Tyramine production of technological important strains of <Emphasis Type="Italic">Lactobacillus</Emphasis>, <Emphasis Type="Italic">Lactococcus</Emphasis> and <Emphasis Type="Italic">Streptococcus</Emphasis>

The aim of this paper was to study the biogenic amines (histamine, tyramine, putrescine, cadaverine, agmatine, spermine and spermidine) production of selected technological important lactic acid bacteria (strains of the genera Lactococcus, Lactobacillus and Streptococcus). Three methods (ion-exchange chromatography (IEC), PCR and cultivation method with pH indicator) were used. Within the 39 strains of lactic acid bacteria tested, the production of tyramine (formed by tyrosine decarboxylase) was detected in eight strains (3 strains of Lactococcus lactis subsp. lactis, three strains of Lactococcus lactis subsp. cremoris, 1 strain of Streptococcus thermophilus and 1 strain of Lactobacillus delbrueckii subsp. bulgaricus). The other tested biogenic amines were not detected. Cultivation in decarboxylation broth seems to be the least accurate method for the detection of biogenic amines due to enhanced risk of false-positive reactions. Therefore, in order to detect bacteria producing biogenic amines, the combination of PCR and chromatographic methods (e.g. IEC) can be recommended.     

Selection of LAB strains for fermented red beet juice production

Authentic LAB strains were investigated for their lactic acid, H₂O₂, and biogenic amine production, and their effect on red beet betalains. There was no significant difference between the strains in respect of cell propagation in beet juice. Lb. curvatus 2770 provided the best pH drop. None of the investigated LAB excreted either histamine or tyramine, but in the spontaneously fermented juice both amines were present and the total amine level was high. H₂O₂ synthesis was similar and not higher than 1 mg l^–1. The betalain content showed greatest reduction in the case of spontaneous fermentation and similar values for the LAB strains.     

Reduced biogenic amine contents in sauerkraut via addition of selected lactic acid bacteria

The contents of free amino acids and biogenic amines in spontaneously fermented sauerkraut, inoculated or not with specific lactic acid bacterium strains, were monitored throughout 45 d of storage. The strains tested were Lactobacillus plantarum 2142, Lactobacillus casei subsp. casei 2763 and Lactobacillus curvatus 2771. In both the control and the experiments, the total amino acid contents increased with time – and the predominant ones were aspartic acid, arginine and glutamic acid. However, upon inoculation with either strain, the total biogenic amine contents remained considerably lower than those of the control (especially in the cases of L. casei subsp. casei and L. curvatus); every single biogenic amine was actually below the 100 ppm-threshold. The dominant biogenic amines in the control were putrescine – and tyramine and histamine, to a lesser extent; the putrescine content was 10-fold lower if inoculation had taken place with either lactobacillus tested; and histamine and tyramine were essentially absent during storage, whereas they ranked above 200 ppm in the control by 45 d. Hence, an efficient food-grade biological tool was made available that constrains buildup of dangerous biogenic amines in fermented vegetables during storage.     

Reduction of soybean oligosaccharides and properties of alpha-D-galactosidase from Lactobacillus curvatus R08 and Leuconostoc mesenteroides [corrected] JK55

This study was undertaken to investigate the potential for reducing non-digestive oligosaccharides (NDO) in soy foods, as well as the influence of exogenous conditions on intracellular α-galactosidase (α-Gal) producing lactic acid bacteria. Two strains, Lactobacillus curvatus R08 and Leuconostoc mesenteriodes JK55, showed the highest levels of raffinose degrading activity at over 40 U mL⁻¹, and presented maximum activities during the stationary phase in a medium where raffinose was the only carbon source. Raffinose was the most effective inducer, followed by melibiose, and galactose; the enzymes were partially inhibited by fructose and sucrose. On the other hand, limited activity was observed in glucose. The strains displayed optimum activity levels at neutral pH and a 35–37 °C temperature range. The α-Gal activities of L. curvatus R08 and Leu. mesenteriodes JK55 were maintained at pH 6.5–10.0. The activity of the α-Gal enzyme was stable in a relatively broad range of temperatures from 0 to 40 °C for 3 h. In soymilk, Leu. mesenteriodes JK55 and L. curvatus R08 completely hydrolyzed the NDO after 18–24 h of fermentation. The abilities of L. curvatus R08 and Leu. mesenteriodes JK55 to degrade raffinose sugars and, particularly, to produce organic acids from sugar, could contribute to reductions in the anti-nutritional properties of soy, and to the accumulation of compounds with beneficial properties during food processing. Furthermore, this study provides the optimum conditions to induce α-Gal from these strains.     

Stimulation of Cadaverine Production by Foodborne Pathogens in the Presence of Lactobacillus,Lactococcus, and Streptococcus spp.

Abstract: The effect of Lactobacillus plantarum (FI8595), Lactococcus lactis subsp. cremoris MG 1363), Lactococcus lactis subsp. lactis (IL 1403), and Streptococcus thermophilus on cadaverine and other biogenic amine production by foodborne pathogens was investigated lysine decarboxylase broth. Both of lactic acid bacteria and foodborne pathogens used (especially Staphylococcus aureus, E. coli, Lc. lactis subsp. lactis and Lb. plantarum) had an ability to convert aminoacids into biogenic amine. The conversion of lysine into cadaverine was the highest (167.11 mg/L) by Lactobacillus spp. Gram‐positive bacteria generally had a greater ability to produce cadaverine with corresponding value of 46.26, 53.76, and 154.54 mg/L for Enterococcus faecalis, S. aureus, and Listeria monocytogenes, respectively. Significant variations on biogenic amine production were observed in the presence of lactic acid bacteria strains (P < 0.05). The role of lactic acid bacteria on biogenic amine production by foodborne pathogens varied depending on strains and specific amine. Cadaverine accumulation by Enterobactericeae was increased in the presence of lactic acid bacteria strains except for St. thermophilus, which induced 2‐fold lower cadaverine production by S. Paratyphi A. Lc. lactis subsp. lactis and Lc. lactis subsp. cremoris induced 10‐fold higher increases in histamine for E. coli and K. pneumoniae, respectively. Lactic acid bacteria resulted in strong increases in cadaverine production by P. aeruginosa, although remarkable decreases were observed for histamine, spermidine, dopamine, agmatine, and TMA in the presence of lactic acid bacteria in lysine decarboxylase broth . The result of the study showed that amine positive lactic acid bacteria strains in fermented food led to significant amine accumulation by contaminant bacteria and their accumulation in food product may be controlled by the use of proper starters with amine‐negative activity. Practical Application: Foodborne pathogens and certain lactic acid bacteria are particularly active in the production of biogenic amines. Most of the strains of bacteria possess more than 1 amino acid decarboxylase activity under lysine enrichment culture conditions. Lactic acid bacteria strains had a significant role on increase putrescine accumulation by foodborne pathogens. The increased production of biogenic amines in mixed culture is the result of presence of amine positive lactic acid bacteria strains. The addition of a proper selected starter culture with amine‐negative activity is advisable to produce safer fermented food with low contents of biogenic amines.     

The effect of ripening and storage conditions on the distribution of tyramine,putrescine and cadaverine in Edam-cheese

The aim of the work was to describe the development of selected biogenic amines (histamine, tyramine, putrescine and cadaverine) in 4 layers of Dutch-type cheese (Edam-cheese) depending on 3 ripening/storage regimes during a 98-day period. Biogenic amines were analysed by means of ion-exchange chromatography. A further goal was to identify microbial sources of biogenic amines in the material analysed. Phenotype characterization and repetitive sequence-based PCR fingerprinting were used to identify the isolated bacteria. The highest content of tyramine, putrescine and cadaverine was determined in cheeses stored in a ripening cellar at a temperature of 10 °C during the whole observation period. Lower biogenic amines content was determined in samples which were moved into a cold storage device (5 °C) after 38 days of storage in a ripening cellar (10 °C). The lowest concentrations of biogenic amines were detected in cheeses which were moved into a cold storage device (5 °C) after 23 days of storage in a ripening cellar (10 °C). During the 98-day period, histamine was not detected in any of the regimes. Within the cheeses analysed, non-starter lactic acid bacteria Lactobacillus curvatus, Lactobacillus casei/paracasei and Lactobacillus plantarum were detected as the main producers of the biogenic amines tested. In starter bacteria Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris the decarboxylase activity tested was not detected.     

Screening of biogenic amine production by lactic acid bacteria isolated from grape must and wine

The potential to produce the biogenic amines tyramine, histamine and putrescine, was investigated for lactic acid bacteria (LAB) of various origin, including commercial malolactic starter cultures, type strains and 78 strains isolated from Spanish grape must and wine. The presence of biogenic amines in a decarboxylase synthetic broth was determined by reverse-phase high performance liquid chromatography (RP-HPLC). Tyramine was the main amine formed by the LAB strains investigated. Leuconostoc strains were the most intensive tyramine formers. No potential to form biogenic amines was observed in Oenococcus oeni strains. Two strains of Latobacillus buchneri were associated with putrescine formation. None of the lactic acid bacteria produced histamine. According to these in vitro results, the commercial starter bacteria analyzed did not produce histamine, tyramine and putrescine.     

Effects of specific lactic acid bacteria species on biogenic amine production by foodborne pathogen

The influences of lactic acid bacteria (LAB) on biogenic amines formation by foodborne pathogens (FBP) were investigated. Biogenic amines production by single and mix cultures was tested in histidine decarboxylase broth. All of the mix cultures (LAB with FBP) inhibited significantly (P < 0.05) the ammonia accumulation except for Escherichia coli with Lactococcus lactic subsp. lactic and Lactobacillus plantarum. Although LAB with most of the pathogen showed considerably stimulation effects on putrescine, cadaverine, spermine, 2‐phenylethylamine, histamine (HIS), tyramine, trimethylamine, dopamine and agmatine (AGM), some of the LAB with pathogens showed poor inhibition effect on different amines formation. HIS production by Klebiella pneumonia was 0.16 mg L^?1, whereas HIS value in presence of Lb. plantarum increased to 30.92 mg L^?1. Consequently, LAB inhibition of the ammonium production by FBP is favourable, while the stimulation effects of LAB on biogenic amines formation by FBPs are not desirable for food industry.     

Screening selected strains of probiotic lactic acid bacteria for their ability to produce biogenic amines (histamine and tyramine)

The aim of this study was to investigate the production of biogenic amines (BA), histamine and tyramine by some probiotic lactic acid bacteria (LAB). Fifteen strains representing six LAB species were screened qualitatively by growing them in a decarboxylase medium. Quantitative analysis was carried out by HPLC analysis with direct derivatization of acid extracts. Lactobacillus casei (TISTR 389) and Lactobacillus delbrueckii subsp. bulgaricus (TISTR 895) were found to produce BA. The highest levels of histamine (1820.9 ± 3.5 mg L^?1) and tyramine (5486.99 ± 47.6 mg L^?1) formation were observed for the TISTR 389 strain, while TISTR 895 produced only histamine (459.1 ± 0.63 mg L^?1) in the decarboxylase broth. Biogenic amine potential was not observed for the Lactobacillus acidophilus, Lactobacillus lactis subsp. lactis, Lactococcus lactis subsp. lactis, and Lactobacillus plantarum strains studied. This study confirmed that BA formation is strain dependent and not related to the species. Therefore, careful screening for amino acid decarboxylase activity is recommended before selecting LAB as appropriate starter or probiotic strains in food and dairy industry.     

Biogenic amine production in Spanish dry-cured "chorizo" sausage treated with high-pressure and kept in chilled storage

Biogenic amine formation and microbiota evolution were assessed in Spanish dry-cured "chorizo" sausage treated with high-pressure (HP) and kept at 2°C. High-pressures did not affect (p<0.05) pH or water activity (a(w)). However, HP treatment did significantly reduce the level of lactic acid bacteria, by <1 logarithmic unit. Microorganism levels remained low throughout storage and the only significant reduction was in the HP treated lot at 160 days. The HP treatment caused a reduction (p<0.05) of tyramine, putrescine, and cadaverine levels, while there was a significant increase in spermidine. Amine levels increased (p<0.05) in the course of storage, although unrelated to increased microorganism levels, possibly because decarboxylase activity continued in the substrate during storage. HP seems to be effective for reducing the formation of biogenic amines in this kind of product.     

Biogenic amines in meat inoculated with Lactobacillus sake starter strains and an amine-positive lactic acid bacterium

 Formation of biogenic amines in minced meat inoculated with two Lactobacillus sake starter culture strains and an amine-positive lactic acid bacterium (G-106) was studied. The effects of these starter cultures against the formation of biogenic amines were dependent on the kind of decarboxylating microorganisms present in the raw material and the effects were different for each amine. Starter strains maintained the microbiological quality of the meat kept at 20°C for 7 days, and inhibited the formation of putrescine and cadaverine. They also inhibited the formation of phenylethylamine caused by G-106 when it was present at an initial level of 10² colony-forming units (CFU)/g. However, the formation of tyramine was not affected and the formation of histamine was increased when starters were used in samples inoculated with G-106. Received: 4 December 1996 / Revised form: 27 January 1997     

Biogenic amines in meat inoculated with Lactobacillus sake starter strains and an amine-positive lactic acid bacterium

 Formation of biogenic amines in minced meat inoculated with two Lactobacillus sake starter culture strains and an amine-positive lactic acid bacterium (G-106) was studied. The effects of these starter cultures against the formation of biogenic amines were dependent on the kind of decarboxylating microorganisms present in the raw material and the effects were different for each amine. Starter strains maintained the microbiological quality of the meat kept at 20°C for 7 days, and inhibited the formation of putrescine and cadaverine. They also inhibited the formation of phenylethylamine caused by G-106 when it was present at an initial level of 10² colony-forming units (CFU)/g. However, the formation of tyramine was not affected and the formation of histamine was increased when starters were used in samples inoculated with G-106. Received: 4 December 1996 / Revised form: 27 January 1997     

Phenotypic and genetic evaluations of biogenic amine production by lactic acid bacteria isolated from fish and fish products

In this work, biogenic amine production (histamine, tyramine and putrescine) by a collection of 74 lactic acid bacteria of aquatic origin has been investigated by means of amino acid decarboxylation by growth on decarboxylase differential medium, biogenic amine detection by thin-layer chromatography (TLC) and decarboxylase gene detection by PCR. None of the evaluated strains showed neither production of histamine and putrescine, nor presence of the genetic determinants encoding the corresponding decarboxylase activities. However, the tyrosine decarboxylase gene (tdc) was present in all the enterococcal strains, and tyramine production was detected by TLC in all of them but Enterococcus faecium BCS59 and MV5. Analysis of the tyrosine decarboxylase operon of these strains revealed the presence of an insertion sequence upstream tdc that could be responsible for their lack of tyrosine decarboxylase activity.     

Potential of wine-associated lactic acid bacteria to degrade biogenic amines

Some lactic acid bacteria (LAB) isolated from fermented foods have been proven to degrade biogenic amines through the production of amine oxidase enzymes. Since little is known about this in relation to wine micro-organisms, this work examined the ability of LAB strains (n = 85) isolated from wines and other related enological sources, as well as commercial malolactic starter cultures (n = 3) and type strains (n = 2), to degrade histamine, tyramine and putrescine. The biogenic amine-degrading ability of the strains was evaluated by RP-HPLC in culture media and wine malolactic fermentation laboratory experiments. Although at different extent, 25% of the LAB isolates were able to degrade histamine, 18% tyramine and 18% putrescine, whereas none of the commercial malolactic starter cultures or type strains were able to degrade any of the tested amines. The greatest biogenic amine-degrading ability was exhibited by 9 strains belonging to the Lactobacillus and Pediococcus groups, and most of them were able to simultaneously degrade at least two of the three studied biogenic amines. Further experiments with one of the strains that showed high biogenic amine-degrading ability (L. casei IFI-CA 52) revealed that cell-free extracts maintained this ability in comparison to the cell suspensions at pH 4.6, indicating that amine-degrading enzymes were effectively extracted from the cells and their action was optimal in the degradation of biogenic amines. In addition, it was confirmed that wine components such as ethanol (12%) and polyphenols (75 mg/L), and wine additives such as SO₂ (30 mg/L), reduced the histamine-degrading ability of L. casei IFI-CA 52 at pH 4.6 by 80%, 85% and 11%, respectively, in cell suspensions, whereas the reduction was 91%, 67% and 50%, respectively, in cell-free extracts. In spite of this adverse influence of the wine matrix, our results proved the potential of wine-associated LAB as a promising strategy to reduce biogenic amines in wine.     

Biogenic Amines in Dry Sausages as Affected by Starter Culture and Contaminant Amine-Positive Lactobacillus

The effect of an amine-negative starter culture, containing Pediococcus pentosaceus and Staphylococcus carnosus, on the growth and amine formation of an amine-positive contaminant lactic acid bacterium (G 106) was studied in dry sausages. Levels of biogenic amines, precursor amino acids, pH, water activity and microbial counts were measured. Levels of phenylethylamine and tyramine increased in the sausages inoculated with the amine-positive strain. The starter culture did not prevent growth of G 106 or its amine formation capability. However, levels of histamine remained low although G 106 could produce histamine in vitro.     

Production of biogenic amines by lactic acid bacteria and bifidobacteria isolated from dairy products and beer

The aim was to monitor production of eight biogenic amines (BAs) (histamine, tyramine (TYR), tryptamine, putrescine, cadaverine (CAD), phenylethylamine, spermine and spermidine) by selected 81 lactic acid bacteria (LAB) strains: Lactobacillus, Lactococcus, Leuconostoc, Enterococcus, Pediococcus, Tetragenococcus and Bifidobacterium. The tested LAB and bifidobacteria were isolated from dairy products and beer. The decarboxylase activity of the micro‐organisms was studied in growth medium after cultivation. The activity was monitored by HPLC after the pre‐column derivatisation with dansylchloride. Fifty LAB showed decarboxylase activity. Thirty‐one strains produced low concentrations of CAD (≤10 mg L^?1). Almost 70% of beer isolates generated higher amounts of TYR (≤3000 mg L^?1). Most of the tested LAB demonstrated decarboxylase activity. The above micro‐organisms can contribute to the increase of content of BAs in dairy products or beer and thereby threaten food safety and health of consumers. Production of BAs even by the representatives of some probiotic strains (Bifidobacterium and Lactobacillus rhamnosus) was detected in this research. This study has also proved that contaminating LAB can act as sources of higher amounts of CAD and TYR in beer.     

发酵剂对牦牛乳硬质干酪成熟过程中生物胺的影响

乳酸菌产生物胺的能力具有菌株特异性,因此,为了探究不同种类发酵剂对牦牛乳硬质干酪中生物胺形成的影响,该试验利用高效液相色谱对3种不同发酵剂制作的硬质干酪成熟过程中生物胺进行了测定和分析。结果表明,嗜热和嗜温发酵剂牦牛乳硬质干酪中检测出2-苯乙胺、腐胺、尸胺、组胺和酪胺,混合发酵剂干酪中检测出腐胺、2-苯乙胺、尸胺和酪胺。各生物胺之间呈现正相关性。3种不同发酵剂干酪在1～6个月成熟过程中,其各生物胺整体呈现增加趋势,嗜热、嗜温和混合发酵剂干酪中总生物胺最高含量分别为(448.3±9.6)、(456.8±58.4)、(293±24.5)mg/kg。组胺和酪胺是2种毒性相对高的生物胺,嗜热发酵剂干酪中组胺和嗜温发酵剂干酪中酪胺最高,其最高含量分别为(20.8±7.9)、(92.9±6.7)mg/kg,混合发酵干酪中未检测出组胺,酪胺含量次之,3种不同发酵剂干酪中组胺、酪胺含量均低于推荐安全剂量50 mg/kg和100 mg/kg。这为合理选择发酵剂和控制干酪中生物胺形成提供了依据。     

Effects of heat treatment and storage temperature on the biogenic amine content of straw mushroom (Volvariella volvacea)

The changes in the biogenic amines tryptamine, 2-phenylethylamine, putrescine, cadaverine, histamine and tyramine of straw mushroom Volvariella volvacea (Bull, ex Fr) Sing produced by heat treatment and during storage at different temperatures were studied. About 80% of the original content of these amines was lost during cooking. Commercial canned straw mushroom contained low amounts of amines. The amine contents of straw mushroom increased during storage at 4°C, with particularly notable increases in the concentrations of 2-phenylethylamine and tyramine after 5 days' storage. However, the levels of all amines increased more markedly during storage at 25°C, and the increases in the putrescine and cadaverine concentrations were much greater than those reached at 4°C.     

Influence of technological conditions of sausage fermentation on the aminogenic activity of L. curvatus CTC273

The influence of technological factors (temperature and relative humidity of the manufacturing process and the diameter of the sausage) on the aminogenic activity of the strain Lactobacillus curvatus CTC273 was evaluated. Inoculation of sausages with L. curvatus CTC273 resulted in the accumulation of large amounts of biogenic amines (higher than 1000 mg/kg dry matter in some samples) during the manufacture of fuet and llonganissa sausages. Sausages produced via process ‘A’ (3 days at 20-23 °C and 90-95% RH followed by 20 days at 12-14 °C and 70% RH) contained significantly higher amounts (p < 0.05) of biogenic amines than those manufactured via process ‘B’ (23 days at 12-13 °C and 70-90% RH), specifically tyramine, cadaverine and phenylethylamine in llonganissa and phenylethylamine in fuet. The higher fermentation temperature and relative humidity during the fermentation stage in process ‘A’ promoted decarboxylase activity in L. curvatus CTC273 and thus favoured amine accumulation. The diameter of the sausages also influenced biogenic amine production. Higher amine levels were found (p < 0.05) in llonganissa than in fuet, regardless of the manufacturing conditions. The effect of the factors considered on the modulation of aminogenic activity is not necessarily linked to the effect of strain growth, but chiefly favouring proteolytic and decarboxylase reactions.     

Production of Biogenic Amines by Lactic Acid Bacteria Isolated from Uzicka Sausages

The aim of this study was to monitor production of seven biogenic amines (Cadaverine – CAD, Putrescine – PUT, Spermine – SPE, Spermidine – SPD, Histamine – HIS, Tyramine – TYR and Tryptamine – TRY) in selected 24 lactic acid bacteria (LAB) strains. The decarboxylase activity of the microorganisms was studied in growth medium after 24 h cultivation. The ability of 24 LAB isolates cultivated in MRS broth and M17 broth supplement with 0.5% glucose to produce biogenic amines was assessed using liquid chromatography tandem mass spectrometry (LC-MS/MS). The investigation showed that LAB isolated from Uzicka sausage are not significant producers of biogenic amines in vitro.