Production of organic acids by Lactobacillus strains in three different media

Ten strains of Lactobacillus (Lb). casei, Lb. rhamnosus, Lb. plantarum, Lb. paracasei and Lb. curvatus species were chosen to determine the production of organic acids after cultivation in skimmed milk, MRS broth and Jerusalem artichoke (JA) medium. The highest acidity was obtained in MRS broth and the weakest acidification was found in skimmed milk. Lb. casei Shirota produced the highest amount and Lb. rhamnosus VT1 the lowest amount of substances being estimated as titratable acidity. All strains produced lactic acid in the investigated broth and most of the strains produced acetic acid in MRS broth except Lb. curvatus 2768 and Lb. casei Shirota, in JA broth except Lb. paracasei SF1 and in skimmed milk except Lb. casei 2750, Lb. curvatus 2768, Lb. curvatus 2775 and Lb. casei Shirota. All strains, except Lb. plantarum 01, produced butyric acid in MRS broth. Beside the lactic and acetic acids, formic, citric, succinic and glutamic acids were also produced in MRS broth; formic and succinic acids were produced in skimmed milk and succinic acid in JA broth. Some strains showed change in their fermentation profile from homofermentative to mix-acid fermentation in milk. The antifungal efficiency of the lactic and acetic acid in the amount produced by lactobacilli was investigated. None of the investigated aspergilli were inhibited. The inhibitory effect of acids against Fusarium increased unequivocally with the increasing concentration. The study pointed at the dissimilarity of organic acid production of Lactobacillus strains, which was considerably influenced by the media.     

Antifungal effect of organic acids from lactic acid bacteria on Penicillium nordicum

The control of fungal contamination is particularly important to avoid both spoilage of food and feed products and the occurrence of toxic compounds, known as mycotoxins. Some lactic acid bacteria (LAB) strains have shown the capacity to inhibit fungal growth and the production of mycotoxins. In this work, cell-free supernatants (CFS) of Lactobacillus plantarum UM55 and Lactobacillus buchneri UTAD104 were tested against Penicillium nordicum radial growth and OTA production. When CFS of these strains were used, the radial growth of the fungus was inhibited by less than 20%, but the production of OTA was reduced by approx. 60%. These antifungal effects resulted from organic acids produced by LAB. The CFS of L. plantarum UM55 contained lactic acid, phenyllactic acid (PLA), hydroxyphenyllactic acid (OH-PLA) and indole lactic acid (ILA), while L. buchneri UTAD104 CFS contained acetic acid, lactic acid and PLA. These organic acids were further tested individually for their inhibitory capacity. Calculation of the inhibitory concentrations (ICs) showed that acetic acid, ILA and PLA were the most effective in inhibiting P. nordicum growth and OTA production. When the inhibitory activity of LAB cells incorporated into the culture medium was tested, L. buchneri UTAD104 inhibited the production of OTA entirely in all conditions tested, but fungal growth was only inhibited completely by the highest concentrations of cells. Acetic acid production was primarily responsible for this effect. In conclusion, the ability of LAB to inhibit mycotoxigenic fungi depends on strain capability to produce specific organic acids, and those acids may differ from strain to strain. Also, the use of LAB cells, especially from L. buchneri, in food products prone to contamination with P. nordicum (e.g. dry-cured meats and cheeses) may be an alternative solution to control fungal growth and OTA production.     

SELECTION AND BIOCHEMICAL CHARACTERISATION OF SACCHAROMYCES CEREVISIAE AND KLOECKERA APICULATA STRAINS ISOLATED FROM SPANISH CIDER

The behaviour of different strains of Saccharomyces cerevisiae and Kloeckera apiculata in apple juice fermentation was studied. Ethanol production was higher for Saccharomyces strains while residual sugars and ethyl acetate content was higher for Kloeckera strains. Kl. apiculata fermented products showed the lowest amount of higher alcohols and the lowest content in organic acids with the exception of acetic acid, so this yeast produced an increment in volatile acidity. On the basis of ethyl acetate, hydrogen sulfide, and acetic acid production, fermentative ability, potassium metabisulfite resistance and sporulation percentage, one strain from Sacch. cerevisiae could be employed as starter for making cider.     

Phytate degradation by Bifidobacterium on whole wheat fermentation

Whole cereal-based products have a beneficial effect on health, but they also contain high levels of phytate, defined as anti-nutrient. The possible use of different strains of bifidobacterial species (B. catenulatum, B. longum and B. breve) with phytate degrading activity as starters in the fermentation process was investigated. Fermentative parameters of doughs (pH, volume, total titrable acidity [TTA] and acetic and lactic acids production) and the hydrolysis profile of the phytic acid were determined during whole wheat dough fermentation. During fermentation in the presence of different bifidobacterial strains, the concentration of phytic acid showed a progressive decrease, leading to a higher release of hydrolysis products than in the control, within short fermentation time. According to the fermentative parameters bifidobacterial strains showed a good adaptation to the dough ecosystem. The range of TTA and lactic acid production was dependent on the strain. Among the tested bifidobacterial strains, those belonging to the species B. breve and B. longum induced a higher hydrolysis of phytic acid with simultaneous production of lower inositol phosphates. In addition, dough containing those strains had high pH and minor acidity than those containing a commercial starter (L. plantarum). Therefore, the tested bifidobacterial strains could be used as breadmaking starters contributing to different acidification degrees and promoting simultaneously the degradation of phytic acid in the whole wheat dough.     

Effect of phenolic acids on glucose and organic acid metabolism by lactic acid bacteria from wine

The influence of phenolic (p-coumaric, caffeic, ferulic, gallic and protocatechuic) acids on glucose and organic acid metabolism by two strains of wine lactic acid bacteria (Oenococcus oeni VF and Lactobacillus hilgardii 5) was investigated. Cultures were grown in modified MRS medium supplemented with different phenolic acids. Cellular growth was monitored and metabolite concentrations were determined by HPLC-RI. Despite the strong inhibitory effect of most tested phenolic acids on the growth of O. oeni VF, the malolactic activity of this strain was not considerably affected by these compounds. While less affected in its growth, the capacity of L. hilgardii 5 to degrade malic acid was clearly diminished. Except for gallic acid, the addition of phenolic acids delayed the metabolism of glucose and citric acid in both strains tested. It was also found that the presence of hydroxycinnamic acids (p-coumaric, caffeic and ferulic) increased the yield of lactic and acetic acid production from glucose by O. oeni VF and not by L. hilgardii 5. The results show that important oenological characteristics of wine lactic acid bacteria, such as the malolactic activity and the production of volatile organic acids, may be differently affected by the presence of phenolic acids, depending on the bacterial species or strain.     

Effects of Inhibitory Environmental Factors on Growth of Oenococcus oeni CCSYU2068 for Malolactic Fermentation of Cider Production

The effects of several inhibitory factors (sulfur dioxide, pH and ethanol) on the growth of lactic acid bacteria and the subsequent malolactic fermentation (MLF) were studied by inoculation of different culture strains of Oenococcus oeni, the major lactic acid bacteria (LAB) in cider production. After comparing their organoleptic properties, three strains of Oenococcus oeni were selected from indigenous and commercial sources and their inhibitory effects on cell growth and MLF examined. The malolactic bacteria expressed variations in tolerance to the environmental conditions of pH, sulfur and ethanol concentration. Isolated from an indigenous cider production facility, O. oeni L4 had a better capacity with constant growth even when the concentration of SO₂ was 50 ppm, ethanol 10% (v/v) and pH 3.0. O. oeni L4 showed better properties for metabolizing the major acids: malic, lactic and acetic acid. The decomposition mean rate of malic acid was as high as 228.52 mg/L per day with a low acetic acid concentration of 101.78 mg/L under the stress conditions of cider production.     

Verwendung von Kartoffelpülpe und Kartoffelfruchtwasser als Nährmedium zur Anzucht von Laccase produzierenden Pilzen

Utilization of Potato Pulp and Potato Liquor for the Production of Laccase by Selected Basidiomycetes. Potato processing wastes were used for the production of enzyme laccase (polyphenol oxidase) by selected strains of Trametes villosa, T. versicolor and Polyporus brumalis. They were capable of utilizing these wastes as the sole carbon and nitrogen source. Maximum activities of laccase was obtained with the optimized mixtures of 5% potato pulp and 4% of potato liquor. Several basidiomycetes strains, when grown on potato wastes, can synthetize alpha-amylase (e.g. Trametes villosa). The amino acids and trace elements in pulp and liquor were analysed. Asparagine and aspartic acid are the main constitutents of amino acid fraction of the potato liquor under the test. Cultivation of Trametes villosa on potatostarch processing wastes can be used for the inexpensive production of valuable fungal enzymes.     

Inhibition of Citrus Fungal Pathogens by Using Lactic Acid Bacteria

Abstract: The effect of lactic acid bacteria (LAB) on pathogenic fungi was evaluated and the metabolites involved in the antifungal effect were characterized. Penicillium digitatum (INTA 1 to INTA 7) and Geotrichum citri-aurantii (INTA 8) isolated from decayed lemon from commercial packinghouses were treated with imazalil and guazatine to obtain strains resistant to these fungicides. The most resistant strains (4 fungal strains) were selected for evaluating the antifungal activity of 33 LAB strains, among which only 8 strains gave positive results. The antifungal activity of these LAB strains was related to the production of lactic acid, acetic acid, and phenyllactic acid (PLA). A central composite design and the response surface methodology were used to evaluate the inhibitory effect of the organic acids produced by the LAB cultures. The antifungal activity of lactic acid was directly related to its concentration; however, acetic acid and PLA showed a peak of activity at 52.5 and 0.8 mM, respectively, with inhibition rates similar to those obtained with Serenade^® (3.0 ppm) imazalil (50 ppm) and guazatine (50 ppm). Beyond the peak of activity, a reduction in effectiveness of both acetic acid and PLA was observed. Comparing the inhibition rate of the organic acids, PLA was about 66- and 600-fold more effective than acetic acid and lactic acid, respectively. This study presents evidences on the antifungal effect of selected LAB strains and their end products. Studies are currently being undertaken to evaluate the effectiveness in preventing postharvest diseases on citrus fruits.     

Food chain approach to lowering the saturated fat of milk and dairy products

Lactating cow diets were supplemented with high‐oleic acid sunflower oil over two production periods spanning two years, to modify the milk fat, partially replacing saturated fatty acids with cis‐monounsaturated fatty acids. The resulting milk was used for ultrahigh‐temperature milk, butter and Cheddar cheese production, and fatty acid profiles were compared with those of conventionally produced products. Fat from products made with modified milk had lower saturated fatty acids and higher cis‐ and trans‐monounsaturated fatty acid concentrations than that of conventional products. This was consistent over both production periods, demonstrating that this food chain approach could be adopted on a wider scale.     

Utilization of <Emphasis Type="Italic">Echium amoenum</Emphasis> Extract as a Growth Medium for the Production of Organic Acids by Selected Lactic Acid Bacteria

Gol-gavzaban (Echium amoenum Fisch. & Mey.) is an indigenous herbal plant, related to the Boraginaceae family, cultivated historically in Iran. In this study, the suitability of Echium extract as a raw material for production of fermented juice by four strains of lactic acid bacteria (Lactobacillus paracasei, Lactobacillus acidophilus, Lactobacillus plantarum, and Lactobacillus delbrueckii) was examined. Echium extract was inoculated with these bacteria and incubated at 30 °C for 24 h. Changes in microbial population, pH, acidity, sugars, and organic acids metabolism were measured during the fermentation process. Results showed that all of the selected bacteria were capable to grow well in Echium extract without any supplementation. They all metabolized sugars including glucose, fructose and sucrose simultaneously. L. paracasei showed more affinity to sugar consumption at almost 45.2%, 38%, and 21.6% of initial glucose, fructose, and sucrose concentration, respectively. Lactic acid was produced immediately after the fermentation started. L. plantarum, L. delbrueckii, and L. acidophilus produced 6.8, 6.6, and 6.4 g/l lactic acid, respectively, which were significantly higher than that produced by L. paracasei (5 g/l). On the other hand in the case of acetic acid, L. paracasei produced significantly greater quantities in comparison with other strains (4.48 g/l). It was proved that E. amoenum was a desirable media for the production of some organic acids by all these bacterial strains.     

Genetic and enological analysis of selected Saccharomyces cerevisiae strains for wine production

The non‐wine Saccharomyces cerevisiae strain of 96581 was found to be a promising candidate for the production of white wine. It produced wines with fusel alcohols that were 57% higher than those produced by the wine yeasts studied and was also more efficient in the production of 2‐phenethyl acetate and 3‐methyl‐1‐butanol acetate. This study also shows that there is a difference in the ester‐formation efficiency for acetates relative to C6, C8 and C10 fatty acid esters for all the studied yeast strains. Therefore, it supports the view that other unidentified enzymes besides those regulated by ATF1 and ATF2 genes are involved in the production of ethyl esters of C6–C10 fatty acids. DNA analysis of the 25S, 18S, 5.8S and 5S ribosomal DNA genes in these strains showed high conservation. Despite the closely related nature of these yeast strains, the chemical profiles of the wines produced were significantly different.     

Isolation and characterisation of lactic acid bacteria strains from raw camel milk for potential use in the production of fermented Tunisian dairy products

The aim of this work was to study the suitability of camel milk for the production of dairy products by lactic acid fermentation. Sixty strains of lactic acid bacteria (LAB) were isolated from camel milk. The strains were tested for their acidification activity, ability to use citrate, exopolysaccharide production, lipolytic, proteolytic activities and resistance to antibiotics. Ten strains were investigated for their ability to metabolize carbohydrates and that resulted in the identification of 5 Lactococcus lactis, 1 Lactobacillus pentosus, 2 Lactobacillus plantarum, 1 Lactobacillus brevis and 1 Pediococcus pentosaceus strains. Two strains of Lactococcus lactis SCC133 and SLch14 were selected to produce traditional Tunisian fermented dairy products (Lben, Raib, Jben cheese and Smen). These strains were chosen based on their acid production capacity and their ability to produce a high yield of biomass.     

CONTROL OF FERULIC ACID AND 4-VINYL GUAIACOL IN BREWING

Phenolic acids in beer are important because they can be decarboxylated to phenols, which usually impart off-flavours. An improved high performance liquid chromatographic system was used to monitor phenolic acids and phenols during the brewing process. Ferulic acid was the most significant phenolic acid found in beers prepared from malted barley. Extraction of ferulic acid from malt involved an enzymatic release mechanism with an optimum temperature about 45°C. Mashing-in at 65°C significantly decreased the release of free ferulic acid into the wort. Wort boiling produced 4-vinyl guaiacol by thermal decarboxylation, in amounts (0.3 mg/L) close to its taste threshold, from worts that contained high contents of free ferulic acid (> 6 mg/L). The capacity of yeasts to decarboxylate phenolic acids (Pof⁺ phenotype) was strong in wild strains of Saccharomyces and absent in all lager brewing yeast and most ale brewing yeasts. Some top-fermenting strains, especially those used in wheat beer production, possessed a weak decarboxylating activity (i.e. Pof^δ). During storage of beers there were appreciable temperature-dependent losses of 4-vinyl guaiacol. These results indicated that the production of 4-vinyl guaiacol is amenable to close technological control.     

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.     

Differentiation between fermenting and spoilage yeasts in wine by total free fatty acid analysis

Differentiation between fermenting and spoilage yeasts in wine was estimated by cellular fatty acid profiles. Forty-two strains of yeasts representing 17 genera were grown on a defined liquid medium for 48 h at 25°C in a rotary shaker. After saponification of yeast cells, free fatty acid extracts were analysed by gas chromatography. Multivariate analysis was performed by Principal Components Analysis (PCA) to define clusters of fatty acids and yeasts. Strains were characterised especially by long-chain fatty acids, palmitic (C16) to linolenic (C18:3) acid under aerobic culture. Nevertheless, most of Saccharomyces cerevisiae and also Dekkera bruxellensis (former names D intermedia and Brettanomyces lumbicus) synthesized medium-chain fatty acids, octanoic (C8) acid to dodecanoic (C12) acid. With this method it was possible to differentiate fermenting grape yeasts such as S cerevisiae from spoilage yeasts on the basis of the absence of linoleic (C18:2) and linolenic (C18:3) acids. However, the method seemed unreliable for the identification of strains, more particularly those of species of S cerevisiae.     

Impact of two Williopsis yeast strains on the volatile composition of mango wine

The aim of the research was to study the volatile composition of mango wine fermented with two Williopsis yeast strains: Williopsis saturnus var. mrakii NCYC500 and W. staurnus var. suaveolens NCYC2586. Thirty terpenoids, twenty‐six esters, ten alcohols, nine acids, seven aldehydes and ketones, two ethers, two phenols and one sulphur compound were identified in the mango wine fermented with strain NCYC500, while twenty‐seven terpenoids, thirty esters, eleven alcohols, eight acids, eight aldehydes and ketones, three ethers, two phenols, one sulphur compound and one furan were detected in the mango wine fermented with the strain NCYC2586. The kinetic changes, final concentrations and odour activity values of major volatiles were compared between the two Williopsis yeast strains and also with other yeast reported in the literature. The results showed that Williopsis yeast strains NCYC500 and NCYC2586 were high producers of acetic acid and acetate esters, but low producers of medium‐ to long‐chain fatty acids and their corresponding ethyl esters. Unlike mango wine fermented with Saccharomyces cerevisiae, most terpenoids derived from mango juice were retained in the resultant mango wine fermented with the two Williopsis yeast strains, suggesting the mango wine could retain the aromatic hints of fresh mango.     

Sensorically and antimicrobially active metabolite production of Lactobacillus strains on Jerusalem artichoke juice

BACKGROUND: In the tubers of Jerusalem artichoke (Helianthus tuberosus L.) the main carbohydrate is the well‐known prebiotic inulin, which is a good growth substrate for gut microorganisms. Jerusalem artichoke tuber is traditionally consumed boiled or pickled rather than in fermented form. Lactic acid bacteria are traditionally used in the production of fermented foods; nevertheless their behavior and metabolite production are considerably influenced by the substrate. The purpose of this study was to investigate the growth and production of the most important sensorically and antimicrobially active metabolites of different Lactobacillus strains on Jerusalem artichoke juice. RESULTS: All investigated strains grew well (in the range 10⁹ cfu mL^?1) in the media. The organic acids (lactic acid, 110–337 mmol L^?1; acetic acid, 0–180 mmol L^?1; and succinic acid, 0–79 mmol L^?1), hydrogen peroxide (0.25–1.77 mg L^?1), mannitol (0.06–3.24 g L^?1), acetoin and diacetyl production of strains varies not only according to the species but also from strain to strain, which will be demonstrated and discussed in the paper. CONCLUSION: Our results showed that lactobacilli can be used for the fermentation of Jerusalem artichoke, which in this form could be used, alone or mixed with other raw food material, as a new synbiotic functional food. Copyright © 2011 Society of Chemical Industry     

Fungal metabolism of butter and shea oils by Penicillium roquefortii in solid state cultures

Monoacylglycerols (monoglycerides, MAGs) were produced from butter oil and shea stearin fraction (shea oil) by two strains of Penicillium roquefortii, FRR 2456 (isolated from a spoilt melon) and Wisbey PJ (a commercial dairy strain), at pH 7.0 at 10 and 25 °C. The system was designed as a model of cheese using modified Czapek medium in solid state cultures. Shea oil with its unique fatty acid profile (stearic, oleic and palmitic acids) was used for comparison with butter oil. Yields of MAGs, which ranged from 3 to 14 g kg^?1 oil, were higher with butter than with shea oil and higher when the spoilage strain FRR 2456 was used. Monoacylglycerols produced by mycelium‐bound lipases from both fungal strains were mainly sn‐1(3) or α isomers (60–70 mol%). Monopalmitin was the major MAG produced from both butter and shea oils. The production and use of MAGs alone or in combination with free fatty acids (FFAs) as food preservatives are discussed. It is implied that sn‐1(3) MAGs together with free fatty acids may be part of a natural antimicrobial system in relatively high‐pH foods such as blue mould‐ripened cheese where growth of foodborne pathogens such as Listeria monocytogenes can be a problem. Copyright © 2003 Society of Chemical Industry     

Phenolic compounds in peanut seeds: Enhanced elicitation by chitosan and effects on growth and aflatoxin B1 production by Aspergillus flavus

Effects of chitosan and Aspergillus flavus to enhance elicitation of phenolic compounds in viable peanut seeds were conducted at two water activity levels. In vitro effects of phenolic acids on A. flavus growth and aflatoxin B₁ production were also studied. Chitosan enhanced elicitation of free phenolic compounds (FPC) at Aw .85 and .95 levels. A. flavus initially decreased and subsequently increased FPC content, but bound phenolic compounds (BPC) decreased during incubation. Chitosan + A. flavus treatment caused an increase in FPC reaching a plateau between 24–48 h at Aw .85 while BPC levels increased over the same period at both Aw levels. Major free and bound phenolic acids detected were p‐coumaric, ferulic and an unknown phenolic acid eluting at a retention time of 22 min. Generally, chitosan significantly enhanced elicitation of free ferulic and p‐coumaric acids and bound p‐coumaric acid at Aw .95. Free unknown phenolic and bound ferulic acids at Aw .85 were enhanced by chitosan. A. flavus caused significant induction of bound p‐coumaric and ferulic acids and free unknown phenol at Aw .85. Chitosan + A. flavus enhanced free p‐coumaric (3 h) and unknown phenolic acids and bound p‐coumaric acid at Aw .95 while bound ferulic acid was enhanced at Aw .85. Chitosan limited A. flavus growth and subsequent aflatoxin production by inducing susceptible tissues to produce more preformed phenolic compounds.

Analysis of liquid cultures of A. flavus revealed that p‐coumaric, ferulic, and vanillic acids and a mixture of these phenolic acids slightly inhibited mycelial growth. Production of aflatoxin B₁ by A. flavus was completely inhibited at 1 mM and 10 mM concentrations of the phenolic acids and their mixture on four days of incubation. Mode of action of phenolic acids is likely on the secondary pathway for aflatoxin B₁ production and not on the primary metabolism for fungal growth.