Phenotypic and molecular identification and clustering of lactic acid bacteria and yeasts from wheat (species Triticum durum and Triticum aestivum) sourdoughs of Southern Italy

The microflora of 25 wheat sourdoughs from the Apulia region, Southern Italy, was characterized. The sourdoughs were mainly produced from Triticum durum wheat. The number of lactic acid bacteria and yeasts ranged from ca. log 7.5 to log 9.3 colony forming units (cfu)/g and from log 5.5 to log 8.4 cfu/g, respectively. About 38% of the 317 isolates of lactic acid bacteria were identified by conventional physiological and biochemical tests. Phenotypic identification was confirmed by 16S rDNA and 16S/23S rRNA spacer region PCR. Overall, 30% of the isolates were identified as Lactobacillus sanfranciscensis, 20% as Lb. alimentarius, 14% as Lb. brevis, 12% as Leuconostoc citreum, 7% as Lb. plantarum, 6% as Lactococcus lactis subsp. lactis, 4% as Lb. fermentum and Lb. acidophilus, 2% as Weissella confusa and 1% as Lb. delbrueckii subsp. delbrueckii. Some of these species have not been previously isolated from sourdoughs. Since bakers yeast is widely used in sourdough production, Saccharomyces cerevisiae was largely found. The phenotypical relationships within the main lactic acid bacteria identified were established by using cluster analysis. A microbial map of the 25 sourdoughs was plotted showing characteristic associations among lactic acid bacteria and differences in the lactic acid bacteria species which were mainly due to the species of wheat flour, use of bakers yeast and type of bread.     

Molecular and technological characterization of lactic acid bacteria from traditional fermented sausages of Basilicata region (Southern Italy)

Lactic acid bacteria (LAB) from traditional fermented sausages of the Basilicata region were investigated by ARDRA-PCR and RAPD-PCR for taxonomic identification at species and strain level and characterized on the basis of the growth and acidification at different temperatures, incubation times, levels of NaCl and KNO₂, hydrolysis of sarcoplasmatic and myofibrillar proteins and antimicrobial, peptide/amino acid release and nitrate reductase activities.Lactobacillus sakei was the predominant species (67%) followed by Pediococcus pentosaceus (16%), Leuconostoc carnosum (8%), Lactobacillus plantarum (4%), Lactobacillus brevis (2%) and Leuconostoc pseudomesenteroides (2%). The technological characterization revealed that most of the isolates had good acidifying and proteolytic properties. Moreover, Lb. sakei strains showed antimicrobial ability, while Leuconostoc strains the highest reduction of nitrates.This work was a preliminary study in the formulation of autochthonous starter cultures in order to standardize the production process of sausages, to preserve their typical organoleptic and sensory characteristics and to improve the quality of final product.     

Volatile compounds in wheat sourdoughs produced by lactic acid bacteria and sourdough yeasts

The content of volatile compounds in wheat sourdoughs fermented with four different starter cultures of the genusLactobacillus was compared with those in chemically acidified doughs. Sourdoughs as well as chemically acidified doughs were also combined with four different sourdough yeasts to investigate the effect of adding yeasts on the production of volatiles. The volatile compounds in the doughs were isolated by a dynamic headspace technique, analysed by gas chromatography (GC), and identified on the basis of GC retention times for reference compounds and mass spectrometry. Few volatile compounds were identified in the chemically acidified doughs compared to the sourdoughs. The content of volatile compounds in the sourdoughs varied with the starter culture used. Ethanol and ethyl acetate were produced in the highest amounts in sourdoughs fermented with heterofermentative cultures, especiallyL. sanfrancisco. Apart from ethanol,n-hexanol was the dominating alcohol in all the sourdoughs. The content of other compounds was low. When sourdough yeasts were added to the sourdoughs, the number and amount of volatile compounds increased. The content of ethanol, 2-methyl-propanol and 2/3-methyl-1-butanol was highest in the sourdoughs with addedSaccharomyces cerevisiae, whereas the ethyl acetate content was highest in the sourdoughs with added strain A. The content of esters other than ethyl acetate was low. The carbonyls acetoin and diacetyl were produced in small amounts in the sourdoughs with the addition of strain A.

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Flüchtige Aromastoffe in Weizensauerteigen bei der Herstellung unter Zugabe vonLactobacillus und Sauerteighefe

Zusammenfassung Der Gehalt flüchtiger Aromastoffe in Weizensauerteigen, mit 4 verschiedenen Starterkulturen desLactobacillus fermentiert, wurde mit dem Gehalt an flüchtigen Aromastoffen in chemisch gesäuerten Teigen verglichen. Sauerteige sowie chemisch gesäuerte Teige wurden außerdem mit Zugabe von 4 verschiedenen Sauerteighefen hergestellt, um den Einfluß einer Hefezugabe auf die Bildung von flüchtigen Aromastoffen zu untersuchen. Die flüchtigen Aromastoffe der Teige wurden durch eine dynamische Headspace-Technik isoliert, gaschromatographisch analysiert, und auf Grund von Retentionszeiten, Referenzstoffen und Massenspektrometrie identifiziert. Wenige flüchtige Aromastoffe wurden von den chemisch gesäuerten Teigen isoliert im Vergleich zu den Sauerteigen. Der Gehalt an flüchtigen Aromastoffen änderte sich mit der verwendeten Starterkultur. Der höchste Gehalt an Ethanol und Ethylacetat bildete sich in Sauerteigen mit heterofermentativen Kulturen, insbesondere bei der Kultur mitL. sanfrancisco. Neben Ethanol warn-Hexanol der dominierende Alkohol in sämtlichen Sauerteigen. Der Gehalt anderer Aromastoffe war gering. Bei den mit Sauerteighefen angesetzten Sauerteigen erhöhte sich die Anzahl und der Gehalt an Aromastoffen. Ein Höchstgehalt an Ethanol, 2-Methyl-1-Propanol und 2/3-Methyl-1-Butanol erwiesen sich in Sauerteigen mitS. cerevisiae, während der höchste Gehalt an Ethylacetat sich in mit dem Stamm A angesetzten Sauerteigen befand. Der Gehalt an Ester, außer Ethylacetat, war gering. Die Carbonyle Acetoin und Diacetyl bildeten sich in kleinen Mengen in Sauerteigen unter Zugabe von Stamm A.

     

Identification of lactic acid bacteria isolated from oat sourdoughs and investigation into their potential for the improvement of oat bread quality

The use of sourdough in wheat and rye breads has been extensively studied; however, little is known about its potential effect when baking oat bread. Consequently, the impact of sourdough on oat bread quality was investigated. Two different sourdoughs were prepared from wholegrain oat flour without the addition of starter cultures, by continuous propagation at 28 (SD 28) or 37 °C (SD 37) until the composition of the lactic acid bacteria remained stable. The dominant LAB were identified by sequence analysis of the 16S rDNA isolated from pure cultures. LAB from SD 28 belonged to the species Leuconostoc argentinum, Pedicoccus pentosaceus and Weissella cibaria, while Lactobacillus coryniformis dominated SD 37. The isolated LAB were further used as starter cultures for the production of oat sourdoughs. Fundamental rheology revealed softening of the sourdoughs compared to non-acidified and chemically acidified controls, which could not be attributed to proteolytic activity. Incorporation of oat sourdough into an oat bread recipe resulted in significantly increased loaf-specific volume as well as improved texture, independent of addition level or sourdough type. Overall, the results of this study show that sourdoughs containing lactic acid bacteria isolated from oats have the potential to enhance oat bread quality.     

Enzymatic activities of lactic acid bacteria isolated from Cornetto di Matera sourdoughs

Forty-one strains of lactic acid bacteria (LAB) isolated from Cornetto di Matera sourdoughs were screened for their enzymatic activities, to elucidate their possible roles during the fermentation process. Urease, peptidase, phytase, phosphatase and beta-glucosidase activities were measured spectrophotometrically using synthetic substrates. Proteolytic activities were examined in model doughs, using neutral and acidified sterile doughs as controls. All strains had low urease, glutamyl aminopeptidase and iminopeptidase activities, whereas differences within species were observed for the other enzymatic activities. Leuconostoc mesenteroides and Lactobacillus curvatus strains generally showed high aminopeptidase, X-prolyl dipeptidyl aminopeptidase, beta-glucosidase and phytase activities, while the enzymatic activities of Lactobacillus plantarum, Lactobacillus pentosus and Weissella cibaria varied between strains. In order to classify the strains on the basis of similar enzymatic profiles, a hierarchical cluster analysis was carried out. Several strains of L. plantarum, L. curvatus and Leuc. mesenteroides showed an interesting combination of proteolytic, peptidase, beta-glucosidase and phytase activities, suggesting their possible usefulness as a mixed starter culture in bread-making processes.     

Identification and functional traits of lactic acid bacteria isolated from Ciauscolo salami produced in Central Italy

Lactic acid bacteria (LAB) from Ciauscolo salami produced in Marche Region of Central Italy, and LAB strains belonging to our laboratory collection were examined for their capability to survive at low pH and bile, to adhere to Caco-2 cells, and for antibiotic resistance. LAB from Ciauscolo were identified by ARDRA and RAPD-PCR. Our study showed that all LAB strains had good adaptation to gastric juice and moderate tolerance to bile. The adhesiveness was variable among strains but significantly lower in LAB from food. Antibiotic resistance was broadly spread among food strains, with level of resistance exceeding 15% for all the antibiotics tested. The resistance determinants erm(B) and tet(M) were found in nine strains of food origin (21.4%) while tet(L) in one strain of our collection (5%). Our work suggests that fermented foods are valuable sources of bacterial strains with functional traits of intestinal lactobacilli. These bacteria may be further studied for their use in probiotic applications.     

Quantitative detection of lactic acid bacteria in dried sourdoughs using real-time PCR

A real-time PCR system with 16S rRNA gene-targeted group-specific primers was developed to quantitatively detect lactic acid bacteria (LAB) of the genera Lactobacillus, Leuconostoc, Pediococcus, and Weissella in different types of commercially available dried sourdoughs. Despite a high degree of degradation in the DNA isolated from the doughs, the 341-bp 16S rRNA gene fragment of the sourdough LAB biota could specifically be amplified. For dried sourdoughs, the resulting calculated LAB cell counts were determined to be up to 3.7 × 10⁷ cells/g fresh dough, whereas in non-fermented dough acidifiers, used as a control, the calculated LAB cell counts did not exceed 3.6 × 10⁴ cells/g fresh dough. Moreover, the effect of low pH and/or high lactic acid concentrations prevailing in the doughs on the detectability of LAB cells in spray- and roller-dried sourdoughs by PCR was investigated. Drying of non-acidified sourdoughs still permitted to detect the LAB cells by PCR, whereas drying of acidified doughs reduced the detectable cell counts by approximately 5 (spray dried) and 2 (roller dried) orders of magnitudes. Incubation of acidified doughs for 24 h did not affect the detectability of LAB cells in spray-dried doughs but further reduced the detectable cell counts in roller-dried doughs by additional 2 orders of magnitude.     

Phenotypic and genotypic aspects of Lactobacillus sanfranciscensis strains isolated from sourdoughs in Italy

Physiological properties and genetic characteristics of 44 strains of Lactobacillus sanfranciscensis, isolated in Italy from sourdough samples for salted and sweet baked products, were investigated. Data were compared with those obtained for reference strains belonging to the same and to related species. Phenotypic analysis showed the presence of two main groups, the first including strains able to ferment glucose and maltose, the second including strains fermenting only maltose. Other few isolates exhibited individual different fermentation patterns. Genetic diversity was evaluated by using Amplified Ribosomal 16S rDNA Restriction Analysis and Amplified 16S-23S rDNA Spacer Region Analysis. Electrophoretical patterns of fragments generated by Hinf I indicated a polymorphism within 16S rDNA gene that allowed the subdivision ofLb. sanfranciscensis strains in two groups. The reliability to use Dde I and Hinf I 16S rDNA restriction analysis for differentiation of Lb. sanfranciscensis strains from the other phylogenetically related Lactic Acid Bacteria was demonstrated. The amplification of the intergenic spacer region produced a unique electrophoretical profile of three bands (approximately 300, 390 and 580 bp) for all Lb. sanfranciscensis strains; this analysis proved to be helpful for the discrimination of sourdough lactobacilli at species level, but was unable to distinguish among the isolates belonging to the same species. Results of grouping, obtained by genetic analyses, did not agree with those attained with phenotypic analysis, and no relation with the geographic area of isolation or type of product was found.     

Biodiversity of lactic acid bacteria and yeasts in spontaneously-fermented buckwheat and teff sourdoughs

In this study, four different laboratory scale gluten-free (GF) sourdoughs were developed from buckwheat or teff flours. The fermentations were initiated by the spontaneous biota of the flours and developed under two technological conditions (A and B). Sourdoughs were propagated by continuous back-slopping until the stability was reached. The composition of the stable biota occurring in each sourdough was assessed using both culture-dependent and -independent techniques. Overall, a broad spectrum of lactic acid bacteria (LAB) and yeasts species, belonging mainly to the genera Lactobacillus, Pediococcus, Leuconostoc, Kazachstania and Candida, were identified in the stable sourdoughs. Buckwheat and teff sourdoughs were dominated mainly by obligate or facultative heterofermentative LAB, which are commonly associated with traditional wheat or rye sourdoughs. However, the spontaneous fermentation of the GF flours resulted also in the selection of species which are not consider endemic to traditional sourdoughs, i.e. Pediococcus pentosaceus, Leuconostoc holzapfelii, Lactobacillus gallinarum, Lactobacillus vaginalis, Lactobacillus sakei, Lactobacillus graminis and Weissella cibaria. In general, the composition of the stable biota was strongly affected by the fermentation conditions, whilst Lactobacillus plantarum dominated in all buckwheat sourdoughs. Lactobacillus pontis is described for the first time as dominant species in teff sourdough. Among yeasts, Saccharomyces cerevisiae and Candida glabrata dominated teff sourdoughs, whereas the solely Kazachstania barnetti was isolated in buckwheat sourdough developed under condition A. This study allowed the identification and isolation of LAB and yeasts species which are highly competitive during fermentation of buckwheat or teff flours. Representatives of these species can be selected as starters for the production of sourdough destined to GF bread production.     

Shelf-life stability of artisanally and industrially produced durum wheat sourdough bread (“Altamura bread”)

Physico-chemical properties and volatile compounds of three commercial Altamura breads were evaluated during storage at 25 °C. Two protected denomination of origin (PDO) artisanally produced Altamura breads (Bari, Italy), characterized either by high (High A) or low (LowA) loaf, and an industrial product, commercialized as “Altamura like” (IndA), were studied.HighA and LowA breads had a tick crust that was also detached from the crumb creating an air cushion between crust and crumb. IndA products had a thinner crust, a more homogeneous crumb structure as well as a more homogeneous water distribution among the different portion of the bread loaf than HighA and LowA. A more pronounced water gradient characterized the artisanal breads. Crust and under crust portion of all breads, and crumb for IndA product, underwent a significant reduction of moisture content and a_w during storage. Both artisanal breads were subjected to a more significant crumb hardening than IndA sample. Fresh crusts of artisanally produced breads were also significantly harder than IndA. Fresh IndA samples were significantly less cohesive and less springy than artisanal products; cohesiveness significantly decreased in all samples during storage. A more complex gas chromatographic profile was found in the artisanal bread as a larger amount of volatile compounds was present as compared to the IndA bread. Volatile compounds originated both from microbial activity and non-enzymatic browning. Larger amount of volatile compounds characteristics of yeast fermentation was found in IndA. Volatiles decreased over storage in both samples, more significant in the IndA product.     

乳酸菌胞外多糖研究新进展

综述了产EPS乳酸菌的筛选方法、已报道的EPS化学结构、分离纯化和结构鉴定的方法、EPS的生理功能、EPS的生物合成及EPS的遗传学和调控方面的研究进展.     

乳酸菌胞外多糖的研究进展

乳酸菌胞外多糖是乳酸菌在生长代谢过程中分泌到细胞壁外、常渗于培养基中的一类天然的生物高分子聚合物。由于胞外多糖具有降低胆固醇、抗肿瘤、促进菌体在肠粘膜上的粘附和免疫应答以及可作为食品增稠剂和稳定剂等重要的生理和物理特性而成为近年来的研究热点。了解乳酸菌胞外多糖的合成途径、结构以及构效关系,对于提高乳酸菌胞外多糖产量、深入了解其生物活性相关机制具有重要的意义。综述了乳酸菌胞外多糖的分类、构效关系、生物合成机制以及生物活性。     

Diversity of bacteriocins produced by lactic acid bacteria isolated from raw milk

Bacteriocin-producing lactic acid bacteria were isolated from 298 samples of raw ewes', goats’ or cows’ milk. Eighty-two bacteriocin producers were phenotypically and genotypically identified as L. lactis subsp. lactis (59 isolates), L. lactis subsp. cremoris (2 isolates), L. lactis subsp. lactis biovar diacetylactis (6 isolates), E. faecalis (7 isolates), E. faecium (1 isolate), L. paracasei subsp. paracasei (4 isolates), L. plantarum (1 isolate) and Leuconostoc spp. (2 isolates). By means of PCR-techniques, nisin was characterized in 39 of the 67 bacteriocin-producing lactococci and lacticin 481 in 23 isolates, some of which presented antilisterial activity. Enterocin AS-48 was produced by four enterococcal isolates. Four non-identified bacteriocins produced by 16 isolates showed a broad inhibitory spectrum. Nisin-producing lactic acid bacteria were the most abundant, but lacticin 481-producing lactococci and AS-48-producing enterococci were found at relatively high rates.     

乳酸菌发酵剂对直接发酵法生产面包品质的影响

在直接发酵法生产面包中应用乳酸菌发酵剂,测定发酵面团的pH、滴定酸和氨基酸态氮含量,探讨发酵过程中乳酸菌和酵母菌的相互作用及其对面团发酵的影响。此外,还对乳酸菌发酵剂添加量对面包比容、硬度、弹性、咀嚼性及感官的影响进行测定和研究,发现乳酸菌发酵剂添加量为10%～30%时有利于改善面包品质,添加量为20%时效果最佳。      

Phenotypic and genetic heterogeneity of lactic acid bacteria isolated from “Alheira”, a traditional fermented sausage produced in Portugal

The aim of this study was to evaluate the phenotypic and genetic heterogeneity of lactic acid bacteria (LAB) isolated from “Alheira”, a fermented sausage produced in Portugal.     

发酵香肠乳酸菌生理生化特性的研究

采用中国传统发酵香肠作为样品,从中分离纯化出乳酸菌,且对其做生理生化特性鉴定,以对发酵香肠的菌株进一步的筛选,从中选出不产黏液、发酵葡萄糖不产气、不产色素、不产H2S、不具有氨基酸脱羧酶、不产氨、产酸能力强的菌株作为发酵香肠中的优良的乳酸菌,最后测定优势菌株24h的生长曲线和pH值变化曲线.     

乳酸菌对中国传统主食馒头质量的影响

传统馒头发酵剂由多种微生物群体组成,乳酸菌是馒头传统发酵剂中的重要微生物类群,其种类和数量与馒头的品质存在内在联系。该文针对馒头传统发酵剂中乳酸菌菌群的发展过程以及乳酸菌发酵与馒头品质的关系进行了综述,为馒头发酵剂的研究提供参考。 关键词：中图分类号：TS211.4 文献标识码：A 文章编号：0254-5071（20１5）11-0010-04 doi:     

Characterization of lactic acid bacteria isolated from raw cows’ milk cheeses currently produced in Galicia (NW Spain)

Twenty cheeses belonging to the four Protected Designations of Origin manufactured in Galicia (NW Spain) (6 Arzúa-Ulloa, 4 Tetilla, 6 Cebreiro and 4 San Simón da Costa) were selected from a total of 60 cheeses on basis of their (typical) sensorial profiles. A total of 218 lactic acid bacteria (LAB) isolates were obtained from the predominant microflora of the selected cheeses and were identified as Lactococci (98 isolates), Leuconostocs (56), Mesophilic Lactobacilli (54), Pediococci (8) and Enterococci (2). Eighty-four of the isolates produced mainly malty, spicy or sulfide flavours in pasteurised whole milk, and were not characterized further. Some good producers of diacetyl-acetoin in milk (>100 mg/L) were found among a total of 129 LAB selected, although the isolates were generally less acidifying and less proteolytic than many of those obtained 10-15 years ago. The results suggest that the microflora in cheese-making environments have undergone changes, with the most evident difference being the practical absence of Enterococcal strains among the current isolates.