Evidence for formation of heterooligosaccharides by Lactobacillus sanfranciscensis during growth in wheat sourdough

Lactobacillus sanfranciscensis is a key organism of the lactic microflora in traditional and industrial sourdough fermentations. In this paper we provide evidence for the formation of heterooligosaccharides (HeOS) by L. sanfranciscensis during growth in sourdough. To identify the HeOS based on HPAEC-PAD analysis, HeOS standards were synthesized by enzymatic reactions with L. sanfranciscensis levansucrase in a chemically defined system in the presence of raffinose, maltotriose, maltose, xylose, or arabinose as acceptor carbohydrates. The oligosaccharides known to originate from the corresponding acceptor reactions, 1(F)-beta-fructosylraffinose, 1(F)-beta-fructofuranosylmaltotriose, erlose (1(F)-beta-fructofuranosylmaltose), xylsucrose, 1(F)-beta-fructosylxylsucrose, and arabsucrose, were identified by HPAEC-PAD. Evidence for the formation of further tri-, tetra-, and pentasaccharides was provided. Wheat doughs with sucrose were fermented with L. sanfranciscesis TMW 1.392 or the isogenic, levansucrase-negative strain TMW 1.392Deltalev, and the analysis of dough extracts or invertase-treated dough extracts provided evidence for the formation of arabsucrose and erlose in sourdough in addition to 1-kestose and nystose.     

Influence of thiol metabolism of lactobacilli on egg white proteins in wheat sourdoughs

In wheat sourdoughs, the degradation of gluten proteins is favored by acidification and reducing conditions. This study aimed to determine the proteolytic degradation of egg white proteins in wheat sourdoughs acidified with lactobacilli differing in their thiol metabolism. Ovotransferrin was the only major egg white protein that degraded during sourdough fermentations. An extensive degradation of ovotransferrin required a heterofermentative lactobacilli starter, Lactobacillus sanfranciscensis, with glutathione reductase activity. Ovotransferrin was more resistant to breakdown when sourdoughs were acidified with homofermentative lactobacilli or a mutant strain of L. sanfranciscensis lacking the glutathione reductase. Its susceptibility to proteolysis in L. sanfranciscensis sourdoughs is thus attributable to thiol accumulation by L. sanfranciscensis, which apparently altered the structure of ovotransferrin through a reduction of disulfide bonds. Proteolytic degradation of ovotransferrin was attributable to wheat aspartic proteinases. In addition to the susceptibility to proteolysis, other functional properties of egg proteins may be influenced by thiol-exchange reactions.     

Influence of peptide supply and cosubstrates on phenylalanine metabolism of Lactobacillus sanfranciscensis DSM20451(T) and Lactobacillus plantarum TMW1.468

Bread spoilage is mainly due to the growth of filamentous fungi, and metabolites produced during sourdough fermentation by lactobacilli can inhibit fungal growth. One of these metabolites is phenyllactic acid (PLA), which is a catabolite from phenylalanine. In this work, the influence of peptide supply and cosubstrates was determined on PLA formation from phenylalanine by Lactobacillus plantarum TMW1.468 and Lactobacillus sanfranciscensis DSM20451(T). Transport of single amino acids is not efficient in lactobacilli, and only 1% of the offered phenylalanine was converted to PLA. PLA yields were increased 2-4-fold when peptides instead of single amino acids were used as a substrate. The accumulation of phenylalanine after peptide addition indicated that, after transport, transamination was the second limiting factor. In L. plantarum TMW1.468, PLA yields were increased from 5 to >30% upon the addition of alpha-ketoglutarate. In L. sanfranciscensis DSM20451, a combination of both citric acid and alpha-ketoglutarate increased PLA formation. The combined effect of citric acid and alpha-ketoglutarate can be attributed to changes in the NAD/NADH ratio.     

Prolamin hydrolysis in wheat sourdoughs with differing proteolytic activities

The prolamins of wheat, rye, and barley contain structures that are harmful to gluten-sensitive people, and an extensive degradation of these prolamins during food processing might eliminate this problem. Sourdough fermentation is a cereal food process during which some protein degradation occurs. In this study, the prolamin hydrolysis that occurred in a high-proteolytic-activity germinated-wheat sourdough (GWSD) was compared with that of wheat sourdough systems which contained moderate or no proteolytic activities. Virtually all of the wheat prolamins (gliadins and glutenins) were degraded during the GWSD fermentation. Quantification of its prolamin levels confirmed that extensive prolamin hydrolysis had occurred in the GWSD. This hydrolysis was attributed to the cysteine proteinase activities of the germinated wheat. The use of high-proteolytic sourdoughs in baking could make it possible to prepare new low-prolamin cereal-based products for use by gluten-sensitive people, who could then diversify their diets by including these whole-grain containing products into their every-day diets.     

LC-MS/MS quantification of bioactive angiotensin I-converting enzyme inhibitory peptides in rye malt sourdoughs

This study quantified antiotensin I-converting enzyme (ACE) inhibitory peptides in rye malt sourdoughs supplemented with gluten proteins and fermented with six strains of Lactobacillus spp. Bioinformatic analysis of prolamins from barley, rye, and wheat demonstrated that the ACE inhibitory peptides LQP, LLP, VPP, and IPP are frequently encrypted in their primary sequence. These tripeptides were quantified by liquid chromatography-tandem mass spectrometry. Tripeptide levels in sourdoughs were generally higher as compared to the chemically acidified controls. Sourdoughs fermented with different strains showed different concentrations of LQP and LLP. These differences corresponded to strain-specific differences in PepO and PepN activities. The highest levels of peptides VPP, IPP, LQP, and LLP, 0.23, 0.71, 1.09, and 0.09 mmol (kg DM)(-1), respectively, were observed in rye malt: gluten sourdoughs fermented with Lactobacillus reuteri TMW 1.106 and added protease. These concentrations were 6-7 times higher as compared to sourdough without fungal protease and exceed the IC(50) by 100-1000-fold.     

Aluminum tolerance variability in rye and wheat Portuguese germplasm

Regional rye populations and wheat cultivars/lines were screened for aluminum tolerance using the hematoxylin staining method. Portuguese regional rye populations showed better tolerance than the Polish cv. Dank. Zlote, used as tolerant tester. In the group of bread wheats, EPM 305/81, a Barbela reselection, was the most tolerant genotype with the same behaviour as the cv. BH 1146, a tolerant tester. In a study with lines selected from a local Barbela landrace, aluminum tolerance variability was detected. Some lines were as tolerant, or higher, as wheat tester. As Portuguese rye populations and the Barbela wheat landrace have grown for centuries on an acid soil region, the data supports the idea that natural biotic or abiotic stresses associated to man selection, lead to the adaptation of genotypes to specific regional conditions and, in this case, to acid soils where aluminum toxicity occurs.     

Climate-induced overwintering challenges for wheat and rye in northern agriculture

Abstract

Winters are typically harsh in the northernmost agricultural areas of Europe, and winter rye (Secale cereale L.) and wheat (Triticum aestivum L.) are the only winter grain crops that can be grown. However, climate change is projected to result in milder winters, which may enable cultivation of winter crops to a greater extent in the future than is possible today. In this study we aimed at identifying main temperature, precipitation events and characteristics that have resulted in past poor overwintering of rye and wheat in their current production areas in Finland. Using long-term (1970–2006), multi-location datasets, we compared our findings with the projected major changes attributable to climate change. Mixed models were used to estimate mutually comparable overwintering damage to all experiments and logistic regression was used to determine whether climatic parameters are related to high levels of overwintering damage. Severity of overwintering damage, and associated yield penalties, fluctuate considerably on a year-to-year basis and no consistent reduction in variability was recorded during the study period. Particularly for wheat, severity of winter damage in any one year was associated negatively with area sown in the following year. There was no evidence of consistent genetic improvements in winter hardiness, but rye was more winter hardy than wheat. Current risks associated with rye production related to low temperatures could be alleviated in the future, although overwintering damage currently enhanced by high autumn precipitation could increase due to climate change. For wheat, fluctuating conditions hampered overwintering, which may be an even harder challenge in future when weather variation is projected to increase and extreme weather events are projected to become more common.     

Qualitative characterization of benzoxazinoid derivatives in whole grain rye and wheat by LC-MS metabolite profiling

Benzoxazinoids are metabolites occurring in a restricted group of plant species including crops such as rye, wheat, and maize. Focus on the analysis of benzoxazinoid metabolites has typically been due to their importance to plant biochemistry and physiology as highly bioactive molecules that plants use as alleochemicals to defend themselves against predators and infections. However, the potential dietary contribution of these compounds has not been addressed. This study conducted a detailed qualitative characterization of benzoxazinoid metabolites present in the whole grain rye and processed fractions of rye bran, and their presence was also detected in whole grain wheat samples. Several novel benzoxazinoid metabolites of the hydroxamic acids (2,4-dihydroxy-1,4-benzoxazin-3-one, DIBOA; 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one, DIMBOA), lactams (2-hydroxy-1,4-benzoxazin-3-one, HBOA), and benzoxazolinones (1,3-benzoxazol-2-one, BOA) were identified, including double-hexose derivatives of DIBOA, DIMBOA, and HBOA. This paper presents an important addition to the information on the phytochemical composition of rye and wheat grains, which deserves attention in the discussion of the potential health-promoting effects of these grains.     

Alkylresorcinols in selected Polish rye and wheat cereals and whole-grain cereal products

The alkylresorcinol content and homologue composition in selected Polish rye and wheat cultivars and selected whole-grain cereal products were determined in this study. Cereal grains and whole-grain cereal products were extracted with acetone, whereas bread types were extracted with hot 1-propanol. The average alkylresorcinol content in tested rye (approximately 1100 mg/kg DM) and wheat (approximately 800 mg/kg DM) grains harvested in Poland was within the range previously reported in Swedish and Finnish samples. The total alkylresorcinol content in tested cereal products available on the Polish market varied from very low levels in barley grain-based foods up to 3000 mg/kg DM in wheat bran. The total alkylresorcinol content in 14 bread samples extracted with hot 1-propanol varied from approximately 100 mg/kg DM in whole bread made with honey up to approximately 650 mg/kg DM in whole-rye bread. Calculated ratios of C17:0 to C21:0 homologues, a useful parameter previously used to distinguish between rye and wheat cereals and their derived products, was about 1.2-1.4 in rye products, about 0.2 in wheat products, and varied between 0.2 and 0.6 in cereal-derived products containing a mixture of whole rye and/or wheat. The data set obtained were subsequently compared using cluster and principal component analysis, which allowed the tested cereal products to be classified into two major groups consisting of whole-rye or whole-wheat products, respectively. On the basis of that approach, mixed cereal products containing rye and wheat bran or whole rye and wheat flour were grouped between those two well-defined clusters. Our work not only provides a detailed examination of alkylresorcinols in selected Polish rye and wheat cultivars and selected whole-grain cereal products, but also demonstrates that this type of analysis accompanied by the use of proper statistical algorithms offers an objective way to evaluate the quality of whole-grain rye and/or wheat and their derived products.     

Alkylresorcinols as markers of whole grain wheat and rye in cereal products

The alkylresorcinol (AR) content of Swedish wheat grain samples, as well as of cereal ingredients and cereal foods containing wheat and rye, was determined. The average total AR content in Swedish wheat was 412 microg/g (ranging between 227 and 639 microg/g), which is lower than that in Swedish rye analyzed in a previous study. The relative composition of AR homologues was consistent for wheat samples and differed markedly from that of rye. Notably, the ratio of the homologues C17:0/C21:0 was approximately 0.1 in wheat and approximately 1.0 in rye, indicating that it can be used to distinguish between those two cereals. The AR content in cereal foods commonly consumed in Sweden varied widely, from nondetectable levels in white wheat flour and products not containing the outer parts of wheat and/or rye to >900 microg/g in some whole grain rye products. AR content in cereal foods was calculated from their recipes using average AR values for the cereal ingredients determined in this study. As there was a good correlation between calculated and analyzed AR levels in cereal foods (R2 = 0.91), it is possible to estimate the proportion of whole grain wheat and/or rye in a given cereal product on the basis of AR content and C17:0/C21:0 ratio. ARs appear to be good markers of whole grain wheat and rye in foods, and their analysis may be an objective way to identify foods rich in whole grain wheat and/or rye or brans thereof.     

Antioxidant activity of steryl ferulate extracts from rye and wheat bran

Antioxidant activity of steryl ferulates from other sources than rice have not yet been studied much, despite the fact that rice steryl ferulates (gamma-oryzanol) have been shown to possess good antioxidant activity. In this study, steryl ferulate extracts from wheat or rye bran were studied for their capability to inhibit hydroperoxide formation in bulk methyl linoleate and methyl linoleate emulsion. Further, their activity to scavenge DPPH radicals was analyzed. The activities were compared to synthetic steryl ferulates, rice steryl ferulates, ferulic acid, and alpha-tocopherol. Nonrice cereal extracts of steryl ferulates exhibited good antioxidant activity, especially in the bulk lipid system. The radical scavenging activity was similar to that of nonesterified ferulic acid, indicating that the ferulic acid moiety is responsible for the antioxidant properties. This study illustrates a new aspect to the health-promoting properties of rye and wheat.     

Fate of benzoxazinone allelochemicals in soil after incorporation of wheat and rye sprouts

Growing cereals (especially rye), which are incorporated into the soil to increase soil fertility or organic matter content, is a common practice in crop rotation. The additional sanitizing effect of this incorporation has often been appreciated and is said to be due to leaching of benzoxazinones and subsequent formation of benzoxazolinones. In this study wheat (Stakado) and rye (Hacada) sprouts were incorporated into soil in amounts that simulated agricultural practice. By extraction and subsequent LC-MS analysis the disappearance and appearance of benzoxazinones, benzoxazolinones, and phenoxazinones in soil were followed. In the wheat experiments 6-methoxybenzoxazolin-2-one (MBOA) was detected as the main compound. 2-Hydroxy-7-methoxy-1,4-benzoxazin-3-one (HMBOA) and 2-hydroxy-1,4-benzoxazin-3-one (HBOA) were detected as well. No phenoxazinones were detected. For the rye experiment the picture was more complex. In the first 2 days of incubation MBOA and 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA) were detected as the main allelochemicals along with HBOA, HMBOA, and benzoxazolin-2-one (BOA), in decreasing order. Later in the incubation period some 2-amino-3H-phenoxazin-3-one (APO) was detected and the amount of HBOA increased considerably and decreased again. The profiling of the benzoxazinone metabolites and their derivates in soil was dynamic and time-dependent. The highest concentrations of most of the compounds were seen at day 1 after incorporation. A maximum concentration was reached at day 4 for a few of the compounds. This study is the first of its kind that shows the dynamic pattern of biologically active benzoxazinone derivates in soil after incorporation of wheat and rye sprouts. Methods for organic synthesis of HBOA and HMBOA were developed as part of the study.     

Response of triticale,wheat, and rye to N in a greenhouse

Abstract

Triticales (X Triticosecale, Wittmack), 6TA 131, 6TA 298 and 6TA 298 and 6TA 385, Arthur wheat (Triticum aestivum L.), and Abruzzi rye (Secale cereale L.) were grown in a soil treated with 0, 50, 100, and 200 ppm N. The objective of this study was to determine the influence of N rate on dry matter yields, N uptake, and N utilization efficiency.

The dry matter and N uptake increased while the N utilization efficiency decreased with increasing N fertilizer applied to the soil for all the cultivars. At high N rate, rye had higher dry matter and N uptake than the triticales and wheat; no consistent differences were obtained between triticale and wheat. However, rye and triticale tended to be more efficient in utilizing absorbed N than wheat at the lower N rates.     

Glucan and fructan production by sourdough Weissella cibaria and Lactobacillus plantarum

After a large screening on sourdough lactic acid bacteria, exopolysaccharide (EPS)-forming strains of Weissella cibaria, Lactobacillus plantarum, and Pediococcus pentosaceus were selected. After 6 days of incubation at 30 degrees C, the synthesis of EPS in MRS-based broth ranged from 5.54 to 7.88 mg mL-1. EPS had an apparent molecular mass of ca. 104 Da. As shown by carbohydrate consumption, the synthesis of EPS was found from sucrose only. Two types of homopolysaccharides were synthesized: glucans simultaneously with growth and fructans after 1 day of incubation. Two protein bands of ca. 180-200 kDa were in situ detected on SDS-PAGE gels incubated with sucrose. PCR products of ca. 220 bp were found for L. plantarum PL9 (100% of identity to putative priming glycosyltransferase of L. plantarum WCFS1) and W. cibaria WC4 (80% of identity to putative glycosyltransferase, epsD, of Bacillus cereus G9241) by using hybrid primers for the priming gtf genes. Degenerated primers DexreuR and DexreuV showed a unique PCR product, and the predicted amino acid sequences were identical for W. cibaria WC4 and L. plantarum PL9. The sequence had similarity with polysaccharide biosynthesis glycosyltransferases. W. cibaria WC4 or L. plantarum LP9 synthesized ca. 2.5 g kg-1 EPS during sourdough fermentation with sucrose added. Compared to the sourdough started with an EPS-negative strain, the sourdough started with W. cibaria WC4 or L. plantarum LP9 increased the viscosity, and the resulting bread had higher specific volume and lower firmness. The synthesis of EPS by selected sourdough lactic acid bacteria could be considered as a useful tool to replace the additives for improving the textural properties of baked goods.     

Structural characterization and biological activities of an exopolysaccharide kefiran produced by Lactobacillus kefiranofaciens WT-2B(T)

Lactobacillus kefiranofaciens, isolated from kefir grains, produces an extracellular polysaccharide when cultured, not only in PYG10 medium but also in a liquid medium containing a rice hydrolysate that had been previously degraded by treatment with a glucoamylase. The maximum yield of the polysaccharide, using the rice hydrolysate as the medium, was 2.5 g/L after a 7-day culture period at pH 5.0 and 33 degrees C. Compositional analysis, methylation analysis, specific rotation, and (1)H and (13)C NMR spectroscopy revealed that the structures of polysaccharides obtained from these two different culture media are essentially identical. The polysaccharide is composed of a hexasaccharide repeating unit and, thus, is known as kefiran. The weight-average molecular weight and the z-average radius of gyration of a sample, purified from the rice hydrolysate medium, were determined to be 7.6 x 10(5) g/mol and 39.9 nm, respectively, by gel permeation chromatography equipped with a multiangle laser-light-scattering photometer. Changes in blood pressure and serum components were examined in SHRSP/Hos rats, using doses of 100 and 300 mg of kefiran/kg of rat. A suppression in the increase in blood pressure was observed in these rats after 30 days. This activity is discussed in terms of the concentration of serum components of the rat, with emphasis on lipid components such as cholesterols, triglycerides, and free fatty acids.     

Preliminary studies on triticale,wheat, barley,and rye responses to lime and N.

Abstract

The lime and N requirements for triticale (X Triticosecale Wittmack) have not been established because of the relatively short history of the crop. This study was designed to evaluate the effects of lime and high N rates on triticale, wheat (Triticum aestivum L.), barley (Hordeum vulgare L.), and rye (Secale cereale L.) on Dickson silt loam (Typic Paleudult) and Decatur silty clay loam (Rhodic Paleudult) in 1974–1976. The soils had pH values of 4.9 and 5.5 with no lime and 5.4 and 5.8, respectively, when limed as recommended. The fertilizer rates were 112, 140, and 170 kg N/ha. Yields and N, P, K, Ca, Mg, Mn, Fe, Al, Zn, Cu, and B were determined in straw and grain. Liming the Dickson soil increased the straw yields of barley at 112 kg N/ha and grain yields of the cultivars generally at the 170 kg N/ha rate. Liming the Decatur soil did not have consistent effects on straw yields but increased the grain yields of the wheat and rye cultivars. Increasing N rate increased the straw yields of wheat on Dickson but decreased the grain yields of barley in the same soil with no lime. Nitrogen fertilization did not have consistent effects on the Decatur soil. The N, P, K, Ca, Mg, and Mn compositions suggested that more differences occured at the species level than at the cultivar level.     

Studies on rye (Secale cereale L.) lines exhibiting a range of extract viscosities. 2. Rheological and baking characteristics of rye and rye/wheat blends and feeding value for chicks of wholemeals and breads

Five rye lines exhibiting a wide range of extract viscosities were evaluated for the rheological and baking properties of their flours, individually and in blends with hard red spring wheat flour. Commercial cultivars of rye and triticale were included in the study as controls. Extract viscosities of rye flours were higher than those of corresponding wholemeals, indicating shifting of water-extractable arabinoxylan into flour during roller milling. Falling numbers of the rye flours correlated positively with their extract viscosities in the presence (r = 0.73, p < 0.05) or absence (r = 0.65, p < 0.05) of an enzyme inhibitor. Farinograms revealed the weakness of rye and triticale flours compared to wheat flour. Extract viscosities of rye flours were negatively correlated (r = -0.65, p < 0.05) with mixing tolerance index and positively correlated (r = 0.64, p < 0.05) with dough stability, suggesting a positive impact of extract viscosity on dough strength. Extract viscosity was negatively correlated (r = -0.74, p < 0.05) with loaf volume and specific volume (r = -0.73, p < 0.05) and positively correlated (r = 0.73, p < 0.05) with loaf weight of rye/wheat bread. Overall, the results indicated that 30% of flour from high or low extract viscosity rye could be incorporated into rye/wheat breads without seriously compromising bread quality. Inclusion of rye, particularly high extract viscosity rye, in chick diets seriously impeded growth performance and feed efficiency. Part of the arabinoxylan survived bread-making and exerted an effect on chicks, although substantially lower digesta viscosities were observed in chicks fed rye bread diets than in those fed rye wholemeals.     

Measurement of acrylamide and its precursors in potato, wheat, and rye model systems

The relationship between acrylamide and its precursors, namely, free asparagine and reducing sugars, was studied in cakes made from potato flake, wholemeal wheat, and wholemeal rye, cooked at 180 degrees C, from 5 to 60 min. Between 5 and 20 min, major losses of asparagine, water, and total reducing sugars were accompanied by large increases in acrylamide, which maximized in all three products between 25 and 30 min, followed by a slow linear reduction. Acrylamide formation did not occur to a large degree until the moisture contents of the cakes fell below 5%. Linear relationships were observed for acrylamide formation with the residual levels of asparagine and reducing sugars for all three food materials.     

Evaluation of biosurfactant production from various agricultural residues by Lactobacillus pentosus

The cost of biosurfactant production may be significantly decreased by using inexpensive carbon substrates like agricultural residues. However, scarce information can be found in the literature about the utilization of lignocellulosic residues for obtaining biosurfactants. Usually agricultural residues are field burned, producing various toxic compounds to the atmosphere; so, as an interesting alternative to the traditional field burning of this kind of residue, this work proposes the utilization of agricultural wastes (barley bran, trimming vine shoots, corn cobs, and Eucalyptus globulus chips) for simultaneous lactic acid and biosurfactant production. Previous to this biotechnological process, lignocellulosic residues were hydrolyzed, using H2SO4, under selected conditions and neutralized with CaCO3. Following, Lactobacillus pentosus was employed for the fermentation of hemicellulosic hydrolyzates after nutrient supplementation. Biosurfactants were measured by taking into account the surface tension reduction. The highest value of reduction (21.3 units) was found when using hemicellulosic sugar hydrolyzates obtained from trimming vine shoots, corresponding to 0.71 g of biosurfactant per g of biomass and 25.6 g of lactic acid/L. On the contrary, barley bran husk hydrolyzates only produced 0.28 g of biosurfactant per g of biomass and 33.2 g of lactic acid/L. The differences between biosurfactant production can be attributed to the different compositions of the hydrolyzates.     

重金属对小麦和黑麦种子萌发及幼苗生长的影响

小麦（Triticum aetivum）作为世界上第一大粮食作物,是人类最主要的食物来源,在全球粮食贸易中占有十分重要的地位。随着人口增加、土地减少和农业生产环境条件改变及生活水平提高,对小麦品种的要求也越来越严格,单纯依靠小麦种内遗传资源已不能满足育种的需要;同时,由于小麦育种的定向选择,使其遗传多样性丢失,导致遗传基础日益狭窄,不仅限制了产量和品质的进一步改良,而且使小麦对生物性和非生物性环境胁迫的脆弱性增加,所以有必要开发新的基因资源,