The sourdough microflora Microbiological, biochemical and breadmaking characteristics of doughs fermented with freeze-dried mixed starters, freeze-dried wheat sourdough and mixed fresh-cell starters

Freeze-dried mixed starters, freeze-dried wheat sourdough and mixed fresh-cell starters made withLactobacillus sanfrancisco CBI,L. plantarum DC400 andSaccharomyces cerevisiae 141 and/orS. exiguus M14 were used for leavening wheat doughs, and their microbiological, biochemical and breadmaking characteristics were compared with those of Italian traditional doughs produced by baker's yeast. All the doughs fermented with starters had more balanced microbiological and biochemical characteristics than dough started with baker's yeast in which alcoholic fermentation end-products largely predominated. By using starters, the greatest lactic acid bacteria cell number and acetic acid production, were achieved, along with more complete profiles of volatile compounds and greater structural stability of fermented doughs. Fresh-cell starters showed higher microbial functionality and represented the only way to enrich the doughs withS. exiguus M14, some of which survived the freeze-drying process. No differences were detected between the two different types of freeze-dried starters and the subsequent use (10 times) of doughs initially produced with freezedried starters eliminated initial differences in the microbial functionality with respect to fresh-cell starters.     

Proteolytic activities in togwa,a Tanzanian fermented food

Proteolytic activities were investigated in sorghum-based togwa prepared by natural fermentation and using starter cultures previously isolated from the native product, i.e., Lactobacillus brevis, Lactobacillus cellobiosus, Lactobacillus fermentum, Lactobacillus plantarum, Pediococcus pentosaceus, and Issatchenkia orientalis in coculture with either L. brevis or L. plantarum.Both proteinase and aminopeptidase activities were substantially higher in naturally fermented togwa than in those with starters (14-30%, 12-70%, respectively). A variable but substantial part of the proteinase activity followed the particulate fraction of togwa; aminopeptidase activity was mainly in that fraction. The breakdown of relatively high molecular mass protein (64 kDa) in togwa was detected by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE); the products were mainly in the 14-30 kDa range. Reversed-phase fast-protein liquid chromatography (RP-FPLC)-protein/peptide patterns changed during fermentation with some variation between togwa of different cultures. Supplementation of gruel with malt increased the concentration of total protein [from 9.5% to 11.0% (w/w) on dry weight basis)] and of most of the free amino acids. Fermentation had no effect on total protein content; however, the concentration of most of the amino acids was reduced, except for the proline content that increased. Natural fermentation also increased the concentration of glutamic acid and ornithine. Fermentation by P. pentosaceus increased aspartic acid, while L. cellobiosus, L. fermentum, and L. brevis in coculture with I. orientalis increased the concentration of citrulline.     

Effects of two different sourdoughs on the characteristics of Pandoro, a typical Italian sweet leavened baked good

Pandoro is a traditional Italian sweet-leavened baked product, usually consumed at Christmas. It is manufactured according to specific procedures and preparation starts from a sourdough called ‘‘madre’’ (mother sponge) continuously refreshed. This sourdough is the result of a complex microbial association including yeasts and lactic acid bacteria (LAB) and its use can improve sensory quality and shelf-life of the resulting products. The aim of this work was to evaluate the effect of the use of two different sourdoughs matured at different temperature (13 and 19 °C) on some metabolites, which can affect the organoleptic characteristics of Pandoro. Different samples, taken throughout the 19 h of process, were analysed for the determination of yeast and LAB counts, pH, a_w, carbohydrates, organic acids content and volatile profile. The results showed that, at the end of fermentation process, the sourdough propagated at 19 °C reached lower pH values than that at 13 °C. This was probably due to higher LAB counts (1 log unit higher), resulting also in higher lactic acid concentration and faster sugars depletion. On the contrary, temperature of dough during maturation did not affected yeast concentrations. Different production processes strongly affected also the volatile profile, both of sourdough at the end of fermentation and of the final products.     

发酵条件对酸樱桃发酵汁挥发性风味物质的影响探究

目的 考察发酵条件对酸樱桃发酵汁中风味物质的影响。方法 利用3株乳酸菌发酵酸樱桃汁, 以pH为指标明确最佳发酵菌种和稀释比例。随后利用气相色谱-离子迁移谱法(gas chromatography-ion mobility spectrometry, GC-IMS)探究糖添加量、pH、氮源、稀释比例对发酵挥发性风味物质的影响。结果 最佳发酵菌为鼠李糖乳杆菌(Lactobacillus GG, LGG), 最佳稀释比例为1:3, 氮源对挥发性风味物质几乎没有影响; 而不同添加量的碳源(葡萄糖)导致发酵液乙偶姻、双乙酰等含量差异; 随着pH增加, 乙偶姻、双乙酰、2-戊酮等含量显著升高; 稀释比例通过影响发酵原汁的浓度导致发酵后风味物质乙偶姻含量显著增加、2-戊酮含量显著降低。进一步差异比对3株乳酸菌酸樱桃发酵汁的风味, 发现LGG代谢脂肪酸可产生较多的2-庚酮及2-戊酮。结论 菌种、糖添加量、稀释比例、起始pH均可导致发酵液中乙偶姻、2-庚酮及2-戊酮等挥发性风味物质产生显著影响, 本研究为酸樱桃的加工利用提供理论指导。     

Manufacture and characterization of functional emmer beverages fermented by selected lactic acid bacteria

Autochthonous lactic acid bacteria from emmer flour were screened based on the kinetic of acidification and used to ferment beverages containing emmer flour, emmer gelatinized flour, and emmer malt at percentages ranging 5-30% (wt/wt). Preliminarily, the concentration of raw flour and malt was selected based on sensory analysis. Different protocols were set up for the manufacture of four different beverages which used Lactobacillus plantarum 6E as the starter. Emmer beverages were mainly differentiated based on the concentration of organic acids, carbohydrates, amino acids, dietary fibers, vitamins, antioxidant and phytase activities, and volatiles and sensory profiles. Wheat flour bread was used as the control to determine the hydrolysis index (HI = 100), as an indirect estimation of the glycemic index. The beverage made with 30% (wt/wt) of gelatinized flour showed an HI of 56%, its viscosity was improved by using an EPS-producing strain and it allowed the survival of the potential probiotic Lactobacillus rhamnosus SP1 at cell density of ca. 5 × 10⁸ cfu/ml throughout storage at 4 °C. Among the exploited biotechnological options, this latter beverage could be considered as a promising novel functional food.     

Molecular characterization of lactic acid bacteria and in situ amylase expression during traditional fermentation of cereal foods

Lactic acid bacteria play an important role in traditional fermented foods consumed in different countries. Study of their taxonomic structure and diversity is necessary for starter culture selection, improved safety and nutritional enhancement. To achieve these objectives, microbial genomic typing methods were used to study genetic differences of autochthonous bacteria and their distribution in two traditional African fermented cereal foods. A total of 85 predominant bacterial species were isolated from ogi and kunu-zaki obtained from Northern and Southern geographical region of Nigeria. They were identified using combination of 16S rRNA gene sequencing, multilocus sequence analysis (MLSA) based on rpoA, pheS and atpA genes as well as M13-PCR gel fingerprints. The results showed that Lactobacillus fermentum was the most frequently isolated species in ogi (71.4%) and kunu-zaki (84.5%). Other species of lactic acid bacteria (LAB) identified were Lactobacillus plantarum, Streptococcus gallolyticus subsp. macedonicus and Pediococcus pentosaceus. Non lactic acid bacteria isolated from these foods were species belonging to the Bacillus and Staphylococcus. Non-metric multidimensional scaling (nMDS) analysis of the M13-PCR fingerprints for LAB strains showed clonal diversity among strains of the same species. In vitro and in situ expression of amylase gene during fermentation by amylolytic L. plantarum ULAG11 was detected, indicating the potential usefulness of such species for development of starter cultures and for controlled fermentation processes.     

Diversity of lactic acid bacteria from Hussuwa, a traditional African fermented sorghum food

The diversity of lactic acid bacteria associated with Hussuwa fermentation, a Sudanese fermented sorghum food, was studied using a polyphasic taxonomical approach. Predominant strains could be well characterised based on a combination of phenotypic tests and genotypic methods such as ARDRA, rep-PCR and RAPD-PCR, as well as 16S rRNA gene sequencing of representative strains. Thus, the majority (128 of 220, 58.3%) of strains exhibited phenotypic properties typical of heterofermentative lactobacilli and of these, 100 strains were characterised more closely using the genotyping methods. The majority (97/100) strains could be characterised as Lactobacillus fermentum strains. Seventy-two of 220 strains (32.7%) showed phenotypic properties that are characteristic of pediococci. Of 41 selected strains investigated by genotyping techniques, 38 (92.7%) could be characterised as Pediococcus acidilactici strains, while three (7.3%) could be characterised as Pediococcus pentosaceus strains. The Hussuwa fermentation thus appears to be dominated by L. fermentum strains and P. acidilactici strains. For this reason, we selected representative and predominant strains as potential starter cultures for Hussuwa fermentation. These strains, L. fermentum strains BFE 2442 and BFE 2282 and P. acidilactici strain BFE 2300, were shown on the basis of RAPD-PCR fingerprinting to predominate in a model fermentation when used as starter cultures inoculated at 1 × 10⁶ CFU/g and to lower the pH of the fermentation to below pH 4.0 within 48 h. These cultures should be studied for further development as starter preparations in pilot scale studies in actual field fermentations.     

四川传统发酵肉中乳酸菌的分离及发酵特性研究

对四川传统发酵肉中乳酸菌进行分离鉴定,并对其发酵特性进行研究。以四川腊肉为研究对象,采用平板划线法对乳酸菌进行分离纯化,利用16S r DNA的序列测定技术对其鉴定,并对分离菌株在不同温度、p H、Na Cl浓度、亚硝酸盐浓度条件下的生长代谢、产酸能力进行研究,并对发酵产物中游离氨基酸进行测定分析。结果表明:筛选出的一株优势乳酸菌经分子生物学鉴定为戊糖片球菌(Pediococcus pentosaceus)。分离菌株对数生长期为6~18 h;最适发酵温度30℃,发酵p H5.5;对Na Cl及Na NO2耐受能力分别高达10%和150 mg/L;其代谢产物中共测出16种游离氨基酸,共包含5种人体必需氨基酸。      

乳酸菌细胞固定化发酵的研究进展

乳酸菌足能利用糖发酵产生乳酸的一类细菌,在食品、医药、工业、农业和科研等领域均有广泛的应用。本文综述了利用乳酸菌进行细胞固定化发酵生产乳酸、乳制品以及Nisin的国内外研究进展情况。并对乳酸菌固定发酵技术的应用前景进行了简要论述。     

A review on traditional Turkish fermented non-alcoholic beverages: Microbiota,fermentation process and quality characteristics

Shalgam juice, hardaliye, boza, ayran (yoghurt drink) and kefir are the most known traditional Turkish fermented non-alcoholic beverages. The first three are obtained from vegetables, fruits and cereals, and the last two ones are made of milk. Shalgam juice, hardaliye and ayran are produced by lactic acid fermentation. Their microbiota is mainly composed of lactic acid bacteria (LAB). Lactobacillus plantarum, Lactobacillus brevis and Lactobacillus paracasei subsp. paracasei in shalgam fermentation and L. paracasei subsp. paracasei and Lactobacillus casei subsp. pseudoplantarum in hardaliye fermentation are predominant. Ayran is traditionally prepared by mixing yoghurt with water and salt. Yoghurt starter cultures are used in industrial ayran production. On the other hand, both alcohol and lactic acid fermentation occur in boza and kefir. Boza is prepared by using a mixture of maize, wheat and rice or their flours and water. Generally previously produced boza or sourdough/yoghurt are used as starter culture which is rich in Lactobacillus spp. and yeasts. Kefir is prepared by inoculation of raw milk with kefir grains which consists of different species of yeasts, LAB, acetic acid bacteria in a protein and polysaccharide matrix. The microbiota of boza and kefir is affected from raw materials, the origin and the production methods.     

Isolation and characterization of the microbial population of different South African kefir grains

Kefir, a slightly acidic fermented milk, is produced by adding lactic acid bacteria and yeasts, in the form of grains, to milk. The bacteria and yeasts present in the kefir grains are known to vary widely. Selective growth media and morphological and biochemical characteristics were used for the isolation and identification of the microbes present in the grains from eight different sources in South Africa. The kefir grains were activated in milk for only 24 h to prevent any changes in the microbial population of the grains. The microbial numbers varied between 6.4 × 10⁴ and 8.5 × 10 ⁸  cfu/g on the media selective for the bacterial species and between 1.5 × 10 ⁵ and 3.7 × 10 ⁸   cfu/g on the media selective for the yeast species. The bacterial genera that were identified included Lactobacillus , Leuconostoc and Lactococcus and the yeast genera included Zygosaccharomyces , Candida and Saccharomyces . The distribution frequencies of the microbes in the different grains were determined and most of the grains were dominated by two microbial species. No pediococci, acetic acid bacteria or propionibacteria were detected.     

发酵肉制品中乳酸菌的主要发酵特性

从发酵肉制品中分离、纯化乳酸菌132株,对它们的主要发酵特性如耐盐性、耐硝性、产黏液、产气等进行了测定,并比较了它们在不同培养基中的产酸情况。     

Chemical and biological characteristics of protein hydrolysates from fermented shrimp by-products

Protein hydrolysates were prepared through lactic acid fermentation of the inedible portions of shrimp (cephalothorax and exoskeleton), the by-products of shrimp processing operations. The protein-rich liquid hydrolysate was further processed into a concentrated paste via vacuum evaporation at 80 °C or was also processed into a dry powder using a spray drying method at 180 °C/140 °C (inlet/outlet temp). The laboratory compared the composition of the three forms of shrimp protein hydrolysates. The protein and ash content of the hydrolysates ranged from 8.43 ± 0.22 to 46.73 ± 1.29 and 2.03 ± 0.52 to 8.25 ± 0.14 g/100 g of wet weight. All the samples were analyzed for fifteen amino acids; the powder form was analyzed for colour, microbial content, and for heavy metal occurrence. The shrimp by-products were successfully converted into micro-nutrient by-products rich in amino acids for potential recommendations in the supplementation of animal and human diets.     

On-farm implementation of a starter culture for improved cocoa bean fermentation and its influence on the flavour of chocolates produced thereof

Cocoa bean fermentations controlled by means of starter cultures were introduced on several farms in two different cocoa-producing regions (West Africa and Southeast Asia). Two starter culture mixtures were tested, namely one composed of Saccharomyces cerevisiae H5S5K23, Lactobacillus fermentum 222, and Acetobacter pasteurianus 386B (three heaps and one box), and another composed of L. fermentum 222 and A. pasteurianus 386B (seven heaps and one box). In all starter culture-added cocoa bean fermentation processes, the inoculated starter culture species were able to outgrow the natural contamination of the cocoa pulp-bean mass and they prevailed during cocoa bean fermentation. The application of both added starter cultures resulted in fermented dry cocoa beans that gave concomitant milk and dark chocolates with a reliable flavour, independent of cocoa-producing region or fermentation method. The addition of the lactic acid bacterium (LAB)/acetic acid bacterium (AAB) starter culture to the fermenting cocoa pulp-bean mass accelerated the cocoa bean fermentation process regarding citric acid conversion and lactic acid production through carbohydrate fermentation. For the production of a standard bulk chocolate, the addition of a yeast/LAB/AAB starter culture was necessary. This enabled an enhanced and consistent ethanol production by yeasts for a successful starter culture-added cocoa bean fermentation process. This study showed possibilities for the use of starter cultures in cocoa bean fermentation processing to achieve a reliably improved fermentation of cocoa pulp-bean mass that can consistently produce high-quality fermented dry cocoa beans and flavourful chocolates produced thereof.     

Potential of amylolytic lactic acid bacteria to replace the use of malt for partial starch hydrolysis to produce African fermented pearl millet gruel fortified with groundnut

Fermentation and starch hydrolysis of a pre-cooked pearl millet–groundnut (MG) slurry inoculated with amylolytic lactic acid bacteria (ALAB) or by back slopping was investigated as a substitute for the addition of malt to prepare infant gruels. The ALAB collection strain Lb. plantarum A6, and the endogenous microflora provided by back slopping were more efficient in acidifying and partially hydrolysing starch in the MG slurry than Lb. plantarum 6.1, isolated from the traditional process in Burkina Faso. Large amounts of maltotriose and maltotetraose accumulated in slurry fermented by strain A6. No accumulation of maltose was observed, which could be an advantage to prevent the growth of microbial contaminants such as yeasts. Starch hydrolysis in the MG slurry inoculated with strain A6 or by back slopping enabled preparation of high-energy density gruels (84.7 ± 4.4 and 80.4 ± 23.8 kcal/100 g of gruel, respectively) of liquid consistency. However variability was higher with back slopping.     

Influence of bacterial dynamics upon the final characteristics of model Portuguese traditional cheeses

The microbiological profile in raw milk cheeses is typically characterized by a multitude of microbial groups, with interactions among them throughout ripening that are not fully understood to date. Incidence of undesired microorganisms in raw cheesemaking milk, as is the case of either spoilage or even pathogenic ones, is a common trait in Portuguese traditional cheeses. Hence, they will likely contribute to the physicochemical changes occurring therein and, consequently, to the characteristics of the final product. In order to gain insight into their role, model cheese systems, manufactured as far as possible according to artisanal practices (except that the initial microbial load and biodiversity were controlled), were experimentally tested. Single contaminants, or a consortium thereof, were inoculated at two levels in sterilized raw ewe's milk, and duly combined with inocula containing one or two lactic acid bacteria normally found in those traditional cheeses. The physicochemical composition, organic acid profile, and evolution of both protein breakdown and rheology were monitored throughout a 60 d-ripening period. Modifications brought about within the cheese matrix as a result of microbial metabolism, especially those arising from the interaction between lactic acid bacteria and unwanted microorganisms, included the enhanced release of peptides and free amino acids, which in turn led to higher viscoelastic moduli. The final model cheeses could be well discriminated, based on the impact of the various inocula considered upon the levels of organic acids. Conversely, proteolysis and viscoelastic properties appeared to be essentially independent of the initial microflora.     

清香型小曲白酒酒醅中乳酸菌的筛选鉴定及其发酵特性研究

该研究采用传统分离方法从不同季节清香型小曲白酒酒醅中分离乳酸菌,通过形态及分子生物学技术对其进行菌种鉴定,并探究其发酵特性。结果表明,共分离纯化出225株乳酸菌,经鉴定,归属于3属15种。通过高粱汁培养基初筛、固态发酵小试实验复筛,得到两株增酯降杂效果较好的乳酸菌,分别为纳格里乳杆菌（Lactobacillus nagelii）X29和布氏乳杆菌（Lactobacillus buchneri）X9。将强化乳酸菌X29和X9麸皮种加入桂花曲,最佳添加量为20%,此添加量下酒样中乙酸乙酯含量分别提升32.84%和16.79%,乳酸乙酯含量分别提升37.38%和36.44%,正丙醇含量分别降低2.61%和5.40%,杂醇油含量分别降低21.33%和26.71%。乳酸菌强化麸皮种糖化前加入能明显提升酒体中乙酸乙酯含量;糖化后加入则对乳酸乙酯含量和出酒率提升效果明显;延长发酵时间也可提升乙酸乙酯和乳酸乙酯含量。     

Lactic acid fermentation of<Emphasis Type="Italic"> Brassica rapa</Emphasis>: chemical and microbial evaluation of a typical Italian product (<Emphasis Type="Italic">brovada</Emphasis>)

Brovada is an ancient traditional product from the northeast Italian region, included in any list of typical Italian products, which is obtained by natural fermentation of turnips (Brassica rapa). Turnips are cleaned and put in vats, alternating them with a layer of grape skins. Before covering the vat a mix of water and salt or water only is added. The vegetable fermentation is a spontaneous process caused by the microorganisms present on the various components (turnip and grape skins). A total 225 lactic acid bacteria (LAB) and 63 yeasts were isolated during this natural fermentation process. The main heterofermentative population was identified as Lactobacillus spp. while the major homofermentative LAB were Pediococcus spp., Candida spp. was the main yeast present. During the first 24 h of fermentation, the pH decreased rapidly to 3.7. At different steps of fermentation the headspace aroma compounds of brovada were analyzed by solid-phase-micro-extraction/gas chromatography/flame ionization detection to define the characteristic aroma profile of this particular product.     

高浓度培养乳酸菌发酵培养基的优化

对一株已经证实具有耐酸、耐胆汁的乳杆菌R8菌株进行培养基的优化研究,以菌体密度为检测指标,采用单因素实验和正交设计实验优化发酵培养基组分,为该益生菌应用于改善胃肠道的健康食品奠定基础。结果显示,该菌最佳发酵配方为:葡萄糖20.0 g,酵母粉30.0 g,MnSO₄·4H₂O 0.075 g,MgSO₄·7H₂O 2.0 g,KH₂PO₄ 2.5 g,柠檬酸铵2.5 g,CH₃COONa·3H₂O 6.25 g,Tween80 1.0 mL,pH6.2。培养基配方经过系统筛选和优化后,菌体浓度(OD_{600 nm})达到2.38。