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The presence of Class II hydrophobins produced by fungi on barley results in primary gushing of beer. Gushing is the spontaneous overfoaming of carbonated beverages by opening of bottles. Solving gushing problems caused by brewing raw materials has received much scientific attention. Lipophilic extract of hops are introduced to brewers as foam suppressor in fermenters. We studied the effects of hop extract on gushing and found that lipophilic hop extract could reduce gushing. The effects are different when hop extract is added before mashing than when it is added after mashing. Hop extract contains fats and waxes and the effects on gushing are explained by a change in the physical state of its components during mashing which are due to temperature effects. Especially the effect of saturated fatty acids and waxes becomes apparent which are known as gushing inducers. This indicates that with respect to gushing potential of the hop extract's components, it is better to be added to cold wort (after mashing and filtration). Our study also showed an important effect of the filtration step on the amount of gushing.

Since their discovery in 1997, work on proteinaceous wheat xylanase inhibitors (XIs) led to the identification and thorough biochemical and structural characterization of three classes of XIs, namely TAXI-type (Triticum aestivum xylanase inhibitor), XIP-type (xylanase inhibiting protein), and TLXI-type (thaumatin-like xylanase inhibitor) XIs. Already early on, a role in plant defense has been put forward for these proteins, mainly based on the observation that these XIs are only active against xylanases of microbial origin and can not inhibit the plant endogenous xylanases known so far. Considerable effort has been devoted to substantiate this plant defense hypothesis. Data at the genomic, transcriptomic and proteomic levels increasingly provide evidence that XIs, occurring as large polymorphic families, do indeed participate in plant defense. This review summarizes the current knowledge on XIs and the accumulated evidence on their role in plant defense. TLXI and XIP can be classified as pathogenesis-rela...

The effects of germination time and temperature on the quality of rye malt and worts derived thereof were investigated using Response Surface Methodology. Amylolytic and proteolytic enzyme activities were increased by long germination periods, while β-glucanase activity was not influenced. Total and Soluble Nitrogen content were also not significantly affected by the variations in germination conditions. Free Amino Nitrogen (FAN) was found in higher amounts in worts prepared from rye malts with long germination times. Extract contents were higher in rye malt than in the control barley malt and could be increased by a favourable germination regime, while no such impact on wort fermentability was found. High wort viscosities could be significantly reduced by a long germination period at low temperatures, but were still unacceptably high. The same conditions favoured the development of endoxylanase activity. Arabinoxylan (AX) accumulated during the germination process and their extractability increased. The results suggest that longer germination periods resulted in an increased number of AX molecules with lower molecular mass. Optimal rye malt qualities within the limits of this study were found for a germination time of 144 h at 10 °C, which resulted in an acceptable FAN content and the lowest measured viscosity.

Within the HEALTHGRAIN diversity screen, the variability of the contents of dietary fiber (DF) and components thereof was studied in wheat. Furthermore, the contribution of genotype and environment to this variability was estimated. The levels of total DF (TDF), total nonstarch polysaccharide (TOTNSP), water-extractable nonstarch polysaccharide (WENSP), total arabinoxylan (TOTAX), lignin, and beta-glucan in whole meal, flour, and/or bran varied approximately 1.8-fold. The highest variability was observed for the water-extractable arabinoxylan (WEAX) level in flour and bran (approximately 3.7-fold). Genotype and environment contributed to a similar extent to the variability in TDF, TOTNSP, and TOTAX content in wheat. The observed relatively high impact of genotype-environment interaction suggests that the levels of these constituents are weak breeding parameters. The WENSP level is a more stable parameter as the effect of the interaction term was much less than the impact of genotype. For TOTAX and WEAX in flour, WEAX in bran, beta-glucan in whole meal, and extract viscosity, wheat genotype determined approximately 50% or higher of the variation observed, whereas the impact of the genotype-environment interaction was relatively low. These findings suggest that the health-related and technological functionality of wheat can be directed to a certain extent by selection of appropriate wheat varieties.

Endo-1,4-β- d -xylanase (EC 3.2.1.8, β- d -xylanase) activity, and arabinoxylan (AX) level and extractability were monitored for the first time simultaneously in wheat kernels (Triticum aestivum cv. Glasgow) up to 24 days post-imbibition (DPI), both in the absence and presence of added gibberellic acid (GA). Roughly three different stages (early, intermediate and late) can be discriminated. Addition of GA resulted in a faster increase of water extractable arabinoxylan (WEAX) level in the early stage (up to 3–4 DPI). This increase was not accompanied by the discernible presence of homologues of the barley X-I β- d -xylanase as established by immunodetection. This suggests that other, yet unidentified β- d -xylanases operate in this early time window. The intermediate stage (up to 13 DPI) was characterized by the presence of unprocessed 67 kDa X-I like β- d -xylanase, which was much more abundant in the presence of GA. The occurrence of higher levels of the unprocessed enzyme was related with higher β- d -xylanase activities and a further increase in WEAX level, pointing to in vivo activity of the unprocessed 67 kDa β- d -xylanase. During the late stage (up to 24 DPI) gradual processing of the 67 kDa β- d -xylanase occurred and was associated with a drastic increase in β- d -xylanase activity. Up to 120-fold higher activity was recorded at 24 DPI, with approx. 85% thereof originating from the kernel remnants. The WEAX level decreased during the late stage, suggesting that the β- d -xylanase is processed into more active forms to achieve extensive AX breakdown.

Xylanases (EC 3.2.1.8) hydrolyse the backbone of cereal cell wall arabinoxylans and often have a significant impact on cereal-based processes and end-products. The use of microbial xylanases as processing aids in this respect is well established and has been extensively studied. Much less research has focused on inherently present cereal-associated xylanases and their possible impact. Cereals produce xylanases for re-modeling and expansion of cereal cell walls during normal cell growth and for more drastic cell wall degradation during seed germination. Besides these endogenous xylanases, cereals also contain microbial xylanases from micro-organisms populating the outer grain kernels layers. Unfortunately, these microbial xylanases are often inhibited by wheat proteinaceous xylanase inhibitors and they hence escape standard xylanase activity measurements. It is more correct to refer to these activity levels as ‘apparent’ xylanase activity levels. As a result, the occurrence of cereal-associated xylanases might have been largely underestimated in the past and hence unjustly been disregarded. The levels and the types of cereal-associated xylanases differ strongly between grain species, varieties, and tissues, and are largely affected by grain growing conditions. These variations in the levels of grain-associated xylanase activity affect several cereal-based food and feed applications. This paper provides an overview of the occurrence and variability of cereal-associated xylanases and of their potential impact on bread making, shelf life of refrigerated doughs, brewing, animal feed efficiency, pasta production, and wheat gluten–starch separation.

The prolamins in seven Algerian Sahara sorghum cultivars of varying seed shape and color were investigated. Protein contents ranged from 12 to 16%. Prolamins were the major protein fraction. They could be separated according to degree of disulfide cross-linking. Kafirin monomers and low molecular weight polymers could be extracted with 70% ethanol, whereas highly cross-linked kafirins additionally needed a reducing agent to become extractable. Kafirin monomers of α-, β- and γ-type were purified and N-terminally sequenced. For the first time, δ-kafirin was identified at the protein level. The study clearly revealed intercultivar differences between protein levels. The joint use of SDS-PAGE, SE-HPLC, and RP-HPLC allowed discriminating among cultivars based on the differences in prolamin levels and composition.

Wheat contains high levels of the three classes of xylanase inhibitors (XIs), Triticum aestivum xylanase inhibitor (TAXI), xylanase-inhibiting protein (XIP) and thaumatin-like xylanase inhibitor (TLXI). These proteins have been linked to plant defense. In this study, expression of XIs during wheat ear development and germination was examined using immunoblotting. The three types of XIs accumulated at high levels between the milky and the soft dough stages of ear development, and reached the highest levels at the hard kernel stage. From the hard kernel stage to harvest ripeness, a slight drop in inhibitor levels was observed, which was more marked for TAXI and TLXI than for XIP. During germination, the levels of the three types of XIs initially decreased, but XIs accumulated again after 1-2d, reaching maximum levels between 5 and 9d after imbibition. The levels of TAXI, XIP and TLXI in the seedlings then gradually and continuously declined as a function of time. 1D- and 2D-immunoblotting indicated that the three types of XIs occur in a wide variety of forms. This polymorphism is maintained throughout ear development and germination, although the proportions of the different (iso)forms vary with time. A differential temporal profile was observed for the unprocessed and processed forms of TAXI-type proteins. Finally, the occurrence of TAXI and XIP, but not TLXI, in roots and shoots of young seedlings was demonstrated. No XIs were detected in roots, leaves or stems at later stages of ear development. Overall, the three classes of XIs show remarkable similarities in their temporal distribution, indicating a related function within the wheat plant.

An XIP (xylanase inhibiting protein) type xylanase inhibitor was purified from Algerian pearl millet ( Pennisetum glaucum L.) grains and characterized for the first time. Cation exchange and affinity chromatography with immobilized Trichoderma longibrachiatum glycoside hydrolase (GH) family 11 xylanase resulted in electrophoretically pure protein with a molecular mass of 27-29 kDa and a pI value of 6.7. The experimentally determined N-terminal amino acid sequence of the purified XIP protein is 87.5%, identical to that of sorghum ( Sorghum bicolor L.) XIP and 79.2% identical to that of wheat ( Triticum aestivum L.) XIP-I. The biochemical properties of pearl millet XIP are comparable to those described earlier for sorghum XIP, except for the higher specific activity toward a T. longibrachiatum GH family 11 xylanase. On the basis of immunoblot neither TAXI nor TLXI type xylanase inhibitors were detected in pearl millet grains.

Within the HEALTHGRAIN diversity screening program, the variation in the content of dietary fiber and components thereof in different types of wheat was studied. The wheat types were winter (131 varieties) and spring (20 varieties) wheats (both Triticum aestivum L., also referred to as common wheats), durum wheat (Triticum durum Desf., 10 varieties), spelt wheat (Triticum spelta L., 5 varieties), einkorn wheat (T. monococcum L., 5 varieties), and emmer wheat (Triticum dicoccum Schubler, 5 varieties). Common wheats contained, on average, the highest level of dietary fiber [11.5-18.3% of dry matter (dm)], whereas einkorn and emmer wheats contained the lowest level (7.2-12.8% of dm). Intermediate levels were measured in durum and spelt wheats (10.7-15.5% of dm). Also, on the basis of the arabinoxylan levels in bran, the different wheat types could be divided this way, with ranges of 12.7-22.1% of dm for common wheats, 6.1-14.4% of dm for einkorn and emmer wheats, and 10.9-13.9% of dm for durum and spelt wheats. On average, bran arabinoxylan made up ca. 29% of the total dietary fiber content of wheat. In contrast to what was the case for bran, the arabinoxylan levels in flour were comparable between the different types of wheat. For wheat, in general, they varied between 1.35 and 2.75% of dm. Einkorn, emmer, and durum wheats contained about half the level of mixed-linkage beta-glucan (0.25-0.45% of dm) present in winter, spring, and spelt wheats (0.50-0.95% of dm). All wheat types had Klason lignin, the levels of which varied from 1.40 to 3.25% of dm. The arabinoxylan contents in bran and the dietary fiber contents in wholemeal were inversely and positively related with bran yield, respectively. Aqueous wholemeal extract viscosity, a measure for the level of soluble dietary fiber, was determined to large extent by the level of water-extractable arabinoxylan. In conclusion, the present study revealed substantial variation in the contents of dietary fiber and constituents thereof between different wheat types and varieties.

One hundred and fifty bread wheat lines and 50 other lines of small-grain cereals (spelt, durum wheat, Triticum monococcum, Triticum dicoccum, oats, rye, and barley) were selected for diversity in their geographical origin, age, and characteristics. They were grown on a single site in Hungary in 2004−2005, harvested, milled, and analyzed for a range of phytochemicals (tocols, sterols, phenolic acids, folates, alkylresorcinols) and fiber components that are considered to have health benefits. Detailed analyses of these components in the different species are reported in a series of accompanying papers. The present paper discusses the comparative levels of the bioactive components in the different species, showing differences in both ranges and mean amounts. Furthermore, detailed comparisons of the bread wheat lines show that it is possible to identify lines in which high levels of phytochemicals and dietary fiber components are combined with good yield and processing quality. This means that commercially competitive lines with high levels of bioactive components are a realistic goal for plant breeders.

A xylanase inhibitor of the xylanase-inhibiting protein (XIP) type was detected in sorghum (Sorghum bicolor L. Moench) whole meal using Western blot and immunoprobing with polyclonal anti-XIP antibodies. No detectable levels of Triticum aestivum xylanase inhibitor (TAXI) and thaumatin-like xylanase inhibitor (TLXI) type xylanase inhibitors were present. Trichoderma longibrachiatum xylanase affinity chromatography (AC) was used for the purification of sorghum XIP. Biochemical characteristics and protein sequence data show that sorghum XIP strongly corresponds to wheat (T. aestivum L.) XIP-I. Like wheat XIP-I, it inhibits both glycoside hydrolase family (GH) 10 and 11 xylanases, indicating that the XIP-I active site residues are well conserved in sorghum XIP. However, in contrast to wheat XIP-I, the inhibitor is unable to affect Aspergillus niger GH 11 xylanase activity. Its specific inhibition activities towards the other xylanases tested are comparable to those of wheat XIP-I. Based on the available sorghum expressed sequence tag (EST) database, XIP is expressed in sorghum in different tissues and developmental stages. Also expression in the presence of several plant hormones and under biotic as well as abiotic stress conditions is suggested.

Wheat kernel associated endoxylanases consist of a majority of microbial endoxylanases and a minority of endogenous endoxylanases. At least part of these enzymes can be expected to end up in wheat flour upon milling. In this study, the contribution of both types of these endoxylanases to changes in the arabinoxylan (AX) population during wheat flour breadmaking was assessed. To this end, wheat flour produced from two wheat varieties with different endoxylanase activity levels, both before and after sodium hypochlorite surface treatment of the wheat kernels, was used in a straight dough breadmaking procedure. Monitoring of the AX population during the breadmaking process showed that changes in AX are to a large extent caused by endogenous endoxylanases, whereas the contribution of microbial endoxylanases to these changes was generally very low. The latter points to a limited contamination of wheat flour with microbial enzymes during milling or to an extensive inactivation of these wheat flour associated microbial endoxylanases by endoxylanase inhibitors, present in wheat flour. When all wheat kernel associated microbial endoxylanases were first washed from the kernels and then added to the bread recipe, they drastically affected the AX population, suggesting that they can have a large impact on whole meal breadmaking.

Polyclonal antibodies (PAbs) were raised against wheat (Triticum aestivum L.) TAXI- and XIP-type xylanase inhibitors by rabbit immunization. A small contaminant in both antigens, not detected by SDS-PAGE and later identified through Western blot as a recently discovered third type of xylanase inhibitor from wheat, i.e. thaumatin-like xylanase inhibitor (TL-XI), led to the coproduction of PAbs against this protein in the rabbits. To obtain inhibitor-specific PAbs, the PAbs against TAXI, XIP and TL-XI were separated by affinity chromatography using immobilised recombinant and native xylanase inhibitors. The purified PAbs allowed the immunoquantification of each type of wheat xylanase inhibitor using Western blot and densitometric analysis against purified inhibitor standards. The method allowed the detection of the purified inhibitors at the 20 ng level. As the PAbs against the wheat xylanase inhibitors cross-reacted with their homologous targets from other cereals, immunoprobing allowed identification of XIP homologues in oats (Avena sativa L.) and TL-XI homologues in durum wheat (Triticum durum Desf.) and rye (Secale cereale L.).

The endoxylanases associated with wheat kernels consist of wheat endogenous endoxylanases on one hand and kernel-associated microbial endoxylanases on the other hand. Assessment of their presence, based on analysis of their enzymic activity, can be expected to be hampered by the presence in wheat of high levels of endogenous endoxylanase inhibitors, which are able to inhibit the wheat-kernel-associated microbial endoxylanases. On the basis of preliminary experiments aimed at clarifying the distribution of the wheat-associated endoxylanases, a method to estimate total endoxylanase activities in wheat kernels was developed. Extensive washing of wheat kernels with universal buffer of pH 8.0 provided near-quantitative separation of the microbial endoxylanases located on the surface of wheat kernels from the endogenous endoxylanases and endoxylanase inhibitors located in such kernels. The microbial or endogenous nature of the endoxylanases was confirmed by making use of the inhibition specificity of endoxylanase inhibitors. Determination of the endoxylanase activity in the washing liquid, corresponding to the microbial endoxylanase population, and the washed kernels, corresponding to the endogenous endoxylanase population, allowed estimation of the total endoxylanase activities associated with the wheat kernel. Results showed that microbial endoxylanases can account for over 90% of the total wheat-associated endoxylanase activity and that the latter can be at least 5 times higher than the apparent endoxylanase activity.

The influence of wheat variety, harvest year and harvest date on the levels of wheat kernel-associated endogenous and microbial endoxylanases and wheat endogenous endoxylanase inhibitors was investigated using ten winter wheat varieties grown in 2002 and 2003. Three of the varieties were harvested on five different dates in a 1 month period spanning the optimal harvest period. Introductory experiments indicated that endoxylanase activity was very heterogeneously distributed among the different wheat kernels, necessitating the use of relatively large amounts of kernels, i.e. 100 g, to obtain representative wholemeal samples. Apparent endoxylanase activities in these samples largely depended on variety and weather conditions prior to harvesting. In contrast, apparent endoxylanase inhibitor levels were less dependent on climatological circumstances and were mainly determined by genetic factors. The data derived from this study may contribute to explaining the differences observed in functionality between wheats.

A recent approach based on affinity chromatography with immobilised endoxylanase inhibitors was used to isolate two endoxylanases (EC 3.2.1.8) with different bread-making functionalities from an Aspergillus niger fermentation broth. TAXI ( Triticum aestivum endoxylanase inhibitor) affinity chromatography yielded a TAXI- and XIP (endoxylanase inhibiting protein)-sensitive family 11 endoxylanase (24 kDa, p I 3.5), XIP affinity chromatography subsequently yielded a family 10 endoxylanase (36 kDa), only inhibited by XIP. While the first enzyme improves bread volume, the latter enzyme has no effect on bread quality whatsoever. The bread-making positive endoxylanase rather selectively hydrolyses water-unextractable arabinoxylan in an in vitro screening method, still performs/is active during bread-making and produces soluble arabinoxylan of high (>11.2 × 10 4 Da) and low molecular mass (≤11.2 × 10 4 Da). In contrast, the bread-making neutral endoxylanase in the in vitro assay displays a bias for water-extractable arabinoxylan and is immediately and almost completely inhibited during the early stages of bread-making. The results show that the functionalities of the purified A. niger endoxylanases in wheat bread-making are strongly dictated by their sensitivities towards wheat endoxylanase inhibitors.

The vast majority of bread is traditionally produced from wheat flour. Apart from its major constituent starch, wheat flour also contains many other types of substances of which the gluten, the non-starch polysaccharides as well as the lipids are the most important in terms of their impact on the processability of the raw material and in terms of the quality of the final products. We here provide the basics on the processability and quality determining wheat flour constituents and present common concepts on their fate during the breadmaking process as well as on approaches targeted to influence their functionality.

Debranning of wheat to remove the outer 7% of the kernel, prior to grinding or milling reduced xylanase activity in wheat wholemeal and wheat endosperm flour by up to 80 and 60%, respectively, whereas there was no significant reduction of xylanase inhibiting activity. Flours obtained after debranning and milling showed no major differences in moisture content, whereas ash content decreased and protein and arabinoxylan content decreased slightly with increasing debranning degree. Part of the xylanase activity in the flour was lost on addition of Triticum aestivum xylanase inhibitor (TAXI). Since TAXI specifically inhibits glycosyl hydrolase family 11 xylanases and since endogenous cereal xylanases belong exclusively to family 10, part of the xylanase activity in the flour is most likely of microbial origin. Debranning also significantly reduced alpha-amylase activities in wheat wholemeal and wheat flour. Debranning prior to milling can, therefore, impact on flour functionality.

Cereals contain proteinaceous inhibitors of endoxylanases, which affect the efficiency and functionality of these enzymes in cereal processing. This review relates their first discovery in wheat and the subsequent purification of two distinct classes of endoxylanase inhibitors, namely Triticum aestivum xylanase inhibitor (TAXI)-type and xylanase inhibitor protein (XIP)-type inhibitors in cereals. Both inhibitor classes occur in monocots as multi-isoform families. The reported data provide an overview of the relative quantitative and qualitative variation of these inhibitors in cereals. Wheat and rye are particularly rich in TAXI-type and XIP-type inhibitors with the latter inhibitors being more abundant. Lower inhibitor levels are present in durum wheat and barley, while maize contains solely XIP-type inhibitors. No inhibitors have been isolated from rice, oats and buckwheat.

Highly pure XIP-type (for Xylanase Inhibiting Protein) endoxylanase inhibitor fractions were selectively obtained with a high yield from rye, durum wheat, barley, and maize extracts by affinity chromatography with immobilised Aspergillus niger endoxylanase Xyn1 following removal of the TAXI-type (for Triticum aestivum xylanase inhibitor) endoxylanase inhibitors by affinity chromatography with immobilised Bacillus subtilis endoxylanase XynA. No inhibitors belonging to the XIP family occur in rice, oats, and buckwheat. N-terminal amino acid sequences of the non-wheat XIP-type inhibitors were very similar or identical to those of wheat XIP-1, a chitinase homologue. The isolated inhibitors are basic, monomeric proteins of ca. 30 kDa with pI values of at least 8.5 (rye, durum wheat, and barley XIP) and ca. 7.0 (maize XIP). They are, in general, active against fungal endoxylanases and do not hydrolyse chitin. SDS–PAGE analysis and high-resolution cation exchange chromatography suggest the presence of multiple XIP-type isoinhibitors in the different cereals.

An affinity-based purification procedure allowed the resolution of two distinct groups of endoxylanase inhibitors with different molecular structures and endoxylanase specificities from wheat wholemeal. The first group comprises the so-called Triticum aestivum L. Endoxylanase inhibitor (TAXI)-type proteins which are of approx. M r 40 000 and occur in two different molecular forms. These inhibitors were removed from a concentrated cation exchange chromatography fraction from wheat wholemeal on a Bacillus subtilis endoxylanase affinity column. The second group of structurally different endoxylanase inhibitors, the so-called xylanase inhibiting protein (XIP)-type, of approx. M r 29 000–32 000, with pI values varying between 8·8 and 9·2, was purified from the unbound fraction from the B. subtilis endoxylanase affinity column by chromatography on an Aspergillus niger endoxylanase affinity column followed by gel permeation chromatography. The XIP-type inhibitors are not active against the B. subtilis endoxylanase and were consequently not retained on the B. subtilis endoxylanase column. Further analysis of the XIP-type proteins by high-resolution cation exchange chromatography, SDS-PAGE and iso-electrofocusing, revealed several forms. They had similar endoxylanase specificities and N-terminal amino acid sequences.

Four Secale cereale L. xylanase inhibitors (SCXI) from rye were purified to homogeneity. The inhibitor proteins all occur in two molecular forms, i.e. a c. 40 kDa monomeric protein and, presumably following proteolytic modification, a heterodimer consisting of two disulphide linked subunits of c. 30 and c. 10 kDa. These basic proteins all have isoelectric points of at least 9·0 and a highly homologous N -terminal amino acid sequence. The isolated proteins strongly inhibited the activity of Aspergillus niger, Bacillus subtilis and Trichoderma viride family 11 endoxylanases, but showed no activity towards an Aspergillus aculeatus family 10 endoxylanase. These characteristics demonstrate that rye contains a family of isoinhibitors with similar structures and specificities, that are homologous with Triticum aestivum L. xylanase inhibitor I (TAXI I) from wheat andHordeum vulgare L. xylanase inhibitor (HVXI) from barley.

Twenty-three wheat samples from 19 different European wheat cultivars (Triticum aestivum L.) were tested for their quantitative and qualitative variation in inhibition activity against family 11 endoxylanases of Aspergillus niger, Bacillus subtilis, and Trichoderma viride and a family 10 endoxylanase of A. aculeatus. Under the experimental conditions, the A. aculeatus enzyme was not inhibited by the wheat extracts, the A. niger and B. subtilis endoxylanases were affected to a similar extent, while the T. viride enzyme was much more inhibited. The inhibition activities in the different wheat samples against the A. niger, B. subtilis, and T. viride endoxylanases varied between 36.0 and 11.7, 34.0 and 12.9, and 86.2 and 46.6 IU/100 mg of dry whole meal, respectively. One IU (inhibition unit) corresponds to the amount of inhibitor resulting in 50% inhibition of endoxylanase activity under the conditions of the assay. The inhibitor activities were linearly related, indicating that the levels of differ...

Steady-state kinetic approaches were used to investigate the binding of a novel Penicillium funiculosum xylanase, XYNC, with three known xylanase inhibitor proteins from wheat (Triticum aestivum). The xylanase gene (xynC) was cloned from a P. funiculosum genomic library and the deduced amino acid sequence of XYNC exhibited high sequence similarity with fungal family 11 xylanases. xynC was overexpressed in P. funiculosum and the product (XYNC: M(r)=23.6 kDa; pI=3.7) purified and shown to efficiently degrade birchwood xylan [K(m)=0.47% w/v, Vmax=2540 micromol xylose min(-1) (mg protein)(-1) at pH 5.5 and 30 degrees C] and soluble wheat arabinoxylans [K(m)=1.45% w/v, Vmax=7190 micromol xylose min(-1) mg protein)(-1) at pH 5.5 and 30 degrees C]. The xylanase activity of XYNC was inhibited strongly by three xylanase inhibitor proteins from wheat; XIP-I, TAXI I and TAXI II. The inhibition for each was competitive, with very tight binding (K(i)=3.4, 16 and 17 nM, respectively) equivalent to free energy changes (deltaG degrees ) of -49, -45 and -45 kJ mol(-1). This is the first report describing a xylanase that is inhibited by all three wheat xylanase inhibitor proteins described to date.

Hordeum vulgare L. xylanase inhibitor (HVXI), an endoxylanase inhibitor with a protein structure, was purified to homogeneity from barley (Hordeum vulgare L.). HVXI is a nonglycosylated monomeric protein, with a molecular weight of ≈40,000 and a pI ≥ 9.3. Although it inhibits different endoxylanases to a varying degree, the activities of an α-L-arabinofuranosidase and a β-d-xylosidase were not inhibited. Apparently, HVXI occurs in two molecular forms. These characteristics and the N-terminal sequences of the composing polypeptides show that HVXI is homologous with Triticum aestivum L. xylanase inhibitor I, an endoxylanase inhibitor from wheat flour.

J. Am. Soc. Brew. Chem. 72(1):12-21, 2014 Gushing is the vigorous overfoaming of carbonated beverages when the bottle is opened. Primary gushing in beer is mostly caused by a group of proteins called hydrophobins secreted by filamentous fungi, which contaminate CO2 gaseous molecules during carbonation and form nanobubbles. The influence of hop oil antifoam on primary gushing showed a complete suppressing effect in sparkling water, a decreasing effect in wort, and no influence on gushing-positive beers. This shows the importance of the critical point of addition of this product in the brewing process. GC and GC-MS analysis show that commercial available lipophilic hop extract comprises fatty acids, either saturated fatty acids (SFA) or unsaturated fatty acids (UFA) (both free and incorporated in mono-, di-, and triglycerides), waxes (long-chain alkanes), and steroid compounds. Gushing analysis of each compound showed that SFA and UFA behave in a different manner regarding gushing. In contrast to SFA and trans-form UFA, cis-form UFAs do not induce gushing. Long-chain alkanes provide sufficient hydrophobic structures to interact with gaseous CO2 molecules and induce gushing. Because hop antifoam molecules are hydrophobic, they interact with hydrophobins and prevent interaction with CO2 and, consequently, inhibit the possibilities of development of the explosive nanobubble structure.

The cell wall polysaccharides of wheat affect the quality for food processing, livestock feed and distilling, and also form a major source of dietary fibre (DF) for human health. Multisite field trials of wheat genotypes show extensive variation in the content of the major types of cell wall polysaccharide in wheat, arabinoxylan and β-glucan, much of which is heritable and hence available for exploitation by plant breeders. Furthermore, contents of DF components have not declined as a result of intensive wheat breeding. The identification of candidate genes for DF synthesis using bioinformatics and RNAi suppression in transgenic wheat will allow the content and composition of DF components to be fine-tuned for specific end uses.

The wheat grain comprises three groups of major components, starch, protein, and cell wall polysaccharides (dietary fiber), and a range of minor components that may confer benefits to human health. Detailed analyses of dietary fiber and other bioactive components were carried out under the EU FP6 HEALTHGRAIN program on 150 bread wheat lines grown on a single site, 50 lines of other wheat species and other cereals grown on the same site, and 23−26 bread wheat lines grown in six environments. Principal component analysis allowed the 150 bread wheat lines to be classified on the basis of differences in their contents of bioactive components and wheat species (bread, durum, spelt, emmer, and einkorn wheats) to be clearly separated from related cereals (barley, rye, and oats). Such multivariate analyses could be used to define substantial equivalence when novel (including transgenic) cereals are considered.

Arabinoxylans (AX), xylanase, and xylanase inhibitors have an important role in many cereal food processing applications. The effects of genotype, growing location, and their interaction (G × L) on AX, apparent xylanase activity, and apparent xylanase inhibition activity of Triticum aestivum xylanase inhibitor (TAXI) and xylanase inhibiting protein (XIP) were investigated for six hard red and six hard white spring wheat genotypes grown at three locations. Difference in total AX level among genotypes was not determined to a significant level by genotype. Instead, variability in total AX content was largely dependent on G × L. However, total AX content was significantly different between the two wheat classes. For bran xylanase activity, 25% of the variability could be attributed to G × L interaction. Moreover, there was significant difference between the bran xylanase activities in the two wheat classes. Bran TAXI activity and XIP activity were significantly different among genotypes. Genotype con...

Grain dietary fiber content in wheat not only affects its end use and technological properties including milling, baking and animal feed but is also of great importance for health benefits. In this study, integration of association genetics (seven detected loci on chromosomes 1B, 3A, 3D, 5B, 6B, 7A, 7B) and meta-QTL (three consensus QTL on chromosomes 1B, 3D and 6B) analyses allowed the identification of seven chromosomal regions underlying grain dietary fiber content in bread wheat. Based either on a diversity panel or on bi-parental populations, we clearly demonstrate that this trait is mainly driven by a major locus located on chromosome 1B associated with a log of p value >13 and a LOD score >8, respectively. In parallel, we identified 73 genes differentially expressed during the grain development and between genotypes with contrasting grain fiber contents. Integration of quantitative genetics and transcriptomic data allowed us to propose a short list of candidate genes that are conserved in the rice, sorghum and Brachypodium chromosome regions orthologous to the seven wheat grain fiber content QTL and that can be considered as major candidate genes for future improvement of the grain dietary fiber content in bread wheat breeding programs.

The crystal structure of the thaumatin-like xylanase inhibitor (TLXI) from Triticum aestivum, a non-competitive inhibitor of glycoside hydrolase family 11 xylanases, has been determined at 2.9 Å resolution. It is similar to that of thaumatin except that TLXI lacks domain II of thaumatin, making it the first obtained crystallographic structure in the small thaumatin-like proteins (TLPs) subfamily within the larger family of TLPs. Unlike most other TLPs, TLXI does not contain the acidic surface cleft which is responsible for antifungal activity and binding of carbohydrates. Comparison with other TLPs suggests the loop containing histidine 22 as being important for xylanase inhibition. ispartof: Proteins-structure function and bioinformatics vol:78 issue:10 pages:2391-2394 ispartof: location:United States status: published

Endo-1,4-beta-d-xylanases (EC 3.2.1.8, xylanases) and xylanase inhibitors, that is, TAXI (Triticum aestivum xylanase inhibitor), XIP (xylanase inhibiting protein), and TLXI (thaumatin-like xylanase inhibitor) type xylanase inhibitors, which naturally occur in cereals, are believed to be at the basis of a significant part of the variability in biotechnological functional properties of cereals. Xylanase inhibitors in particular affect grain functionality during processing and in animal feeds when xylanases are used to improve processing parameters and product quality. In the present study the variability of xylanase, TAXI, and XIP activities was quantified in different cereals, including different wheat types [common wheat (Triticum aestivum L.), durum wheat (Triticum durum Desf.), spelt wheat (Triticum spelta L.), einkorn wheat (Triticum monococcum L.), and emmer wheat (Triticum dicoccum Schubler)], barley (Hordeum vulgare L.), rye (Secale cereale L.), and oat (Avena sativa L.), and the contribution of genotype and environment to this variability in common wheat was estimated. Substantial differences in xylanase, TAXI, and XIP activities exist between the different cereal types and varieties. Under the experimental conditions of this study, the durum wheat samples show very high xylanase activities compared to the other cereals. High TAXI and XIP activities were measured in, for example, common wheat, spelt wheat, and rye, whereas low activities occur in barley and oat. For wheat, a significant part of the variability in inhibitor levels can be explained by genotype, whereas xylanase activity is most strongly determined by environment. The results obtained suggest that plant breeders and industry to certain extent can select for wheat varieties with high or low xylanase inhibition activities, but the relatively high contribution of the genotype-environment interaction term to the total variability in inhibition activities indicates that TAXI and XIP activities are not very stable breeding parameters.

Wheat contains three different classes of proteinaceous xylanase inhibitors (XIs), i.e. Triticum aestivum xylanase inhibitors (TAXIs) xylanase-inhibiting proteins (XIPs), and thaumatin-like xylanase inhibitors (TLXIs) which are believed to act as a defensive barrier against phytopathogenic attack. In the absence of relevant data in wheat kernels, we here examined the response of the different members of the XI protein population to infection with a DeltaTri5 mutant of Fusarium graminearum, the wild type of which is one of the most important wheat ear pathogens, in early developing wheat grain. Wheat ears were inoculated at anthesis, analyzed using 2-D DIGE and multivariate analysis at 5, 15, and 25 days post anthesis (DPA), and compared with control samples. Distinct abundance patterns could be distinguished for different XI forms in response to infection with F. graminearum DeltaTri5. Some (iso)forms were up-regulated, whereas others were down-regulated. This pathogen-specific regulation of proteins was mostly visible at five DPA and levelled off in the samples situated further from the inoculation point. Furthermore, it was shown that most identified TAXI- and XIP-type XI (iso)forms significantly increased in abundance from the milky (15 DPA) to the soft dough stages (25 DPA) on a per kernel basis, although the extent of increase differed greatly. Non-glycosylated XIP forms increased more strongly than their glycosylated counterparts.

Analysis of the contents of bioactive components (tocols, sterols, alkylresorcinols, folates, phenolic acids, and fiber components) in 26 wheat cultivars grown in six site × year combinations showed that the extent of variation due to variety and environment differed significantly between components. The total contents of tocols, sterols, and arabinoxylan fiber were highly heritable and hence an appropriate target for plant breeding. However, significant correlations between the contents of bioactive components and environmental factors (precipitation and temperature) during grain development also occurred, with even highly heritable components differing in amount between grain samples grown in different years on different sites.

In wheat ( Triticum aestivum ) grains, TAXI- (T. aestivum xylanase inhibitor), XIP- (xylanase inhibiting protein), and TLXI-type (thaumatin-like xylanase inhibitor) xylanase inhibitors (XIs) are expressed in considerable levels and under different forms. As these proteins have a significant impact on microbial xylanases frequently used in cereal-based biotechnological processes, knowledge of their quantitative and qualitative variability in wheat is of great interest. This paper reports the successful use of immunoquantification by Western blotting to determine the intercultivar variation in the three structurally different classes of XIs, as well as their distribution among various industrial milling fractions. TAXI and XIP protein levels in eight wheat cultivars ranged from 81 to 190 ppm and from 156 to 371 ppm, with average values of 133 and 235 ppm, respectively. Using immunoblotting, TLXI protein levels could be measured directly for the first time. They ranged from 51 to 150 ppm and amounted to 112 ppm on average. The three classes of XIs were distributed among different wheat milling fractions in a similar way, with 4 and 10 times higher concentrations in the aleurone-enriched fraction than in white flour and pericarp fractions, respectively. Immunoblot patterns suggested that the observed intercultivar and spatial variabilities within the wheat grain are not due to the presence or absence of specific members of the large polymorphic XI families but to differences in the overall level and/or proportions of the specific members.

Wheat grains contain three classes of xylanase inhibitors (XIs), i.e. TAXI (Triticum aestivum xylanase inhibitor), XIP (xylanase inhibiting protein) and TLXI (thaumatin-like xylanase inhibitor). These proteins are involved in plant defence and strongly affect cereal based processes in which inhibitor-sensitive xylanases are used. This paper reports on the successful use of 2D-DIGE and tandem MS to discriminate XI (iso)forms and measure their qualitative and quantitative variation in six different wheat cultivars. In total, 18 TAXI-, 27 XIP- and 3 TLXI-type XI spots were identified. The multiple members of the large TAXI-gene family make a considerable contribution to the total TAXI population. For XIP-type XIs, XIP-I is expressed as the predominant form, albeit under variable degrees of PTMs. Only one TLXI genetic variant was identified, showing different degrees of glycosylation. Multiple comparison analysis revealed up to 5-fold intercultivar differences in protein level of XI (iso)forms. Evaluation of abundance patterns using multivariate statistical tools revealed highly distinctive as well as correlated levels of different XI forms among the six cultivars. As the constitutive (and induced) levels of the different XI (iso)forms, which are differentially regulated in response to various forms of stress in other wheat plant parts, considerably vary between the cultivars, it can be assumed that their degree of resistance against pathogenic attack differs. Similarities in abundance profiles between XI (iso)forms and pathogenesis-related chitinases are also in line with a role in plant defence.

Thaumatin-like xylanase inhibitors (TLXI) are recently discovered wheat proteins. They belong to the family of the thaumatin-like proteins and inhibit glycoside hydrolase family 11 endoxylanases commonly used in different cereal based (bio)technological processes. We here report on the biochemical characterisation of TLXI. Its inhibition activity is temperature- and pH-dependent and shows a maximum at approximately 40 degrees C and pH 5.0. The TLXI structure model, generated with the crystal structure of thaumatin as template, shows the occurrence of five disulfide bridges and three beta-sheets. Much as in the structures of other short-chain thaumatin-like proteins, no alpha-helix is present. The circular dichroism spectrum of TLXI confirms the absence of alpha-helices and the presence of antiparallel beta-sheets. All ten cysteine residues in TLXI are involved in disulfide bridges. TLXI is stable for at least 120 min between pH 1-12 and for at least 2 hours at 100 degrees C, making it much more stable than the other two xylanase inhibitors from wheat, i.e. Triticum aestivum xylanase inhibitor (TAXI) and xylanase inhibitor protein (XIP). This high stability can probably be ascribed to the high number of disulfide bridges, much as seen for other thaumatin-like proteins. ispartof: Journal of Enzyme Inhibition and Medicinal Chemistry vol:24 issue:3 pages:646-654 ispartof: location:England status: published

Recently, a novel wheat thaumatin-like protein, TLXI, which inhibits microbial glycoside hydrolase family (GH) 11 xylanases has been identified. It is the first xylanase inhibitor that exerts its inhibition in a non-competitive way. In the present study we gained insight into the interaction between TLXI and xylanases via combined molecular modeling and mutagenic approaches. More specifically, site-specific mutation of His22, situated on a loop which distinguishes TLXI from other, non-inhibiting, thaumatin-like proteins, and subsequent expression of the mutant in Pichia pastoris resulted in a protein lacking inhibition capacity. The mutant protein was unable to form a complex with GH11 xylanases. Based on these findings, the interaction of TLXI with GH11 xylanases is discussed. ispartof: Journal of enzyme inhibition and medicinal chemistry vol:24 issue:1 pages:38-46 ispartof: location:England status: published

Ten different barley varieties grown in one location were studied for their content of tocols, folate, plant sterols, alkylresorcinols, and phenolic acids, as well as dietary fiber components (arabinoxylan and beta-glucan). The samples included hulled and hull-less barley types and types with normal, high-amylose, and waxy starch. The aim was to study the composition of raw materials, and therefore the hulls were not removed from the hulled barleys. A large variation was observed in the contents of all phytochemicals and dietary fibers. Two varieties from the INRA Clermont Ferrand barley program in France (CFL93-149 and CFL98-398) had high content of tocopherols and alkylresorcinols, whereas the variety Dicktoo was highest in dietary fiber content and phenolics. Positive correlations were found between 1000 kernel weight, alkylresorcinols, and tocols, as well as between dietary fiber content and phenolic compounds. The results demonstrate that the levels of phytochemicals in barley can likely be affected by breeding and that the contents of single phytochemicals may easily be adjusted by a right selection of a genotype.

The impact of wheat flour associated xylanases on the AX population during dough mixing and resting was investigated. The solubilization of AX during mixing was mainly due to mechanical forces, while solubilization of AX during resting was caused by xylanase activity. While large differences were observed for flours derived from wheat kernels with high and low apparent xylanase activity, limited differences were found between flours derived from untreated and surface sterilized wheat kernels, indicating that the changes in AX were mainly caused by endogenous xylanases. This suggests that either the contamination of flour with bran associated microbial xylanases is limited or that these xylanases are inhibited rapidly by xylanase inhibitors, present in large quantities in flour. From the results, it is clear that varying levels of wheat flour associated xylanases may contribute to wheat quality variation.

This study is an in-depth investigation of the interaction between polysaccharides and the proteinaceous xylanase inhibitors, Triticum aestivum xylanase inhibitor (TAXI), xylanase inhibitor protein (XIP), and thaumatin-like xylanase inhibitor (TLXI). The binding affinities of all three known types of xylanase inhibitors from wheat are studied by measuring the residual xylanase inhibition activity after incubation of the inhibitors in the presence of different polysaccharides, such as beta-glucans and (arabino)xylans. The binding affinities of all three xylanase inhibitors for (arabino)xylans increased with a decreasing arabinose/xylose ratio (A/X ratio). This phenomenon was observed both with water-extractable and water-unextractable (arabino)xylans. The inhibitors also interacted with different soluble and insoluble beta-glucans. None of the inhibitors tested had the ability to hydrolyze the polysaccharides investigated. The present findings contribute to the unraveling of the function of xylanase inhibitors in nature and to the prediction of the effect of added xylanases in cereal-based biotechnological processes, such as bread making and gluten-starch separation. ispartof: Journal of Agricultural and Food Chemistry vol:56 issue:2 pages:564-570 ispartof: location:United States status: published

Cereals contain proteinaceous inhibitors of endo-beta-1,4-xylanases (E.C.3.2.1.8, xylanases). Since these xylanase inhibitors (XIs) are only active against xylanases of microbial origin and do not interact with plant endogenous xylanases, they are believed to act as a defensive barrier against phytopathogenic attack. So far, three types of XIs have been identified, i.e. Triticum aestivum XI (TAXI), xylanase inhibiting protein (XIP), and thaumatin-like XI (TLXI) proteins. In this study the variation in XI forms present in wheat grain was elucidated using high-resolution 2-DE in combination with LC-ESI-MS/MS and biochemical techniques. Reproducible 2-DE fingerprints of TAXI-, XIP-, and TLXI-type XIs, selectively purified from whole meal of three European wheat cultivars using cation exchange chromatography followed by affinity chromatography, were obtained using a pH-gradient of 6 to 11 and a molecular mass range of 10 to 60 kDa. Large polymorphic XI families, not known to date, which exhibit different pI- and/or molecular mass values, were visualised by colloidal CBB staining. Identification of distinct genetic variants by MS/MS-analysis provides a partial explanation for the observed XI heterogeneity. Besides genetic diversity, PTMs, such as glycosylation, account for the additional complexity of the 2-DE patterns.

The effect of steaming conditions (mild, intermediate and severe) during parboiling of five different long-grain rice cultivars (brown rice cultivars Puntal, Cocodrie, XL8 and Jacinto, and a red rice) on rice colour, and Maillard precursors and indicators was investigated. Rice colour increased with severity of parboiling conditions. Redness increased more than yellowness when parboiling brown rice. Parboiling turned red rice black. It changed the levels of glucose, fructose, sucrose, and maltose. Losses of the non-reducing sugar, sucrose were caused by both leaching into the soaking water and enzymic conversion, rather than by thermal degradation during steaming. Concentrations of the reducing sugars, glucose and fructose, in intermediately parboiled rice were higher than those of mildly parboiled rice. After severe parboiling, glucose levels were lower than those of intermediately parboiled rice, while fructose levels were higher. These changes were ascribed to the sum of losses in the Maillard reaction (MR), formations as a result of starch degradation and isomerisation of glucose into fructose. It was clear that the e-amino group of protein-bound lysine was more affected by parboiling conditions and loss in MRs, than that of free lysine. Low values of the MR indicators furosine and free 5-hydroxymethyl-2-furaldehyde (HMF) in processed brown and red rices were related to mild parboiling, whereas high furosine and low free HMF levels were indicative of rices being subjected to intermediate processing conditions. High furosine and high free HMF contents corresponded to severe hydrothermal treatments. The strong correlation (r=0.89) between the free HMF levels and the increase in redness of parboiled brown rices suggested that Maillard browning was reflected more in the red than in the yellow colour.

To quantify Triticum aestivum xylanase inhibitor (TAXI) and xylanase inhibiting protein (XIP) type proteins in cereals in general and wheat ( T. aestivum) in particular, a robust enzyme-linked immunosorbent assay (ELISA) using an uncommon enzyme-antibody sandwich format was developed. Bacillus subtilis glycoside hydrolase family (GH) 11 and Aspergillus oryzae GH 10 xylanases were selected for coating ELISA plate wells to capture TAXI and XIP, respectively, prior to probing with antibodies. The detection threshold of the developed ELISA was much lower than that of the currently used xylanase inhibitor assay and the recently described Western blot approach. Because of its broad dynamic range (TAXI, 30-600 ng/mL, and XIP, 3-60 ng/mL), one proper standard extract dilution can be used for analyzing different wheat varieties, whereas for the currently used colorimetric assay, often different dilutions need to be analyzed. The TAXI ELISA for wheat was successfully adapted for barley ( Hordeum vulgare) and could also be used for other cereals.

The impact of varying levels of endoxylanase activity in wheat flour on arabinoxylan (AX) in mixed and rested dough was studied using eight industrially milled wheat flour fractions with varying endoxylanase activity levels. Analysis of the levels of reducing end xylose (RX) and solubilized AX (S-AX) formed during mixing and resting and their correlation with the endoxylanase activity in the flour milling fractions showed that solubilization of AX during the mixing phase is mainly due to mechanical forces, while solubilization of AX during resting is caused by endoxylanase activity. Moreover, solubilization of AX during the dough resting phase is more outspoken than that during the mixing phase. Besides endoxylanase activity, there were significant xylosidase and arabinofuranosidase activities during the dough resting phase. The results indicate that wheat flour-associated endoxylanases can alter part of the AX in dough, thereby changing their functionality in bread making and potentially affecting dough and end product properties.

Wheat (Triticum aestivum) contains a previously unknown type of xylanase (EC 3.2.1.8) inhibitor, which is described in the present paper for the first time. Based on its > 60 % similarity to TLPs (thaumatin-like proteins) and the fact that it contains the Prosite PS00316 thaumatin family signature, it is referred to as TLXI (thaumatin-like xylanase inhibitor). TLXI is a basic (pI >=, 9.3 in isoelectric focusing) protein with a molecular mass of approx. 18 kDa (determined by SDS/PAGE) and it occurs in wheat with varying extents of glycosylation. The TLXI gene sequence encodes a 26-amino-acid signal sequence followed by a 151-amino-acid mature protein with a calculated molecular mass of 15.6 kDa and pl of 8.38. The mature TLXI protein was expressed successfully in Pichia pastoris, resulting in a 21 kDa (determined by SDS/PAGE) recombinant protein (rTLXI). Polyclonal antibodies raised against TLXI purified from wheat react with epitopes of rTLXI as well as with those of thaumatin, demonstrating high structural similarity between these three proteins. TLXI has a unique inhibition specificity. It is a noncompetitive inhibitor of a number of glycoside hydrolase family 11 xylanases, but it is inactive towards glycoside hydrolase family 10 xylanases, Progress curves show that TLXI is a slow tight-binding inhibitor, with a K-i of approx. 60 nM. Except for zeamatin, an alpha-amylase/trypsin inhibitor from maize (Zea mays), no other enzyme inhibitor is currently known among the TLPs. TLXI thus represents a novel type of inhibitor within this group of proteins. ispartof: Biochemical Journal vol:403 issue:3 pages:583-591 ispartof: location:England status: published

To gain insight into the distribution of arabinoxylans (AX), endoxylanases, and endoxylanase inhibitors in industrial wheat roller milling, all streams, that is, 54 flour fractions, 4 bran fractions, and the germ, were analyzed for ash, starch, and protein contents, alpha-amylase activity levels, total (TOT-AX) and water-extractable arabinoxylan (WE-AX) contents, endoxylanase activity levels, and endoxylanase inhibitor (TAXI and XIP) contents. In general, bran fractions were significantly richer in TOT-AX and WE-AX contents, endoxylanase activity levels, and endoxylanase inhibitor contents than germ and, even more so, than flour fractions. In the 54 different flour fractions, minimal and maximal values for TOT-AX and WE-AX contents differed by ca. 2-fold, whereas they differed by ca. 15-fold for endoxylanase activity levels. The latter were positively correlated with ash and negatively correlated with starch content, suggesting that the endoxylanase activity in flour is strongly influenced by the level of bran contamination. TAXI contents in the flour fractions varied ca. 4-fold and were strongly correlated with bran-related parameters such as ash content and enzyme activity levels, whereas XIP contents varied ca. 3-fold and were not correlated with any of the parameters measured in this study. The results can be valuable in blending and optimizing wheat flour fractions to obtain flours with specific technological and nutritional benefits.

A rice XIP-type inhibitor was purified by affinity chromatography with an immobilized Aspergillus aculeatus family 10 endoxylanase. Rice XIP is a monomeric protein, with a molecular mass of ca. 32 kDa and a pI of ca. 5.6. Its N-terminal amino acid sequence was identical to that of a rice chitinase homologue, demonstrating the difficulty when using sequence information to differentiate between endoxylanase inhibitors and (putative) chitinases in rice. Rice XIP inhibited different endoxylanases to a varying degree. In particular, it most strongly inhibited family 10 endoxylanases from A. niger and A. oryzae, while several family 11 enzymes from Bacillus subtilis, A. niger and Trichoderma sp. were not sensitive to inhibition. The above mentioned A. aculeatus endoxylanase was not inhibited either, although gel permeation chromatography revealed that it complexed rice XIP in a 1: 1 molar stoichiometric ratio. ispartof: Journal of enzyme inhibition and medicinal chemistry vol:20 issue:1 pages:95-101 ispartof: location:England status: published

International audience; A xylanase from Aspergillus aculeatus (glycoside hydrolase family 10), uninhibited by TAXI, and a xylanase from Bacillus subtilis (family 11), uninhibited by XIP-I, were selected to quantify the respective apparent levels of XIP-I and TAXI inhibitors, in flours and grains of 20 wheat varieties. The apparent TAXI amount ranged from 0.05 to 0.19 mg/g in flour (mean: 0.11 mg/g) and from 0.07 to 0.2 mg/g in grain (mean: 0.14 mg/g). The range observed for XIP-I was 0.12-0.6 mg/g in flour (mean 0.32 mg/g), and 0.21-0.56 mg/g in grain (mean: 0.41 mg/g). The inhibition profile of the xylanase from A. aculeatus by a crude inhibitor preparation suggested the presence of an additional component in wheat flour, responsible for an increase in inhibition.