Selection of phytate-degrading human bifidobacteria and application in whole wheat dough fermentation

Lately, whole wheat products are highly recommended from their healthy properties. However, the presence of phytic acid (InsP(6)) could partly limit their benefits because it decreases the mineral bioavailability due to its chelating properties. The objective of this work was to select strains with high phytate-degrading activity from human feces, and evaluate their suitability for the bread making process. Twenty-three different bifidobacterial strains (13 from infants and 10 from adults) were isolated, belonging to the species Bifidobacterium longum, Bifidobacterium breve and Bifidobacterium catenulatum. The phosphatase and phytase activities of these strains were evaluated as well as their ability to degrade InsP(6) during growth. Then, the fermentative ability of the strain showing the highest phytate-degrading activity (B. longum. BIF307) was determined in whole wheat breadmaking. The use of the selected bifidobacterial strain as starter during whole wheat fermentation resulted in bread with similar technological quality than the control (in absence of bifidobacteria) and crumb with lower levels of inositol phosphates. Therefore, the used of the selected Bifidobacterium strain in whole wheat breadmaking process could provide potential nutritional benefits by decreasing the antinutrient content of the product.     

Selection of lactic acid bacteria with high phytate degrading activity for application in whole wheat breadmaking

Whole wheat products although highly recommended from the nutritional point of view, contain high levels of phytic acid, an antinutrient that decrease the mineral bioavailability. The objective of this study was to select strains with high phytate-degrading activity from different parts of the gastrointestinal tract of chickens following a phytate-rich diet and to test their suitability for the breadmaking process of whole wheat bread. Different lactobacilli and bifidobacteria strains were isolated and individually assayed for phosphatase and phytase activities, since both enzymes could contribute to the degradation of phytate. The isolates showing the highest phytate degrading activity belonged to the species Bifidobacterium dentium, Lactobacillus reuteri (L-M15) and Lactobacillus salivarius (L-ID15). The two lactobacilli L-M15 and L-ID15 were selected and tested their fermentative ability in whole wheat breadmaking. Whole wheat breads in the presence of the selected lactobacilli had similar technological quality than the control (in absence of lactobacilli) and extended freshness; moreover, their presence resulted in bread crumbs with lower levels of inositol phosphates. Overall, the two intestinal lactobacilli strains showing high phytate degrading activity were proven to have good properties for being used as starters in whole wheat breadmaking process.     

Phytate degradation by human gut isolated Bifidobacterium pseudocatenulatum ATCC27919 and its probiotic potential

The growing awareness of the relationship between diet and health has led to an increasing demand for food products that support health above and beyond providing basic nutrition. Probiotics are live organisms present in foods, which yield health benefits related to their interactions with the gastrointestinal tract. Phytases are a subgroup of phosphatases that catalyse the desphosphorylation of phytate, which reduces its negative impact on mineral bioavailability, and generates lower inositol phosphates. The aims of this investigation were to (i) study the ability of the probiotic candidate Bifidobacterium pseudocatenulatum to degrade phytate in synthetic medium, to (ii) identify the lower inositol phosphates generated, to (iii) study its survival under conditions mimicking gastrointestinal passage and finally to (iv) assess adhesion of the bacteria to Caco-2 cells. The first steps of InsP₆ degradation by B. pseudocatenulatum phytate-degrading enzyme/s were preferentially initiated at the DL-6-position and 5-position of the myo-inositol ring. It suggests that the main InsP₆ degradation pathway by B. pseudocatenulatum by sequential removal of phosphate groups was D/L-Ins(1,2,3,4,5)P₅ or D/L-Ins(1,2,3,4,6)P₅; D/L-Ins(1,2,3,4)P₄; to finally Ins(1,2,3)P₃ and D/L-Ins(1,2,4)P₃/D/L-Ins(1,3,4)P₃. This human strain also showed a notable tolerance to bile as well as a selective adhesion capacity (adhesion to control surfaces was zero), to human intestinal Caco-2 cells comparable to the commercial probiotic B. lactis. The phytate-degrading activity constitutes a novel metabolic trait which could contribute to the improvement of mineral absorption in the intestine as a nutritional probiotic feature with potential trophic effect in human gut.     

Effects of soaking, germination and fermentation on phytic acid, total and in vitro soluble zinc in brown rice

Rice is an important staple food in Asian countries. In rural areas it is also a major source of micronutrients. Unfortunately, the bioavailability of minerals, e.g. zinc from rice, is low because it is present as an insoluble complex with food components such as phytic acid. We investigated the effects of soaking, germination and fermentation with an aim to reduce the content of phytic acid, while maintaining sufficient levels of zinc, in the expectation of increasing its bioavailability. Fermentation treatments were most effective in decreasing phytic acid (56–96% removal), followed by soaking at 10 °C after preheating (42–59%). Steeping of intact kernels for 24 h at 25 °C had the least effect on phytic acid removal (<20%). With increased germination periods at 30 °C, phytic acid removal progressed from 4% to 60%. Most wet processing procedures, except soaking after wet preheating, caused a loss of dry mass and zinc (1–20%). In vitro solubility, as a percentage of total zinc in soaked rice, was significantly higher than in untreated brown rice while, in steeped brown rice, it was lower (p < 0.05). Fermentation and germination did not have significant effects on the solubility of zinc. The expected improvement due to lower phytic acid levels was not confirmed by increasing levels of in vitro soluble zinc. This may result from zinc complexation to other food components.     

Fermentation of rice using amylolytic Bifidobacterium

Twenty-two amylolytic bifidobacteria grown in BHI-starch media were compared for the amylase activity of the intra- and extra-cellular enzymes. The activity of the cells grown in the liquid medium differed considerably. Among the strains tested B. adolescentis Int57 and B. adolescentis ZS8 exhibited higher activities than others. In rice medium containing 0.05% -cysteine·HCl and 0.2% yeast extract, the amylolytic Bifidobacterium strains grew considerably better than non-amylolytic Bifidobacterium strains. B. adolescentis Int57, which showed highest growth and amylase activity in the rice medium, was chosen and rice fermentation was investigated. The titratable acidity (TA) reached 2.43 and pH decreased to 4.4 after 24 h fermentation. The relative ratio of acetic acid to lactic acid gradually decreased during fermentation. The concentration of reducing sugar and amylase activity gradually increased and reached 14 mg maltose equivalent/ml and 35 mU/ml min, respectively, in 24 h. The accumulated reducing sugar was mostly maltotriose. The layer-separation of fermented product was stabilized by the addition of 1% gelatin. It was suggested that amylolytic bifidobacteria may be used for the production of fermented rice products.     

Lactobacillus reuteri CRL 1100 as starter culture for wheat dough fermentation

The effect of sucrose on the fermentation balance of Lactobacillus reuteri CRL 1100 and the invertase activity of this strain in wheat dough and culture medium (MRSs) was evaluated. The enzyme activity was dependent on the environmental pH releasing glucose and fructose from sucrose hydrolysis. Glucose was used as carbon source, while fructose was mainly used as electron acceptor to produce mannitol up to 10h of fermentation. Thereafter, fructose seemed to be metabolized by the heterofermentative pathway, which determined an increase in the concentration of acetate (6 mmol l(-1)), lactate (2 mmol l(-1)) and ethanol (1 mmol l(-1)) and the lack of mannitol formation after glucose depletion. The fermentation balance of Lb. reuteri CRL 1100 during the dough fermentation resulted in lower (63%) ethanol, higher (75%) acetate production and soluble carbohydrates concentrations, like MRSs cultures. This fermentation profile would be important to obtain an optimal growth of yeast and the optimal bread flavor and taste.     

Effects of milk products fermented by Bifidobacterium longum on blood lipids in rats and healthy adult male volunteers

The effects of milk products fermented by Bifidobacterium longum strain BL1, a probiotic strain, on blood lipids in rats and humans were studied. Rats were fed a cholesterol-enriched experimental diet, supplemented with lyophilized powders of 1) acid milk (control), 2) milk fermented with a mixed culture of ordinary yogurt starters composed of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus (SL), and 3) bifidobacterium milk fermented with the probiotic B. longum strain BL1, respectively. The bifidobacterium milk feeding brought about significant lowering of the serum concentrations of total cholesterol, low-density lipoprotein cholesterol, and triglycerides, in comparison with the control, while no change in high-density lipoprotein cholesterol concentration was observed. On the other hand, supplementation with SL milk resulted in only slight, nonsignificant decreases in serum lipid concentrations in comparison with the control. In the human study, 32 subjects with serum total cholesterol ranging from 220 to 280 mg/dl were randomly assigned to two treatments: 1) intake of a low-fat drinking yogurt prepared with ordinary yogurt starters composed of S. thermophilus and L. delbrueckii subsp. bulgaricus (P-group) and 2) intake of a low-fat drinking yogurt prepared with the two ordinary yogurt starters plus B. longum strain BL1 (B-group). After intake for 4 wk at 3 x 100 ml/day, reduction of serum total cholesterol was observed in approximately half of the B-group subjects; a particularly significant decrease in serum total cholesterol was found among subjects with moderate hypercholesterolemia (serum total cholesterol > 240 mg/dl). However, the serum lipid concentrations in the P-group subjects were almost stable during the experimental periods. The present results indicate the potential of the probiotic B. longum strain BL1 in serum lipid improvement.     

Effect of fortification on physico-chemical and microbiological stability of whole wheat flour

Stability of fortified whole wheat flour (WWF) was evaluated using NaFeEDTA, elemental iron, ZnSO₄ and ZnO as fortificants. Fortified WWF was stored in tin boxes and polypropylene bags for 60 days under ambient storage condition (ASC) and controlled storage condition (CSC). Fortification significantly (p ? 0.05) decreased moisture and protein content and increased ash content to 5.44%, 6% and 23%, as compared to control. Fortified WWF, assayed periodically for mould contamination manifested a significant inhibition (∼1 log reduction) in flours containing elemental iron. Low storage temperature and relative humidity (RH) indicated lower level of mould count during extended storage time. Tin boxes, as storage material, exhibited a better protection against mould attack, acting as an effective barrier for moisture. Fortificants exerted a slight deteriorative effect on texture characteristics of the chapattis made of these flours but chapattis were still accepted by the judges. Zinc fortificants seemed like having little or no effect on the quality of the flours and chapattis, made of such flours. Shelf life of fortified flour may be extended by using elemental iron as fortificant and storing the product in tin boxes under relatively low temperature and RH.     

Effect of sourdough fermentation on stabilisation,and chemical and nutritional characteristics of wheat germ

Lactic acid bacteria strains were identified from wheat germ by 16S rRNA partial sequencing, subjected to RAPD-PCR typing and screened. Lactobacillus plantarum LB1 and Lactobacillus rossiae LB5 were used as starters to produce sourdough fermented wheat germ (SFWG). The chemical and nutritional characteristics of SFWG were compared to those of the raw wheat germ (RWG). Lipase activity in SFWG was ca. 2.6-fold lower than that found in RWG. As shown by SPME/GC/MS analysis, most of the volatile compounds derived from lipid oxidation during storage (40 days) were at markedly lower levels in SFWG compared to RWG. Fermentation of wheat germ increased of ca. 50% the concentration of free amino acids. Glu markedly decreased in SFWG, due to its conversion in GABA. The concentration of the anti-nutritional factor raffinose also decreased in SFWG. The in vitro protein digestibility, the concentration of total phenols, phytase and antioxidant activities were increased by fermentation.     

Safety and intestinal microbiota modulation by the exopolysaccharide-producing strains Bifidobacterium animalis IPLA R1 and Bifidobacterium longum IPLA E44 orally administered to Wistar rats

Bifidobacterium animalis subsp. lactis IPLA R1 and Bifidobacterium longum IPLA E44 strains were tested for their safety and ability to modulate the intestinal microbiota in vivo. Chemically simulated gastrointestinal digestion showed considerably lower survival of E44 than R1 strain, the first microorganism also being more sensitive to refrigerated storage in 10% skimmed milk at 4 °C. Harmful glycosidic activities were absent, or at low levels, in the strains R1 and E44. Both strains were sensitive to most antibiotics and resistant to aminoglycosides, a common feature in bifidobacteria. Similar to several other bifidobacteria strains, B. animalis subsp. lactis IPLA R1 displayed a moderate resistance against tetracycline which correlated with the presence of tet(W) gene in its genome. The general parameters indicating well-being status, as well as translocation to different organs and histological examination of the gut tissues, revealed no changes induced by the administration of bifidobacteria to rats. Twelve-week-old male Wistar rats were distributed into three groups, eight rats in each. Two groups were administered daily over 10⁸ cfu of the corresponding strain suspended in 10% skimmed milk for 24 days, whereas rats in the placebo group received skimmed milk without microorganisms added. The microbiota and short chain fatty acids (SCFA) were monitored in faeces at different time points during treatment and in caecum content at the end of the assay. Quantitative PCR (qPCR) showed that faecal and caecal Bifidobacterium levels were higher in bifidobacteria-fed rats than in the placebo rats at the end of the intervention, whereas total anaerobic plate counts did not show significant differences. Quantification of B. animalis and B. longum by qPCR showed that, independent of the microorganism administered, treatment with bifidobacteria resulted in higher levels of B. animalis in the caecum. PCR-DGGE analysis of microbial populations revealed a higher diversity of bands in caecum content of rats fed B. animalis IPLA R1 than in the placebo group and rats fed B. longum IPLA E44. Remarkably, although no variations in the proportion of acetate, propionate and butyrate were found, at the end of the assay the total SCFA concentration in the faeces of rats fed bifidobacteria was significantly higher and those in caecum content significantly lower, than that of the placebo group. This suggests a displacement of the SCFA production to parts of the colon beyond the caecum in rats receiving bifidobacteria. Therefore, the oral administration of B. animalis IPLA R1 and B. longum E44 can be considered safe, these microorganisms having the ability to modulate the intestinal microbiota of rats by influencing SCFA and the bifidobacterial population levels.     

Behaviour ofYersinia enterocolitica andAeromonas hydrophila in skim milk during fermentation by various lactobacilli

In this study, behaviour ofYersinia enterocolitica andAeromonas hydrophila in skim milk during fermentation by various_{lactobacillus} sp. were determined. pH values of the skim milk samples were also examined during fermentation. The amount of produced lactic acid and diacetyl/acetoin productions of theLactobacillus sp. were estimated. Antimicrobial effects of the lactobacilli onY. enterocolitica andA. hydrophila were also determined by an agar diffusion method. WhileY. enterocolitica was not inhibited and grew during fermentation,A. hydrophila was inhibited, in part, and the growth was retarded. Results were supported by the agar diffusion method forY. enterocolitica, whereas inhibition activity was not found forA. hydrophila. The highest lactic acid productions were estimated inL. bulgaricus (7.50 mg/ml) andL. acidophilus (5.63 mg/ml) and four out of sixLactobacillus sp. were found to be diacetyl/acetoin producers.     

One-pot fermentation of pladienolide D by Streptomyces platensis expressing a heterologous cytochrome P450 gene

Pladienolide D is a 16-hydroxylated derivative of pladienolide B, produced by Streptomyces platensis. To facilitate the production of pladienolide D, the gene encoding a pladienolide B 16-hydroxylase from S. bungoensis was introduced into S. platensis. The recombinant produced pladienolide D at a production level comparable to that of pladienolide B.     

Exopolysaccharides produced by Bifidobacterium longum IPLA E44 and Bifidobacterium animalis subsp. lactis IPLA R1 modify the composition and metabolic activity of human faecal microbiota in pH-controlled batch cultures

Exopolysaccharides (EPS) isolated from two Bifidobacterium strains, one of human intestinal origin (Bifidobacterium longum subsp. longum IPLA E44) and the other from dairy origin (Bifidobacterium animalis subsp. lactis IPLA R1), were subjected to in vitro chemically simulated gastrointestinal digestion, which showed the absence of degradation of both polymers in these conditions. Polymers were then used as carbon sources in pH-controlled faecal batch cultures and compared with the non-prebiotic carbohydrate glucose and the prebiotic inulin to determine changes in the composition of faecal bacteria. A set of eight fluorescent in situ hybridisation oligonucleotide probes targeting 16S rRNA sequences was used to quantify specific groups of microorganisms. Growth of the opportunistic pathogen Clostridium histolyticum occurred with all carbohydrates tested similarly to that found in negative control cultures without added carbohydrate and was mainly attributed to the culture conditions used rather than enhancement of growth by these substrates. Polymers E44 and R1 stimulated growth of Lactobacillus/Enterococcus, Bifidobacterium, and Bacteroides/Prevotella in a similar way to that seen with inulin. The EPS R1 also promoted growth of the Atopobium cluster during the first 24 h of fermentation. An increase in acetic and lactic acids was found during early stages of fermentation (first 10–24 h) correlating with increases of Lactobacillus, Bifidobacterium, and Atopobium. Propionic acid concentrations increased in old cultures, which was coincident with the enrichment of Clostridium cluster IX in cultures with EPS R1 and with the increases in Bacteroides in cultures with both microbial EPS (R1 and E44) and inulin. The lowest acetic to propionic acid ratio was obtained for EPS E44. None of the carbohydrates tested supported the growth of microorganisms from Clostridium clusters XIVa+b and IV, results that correlate with the poor butyrate production in the presence of EPS. Thus, EPS synthesized by bifidobacteria from dairy and intestinal origins can modulate the intestinal microbiota in vitro, promoting changes in some numerically and metabolically relevant microbial populations and shifts in the production of short chain fatty acids.     

三株双歧杆菌鉴定及遗传特征分析

目的 对3株双歧杆菌用不同方法进行鉴定,并结合药物敏感性测试结果和全基因组测序（WGS）数据,对菌株获得性耐药元件及其可转移性进行分析。方法 分别采用生化、基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）及WGS方法对3个益生菌产品中分离的3株双歧杆菌进行鉴定,测试菌株对氨苄西林等8种抗生素的敏感性,结合WGS数据,使用在线数据库分析菌种携带的获得性耐药元件,并对其可转移性进行分析。结果 对3株双歧杆菌,生化方法可给出较为可靠的属水平鉴定结果;MALDI-TOF MS方法可给出可靠的种水平鉴定结果;基于WGS结果计算平均核苷酸一致性和DNA-DNA杂交值,可给出可靠的种和亚种水平鉴定结果,结合wgSNP及wgMLST分析,3株菌鉴定结果均为动物双歧杆菌乳亚种。3株菌对四环素和庆大霉素均呈现敏感性下降表型,其中庆大霉素敏感性下降与厌氧菌种转运系统缺乏有关,为固有表型;3株菌染色体上均携带tet（W）,且其下游均存在IS5序列,说明该耐药基因可能为可转移元件,经与GenBank中58个来自不同种属细菌的tet（W）对比发现,动物双歧杆菌乳亚种中的tet（W）序列具有一定的保守性。结论 双歧杆菌的鉴定与遗传特征分析应结合生化、MALDI-TOF MS、抗生素敏感性和WGS等技术综合开展,尤其WGS技术不仅可以对双歧杆菌进行亚种水平的精准鉴定,还可以针对耐药元件及可转移性进行深入分析,为益生菌相关菌种产品的研发和监管、益生菌类保健食品行业的健康发展提供有力的技术支撑。     

Effect of oxygen absorber on accumulation of free fatty acids in brown rice and whole grain wheat during storage

Because rapid rancidities of lipids in unpolished grains, such as brown rice and whole grain wheat, are often problematic, Suppressions of their lipid degradations during postharvest therefore important for their further utilization. In this study, effect of oxygen absorber on lipid deterioration in unpolished grain during storage was investigated. Kernels and flours of brown rice, rice with germ and whole grain wheat were stored for eight weeks at 4 °C and 26 °C, with and without oxygen absorber. Free fatty acids (FFAs) accumulation in the unpolished grains during storage was accelerated by polishing, milling and storage at high temperature (26 °C), because these storage conditions stimulated activities of their lipid hydrolyses. When oxygen absorber was added and the storage system was sealed, additional FFAs accumulations in these unpolished grains. Decreases both in absolute amounts of linoleic acids of the lipids and in total tocochromanols contained in these unpolished grains were depressed by addition of oxygen absorber to the storage system, which was confirmed by an anti-oxidative effect of oxygen absorber. These results suggested that introduction of oxygen absorber into the storage system of unpolished grains would be effective under the condition where their hydrolytic activities of lipids are suppressed.     

Oviposition attractants for Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae) in the volatiles of whole wheat flour

Plodia interpunctella is attracted to whole wheat flour. Volatiles obtained from whole wheat flour by Porapak Q trapping were assayed using pitfall olfactometers, and were attractive to mated females (active at 10⁻¹ gram/day equivalent, gde), but not to males or unmated females, suggesting that the volatiles are oviposition attractants for P. interpunctella. Silicon dioxide column chromatography of the crude volatiles revealed that the fraction recovered with 3% ether in pentane (3% EP) was active and contained at least 27 components, in which alkanals (C₆-C₁₀) and 2E-alkenals (C₇-C₁₁) were active as individual aldehydes or mixtures. Nonanal was most active (the lowest active limit, LAL: 10⁻² μg), followed by 2E-nonenal and 2E-decenal (1 μg each). The synthetic mixture of the 27 components identified from 3% EP was attractive at 1.59×10⁻⁴ μg, which was equal in activity to the crude volatiles at 10⁻¹ gde (LAL) for mated females. The ether fraction (E) showed no activity by itself but synergistically enhanced the activity of the 10% EP fraction (LAL: 10⁻⁴ μg). Alkanols (C₅-C₉), lactones and carboxylic acids (C₅-C₁₈) were identified from the E fraction, in which hexanol and hexanoic acid were the major components. Both hexanol and palmitic acid were synergistic in combination with the aldehyde mixture.     

Comparison of releasing bound phenolic acids from wheat bran by fermentation of three Aspergillus species

Wheat bran was fermented at 28 °C for 7 days under 70% humidity by Aspergillus niger, Aspergillus oryzae and Aspergillus awamori. Total phenolic content (TPC) of the unfermented sample was 1531.5 μg g^?1 wheat bran. After the fermentation of Aspergillus awamori, Aspergillus oryzae and Aspergillus niger, TPC reached 5362.1, 7462.6 and 10 707.5 μg g^?1, respectively. The antioxidant activity in the extractions of fermented wheat bran also increased significantly compared with the unfermented sample (P < 0.05). Aspergillus niger showed the greatest capacity to release bound ferulic acid (416.6 μg g^?1). Aspergillus oryzae and Aspergillus awamori had the advantages of releasing more chlorogenic acid (84.0 μg g^?1) and syringic acid (142.3 μg g^?1), respectively. The destructive effect of Aspergillus niger on wheat bran structure was the strongest, followed by that of Aspergillus oryzae. This effect of Aspergillus niger may be due to its higher cellulase, xylanase, arabinofuranosidase and β‐xylosidase activities. Besides, Aspergillus oryzae possessed higher β‐glucosidase activity, and Aspergillus awamori had higher α‐amylase and feruloyl esterase activities. Aspergillus niger may be the best to release bound phenolic acids in the three Aspergillus species. These will provide the helpful information for understanding mechanism of the fermentation by Aspergillus species releasing bound phenolic in wheat bran.     

Galactooligosaccharides derived from lactose and lactulose: influence of structure on Lactobacillus, Streptococcus and Bifidobacterium growth

The effect of structure on the fermentative properties of potential prebiotic trisaccharides derived from lactulose like 6′-galactosyl-lactulose (β-d-galactopyranosyl-(1 → 6)-β-d-galactopyranosyl-(1 → 4)-β-d-fructopyranose), 4′-galactosyl-lactulose (β-d-galactopyranosyl-(1 → 4)-β-d-galactopyranosyl-(1 → 4)-β-d-fructopyranose), and 1-galactosyl-lactulose (β-d-galactopyranosyl-(1 → 4)-β-d-fructopyranosyl-(1 → 1)-β-d-galactopyranose); and from lactose like 4′-galactosyl-lactose (β-d-galactopyranosyl-(1 → 4)-β-d-galactopyranosyl-(1 → 4)-β-d-glucopyranose) and 6′-galactosyl-lactose (β-d-galactopyranosyl-(1 → 6)-β-d-galactopyranosyl-(1 → 4)-β-d-glucopyranose), has been assessed in vitro. Fermentations with twelve pure strains of Lactobacillus, Streptococcus and Bifidobacterium were carried out using the purified trisaccharides as the sole carbon source, and bacteria growth was evaluated at 600 nm by means of a microplate reader during 48 h. Maximum growth rates (μ_max) and lag phase were calculated. In general, all the strains tested were able to utilize lactulose and pure trisaccharides derived from lactulose and lactose when they were used as sole carbon source. Nonetheless, glycosidic linkage and/or the monosaccharide composition of the trisaccharides affected the individual strains lag phase, cell densities and growth rates. A general preference towards β-galactosyl residues β(1-6) and β(1-1) linked over those β(1-4) linked was observed, and some strains showed higher cell densities and speed of growth on 6′-galactosyl-lactulose than on 6′-galactosyl-lactose. This is the first study of the effect of lactulose-derived oligosaccharides on pure culture growth which shows that transglycosylation of lactulose allows for obtaining galactooligosaccharides with new glycosidic structures and would open new routes to the synthesis of compounds with potential prebiotic effects.     

Removing gluconic acid by using different treatments with a Schizosaccharomyces pombe mutant: Effect on fermentation byproducts

Three different treatments involving inoculation with Schizosaccharomyces pombe YGS-5 and Saccharomyces cerevisiae G1 strains were tested with a view to reducing the amount of gluconic acid in synthetic medium. The treatments involved (a) simultaneous inoculation with S. cerevisiae and S. pombe (SpSc); (b) depletion of gluconic acid with S. pombe and subsequent inoculation with S. cerevisiae following removal of S. pombe from the medium (Sp − Sp + Sc); and (c) as (b) but without removing S. pombe from the medium (Sp + Sc). The results thus obtained were compared with those for a control treatment involving fermentation with S. cerevisiae alone (Sc). The amounts of volatile compounds quantified in the fermented media were similar with the treatments where gluconic acid was previously depleted (viz.Sp − Sp + Sc and Sp + Sc). Amino acids were used in large amounts by S. pombe during removal of gluconic acid; this affected subsequent fermentation by S. cerevisiae and the formation of byproducts. Based on the gluconic acid uptake, fermentation kinetics, volatile composition and absence of off-flavours in the fermented media, both treatments (Sp − Sp + Sc and Sp + Sc) can be effectively used in winemaking processes to remove gluconic acid from must prior to fermentation.     

Co-metabolism of citrate and maltose byLactobacillus brevis subsp.lindneri CB1 citrate-negative strain: effect on growth, end-products and sourdough fermentation

Growth, substrates and end-product formation of the maltose and citrate co-metabolization byLactobacillus brevis subsp.lindneri CB1 citrate-negative strain were initially studied in synthetic medium. Compared to maltose (19 g/l) fermentation, the co-metabolization of maltose (10 g/l) plus citrate (9 g/l) caused faster cell growth, increased the concentrations of lactic acid and especially of acetic acid (from 0.7 g/l to 2.9 g/l), produced succinic acid (0.5 g/l) and reduced ethanol synthesis. Highest activities of acetate kinase, the same of lactate dehydrogenase and a reduced alcohol dehydrogenase activity were detected in cytoplasmic extracts of cells growing on maltose plus citrate. The breakdown of citrate depended upon the continuous presence of maltose in the growth medium. Upon depletion of citrate, the cells continued through the normal maltose fermentation, having a diauxic metabolic curve as shown by impedance measurements. Concentrations of citrate from 3 g/l to 15 g/l led to increases of acetic acid from 1.25 g/l to 5.55 g/l. Since maltose was naturally present during sourdough fermentation, the addition of 9 g citrate per kg wheat dough enabled the co-metabolization of maltose and citrate byL. brevis subsp.lindneri CB1. Compared with traditional sourdough fermentation, faster cell growth, a higher acetic acid concentration and a reduced quotient of fermentation were obtained by co-metabolism.