The effect of starter cultures on proteolytic changes and amino acid content in fermented sausages

The main objective of this work was to examine the effects of using five types of commercial starter cultures in fermented sausages. During the fermentation stage, changes in proteolytic characteristics were observed in fermented sausages. Proteolytic activity was high in Ls_b + Sc:(Lactobacillus sakei + Staphylococcus carnosus) and Pp + Sx:(Pediococcus pentosaceus + Staphylococcus xylosus) starter-inoculated sausages during processing. Moreover, a slight increase in proteolytic activity was detected during storage in both these sausages. Sarcoplasmic and myofibrillar proteins were also affected by this starter culture addition, during the fermentation, ripening and intense proteolysis were observed in both the fermented sausages. The content of free amino acids was similar at the beginning of the fermentation stage for all the studied batches. However, the high differences in the content of free amino acids at the end of the process could be attributed to the starter culture activity.     

THE EFFECT OF PROCESSING METHOD AND STORAGE TIME ON CONSTITUENTS OF TURKISH SAUSAGES (SUCUK)

Turkish sausages ( sucuk ) were produced by two methods (heat processing and traditional). Their moisture, pH and water activity and lipid stability were studied during refrigerated storage (4  ±  2C/90 days). Heat-processed (Therm) sausages had higher moisture content, water activity and pH values, and low thiobarbituric acid (TBA) values, free fatty acids (FFA) and peroxide values. FFA content of Therm sausages increased from 2.5 to 5.5% during storage and was significantly ( P <  0.05) affected by storage time. In the case of traditionally produced (Trad) sausages, there was no significant impact of storage time, but their TBA value increased from 0.38 to 0.48 mg MA/kg, while only a slight increase was observed in Therm sausages, from 0.41 to 0.43 mg MA/kg. Storage time significantly ( P <  0.05) increased the TBA value of Trad sausages.

PRACTICAL APPLICATIONS

In the current study, the effect of processing methods (heat processing and traditional) and storage time on some constituents of Turkish sausages was studied. Processing method significantly affected proximate composition of the sausages. On the other hand, lipolytic and oxidative changes were observed in both heat-processed and traditional sausages during storage. This could be attributed to hydrolytic enzymes and residual oxygen in vacuum packs. In practical applications, low oxygen permeability packing materials or modified gas atmosphere packing are recommended for both processing methods.     

CHANGES IN BIOCHEMICAL AND MICROBIOLOGICAL CHARACTERISTICS OF TURKEY SUCUKS AS AFFECTED BY PROCESSING AND STARTER CULTURE UTILIZATION

The effects of starter culture and processing on the microbiological and biochemical characteristics of turkey sucuk were studied during production and storage. No differences ( P >  0.05) in proximate composition and salt contents were found in sucuks produced either by traditional or heated methodologies. Both traditional ( P <  0.05) and heat-processed ( P >  0.05) sucuks that incorporated S1 had the lowest pH value, and thus the highest titratable acidity. Heating process resulted in about 1 log unit reduction in yeast counts and 1.5 log unit reduction in lactic acid bacteria, total mesophilic aerobic bacteria and micrococci–staphylococci counts of all sucuks. Within the heat-processed sucuks, heating stage did not result in a significant change in nonprotein nitrogen contents ( P >  0.05). Changes in myofibrillar and sarcoplasmic protein were observed because of proteolytic changes during ripening, but, in general, no major difference was observed between sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns as a result of starter culture treatments.

PRACTICAL APPLICATIONS

Results of this study can be applied to improve some biochemical and microbiological characteristics of fermented turkey sausages. The findings showed that utilizing commercial starter culture mix, containing Lactobacillus sake, Staphylococcus carnosus and Staphylococcus xylosus II resulted in lower pH value with a higher titratable acidity than traditional style-fermented turkey sausages. These parameters are responsible for texture development and characteristic taste of fermented sausages.     

Destruction of Yersinia enterocolitica by Lactobacillus sake and Pediococcus acidilactici During Low-temperature Fermentation of Turkish Dry Sausage (sucuk)

Effects of starter cultures, Lactobacillus sake and Pediococcus acidilactici, on the survival of Yersinia enterocolitica were studied in Turkish dry sausage (sucuk). Inoculated Y. enterocolitica (approximately 5.0 log CFU/g) was eliminated completely in sausages by L. sake (about 7.0 log CFU/g) and P. acidilactici (about 7.0 log CFU/g) by the 3^rd d of fermentation but was not totally eliminated in natural fermentation without starter(s) after completion of 4 d of fermentation and 12 d drying (p < 0.05). pH decreased from 6.3 to about 4.7 in starter culture treatments and from 6.3 to 5.6 in natural fermentation after completion fermentation and drying (p < 0.05).     

Biochemical and sensory characteristics of traditional fermented sausages of Vallo di Diano (Southern Italy) as affected by the use of starter cultures

In this study, two strains of Staphylococcus xylosus isolated from traditional fermented sausages of Vallo di Diano (Southern Italy) were used in combination with an acidifying strain of Lactobacillus curvatus as starter culture for the production of fermented sausages. Two starter formulation were developed combining the proteolytic but not lipolytic (prt(+), lip(-)) S. xylosus CVS11 with the L. curvatus AVL3 (starter S1) and the S. xylosus FVS21 (prt(-), lip(+)) with the same strain of L. curvatus (starter S2). Proteolysis and lipolysis were observed during ripening by the increase in total free amino acids (FAA) and free fatty acids (FFA), respectively. Such activities were observed in both started and non started sausages (control). Moreover, the proteolytic and lipolytic activities were detected in products started by both formulations irrespective of the presence of such activities in the strains used. Therefore, it was not possible to conclude whether the effect of proteolysis and lipolysis during ripening of the started fermented sausages was due to the activity of the starter cultures or to the action of meat endogenous enzymes.     

复合发酵剂和天然香辛料对羊肉发酵香肠品质特性的影响

通过接种复合发酵剂和添加天然香辛料,采用相同的工艺条件生产4组羊肉发酵香肠,1组为对照组,2组为接种发酵剂组,3组为发酵剂+黑胡椒组,4组为发酵剂+黑胡椒+孜然组。分别测定4个组羊肉发酵香肠在发酵成熟和贮藏过程中pH值、Aw值、水分含量及微生物的变化。结果表明:在发酵结束(即第1.5天)时,试验组和对照组的pH值、Aw值和水分含量均呈下降趋势,试验组的pH值迅速降到最低值(即5.0以下),与对照组差异显著(P<0.05);在干燥成熟过程中,试验组的pH值、Aw值和水分含量迅速下降,添加香辛料组的pH值迅速回升,同时乳酸菌数维持在108cfu/g以上水平,高于对照组,细菌总数却低于对照组;在贮藏过程中,试验组的pH值和水分含量基本保持不变,且均低于对照组,而Aw值始终呈下降趋势,细菌总数小于对照组。上述结果表明,接种复合发酵剂和添加天然香辛料能迅速降低pH值、Aw值和水分含量,有效抑制腐败菌和致病菌的生长,提高羊肉发酵香肠的品质和保藏性。     

Reduction of biogenic amine formation using a negative amino acid-decarboxylase starter culture for fermentation of Fuet sausages

The ability of Lactobacillus sakei CTC494, a negative amino acid-decarboxylase starter culture, to reduce biogenic amine accumulation during sausage fermentation and storage at 4 and 19 degrees C was studied. The effect on the amine formation of the tyramine producer Lactobacillus curvatus CTC371, as a positive strain, was also examined in comparison to a spontaneous fermentation process without starter culture (control batch). The polyamines spermine, spermidine, and diaminopropane were not influenced by the ripening, and their levels slightly decreased in all the batches throughout the storage. Tyramine, cadaverine, and putrescine were the main amines formed during the ripening. The addition of starter culture resulted in a decrease on the biogenic amine formation, depending on the strain inoculated. A great reduction in tyramine content was achieved when L. sakei CTC494 was inoculated, whereas L. curvatus CTC371 only attenuated tyramine accumulation compared with the control batch. Both starters were able to significantly limit the production of putrescine and cadaverine, and they inhibited tryptamine and phenylethylamine formation by the wild microbial flora. Tyramine levels of the control sausages rose during the storage at both temperatures, whereas those of cadaverine only increased at 19 degrees C. On the contrary, sausages manufactured through the starter controlled fermentation did not show changes of amine contents during the storage. The addition of a proper selected starter culture is advisable to produce safer sausages with low contents of biogenic amines.     

Effect of Starter Culture Addition and Processing Method on Proteolysis and Texture Profile of Traditional Dry-Fermented Sausage Petrovská klobása

Proteolysis and texture profile of Petrovská klobása, made with addition of starter culture, were investigated during the ripening in different conditions (traditional versus industrial). Due to intensive degradation of myofibrillar proteins, significant increases of non-protein nitrogen and free amino acids nitrogen contents were registered in all samples. Similar proteolytic changes were found in both starter and non-starter sausages, but were more pronounced in samples ripened in industrial room. In relation to ripening conditions hardness and chewiness differed significantly, while starter culture had significant effect only on hardness of sausages processed in industrial room.     

Effects of starter cultures and additives on the quality of Turkish style sausage (sucuk)

The effects of starter cultures and additives on the quality of Turkish style sausage (sucuk) were investigated during ripening and storage. Total aerobic plate counts (aerobic bacteria), mould and yeast counts, pH, thiobarbituric acid (TBA) value and biogenic amine formation were followed. Aerobic bacteria increased (P<0.05) during the first 10 days of ripening to 9.26 log CFU/g and they decreased (P<0.05) to 5.70 at the end of the storage. Larger reductions (P<0.05) were observed in sausages made with high levels of potassium sorbate, nitrite and nitrate than in those containing low levels of additives and those without additives at the end of storage. During the first 3 days of ripening, the pH values of all sausages decreased (P<0.05) from 5.98 to about 4.53. Later, the pH values increased slowly, due to decomposition of acids to an approximately constant value of about 5.20. TBA values were higher (P<0.05) in the sausages made without additives than in the others. Sausages prepared using high concentration of additives had lower (P<0.05) concentrations of biogenic amines than the others. Addition of starter culture with high concentration of additives (nitrite, nitrate, α-tocopherol, ascorbic acid, potassium sorbate, potassium pyrophosphate and di-potassium hydrogenphosphate) decreased the formation of biogenic amine.     

Safety improvement and preservation of typical sensory qualities of traditional dry fermented sausages using autochthonous starter cultures

Traditional dry fermented sausages are manufactured without addition of starter cultures in small-scale processing units, their fermentation relying on indigenous microflora. Characterisation and control of these specific bacteria are essential for the sensory quality and the safety of the sausages. The aim of this study was to develop an autochthonous starter culture that improves safety while preserving the typical sensory characteristics of traditional sausages. An autochthonous starter composed of Lactobacillus sakei, Staphylococcus equorum and Staphylococcus succinus isolated from a traditional fermented sausage was developed. These strains were tested for their susceptibility to antibiotics and their production of biogenic amines. This starter was evaluated in situ at the French traditional processing unit where the strains had been isolated. Effects of the autochthonous starter were assessed by analysing the microbial, physico-chemical, biochemical and sensory characteristics of the sausages. Inoculation with the chosen species was confirmed using known species-specific PCR assays for L. sakei and S. equorum and a species-specific PCR assay developed in this study for S. succinus. Strains were monitored by pulse-field gel electrophoresis typing. Addition of autochthonous microbial starter cultures improved safety compared with the traditional natural fermentation of sausages, by inhibiting the pathogen Listeria monocytogenes, decreasing the level of biogenic amines and by limiting fatty acid and cholesterol oxidation. Moreover, autochthonous starter did not affect the typical sensory quality of the traditional sausages. This is the first time to our knowledge that selection, development and validation in situ of autochthonous starter cultures have been carried out, and also the first time that S. equorum together with S. succinus have been used as starter cultures for meat fermentation. Use of autochthonous starter cultures is an effective tool for limiting the formation of unsafe compounds in traditional sausage while preserving their original and specific sensory quality.     

Use of Fourier transform infrared spectroscopy to evaluate the proteolytic activity of Yarrowia lipolytica and its contribution to cheese ripening

The interaction between tyrosine-decarboxylase and proteolytic activities of a Lactobacillus curvatus and Staphylococcus xylosus, respectively, on biogenic amine production during the ripening and the storage of dry fermented sausages was investigated. Water content, pH, proteolysis parameters, microbial counts, and biogenic amine contents were monitored in spontaneously and starter fermented sausages. The use of proteolytic staphylococci as starter resulted in a higher content of non-protein nitrogen and total free amino acids. Tyramine was the main amine produced in all batches. However, tyrosine-decarboxylase activity of the L. curvatus starter strain was weak and yielded lower amounts of tyramine than those produced by the wild mioroflora in the control batch. Association between tyramine production and proteolysis could only be established in a defectively dried batch. Putrescine and cadaverine accumulation was efficiently reduced in the starter-mediated fermentation, in agreement with the lower development of enterobacteria. Phenylethylamine and tryptamine were only detected in the spontaneously fermented sausages, while histamine, spermine and spermidine did not vary during the ripening. Biogenic amine levels and related parameters showed significant changes during the storage of dry sausages depending on the temperature and the batch. As a general rule, changes in the pH, proteolysis, microbial counts, and biogenic amine contents were stronger at 19 degrees C than at 4 degrees C. The results suggest that refrigeration would be advisable for preventing further accumulation of biogenic amines during the storage of dry fermented sausages.     

The use of commercial starter cultures in the production of Turkish semi-dry fermented sausages

 The effects of three different starter cultures (Pediococcus acidilactici, Staphylococcus xylosus plus P. pentosaceus, S. carnosus plus Lactobacillus pentosus) were evaluated during the production of Turkish semi-dry fermented sausages. Sausages were studied during the fermentation phase, after heat processing and after drying for 24 h and 72 h. Chemical and organoleptical results indicated that in the processing of these semi-dry sausages a starter culture of P. acidilactici should be used. The use of this culture significantly reduced the pH, increased the lactic acid content and percentage of total heme pigments converted to the cured pigment and improved the development of the sausages' characteristics, i.e. color, appearance, flavor and general acceptability. Received: 20 February 1998 / Revised version: 7 May 1998     

Effect of autochthonous starter cultures isolated from Siahmazgi cheese on physicochemical,microbiological and volatile compound profiles and sensorial attributes of sucuk,a Turkish dry-fermented sausage

The effect of adding autochthonous starter cultures isolated from Siahmazgi cheese, on the physicochemical parameters and microbial counts of sucuk was investigated during the ripening period. SPME–GC/MS was used in volatile compound analysis and a trained group of panelists carried out sensory analysis of the final product. After preliminary screening, three strains of Lactobacillus plantarum, which possess desirable technological properties, were used to prepare three starter cultures: LBP7, LBP10 and LBP14. The addition of LBP7 and LBP14 starter cultures had a significant effect (P < 0.05) on lightness, leading to higher L values compared to control sausages during the ripening period. Both LBP7 and LBP14 sausages showed higher counts of lactic acid bacteria, lower growth of Enterobacteriaceae and Gram-positive catalase-positive cocci and greatly lowered the pH value compared to control sausages throughout the ripening process. At the end of the ripening process, lactic acid bacteria counts were affected (P < 0.05) by the addition of starter culture since higher counts were observed in sausages prepared with LBP7 (9.14 log CFU/g) and LBP14 (8.96 log CFU/g) batches. The decrease of water activity during the ripening of sausages was not affected by the various starters. The texture profiles of all sausages were similar except for LBP10, which showed lower hardness and gumminess during ripening. Under the conditions of the study, volatile compounds were mainly from spices, and no marked differences were found among inoculated sausages. However, sensory evaluation revealed that most of the sensory attributes were scored higher for inoculated sausages than for the control ones. Therefore, LBP7 and LBP14 could be promising candidates for inclusion as starter cultures for the manufacture of sucuk.     

Microbiological safety of traditional and starter-mediated processes for the manufacture of Italian dry sausage

Microbiological changes occurring during the commercial manufacture of Italian dry sausages (Genoa and salametti) were studied in two urban Canadian centres over a 5 month period. A comparison was made between 6 plants which used bacterial starter cultures and 4 plants where more traditional processes (without starters) were used. A total of 600 samples of raw, fermented and finished products were tested for the presence of coliforms, salmonellae, staphylococci, streptococci, the rate of pH reduction and final water activity (aW). Numbers of total bacteria peaked earlier and were significantly higher in sausages at the fermentation stage produced with starter cultures than in those traditionally manufactured. This corresponded with a more rapid drop in pH of the starter-inoculated products. Staphylococci and streptococci were significantly higher in starter-fermented Genoa sausages at the fermentation stage, but no significant differences were seen in the microbiological content or aW of mature finished sausages manufactured by the two different techniques. Of 128 randomly chosen isolates of coagulase-positive staphylococci, 34.4% were enterotoxin producers and 80% of these produced type A toxin. Enterotoxigenic staphylococci were found in 2 different samples of finished salametti and one sample of finished Genoa made with starter cultures and in one sample of finished Genoa made without added culture. Total numbers of staphylococci in these samples were not greater than 500/g. No correlation between the method of manufacture and presence of enterotoxigenic staphylococci could be made. Five subsamples from one lot of raw Genoa were the only samples positive for Salmonella during this study. Results indicated that low temperature traditional fermentations can yield products which are as safe as those produced by the higher temperature starter-controlled process. One of the most important elements in the traditional process was believed to be the selection and use of raw materials of the highest possible quality.     

Effects of Lactobacillus plantarum and Staphylococcus xylosus on the Quality Characteristics of Dry Fermented Sausage "Sucuk"

ABSTRACT:  The effect of starter culture, containing the strains Lactobacillus plantarum GM77 and Staphylococcus xylosus GM92, isolated from traditional sucuk, on the quality characteristics of dry fermented sausage (sucuk) during ripening period was studied. The microbiological (lactic acid bacteria, Micrococcus/Staphylococcus , Enterobacteriaceae) and physicochemical (pH, a_w, NPN, TBARS) properties and volatile compounds, extracted by solid phase microextraction and analyzed by gas chromatography-mass spectrometry, were determined in sucuk samples with starter culture and in the control group (without starter culture). Both starter culture and ripening period had significant effects on lactic acid bacteria, Micrococcus/Staphylococcus counts, and pH and a_w values ( P  < 0.01). The highest value for nonprotein nitrogen (NPN) was observed to occur in samples with starter culture ( P  < 0.05). TBARS value increased during ripening, the highest value was determined at 14th day in samples with starter culture. Among volatile compounds, terpenes were the major group in sucuk. Other volatile compounds (aldehydes, ketones, sulfur compounds, acids, esters, aliphatic hydrocarbons, aromatic hydrocarbons, and alcohols) can be found in fewer amounts. While the use of starter culture was affecting a few compounds, ripening period had affected most of the compounds.     

功能性发酵剂对发酵香肠氧化稳定性及挥发性风味物质的影响

以功能性发酵剂植物乳杆菌（Lactobacillus plantarum CD101）和模仿葡萄球菌（Staphylococcus simulans NJ201）为混合发酵剂制作发酵香肠,同时以商业发酵剂和自然发酵为对照。结合过氧化值（peroxide value,POV）、硫代巴比妥酸反应物（thiobarbituric reactive substances,TBARS）值、羰基值、巯基值和挥发性风味物质等探究功能性发酵剂对发酵香肠脂肪和蛋白质氧化以及风味的影响。结果表明：在香肠加工和贮藏过程中,随着时间的延长,不同处理组香肠的POV、TBARS值、羰基值总体呈现上升趋势,巯基值呈现下降趋势。接种功能性发酵剂制作发酵香肠在一定程度上能够抑制产品的脂肪氧化和蛋白氧化,显著提升挥发性风味物质种类（P＜0.05）,其抑制脂肪氧化的能力显著优于商业发酵剂（P＜0.05）,抑制蛋白氧化的能力与商业发酵剂效果相当。     

Fermentation of bighead carp (Aristichthys nobilis) surimi and the characteristics of fermented bighead carp surimi products

BACKGROUND: To improve the properties and functionality of bighead carp (Aristichthys nobilis) muscle, bighead carp surimi was inoculated with combinations of Saccharomyces cerevisiae, Lactobacillus and Monascus and fermented, and then bighead carp sausages were prepared. The characteristics of fermented bighead carp surimi and sausages were investigated. RESULTS: During the 24 h fermentation at 30 °C, bighead carp surimi inoculated with mixed starter cultures resulted in a rapid decrease of pH, and suppression in the growth of Enterobacteria. The bighead carp sausages exhibited better colour, taste, flavour and appearance than the control (P < 0.05). The changes in non‐protein nitrogen (NPN), free amino acid in the fermented bighead carp surimi and SDS–PAGE patterns indicated that severe hydrolysis of muscle protein had occurred during fermentation. The sausages inoculated with the mixed starter cultures gained higher scores for flavour and overall acceptability than the control. CONCLUSIONS: The characteristic of fermented bighead carp surimi and sausages can be significantly improved by fermentation with the combinations of yeast, Lactobacillus and Monascus. Copyright © 2008 Society of Chemical Industry     

FORMATIONS OF BIOGENIC AMINES IN TURKISH STYLE SAUSAGE DURING RIPENING AND STORAGE PERIODS

Changes of biogenic amine (histamine, β‐phenylethylamine, serotonin, spermine, spermidine, cadaverine, putresine, 1,7‐diamino heptane, tryptamine and tyramine) concentrations and pH values of Turkish style sausages were investigated during ripening and storage. Cadaverine and β‐phenylethylamine were not detected during the ripening and storage periods for all sausage types. Storage time had a significant (P<0.05) effect on the formation of all biogenic amines. Starter culture usage decreased (P<0.05) biogenic amine formation. Histamine levels were higher in sausages prepared without additives and lower in the sausages made with starter culture than the sausages made without starter culture. There was no effect of starter culture and additives on formation of tryptamine, 1,7‐diamino heptane, serotonin or spermidine. Starter culture and additives affected (P<0.05) the formation histamine, putrescine and spermine. pH values of sausages sharply decreased (P< 0.05) from 5.98 to about 4.53 during 3 days of ripening. After that, pH values increased slowly and were nearly constant during the storage period.     

Influence of starter cultures on the generation of antioxidant nitrogen compounds in Iberian dry‐fermented sausages

The purpose of this work was to investigate the influence of a starter culture on the generation of nitrogen compounds with antioxidant activity during the ripening of Iberian dry‐fermented sausages. Starters P200S34 (P. acidilactici M200 and S. vitulus RS34) and P198S34 (P. acidilactici MS198 and S. vitulus RS34) were used to make the Iberian dry‐fermented sausages ‘salchichón’ and chorizo; then, the physicochemical and microbial properties were determined during the ripening process. The oxygen radical absorbance capacity (ORAC) assay was employed to evaluate the antioxidant potential of the nitrogen extracts obtained during ripening of the sausages. This activity was correlated with the most relevant compounds detected by HPLC‐ESI‐MS in the final ripened extracts. Although a relevant part of the antioxidant activity was attributed to the predominant natural nitrogen fraction, the microbial population found in fermented sausages and the fermentation conditions significantly influenced the low molecular weight nitrogen profile and antioxidant activities. Inoculation with the starter culture P200S34 increased free amino acids and amines, such as methionine and tyramine, but other nitrogen compounds also increased the antioxidant activity of the low molecular weight nitrogen extracts. Thus, these starter cultures in Iberian sausages can contribute to delaying oxidative changes during storage.     

Assessment of high hydrostatic pressure and starter culture on the quality properties of low-acid fermented sausages

The addition of starter culture and high pressure processing after ripening improved the microbial quality of low-acid fermented sausages (fuet and chorizo). The use of Lactobacillus sakei CTC6626 and Staphylococcus xylosus CTC6013 as starter culture significantly reduced Enterobacteriaceae and Enterococcus levels in the finished sausages. Moreover, the addition of starter culture produced sausages of similar quality to traditional low-acid fermented sausages. Slightly lower pH values and higher cohesiveness were obtained for both fuet and chorizo with starter culture. Sensory analysis showed no differences between lots of chorizo whereas starter fuet was more acid and gummy. High pressure induced an additional reduction of Enterobacteriaceae in non-starter sausages. An increase of textural properties was observed after pressurization. No other differences were observed between non-treated and pressurized sausages.