Effect of fermentation with mixed starter cultures on biogenic amines in bighead carp surimi

The effects of one group mixed starter cultures, combined with Lactobacillus casei, Streptococcus lactis, Saccharomyces cerevisiae Hansen, and Monascus anka, and a batch without starter as control on biogenic amines accumulation in the bighead carp surimi during fermentation were investigated. Determination of five different biogenic amines was carried out by reverse‐phase high‐performance liquid chromatography with UV detection. Results showed that mixed starter cultures decreased the pH quickly from initial pH value of 6.4 to 5.2, inhibited the growth of contaminant microorganisms, such as Enterobacteriaceae and Pseudomonas, present in the raw materials, and suppressed the accumulation of histamine, tyramines, spermine and spermidine. Fermentation inoculated with combination of Lactobacillus casei, Streptococcus lactis, Saccharomyces cerevisiae Hansen, and Monascus anka with negative‐decarboxylase activity may prevent biogenic amine formation in the fermented ssurimi and improve hygienic quality of fermented surimi.     

Improvement of the quality and abatement of the biogenic amines of grass carp muscles by fermentation using mixed cultures

BACKGROUND: To improve the quality of processed grass carp (Ctenopharyngodon idellus) products and control the accumulation of hazardous substances therein, minced grass carp slices were salted for 6 h at room temperature and then inoculated with mixed starter cultures of Lactobacillus casei, Streptococcus lactis, Saccharomyces cerevisiae Hansen and Monascus anka and fermented for 12 h at 30 °C. The changes in some characteristics and biogenic amine contents of the fermented muscles were investigated. RESULTS: During the 12 h fermentation at 30 °C, muscles inoculated with mixed starter cultures showed a rapid decrease in pH from 6.0 to 5.1 and suppression of the growth of enterobacteria and pseudomonads. The fermented muscles exhibited better colour, appearance, flavour and overall acceptability than the control (P < 0.05). The changes in non‐protein nitrogen and free amino acid contents of the fermented muscles and in their sodium dodecyl sulfate polyacrylamide gel electrophoresis profiles indicated that severe hydrolysis of muscle proteins occurred during fermentation. The accumulation of biogenic amines in the muscles was efficiently reduced by fermentation with mixed starter cultures. CONCLUSION: Fermentation with mixed starter cultures of L. casei, S. lactis, S. cerevisiae Hansen and M. anka significantly improved the characteristics of grass carp muscles and controlled the accumulation of biogenic amines. Copyright © 2009 Society of Chemical Industry     

Characterization of fermented silver carp sausages inoculated with mixed starter culture

To improve the characteristics and functionality, and increase the use of fish muscle, three groups mixed starter cultures (group one: Lactobacillus plantarum-15, Staphylococcus xylosus-12 and Pediococcus pentosaceus-ATCC33316 [S-PXP]; group two: Lactobacillus planatrum-15, Staphylococcus xylosus-12 and Lactobacillus casei subs casei-1.001 [S-PXC]; and group three: Staphylococcus xylosus-12, Lactobacillus casei subsp. casei-1.001 and P. pentosaceus-ATCC33316 [S-XCP]) were inoculated in minced silver carp muscle to produce a fermented fish product. During the 48 h fermentation at 30 °C, silver carp muscle inoculated with mixed starter cultures resulted in a rapid pH decrease, suppression in the increase of thiobarturic acid (TBARS) values, total volatile base nitrogen (TVB-N), trimethylamine (TMA), and the growth of spoilage bacteria and pathogens, and had higher whiteness than the control (without any starter) (P<0.05). The changes in SDS-PAGE indicated extensive hydrolysis of muscle protein occurred during fermentation. This study showed that the mixed starter cultures could substantially improve the flavor, digestibility, and nutritional value of the silver carp muscle.     

Utilisation possibility of Enterococcus faecalis isolates from neonate's faeces for production of fermented sausages as starter cultures

This study aimed at assessing the utilisation possibility of Enterococcus faecalis as starter cultures for production of fermented sausages. The E. faecalis isolates isolated from faeces samples of 5‐day‐old healthy newborn babies, identified by 16SrDNA sequencing and free from virulence determinants as proven by genomic sequencing, were used for fermented sausages production as starter culture. Six treatments (four inoculated with four different E. faecalis isolates, one with commercial starter culture and one control) were prepared. The inoculation with E. faecalis showed a stronger acidification compared with the control. The treatments with E. faecalis showed significantly higher flavour and acceptability scores in comparison with the control or commercial starter culture. Both of commercial starter culture and E. faecalis were positive with tyramine. Apart from the amine production, the inoculation with E. faecalis generally improved the acidifying activity and flavour of the products, suggesting their application potentials in fermented sausages production as starter cultures.     

Effects of Inoculation of Commercial Starter Cultures on the Quality and Histamine Accumulation in Fermented Sausages

To meet the requirements of high‐quality safe products, starter cultures are used to produce fermented sausages. The effects of 3 commercial starter cultures, namely SM‐194, T‐SPX, and SM‐181, on histamine accumulation and quality parameters including microbial quality, pH, water activity, and total volatile base nitrogen, as well as the color and texture properties, were evaluated during the fermentation and ripening of fermented sausages. Although initial counts of Escherichia coli, Enterobacteriaceae, and Pseudomonas were similar in the 4 batches, the growth of these microorganisms was significantly inhibited (P < 0.05) in batches SM‐194, T‐SPX, and SM‐181 throughout the fermentation and ripening period. The counts of E. coli, Enterobacteriaceae, and Pseudomonas increased to maximum levels of 3.89, 4.41, and 5.15 log₁₀ colony forming units/g in the control sausages, respectively. At the end of ripening, the levels of histamine were 8.85, 0.32, 7.82, and 3.18 mg/kg for batches C, SM‐194, T‐SPX, and SM‐181, respectively. The results revealed that commercial starter cultures, particularly starter cultures SM‐194 and SM‐181, made a great contribution to histamine reduction. In addition, batches inoculated with starter cultures showed a stronger acidification and lower level of total volatile base nitrogen than the control sample during production (P < 0.05). In conclusion, it seems that the inoculation of commercial starter cultures, particularly starter cultures SM‐194 and SM‐181, contributes to improving microbial quality, hygienic quality and food safety of fermented sausages.     

Characterization of Proteolytic Effect of Lactic Acid Bacteria Starter Cultures on Thai Fermented Sausages

The objective of this study was to investigate the effect of starter culture addition on proteolysis of Thai fermented sausages. Sausages inoculated with six different external starter cultures—Pediococcus pentosaceous, Pediococcus acidilactici, Weissella cibaria, Lactobacillus plantarum, Lactobacillus pentosus, and Lactobacillus sakei—were compared with naturally fermented sausages. The results of microbiological analysis indicated that the dominance of lactic acid bacteria (LAB) could inhibit the growth of pathogens and spoilage. Proteolysis was observed during fermentation by the reduction of myofibrillar and sarcoplasmic proteins and the increase in nonprotein nitrogen (NPN) and total free amino acids. The highest increase in concentration of NPN and free amino acids was obtained from sausages inoculated with LAB. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) showed a similar pattern of proteolysis of sarcoplasmic proteins in all sausages, while that of the inoculated sausages with L. plantarum, L. pentsus, and L. sakei exhibited increased degradation of myofibrillar protein bands at 200 and 45 kDa.     

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.     

Use of molecular methods to characterize the bacterial community and to monitor different native starter cultures throughout the ripening of Galician chorizo

The development of Lactobacillus and Staphylococcus strains used as starter cultures throughout the ripening of Galician chorizo, a traditional dry fermented sausage from the north-west of Spain, was monitored combining different molecular-based techniques. The bacterial diversity occurring in the inoculated sausages at the beginning and the end of the ripening was also studied and compared to the indigenous population in an uninoculated control batch.     

Bacterial community succession and biogenic amine changes during fermentation of fish-chili paste inoculated with different commercial starter cultures

The microbial dynamics, diversity and succession characterisation during the fermentation of fish-chili paste, with and without starter cultures by high-throughput sequencing, were investigated to identify the relationship between microbial composition and the accumulation of biogenic amines. Results showed that Firmicutes was the predominant phylum, and Lactobacillus and Weissella were the main genera during fermentation process, regardless of the inclusion of starter cultures. Compared with naturally fermented samples (NS; no starter cultures), Pediococcus and Staphylococcus flourished in samples inoculated with SBM-52 starter culture (Staphylococcus xylosus + Staphylococcus carnosus + Pediococcus pentosaceus + Pediococcus acidilactici). Staphylococcus also accounted for a larger proportion in samples inoculated with WBX-43 starter culture (Staphylococcus xylosus + Staphylococcus carnosus). Furthermore, Enterobacter was inhibited in inoculated samples. Unweighted pair-group method with arithmetic means and principal component analyses revealed that microbiota structures were different among NS, SBM and WBX samples. In case of biogenic amines, the concentration of putrescine, cadaverine and tyramine increased to higher levels in NS samples compared with inoculated samples. Our work also investigated the relationship between core bacterial communities and biogenic amines. This study provided an improved understanding for the effects of starter cultures on the microbial community and the quality of fish-chili paste.     

Effect of selected strains of Debaryomyces hansenii on the volatile compound production of dry fermented sausage “salchichón”

Different biotypes of Debaryomyces hansenii, characterized by mitochondrial DNA (mtDNA) restriction analysis, were inoculated in dry fermented sausages to evaluate their influence as single starter culture on volatile compound generation throughout the ripening process. Similar evolution of physicochemical parameters and microbial population was observed in both uninoculated and inoculated sausages. The tested biotypes modified the volatile compound profile of sausages specially in esters, branched alcohols and aldehydes. The biotype of D. hansenii with the E mtDNA restriction pattern is the most suitable to be used as starter culture since it produced volatile compounds involved in flavour development of dry-cured meat products such as 3-methylbutanol, 3-methylbutanal and 2-propanone. Moreover, the use of D. hansenii strains with the B, C2 and E mtDNA restriction patterns, as a mixed starter culture, should be also considered to generate low amount of sulphur compounds in dry-cured meat products.     

Characterization of flavour and volatile compounds of fermented squid using electronic nose and HPMS in combination with GC-MS

The present study investigated the flavour characteristics and key volatile components of fermented squid (Dosidicus gigas) as well as their formation mechanism by three different starter cultures of lactic acid bacteria (LAB) and natural squid. The sensory assessment, volatile compounds, as well as the amino acids of the mixed fermented squid were detected using pickled brine as samples of fermented squid. A total 88 types of volatile compounds were found in four samples by 0–72 h of fermentation. After fermentation by LAB, the volatile compounds of squid showed that the fermented squid with different started cultures differed mainly in terms of numbers of alcohols, ketones, and esters. Furthermore, the amounts of volatile compounds among three starter cultures were also obviously different. Gas Chromatography-Mass Spectrometer (GC-MS) analysis indicated that the mixed fermentation process contained relatively higher (+)-limonene (11.71%) (lemon scent) and 3-hydroxy-2-butanone (5.72%) (creamy taste). These compounds were not detected in the control group. The main flavour compounds of nonanal and 2, 4-decadienal were gradually reduced. Meanwhile, 2, 3-butanedione, 2-heptanone, 2-pentylfuran, and nonanoic acid are the key flavour components. The total principal component is 94.01%, which were clarified by electronic nose in combination with principal components analysis. After mixed fermentation by LAB, the total amino acid (TAA) content reached 85.36 g/100 g, which was almost twice the amount of the control, especially in aspartate, glutamate, and threonine. Our results indicated that LAB has obvious effects on deodorization and flavour promotion during the fermentation of squid. This study provided an important basis for the further development and utilization of squid.     

Multiple-strain Lactobacillus-fermented soymilk with antioxidant capacity and delicate flavour

Soymilk is a high-quality fermentation substrate, but its flavour is not accepted by many consumers. In this study, soymilk was fermented under different fermentation conditions by different Lactobacillus strains. The antioxidant activity of the soymilk was explored using TOPSIS and the changes in flavour during fermentation were investigated by e-nose using PCA. The results of TOPSIS showed that Lactobacillus plantarum (LP) ranked first (0.734) and L. acidophilus (LA) ranked second (0.651). In addition, L. bulgaricus (LB) was the only strain that could increase ABTS antioxidant activity (by around 11%). The LP, LA and LB mixed fermentation sample (w/w/w, 1 : 1 : 1) provided 10 new kinds of flavour compounds and stronger antioxidant ability than the unfermented sample. In addition, storage at 4 °C after fermentation elicited beneficial changes in the content of key flavour substances. Therefore, multiple-strain Lactobacillus-fermentation of soymilk not only improved the antioxidant activity but also provided a delicate flavour.     

Proteomic and peptidomic insights on myofibrillar protein hydrolysis in a sausage model during fermentation with autochthonous starter cultures

The hydrolysis of myofibrillar proteins during fermentation of sausage models by an autochthonous starter culture was investigated. In order to provide a whole map of the generated products, proteomic and peptidomic were used and complemented with the amino acid profile. Beaker sausages (BS) were used as models which were inoculated or not with Lactobacillus curvatus CRL705 and Staphylococcus vitulinus GV318 as starter cultures. The hydrolysis of actin, myosin light chain 1/3 (MLC 1/3), myosin regulatory light chain-2 (MRLC-2) and myosin heavy chain (MHC) was evidenced by two-dimensional gel electrophoresis (2-DE). In addition, a total of 33 peptides arisen from troponin T, MRLC-2 and particularly from actin were identified by LC–MS/MS. These results showed that the starter culture significantly enhanced the proteolysis of the proteins named above, even when the endogenous enzymes induced a clear breakdown. L. curvatus CRL705 highly enriched both peptide pattern and amino acid concentrations. When the autochthonous starter culture was inoculated, although proteolysis was remarkably reinforced, a reduction in peptide and amino acid composition was observed. Regarding actin primary structure, three regions of this protein were highly susceptible to degradation by the starter culture. Additionally, the essential role of exopeptidases – from meat and bacteria – in diversity of actin peptides during fermentation was shown. This study improved the knowledge of the proteolysis of myofibrillar proteins and the involved enzymes, as well as, completed the previously reported degradation of sarcoplasmic proteins by the same autochthonous starter culture. The singular peptides and amino acids pattern generated might contribute to the uniqueness of produced fermented sausages while they may be used as quality markers.     

Mixed starter cultures to control biogenic amine production in dry fermented sausages

Several combinations of an amine-negative Lactobacillus sakei strain, along with proteolytic Staphylococcus carnosus or Staphylococcus xylosus strains, were used to study the influence of mixed starter cultures on biogenic amine production during the manufacture of dry fermented sausages. Changes in pH, water content, proteolysis, microbial counts, and biogenic amine contents were simultaneously examined in a spontaneously fermented batch and in three mixed starter-mediated batches. A double-controlled microbial charge initially inoculated as mixed starter culture of L. sakei and Staphylococcus spp. (all amine-negative strains) drastically reduced tyramine, cadaverine, and putrescine accumulation. No production of other aromatic amines such as histamine, phenylethylamine, or tryptamine was observed in any batch. The polyamines, spermine and spermidine, were found in raw materials and their levels decreased slightly in the spontaneously fermented batch. No correlation between proteolysis and biogenic amine production was observed. The use of proper technological conditions favoring starter development and the use of the raw materials with good hygienic quality make it possible to produce fermented sausages nearly free of biogenic amines.     

Comparing the differences of characteristic flavour between natural maturation and starter culture for Mucor‐type Douchi

In this research, effects of maturation times on volatiles of Mucor‐type Douchi were investigated. Results showed that 2‐year matured sample exhibited higher concentrations of characteristic flavours including alcohols, esters, phenols, ketones and pyrazines. Additionally, effects of starter cultures addition on volatiles were investigated, and results demonstrated that inoculated fermented samples presented higher amounts of aldehydes, ketones, phenols and pyrazines compared with that of naturally fermented sample. Analysis of odour activity value (OAV) suggested that increased OAVs of eight compounds that mainly contributed to the flavour of Douchi ranged from onefold to 6.5‐fold in the Tetragenococcus halophilus, Zygosaccharomyces rouxii and Candida versatilis co‐cultured sample. Furtherly, by comparing the characteristic flavour of co‐cultured and 2‐year matured samples, no significant difference was observed in the OAV of flavour except rose‐like and soapy flavour, which indicated that co‐culture of the three strains during Douchi fermentation may facilitate the formation of flavours and shorten the fermentation process.     

Impact of starter cultures and fermentation techniques on the volatile aroma and sensory profile of chocolate

The sensory quality of chocolate is widely determined by the qualitative and quantitative composition of volatile compounds resulting from microbial metabolism during fermentation, and Maillard reactions taking place during drying, roasting and conching. The influence of applying mixed starter cultures on the formation of flavour precursors, composition of volatile aroma compounds and sensory profile was investigated in cocoa inoculated with cultures encompassing a highly aromatic strain of Pichia kluyveri or a pectinolytic strain of Kluyveromyces marxianus, and compared to commercially fermented heap and tray cocoa. Although only minor differences in the concentration of free amino acids and reducing sugars was measured, identification and quantification by dynamic headspace gas chromatography–mass spectrometry (HS/GC–MS) revealed pronounced differences in the composition of volatiles in roasted cocoa liquors and finished chocolates. 19 of the 56 volatile compounds identified in the chocolates were found in significantly higher amounts in the tray fermented sample, whilst significantly higher amounts of 2-methoxyphenol was measured in the two inoculated chocolates. The P. kluyveri inoculated chocolate was characterized by a significantly higher concentration of phenylacetaldehyde and the K. marxianus inoculated chocolate by significantly higher amounts of benzyl alcohol, phenethyl alcohol, benzyl acetate and phenethyl acetate compared to a spontaneously fermented control. Sensory profiling described the heap and tray fermented chocolates as sweet with cocoa and caramel flavours, whilst the inoculated chocolates were characterized as fruity, acid and bitter with berry, yoghurt and balsamic flavours. The choice of fermentation technique had the greatest overall impact on the volatile aroma and sensory profile, but whilst the application of starter cultures did affect the volatile aroma profile, differences were too small to significantly change consumer perception of the chocolates as compared to a spontaneously fermented control.     

Improvement of flavour compound synthesis in a mixed culture system at high temperature by solid‐state fermentation

In order to enhance the synthesis of flavour compounds in solid‐state fermentation (SSF) at a high temperature, Bacillus subtilis XJ‐013 and Saccharomyces cerevisiae Z‐06 were used as a mixed culture with Monascus HQ‐3. The culture temperature was enhanced from 37°C to 56°C by the synergetic effect, and the synthesis of esterase was enhanced from 85.43 U/g to 129.65 U/g in the mixed culture system (over 50% higher than that of the culture using a single strain). This resulted in the synthesis of favourable flavour compounds in the solid‐state fermentation. These results signified that a mixed culture of Monascus and S. cerevisiae was favourable for enzyme production. The mixed culture of Monascus and B. subtilis resulted in a high culture temperature that promoted flavour compound synthesis in the solid‐state fermentation system dramatically. These results present a model to explain the synergetic effects between the fungus and the Bacillus in the solid‐state fermentation. Copyright © 2015 The Institute of Brewing & Distilling     

Type of bacterial starter culture,aging and fermentation effects on some characteristics of inoculated beef sausages

Beef sausage mixes were inoculated with either Pediococcus acidilactici with Staphylococcus xylosus or P. acidilactici with S. carnosus, subdivided and then held for 0, 24, 48 or 72 h at 8–10 °C prior to fermentation. After aging (pre-fermentation holding), the mixes were fermented for 16 h ending at 41 °C. Moisture, protein and fat contents of all sausage mixes did not differ due to holding effects over all starter cultures. The pH of mixes followed the same pattern for all mixes, declining (p < 0.05) from approximately 5.8 to pH 5.2–5.3 at 72 h aging and to 4.4–4.5 after fermentation. Total acidity of the mixes followed an inverse pattern to pH, increasing (p < 0.05) after fermentation although there was no effect due to type of starter culture. Aging had no effect on nonprotein nitrogen (NPN) content as ΔNPN among all cultures. After fermentation, however, sausages held 72 h and inoculated with S. carnosus had higher NPN contents compared to P. acidilactici alone (p < 0.05) and with S. xylosus (p < 0.10). The same effects of starter cultures on changes in total amino acid concentration were observed. Concentrations of individual amino acids showed increases depending on pre-fermentation aging period (0 h versus 72 h) followed by fermentation.     

商业发酵剂对发酵香肠成熟过程中理化性质的影响

采用SBM-52、SHI-59、VBM-60复合型商业发酵剂生产发酵香肠,以不添加发酵剂作为对照组,在成熟第0、1、4、7、10天检测4 组香肠的pH值、非蛋白氮（non-protein nitrogen,NPN）含量、酸价、亚硝酸盐含量、水分活度（water activity,aw）、色差等理化指标的变化,考察不同商业发酵剂对发酵香肠成熟过程中理化品质的影响。结果表明：成熟终点时,4 组发酵香肠的pH值均小于5.0,亚硝酸盐含量低于30 mg/kg,充分保证了发酵香肠的安全性;NPN含量、aw和失重率的测定结果表明,添加3 种商业发酵剂均有助于发酵香肠中蛋白质的分解,加快成熟进程;SBM-52和VBM-60组香肠的酸价低于对照组,表明SBM-52和VBM-60具有较好的抑制脂肪氧化的作用。     

Physical,chemical and microbiological characteristics of fermented surimi with Actinomucor elegans

The physical, chemical and microbiological changes of surimi during surimi pehtze preparation stage, with Actinomucor elegans XH-22 (A. elegans XH-22) as fermentation starter, were investigated. Enzyme assay showed that A. elegans XH-22 could continuously produce protease and α-amylase on surimi surface. Texture profile analysis (TPA) indicated that fermentation for 48 h decreased pH and moisture content and modified the texture properties of surimi. Increases in amino nitrogen and trichloroacetic acid-soluble nitrogen (TCA-N) contents revealed obvious surimi protein degradation, which was further confirmed by size-exclusion HPLC analysis of surimi water soluble protein. Fermentation significantly increased the content of free amino acids and enriched special amino acids such as histidine, phenylalanine and glutamic acid, which could influence the nutritional and sensory properties of fermented surimi product. Considerable levels of aerobic plate count, spores and LAB were detected during the fermentation process. It is thus, from a public health point of view, advisable that the fermentation time in the first stage be controlled to be within 36 h. This research gives a new clue for improving the utilization of low-value fish.