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.     

Phytate degradation by Bifidobacterium on whole wheat fermentation

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

Calcium Additives and Sprouted Wheat Effects on Phytate Hydrolysis in Whole Wheat Bread

Whole wheat bread loaves were subjected to treatments of different fermentation periods, different sources and levels of calcium, and the addition of sprouted wheat. Phytate losses increased with increased fermentation time. Increasing the calcium level inhibited phytate hydrolysis when the calcium was provided by nonfat dry milk, CaCl₂, or nonfat yogurt, whereas phytate hydrolysis in loaves supplemented with CaCO₃ remained nearly constant. Milk-derived calcium exerted the greatest inhibition of phytate hydrolysis. The addition of sprouted wheat decreased absolute phytate losses. A comparison of phytate losses in yeasted vs nonyeasted loaves suggested that endogenous wheat phytase was quantitatively more important than yeast phytase during breadmaking.     

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.     

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.     

Nonstarter Lactobacillus strains as adjunct cultures for cheese making: in vitro characterization and performance in two model cheeses

Nonstarter lactic acid bacteria are the main uncontrolled factor in today's industrial cheese making and may be the cause of quality inconsistencies and defects in cheeses. In this context, adjunct cultures of selected lactobacilli from nonstarter lactic acid bacteria origin appear as the best alternative to indirectly control cheese biota. The objective of the present work was to study the technological properties of Lactobacillus strains isolated from cheese by in vitro and in situ assays. Milk acidification kinetics and proteolytic and acidifying activities were assessed, and peptide mapping of trichloroacetic acid 8% soluble fraction of milk cultures was performed by liquid chromatography. In addition, the tolerance to salts (NaCl and KCl) and the phage-resistance were investigated. Four strains were selected for testing as adjunct cultures in cheese making experiments at pilot plant scale. In in vitro assays, most strains acidified milk slowly and showed weak to moderate proteolytic activity. Fast strains decreased milk pH to 4.5 in 8 h, and continued acidification to 3.5 in 12 h or more. This group consisted mostly of Lactobacillus plantarum and Lactobacillus rhamnosus strains. Approximately one-third of the slow strains, which comprised mainly Lactobacillus casei, Lactobacillus fermentum, and Lactobacillus curvatus, were capable to grow when milk was supplemented with glucose and casein hydrolysate. Peptide maps were similar to those of lactic acid bacteria considered to have a moderate proteolytic activity. Most strains showed salt tolerance and resistance to specific phages. The Lactobacillus strains selected as adjunct cultures for cheese making experiments reached 10⁸ cfu/g in soft cheeses at 7 d of ripening, whereas they reached 10⁹ cfu/g in semihard cheeses after 15 d of ripening. In both cheese varieties, the adjunct culture population remained at high counts during all ripening, in some cases overcoming or equaling primary starter. Overall, proximate composition of cheeses with and without added lactobacilli did not differ; however, some of the tested strains continued acidifying during ripening, which was mainly noticed in soft cheeses and affected overall quality of the products. The lactobacilli strains with low acidifying activity showed appropriate technological characteristics for their use as adjunct cultures in soft and semihard cheeses.     

Application of phytases from bifidobacteria in the development of cereal-based products with amaranth

The effects of the inclusion of purified phytases from Bifidobacterium longum spp. infantis ATCC15697 and Bifidobacterium pseudocatenulatum ATCC27919 on phytate (InsP ₆) levels were analyzed during breadmaking process. Two different levels of whole amaranth (Amaranthus cruentus) flour (25 and 50 %) were used in bread dough preparation, and they were compared to control doughs made with 100 % wheat flour and 100 % whole wheat flour. Bread samples made with 50 % of amaranth flour showed a significant decrease in technological quality parameters in comparison with control white breads. However, a 25 % of amaranth flour improved the nutritional value of the bread, with only a slight depreciation in the quality. Addition of bifidobacterial phytases resulted in higher InsP ₆ degradation compared with a commercial fungal phytase, without affecting the bread quality. InsP ₆ reduction was especially efficient in breads with 25 % amaranth, leading to InsP ₆ levels below the threshold of mineral bioavailability inhibition for Fe and Zn in human nutrition.     

Fermentation of commercial soy beverages with lactobacilli and bifidobacteria strains featuring high β-glucosidase activity

An increase of isoflavone aglycone content in soy foods can be attained through fermentation with food-grade bacteria. In this study, two commercial soy beverages with distinctive chemical composition (AS and VS) were fermented by strains of lactobacilli (eight strains) and bifidobacteria (two strains) with a high β-glucosidase activity. Along fermentation, growth of the strains and isoflavone deglycosylation in the soy beverages were monitored. Large differences in growth, aglycone content and chemical parameters in AS and VS beverages fermented by different species and strains were observed. Isoflavone glycosides were completely transformed into their corresponding aglycones by most strains during fermentation of AS beverage, whereas large amounts of undeglycosilated isoflavones were still present in fermented VS. Four strains showing strong deglycosylation activity and appropriate technological properties (Lactobacillus casei LP71, Lactobacillus plantarum E112, Lactobacillus rhamnosus E41 and Bifidobacterium pseudocatenulatum C35) were proposed as industrial starters to improve functionality in soy-based fermented foods.     

Discrimination of Volatiles of Refined and Whole Wheat Bread Containing Red and White Wheat Bran Using an Electronic Nose

Abstract: The principal objective of this study was to evaluate the capability of electronic (E) nose technology to discriminate refined and whole wheat bread made with white or red wheat bran according to their headspace volatiles. Whole wheat flour was formulated with a common refined flour from hard red spring wheat, blended at the 15% replacement level with bran milled from representative samples of one hard red and 2 hard white wheats. A commercial formula was used for breadmaking. Results varied according to the nature of the sample, that is, crust, crumb, or whole slices. Bread crust and crumb were completely discriminated. Crumb of whole wheat bread made with red bran was distinct from other bread types. When misclassified, whole wheat bread crumb with white bran was almost invariably identified as refined flour bread crumb. Using crust as the basis for comparisons, the largest difference in volatiles was between refined flour bread and whole wheat bread as a group. When refined flour bread crust was misclassified, samples tended to be confused with whole white wheat crust. Samples prepared from whole bread slices were poorly discriminated in general. E‐nose results indicated that whole wheat bread formulated with white bran was more similar in volatile makeup to refined flour bread compared to whole wheat bread made with red bran. The E‐nose appears to be very capable to accommodate differentiation of bread volatiles whose composition varies due to differences in flour or bran type. Practical Application: Consumer preference of bread made using refined flour in contrast to whole wheat flour is partly due to the different aroma of whole wheat bread. This study used an electronic nose to analyze bread volatiles, and showed that whole wheat bread incorporating white bran was different from counterpart bread made using red bran, and was closer in volatile makeup to “white” bread made without bran. Commercial millers and bakers can take advantage of these results to formulate whole wheat flour with brans of preferred type in order to foster increased consumption of whole wheat products which confer many favorable health benefits.     

Dough and bread qualities of flours with whole waxy wheat flour substitution

Whole wheat flour has been developed as fiber-rich food for healthy bowel function. In this study, whole waxy wheat flour (WWF) was characterized and applied for breadmaking with the hope that breads made from WWF would improve the quality based on the superior properties of the waxy flour in retardation of bread staling. WWF had significantly higher protein, ash and dietary fiber contents (13.5%, 1.6% and 15.3%, respectively), but lower lipid content (0.8%) than commercial white flour (CWF). Amounts of soluble and insoluble dietary fiber in WWF were 4.1% and 11.2% (% total weight), respectively. WWF had lower pasting temperature and viscosities than CWF. Dough made from WWF increased water absorption and showed lower stability during mixing than that from CWF. The high amount of dietary fiber diluted the gluten protein in dough during mixing to form weak and inextensible dough. Bread made from whole waxy wheat flour had significantly low specific volume and big gal cell distribution as compared to that from CWF. Also, dark-brown color and bitter taste are weak points of whole waxy wheat bread. However, the WWF bread was significantly softer than CWF bread during storage. The qualities of breads were also improved using partial WWF substitution (10%, 30% or 50%) for CWF. As a result, whole waxy wheat flour can be used for breadmaking to improve nutritious quality of bread for health benefits.     

Supplementation of alkaline phytase (Ds11) in whole-wheat bread reduces phytate content and improves mineral solubility

Abstract: In this study, alkaline phytase was added to whole‐wheat bread and the phytate content and mineral profiles were compared to commercially available acidic phytase. At neutral pH, some phytate (approximately 20%) was degraded by endogenous phytase in wheat flour, while 40% of phytate was hydrolyzed by alkaline phytase DS11 and a 35% reduction was observed with acidic phytase. Most of the enzymatic activity occurred during the proofing stage, and the rate of reaction depended on pH. DS11 phytase effectively degraded the phytate level within a 30 min treatment at pH 7; however, at least 60 min was needed with acidic phytase to achieve the same hydrolysis level. Mineral profiles were also dramatically affected by the phytate reduction. The biggest increase was observed in Fe²⁺ by the phytase treatment. The Fe²⁺ content increased 10‐fold at pH 7 and 8‐fold at pH 5 with alkaline phytase DS11. Alkaline phytase DS11 was shown to be effective at phytate reduction in whole‐wheat bread preparation. Additionally, phytate degradation enhanced the mineral availability of bread.     

In vitro evaluation of the suitability potential probiotic of lactobacilli isolates from the gastrointestinal tract of chicken

Probiotic lactobacilli could be used to decrease the colonization of pathogenic bacteria in chicken and therefore decrease the risk of foodborne illness to consumers. The present study was conducted to select appropriate microbial strains for the development of potential probiotic. In experiment 1, 18 strains of lactobacilli isolated from the gastrointestinal tract of chicken were evaluated. The strains were demonstrated for their lactic acid, hydrogen peroxide, and exopolysaccharide productions. For experiment 2, the strains were tested for their acid, bile, antimicrobial activity, and antibiotic resistance levels. Among them, Lactobacillus delbrueckii ssp. delbrueckii BAZ32, Lactobacillus acidophilus BAZ29, BAZ36, BAZ43, and BAZ63 which produced high EPS were selected to aggregation ability. It is concluded that L. delbrueckii ssp. delbrueckii BAZ32, L. acidophilus BAZ29 confer high tolerance to acid, bile, antibiotic resistance, high antimicrobial activity, aggregation ability, and EPS production. These strains may be functional feed additives as potential probiotic in chicken.     

Safety and functional aspects of pre-selected lactobacilli for probiotic use in Iberian dry-fermented sausages

The purpose of this study was to investigate lactobacilli for potential probiotic use in Iberian dry-fermented sausages. A total of 18 strains, previously selected, isolated from different sources were evaluated according to safety and functional characteristics including biogenic amines and d-lactic acid production, antibiotic susceptibility, haemolysis, cell adhesion, and antimicrobial activity against food-borne pathogens. The strains Lactobacillus reuteri PL519, L. reuteri PL542, and Lactobacillus fermentum HL57 were able to establish their self on the intestinal epithelium, and an inhibition of such pathogenic bacteria as Listeria monocytogenesin vitro. These strains were also considered safe to be used with regard to their low aminogenic potential and d-lactic acid production, antibiotic resistance pattern and haemolytic activity. Thus, they were selected as a potential probiotic meat starter culture suitable for manufacture of dry-fermented Iberian sausages.     

Evaluation of Leuconostoc citreum HO12 and Weissella koreensis HO20 isolated from kimchi as a starter culture for whole wheat sourdough

Leuconostoc citreum HO12 and Weissella koreensis HO20 isolated from kimchi were evaluated as starter cultures in the making of whole wheat sourdough bread. After 24 h of fermentation at 25 °C, both lactobacilli grew to the final cell numbers of ca. 10⁹ cfu/g dough, and both doughs had similar pHs and total titratable acidities. In addition, the fermentation quotient of the dough with Lc. citreum HO12 was slightly lower than that of the dough with W. koreensis HO20 (1.6 versus 2.8). Sourdoughs and bread with 50% sourdough produced with the starter cultures exhibited consistent ability to retard the growth of bread spoilage fungi (Penicillium roqueforti and Aspergillus niger) and rope-forming bacterium (Bacillus subtilis). Sourdough breads underwent a significant reduction in bread firming during storage. It seems that both lactobacilli have the potential to improve the shelf-life of wheat bread. The results indicate that the selected lactobacilli have unique fermentation characteristics and produce sourdough breads with overall satisfactory quality.     

Effect of whole amaranth flour on bread properties and nutritive value

This study investigated the effect of replacing wheat flour by whole Amaranthus cruentus flour (up to 40 g/100 g) to evaluate its potential utility as a nutritious breadmaking ingredient. The incorporation of amaranth flour significantly increased protein, lipid, ash, dietary fibre and mineral contents. Breads with amaranth have significantly higher amounts of phytates and lower myo-inositol phosphates, which could predict low mineral bioavailability at high levels of substitution (30–40 g/100 g). An increase in crumb hardness and elasticity was observed, and tristimulus colour values were significantly affected when the amaranth concentration was raised. Mineral contents, both micro- and macroelements, were increased significantly by the wheat flour substitution. Whole amaranth flour could be used as a partial replacement for wheat flour in bread formulations, increasing the product’s nutritional value and raising dietary fibre, mineral and protein levels, with a significant slight depreciation in bread quality when used in proportions between 10 and 20 g/100 g. Thus, the inclusion of amaranth flour could be limited to a maximum proportion of 20 g/100 g, thereby maintaining both product quality as well as the nutritional benefit of this ingredient.     

Enzymatic Reduction of Phytate in Whole Wheat Breads

The presence of phytate in flour may be responsible for reduced bioavailability of iron, magnesium, zinc, and calcium from bread. The effect of various concentrations of commercial phytase or phosphatase added to whole wheat flour-yeast doughs on their phytate and nonphytate phosphorus content has been investigated. By using 2.0% (flour basis) of phytase and 0.11% phosphatase the initial phytate phosphorus concentration of the dough was reduced to 1/8 and 1/12 of its initial values, respectively. Storage of the whole wheat breads for up to 96 hr at room temperature showed further significant reduction of phytate phosphorus. The phytate phosphorus content of yeast leavened whole wheat breads decreased during 2 hours of dough fermentation, baking and the subsequent 48 hours of storage at room temperature from 24 mg/100g dough (dry matter) to 1.7 mg/100g bread (dry matter); the phytate phosphorus continued to decrease and after 96 hours storage it was 0.6 mg/100g bread.     

Phytate dephosphorylation by Lactobacillus pentosus CFR3

Studies on phytate‐degrading enzymes from lactobacilli are scarce, despite its potential in improving the nutritional quality of plant‐based foods. Therefore, the current investigation deals with the phytate‐degrading enzyme produced by a native Lactobacillus pentosus strain. Phytase activity was highest towards the end of the exponential phase. Activity increased in the presence of maltose (381.1%) compared with glucose. The presence of phytate in the media stimulated the enzyme production. The enzyme of interest was a 70 kDa protein with a pH and temperature optima of 5.0 and 55–60 °C, respectively. It retained 46% of activity after exposure to 70 °C for 20 min and also showed broad substrate specificity. It was completely inhibited by Hg²⁺, Fe²⁺ and PMSF while being activated by Co²⁺. This report is the first to show dephytinisation of autoclaved finger millet flour either by fermentation with L. pentosus or by treatment with the corresponding cell‐free extract.     

Sourdough-isolated<Emphasis Type="Italic"> Lactobacillus fermentum</Emphasis> as a potent anti-mould preservative of a traditional Iranian bread

Sourdough bread is an ancient method for making long microbial shelf life and strongly flavoured breads. Traditional sourdough bread-making has been practised for centuries in Iran. Although rural bread-making is still highly reliant on sourdough, urban bakeries usually use bakers yeast and also sodium bicarbonate instead. In this study the anti-mould preservative effects of three lactic acid bacteria isolates were investigated on one of the popular traditional Iranian bread (Lavash). Starter cultures were prepared using 20 h cultures of three sourdough-isolated strains, Lactobacillus casei, Lactobacillus plantarum and Lactobacillus fermentum. Different concentrations (0.5%, 1%, 2%, 4% and 8%) of lactobacillus starter cultures were used to prepare a variety of sourdoughs. Lavash doughs were inoculated with 20% of each lactobacilli-fermented sourdough. After a 20 h incubation at 30 °C, a decrease in pH was observed in the different lactobacilli sourdoughs. However an 8% inoculum of L. fermentum resulted in a significant decrease in pH (3.87 to 2.70). Concentrations of 2% and higher of the three lactobacilli used in Lavash sourdough delayed mould growth during storage. However this preservative effect was more significant when an 8% inoculum of L. fermentum was used. These results tend to suggest that selected strains of lactobacilli have a beneficial role in prevention of bread waste by mould spoilage, and hence could extend bread shelf life.     

Sourdough-type bread from waste bread crumb

Optimal conditions for sourdough production from waste bread containing sugar were studied. Using a dough prepared with 50% white bread crumb in water instead of 35% crumb, high levels of acidity were formed at 35°C with a commercial lyophilized starter containingLactobacillus plantarum. The effect of temperature (25 or 35°C) on titratable acidity development was equivalent to crumb concentration (35 or 50%). Cell growth stopped after 12–24 h but acid production continued for more than 48 h. Best results were obtained with whole wheat bread crumb followed by sweet-type crumb (about 8% sugar, dry basis) and white bread crumb. Two commercial sources ofLactobacillus plantarum, one meat starter and one bread starter, were screened based on acid production in fermented bread crumb.     

Potential of Lactic Acid Bacteria to Inhibit Rope Spoilage in Wheat Sourdough Bread

The ability of selected lactic acid bacteria to inhibit the growth of rope-forming Bacillus strains in laboratory experiments and in wheat bread was investigated. Growth of Bacillus subtilis and Bacillus licheniformis was inhibited by Lactobacillus plantarum VTT E-78076 and Pediococcus pentosaceus VTT E-90390 in an automated turbidometry assay and in test bakings. Rope spoilage of wheat bread was inhibited by adding 20-30 g of sourdough/100 g of wheat dough if the sourdough was fermented with Lactobacillus plantarum VTT E-78076,Pediococcus pentosaceus VTT E-90390 or Lactobacillus brevis (commercial starter culture) and the pH values of sourdoughs were adjusted below 4.0 and the amount of total titratable acidity value was >12. Addition of lactic acid alone in concentrations comparable with those formed in sourdoughs did not prevent rope spoilage.