  以K326为材料,分析烟叶蛋白质、游离氨基酸含量,内肽酶、氨肽酶和羧肽酶活性变化情况。  结果  1）烘烤过程中,烟叶蛋白质含量持续下降;2）蛋白质的降解幅度与内肽酶活性密切相关。与尚熟烟叶相比,适熟和过熟烟叶在变黄前期内肽酶活性最高,蛋白质含量降解幅度大。3）游离氨基酸含量升高幅度与氨肽酶和羧肽酶活性有关。与适熟和过熟烟叶相比,尚熟烟叶在变黄后期,氨肽酶和羧肽酶活性较高。  结论  适当延长变黄期（36~38℃）烘烤时间,有利于保持烟叶内肽酶的活性,减少不同成熟度烘烤烟叶中蛋白质的含量。