传统酸面团中细菌与酵母菌的分离与鉴定

为进一步描述我国传统面食发酵剂的理化性质和菌落组成,收集了北方地区6份发酵剂样品,测定了其酸度和菌落总数,并对分离、纯化、初筛后得到的75株细菌和60株酵母菌进行了测序鉴定。结果显示,样品pH值范围为3.73~5.46,总滴定酸度为8.3~19.8 mL;乳酸菌和酵母菌的计数结果分别为8.35±0.07~9.75±0.12 Log cfu/g,6.31±0.22~8.68±0.04 Log cfu/g。鉴定出包括短乳杆菌(Lactobacillus brevis)、植物乳杆菌(Lactobacillus plantarum)和旧金山乳杆菌(Lactobacillus sanfranciscensis)在内的乳酸菌8种;酵母菌4种,其中优势菌为酿酒酵母(Saccharomyces cerevisiae);其他细菌6种,主要为解淀粉芽孢杆菌(B.amyloliquefaciens)、地衣芽孢杆菌(B.licheniformis)和醋杆菌。结果表明,我国传统面食发酵剂菌相复杂,以酵母菌和乳酸菌为主,还包括芽孢杆菌、醋酸杆菌在内的多种其他细菌,甚至可能含有致病菌。通过比较细菌和酵母在不同酸面团样品中的分布,发现不同来源样品的微生物种类组成存在差异。     

On the microbiological profile of traditional Portuguese sourdough

Traditional manufacture of bread from maize has been noted to play important roles from both economic and social standpoints; however, enforcement of increasingly strict hygiene standards requires thorough knowledge of the adventitious microbiota of the departing dough. To this goal, sourdough as well as maize and rye flours from several geographic locations and in two different periods within the agricultural year were assayed for their microbiota in sequential steps of quantification and identification. More than 400 strains were isolated and taxonomic differentiation between them was via Biomerieux API galleries (375 of which were successfully identified) following preliminary biochemical and morphological screening. The dominant groups were yeasts and lactic acid bacteria (LAB). The most frequently isolated yeasts were Saccharomyces cerevisiae and Candida pelliculosa. The most frequently isolated LAB were (heterofermentative) Leuconostoc spp. and (homofermentative) Lactobacillus spp.; L. brevis, L. curvatus, and L. lactis ssp. lactis were the dominant species for the Lactobacillus genera; Lactococcus lactis ssp. lactis for lactococci; Enterococcus casseliflavus, E. durans, and E. faecium for enterococci; and Streptococcus constellantus and S. equinus for streptococci.     

Study of the behaviour of Lactobacillus plantarum and Leuconostoc starters during a complete wheat sourdough breadmaking process

The acidification properties, metabolic activity and technological performance of four individual Lactobacillus plantarum or Leuconostoc freeze-dried starters were investigated during a complete wheat sourdough breadmaking process including 0.2 g/100 g baker's yeast. Microbiological contents (lactic acid bacteria and yeasts), acidification characteristics (pH and total titratable acidity), soluble carbohydrates (maltose, glucose and fructose) and fermentative end-products (lactic and acetic acids, ethanol) contents were evaluated during both sourdough and corresponding bread dough fermentation. Biochemical and technological analysis of the resulting bread products are also presented. Some differences among strains in acidification properties and soluble carbohydrates availability were outlined both in sourdough and bread dough. Each individual Leuconostoc or Lb. plantarum starter was able to produce a characteristic fermentation and was found to ensure the production of breads with overall satisfactory acceptance.     

The bacteriocin producer Lactobacillus amylovorus DCE 471 is a competitive starter culture for type II sourdough fermentations

Type II sourdoughs were prepared using Lactobacillus amylovorus DCE 471, a producer of the bacteriocin amylovorin L. The strain was used as a starter culture for rye and wheat sourdoughs on laboratory scale (10 L), and in rye sourdough on pilot scale (100 L). The sourdoughs were acidified to a pH of around 3.5 within 15 h (laboratory dough) to 25 h (pilot‐scale dough). Final amylovorin L titres of 0.3–0.4 (laboratory scale) and 0.2 (pilot scale) MAU kg^?1 of sourdough were detected. After baking of wheat dough that was supplemented with the pilot‐scale sourdough, no amylovorin L activity was recovered from the breadcrumbs. On laboratory scale, aeration or the addition of complex carbohydrates hardly affected growth or amylovorin L production. Rye and wheat sourdough fermentation were rather similar despite differences in sugar concentrations. The persistence of L. amylovorus DCE 471 during rye sourdough fermentation, both on laboratory and pilot scale, was confirmed by repetitive sequence‐based polymerase chain reaction (rep‐PCR) and by testing isolates towards an amylovorin L‐sensitive organism. Further, rep‐PCR indicated that the background microbiota of the flour—probably responsible for the production of low amounts of acetic acid—grew poorly and were overgrown by L. amylovorus DCE 471 during the pilot‐scale fermentation. Copyright © 2007 Society of Chemical Industry     

Adaptability of lactic acid bacteria and yeasts to sourdoughs prepared from cereals, pseudocereals and cassava and use of competitive strains as starters

The adaptability of lactic acid bacteria (LAB) and yeasts to sourdoughs prepared from cereals, pseudocereals and cassava was investigated using PCR-DGGE and bacteriological culture combined with rRNA gene sequence analysis. Sourdoughs were prepared either from flours of the cereals wheat, rye, oat, barley, rice, maize, and millet, or from the pseudocereals amaranth, quinoa, and buckwheat, or from cassava, using a starter consisting of various species of LAB and yeasts. Doughs were propagated until a stable microbiota was established. The dominant LAB and yeast species were Lactobacillus fermentum, Lactobacillus helveticus, Lactobacillus paralimentarius, Lactobacillus plantarum, Lactobacillus pontis, Lactobacillus spicheri, Issatchenkia orientalis and Saccharomyces cerevisiae. The proportion of the species within the microbiota varied. L. paralimentarius dominated in the pseudocereal sourdoughs, L. fermentum, L. plantarum and L. spicheri in the cassava sourdough, and L. fermentum, L. helveticus and L. pontis in the cereal sourdoughs. S. cerevisiae constituted the dominating yeast, except for quinoa sourdough, where I. orientalis also reached similar counts, and buckwheat and oat sourdoughs, where no yeasts could be detected. To assess the usefulness of competitive LAB and yeasts as starters, the fermentations were repeated using flours from rice, maize, millet and the pseudocereals, and by starting the dough fermentation with selected dominant strains. At the end of fermentation, most of starter strains belonged to the dominating microbiota. For the rice, millet and quinoa sourdoughs the species composition was similar to that of the prior fermentation, whereas in the other sourdoughs, the composition differed.     

不同酵母菌与植物乳杆菌复合发酵对新会柑酵素品质的影响

为评价不同酵母菌与植物乳杆菌复合发酵对新会柑酵素品质的影响,以新会柑新鲜果浆为原料,采用3种酵母菌种分别与植物乳杆菌进行复合发酵.追踪发酵过程中发酵液的酵母活菌数、总糖含量、pH和总酸的变化趋势,并比较不同发酵体系新会柑酵素的生物活性物质及其体外抗氧化活性.结果显示:三株酵母菌均可在新会柑中正常生长,但酿酒酵母和马克斯...     

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.     

韩国泡菜加工过程中微生物区系的研究

为了解韩国泡菜在制作过程中微生物的区系,分离鉴定其在腌制、发酵过程中的优势微生物。结果表明：韩国泡菜腌制过程中优势乳酸菌经鉴定为短乳杆菌（Lactobacillus brevis）、植物乳杆菌（Lactobacillusplantarum）;优势酵母菌为近平滑假丝酵母（Candida parapsilosis）、粗状假丝酵母（Candida valida）;韩国泡菜在发酵过程中的优势乳酸菌为植物乳杆菌（Lactobacillus plantarum）、弯曲乳杆菌（Lactobacillus curvatus）、短乳杆菌（Lactobacillus brevis）、肠膜明串珠菌肠膜亚种（Leuconostoc mesenteroides subsp. mesenteroides）;优势酵母菌为近平滑假丝酵母（Candida parapsilosis）、粗状假丝酵母（Candida valida）、酿酒酵母（Saccharomyces cerevisiae）。     

The microbial ecology of tape ketan fermentation

The growth of fungi, yeasts and bacteria was followed during the fermentation of tape ketan. The tape was prepared using samples of Indonesian ragi as inoculum. Fermentation was characterized by the dominant growth of Amylomyces rouxii and Candida pelliculosa (10(5)-10(7) cfu/g), and a lesser contribution from Saccharomyces cerevisiae. Hansenula anomala grew to a limited extent during some fermentations. Bacteria of the genera Bacillus and Acetobacter contributed also to the fermentation, producing populations up to 10(5) cfu/g. Ragi was the main source of microorganisms involved in the fermentation. Tape fermented by inoculating pure cultures of Amylomyces rouxii and the yeasts, either individually or as mixtures, was not of typical quality, indicating the importance of bacteria in the overall fermentation.     

The microbial ecology of soybean soaking for tempe production

Soybeans soaked in tap water for 24 to 36 h at 20, 30 or 37 degrees C underwent a natural fermentation that was characterized by the growth of microorganisms to 10(8)-10(10) cfu/ml (depending on temperature) and a reduction of pH from 6.5 to 4.5. Lactobacillus casei, Streptococcus faecium, Staphylococcus epidermidis and Streptococcus dysgalactiae dominated the fermentation but, significant contributions were also made by Klebsiella pneumoniae, Klebsiella ozaenae, Enterobacter cloacae, Enterobacter agglomerans, Citrobacter diversus and Bacillus brevis, and the yeasts Pichia burtonii, Candida didensiae and Rhodotorula rubra. Fermentation of surface-decontaminated beans in sterile water with pure cultures of these isolates showed L. casei, Strep. faecium and Staph. epidermidis to be the main species responsible for the pH reduction. Soybeans were the main source of microorganisms for the fermentation. Boiled beans did not undergo an acid fermentation.     

Propionic acid production by cofermentation of Lactobacillus buchneri and Lactobacillus diolivorans in sourdough

Cooperative metabolism of lactobacilli in silage fermentation converts lactate to propionate. This study aimed to determine whether propionate production by Lactobacillus buchneri and Lactobacillus diolivorans can be applied for bread preservation. Propionate formation was observed in cofermentation with L. buchneri and L. diolivorans in modified MRS broth as well as sourdough with low, medium and high ash contents. 48 mM of propionate was formed in sourdough with medium ash content, but only 9 and 28 mM propionate were formed in sourdoughs prepared from white wheat flour or whole wheat flour, respectively. Acetate levels were comparable in all three sourdoughs and ranged from 160 to 175 mM. Sourdough fermented with L. buchneri and L. diolivorans was used in breadmaking and its effect on fungal spoilage was compared to traditional sourdough or propionate addition to straight doughs. Bread slices were inoculated with Aspergillus clavatus, Cladosporium spp., Mortierella spp. or Penicillium roquefortii. The use of 20% experimental sourdough inhibited growth of three of the four moulds for more than 12 days. The use of 10% experimental sourdough deferred growth of two moulds by one day. Bread from traditional sourdough with added acetate had less effect in inhibiting mould growth.     

Involvement of Lactic Acid Bacteria in the Fermentation of TEF (Eragrosfis tef), an Ethiopian Fermented Food

Tef injera, a pancake-like acidic food is prepared from fermented tef ( Eragrostis tef ), flour. A complex group of microorganisms is involved in the fermentation. Fermentation is initiated by members of the Enterobacteriaceae. Their activities during the first 18 hr of fermentation reduce the pfi of the dough to about 5.8. They are then succeeded by Leuconostoc mesenteroides and Streptococcus faecalis. As the pH is further reduced to about 4.7, Pediococcus cerevisiae, Lactobacillus brevis, Lactobacillus plantarum and Lactobacillus fermentum become the predominating flora and remain so until the fermentation is terminated at 72 hr. These lactic acid bacteria are responsible for the acidic characteristics of the dough. Yeasts only appear in significant numbers at a latter stage of the fermentation.     

植物乳酸菌M616对发酵酸面团发酵特性的影响

本文研究了在酸面团发酵过程中,植物乳酸菌M616(Lactobacillus plantarum M616)对面团p H、TTA以及糖类含量变化的影响,并利用F3流变式发酵仪和吹泡仪对面团发酵力及流变特性的变化进行了研究。结果表明植物乳酸菌M616不仅对酵母菌的生长具有一定抑制作用,而且在面团p H和TTA的变化过程中起主导作用;另外,乳酸菌对淀粉的降解作用大于对还原糖的吸收,从而使面团中还原糖的含量增加,增加面团的甜味。在发酵后期,面团p H降低至3.73,这在一定程度上抑制了体系中霉菌和其它杂菌的生长。植物乳酸菌的生长代谢不仅显著提高了面团黏度、筋力,而且减缓了面团在发酵过程中韧性的降低。其中发酵酸面团对面团黏度和筋力影响最大,发酵过程中酸面团的最大黏度和筋力可以达到酵母发酵面团的2倍。     

HPLC Determination of Folate in Liver and Liver Products

Our objective was to produce reliable data on the contents of folate vitamers in pig, beef and chicken liver, beef kidney and some liver products. Folate contents were determined in monoglutamate form, using reversed-phase HPLC. Pig and beef livers contained 730-1470 mg folates in 100g, tetrahydrofolate being the most abundant form detected. Frozen chicken liver also contained about 1000 μg folates in 100g. Processed foods containing liver, except canned baby foods, were also good folate sources. In processed foods, 5-methyltetrahydrofolate was the predominant folate form. The detected folate contents were high, compared with published data.     

Sourdough in gluten-free bread-making: An ancient technology to solve a novel issue?

The increasing demand for high quality gluten-free (GF) bread, clean labels and natural products is raising the need for new approaches in GF bread-making. Sourdough is the foremost fermentation used for baking purposes and it has been proven to be ideal for improving the texture, palatability, aroma, shelf life and nutritional value of wheat and rye breads. These characteristic features derive from the complex metabolic activities of the sourdough-resident lactic acid bacteria and yeasts, e.g. acidification, production of exopolysaccharides, proteolytic- amylolytic- and phytase activity, and production of antimicrobial substances. These effects have been extensively studied and well described for traditional baking, whereas little is known about the role of sourdough in GF baking. Yet, the microbiological and qualitative characterisation of local GF fermented products indicate an overlap with the microbiota of wheat/rye fermentation and suggest that the positive metabolic activities of the sourdough microbiota are still retained during fermentation of GF crops. Thus, the use of sourdough in GF baking may be the new frontier for improving the quality, safety and acceptability of GF bread.     

酸面团中优势乳酸菌和酵母菌对麦谷蛋白结构的影响

以分离自我国传统酸面团中的优势菌种植物乳杆菌（Lactobacillus plantarum）Sx9、旧金山乳杆菌（Lactobacillus sanfranciscensis）Gm4及酿酒酵母（Saccharomyces cerevisiae）Sq7为发酵菌种,研究其对小麦麦谷蛋白大聚体的溶解性、游离巯基含量及二级结构变化的影响。结果表明,发酵24 h后,植物乳杆菌Sx9、旧金山乳杆菌Gm4及酿酒酵母Sq7使麦谷蛋白大聚体的溶解率分别增加7.06%、2.47%、2.47%。在发酵6～12 h,植物乳杆菌Sx9对二硫键作用明显,发酵液中游离巯基含量从0.29 μmol/mg迅速增加至0.61 μmol/mg。植物乳杆菌Sx9和旧金山乳杆菌Gm4对二级结构的影响明显,发酵6 h后,无规则卷曲含量几乎为零,全部转化为β-折叠和β-转角,而酿酒酵母Sq7影响甚微,仅有极少量的转化。因此,相比于酿酒酵母Sq7,植物乳杆菌Sx9和旧金山乳杆菌Gm4对麦谷蛋白大聚体的降解效果较好,且植物乳杆菌Sx9最佳。     

Effect of Select Parameters of the Sourdough Rye Fermentation on the Activity of Some Mixed Starter Cultures

The effect of select parameters (i.e., rye flour ash content, temperature, and dough yield) of the sourdough fermentation on the fermentation activity of different starter cultures (lactic acid bacteria Lactococcus lactis ssp. Lactis, Weissella confusa, Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus helveticus, and yeast Kluyveromyces marxianus subsp. Marxianus) was determined. The major metabolic end products of fermentation (D, L-lactic acid, acetic acid, ethanol and glycerol) and the evolution of total phenolic content and folic acid during bread making were measured. Lactobacillus helveticus and Kluyveromyces marxianus allowed obtaining sourdoughs with the highest lactic acid/acetic acid ratios. The mixed starter culture with Lactococcus lactis and Saccharomyces cerevisiae generated the most important quantities of D/L lactic acid. The maximum values of ethanol concentration were obtained in case of the sourdoughs from whole rye flour fermented at lower temperature (30°C) with mixed starter cultures containing Sacchomyces cerevisiae. The fermentation process and type of starter culture are also tools to increase the bioactive compounds, enabling the increase of the phenolic content of the sourdough.     

Defined multi-species semi-liquid ready-to-use sourdough starter

This paper aimed at setting up a protocol for the manufacture of a defined multi-species semi-liquid ready-to-use sourdough starter consisting of lactic acid bacteria (LAB) selected on the basis of their capacity of rapid acidification and adaptation to several technological factors. Preliminarily, 56 strains of sourdough LAB were screened based on the acidification kinetics during sourdough fermentation. The influence of temperature (25-37 degrees C), NaCl (2%, w/w, of wheat flour), initial cell number (10(7)-10(9) cfu g(-1)), dough yield (150-180), substrate for cell cultivation (mMRS or SDB media) and phase of cell growth (exponential or stationary phases) on the kinetics of acidification was also determined. Lactobacillus casei DPPMA27, Weissella confusa DPPMA20 and Lactobacillus fructivorans DPPMA8 were selected and used to produce a defined multi-species semi-liquid sourdough starter. Fermentation was carried out in a 2l batch fermentation bioreactor (12 h at 37 degrees C), under controlled pH, by cultivating cells in a medium made of water (60%), flour and other nutrients (40%). The semi-liquid sourdough starter was stored for 30 days under different conditions and used for dough fermentation (dough yield 160). The microbial composition, acidifying activity and fermentation end-products were kept constant during 21 days of storage. The findings of this study may be useful to increase the use of sourdough in bakery industries.     

The microflora of sourdough. XVIII. The protein degrading capabilities of lactic acid bacteria in sourdough

Acidification of the dough by the use of sourdough or acidifiers is necessary not only for good baking quality of rye flour but it is also very important for development of the typical sensory characteristics of rye bread. We confirmed that the lactic acid bacteria of sour dough are proteolytic. Proteolytic effects are observed in the increase of the amino acid content during fermentation. A marked increase was found in the content of leucine, alanine, valine, isoleucine, glutamic acid, glutamine, arginine, lysine, methionine, phenylalanine, tyrosine and serine. Lactobacillus plantarum showed a higher proteolytic activity than L. brevis ssp. lindneri or L. fructivorans.     

东北粘豆包酸面团菌群多样性与自制酸面团中菌群变化规律的研究

东北粘豆包是我国北方地区一种历史悠久、风味独特的民族传统食品,长期以来深受广大北方人民的喜爱,但对东北粘豆包酸面团的研究十分匮乏。本文以不同地区采集的东北粘豆包酸面团和自制的东北粘豆包酸面团为研究对象,利用聚合酶链式反应-变性梯度凝胶电泳(polymerase chain reaction-denaturing gradient gel electrophoresis,PCR-DGGE)技术探究了东北粘豆包酸面团中微生物菌群的多样性及动态变化。实验结果表明:在东北粘豆包酸面团中共鉴定了6个菌属、11个菌种的细菌,推测其中的发酵乳杆菌和植物乳杆菌为细菌优势发酵菌种;同时鉴定了3个菌属、4个菌种的真菌,推测其中的酿酒酵母为真菌优势发酵菌种;自制东北粘豆包酸面团中微生物菌群多样性比较丰富,在不同发酵阶段,菌群多样性与优势发酵菌种都会随时间推移而有所改变。