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1.
Foam stability is an important quality trait of beer. Our previous results of two-dimensional gel electrophoresis (2DE) analyses of beer proteins implied a relationship between barley dimeric alpha-amylase inhibitor-1 (BDAI-1) and beer foam stability as judged by the NIBEM-T analyzer. To develop a novel prediction method of beer foam stability under different conditions of barley cultivar and malt modification, multiple linear regression analysis was applied. The spot intensities of major beer proteins on 2DE gel were quantified and used as explanatory variables. The foam stabilities of 25 beer samples each brewed from malt with different malt modification in one of the three cultivars (cultivars A, B, and C) were explained by the spot intensities of BDAI-1 at the 5% significance level ( r = 0.421). Furthermore, two other major protein spots (b0 and b5) were observed on the 2DE gels of Japanese commercial beer samples with different foam stability. Then, multiple regression for foam stability was calculated using these three spot intensities as explanatory variables. As a result, 72.1% of the beer foam stability in 25 beer samples was explained by a novel multiple regression equation calculated using spot b0 and BDAI-1 as positive explanatory variables and spot b5 as a negative variable. To verify the validity of the multiple regression equation and the explanatory variables, the beer foam stability in practical beer samples was analyzed. As a result, 81.5% of the beer foam stability in 10 Japanese commercial beer samples was also explained by using spot b0 and BDAI-1 as positive explanatory variables and spot b5 as a negative variable. Mass spectrometry analyses followed by database searches revealed that protein spots b0 and b5 were identified as protein Z originated from barley and thioredoxin originated from yeast, respectively. These results confirm that BDAI-1 and protein Z are foam-positive factors and identify yeast thioredoxin as a possible novel foam-negative factor.  相似文献   

2.
Barley lipid transfer protein (LTP1) is a heat-stable and protease-resistant albumin that concentrates in beer, where it participates in the formation and stability of beer foam. Whereas the barley LTP1 does not display any foaming properties, the corresponding beer protein is surface-active. Such an improvement is related to glycation by Maillard reactions on malting, acylation on mashing, and structural unfolding on brewing. The structural stability of purified barley and glycated malt LTP1 toward heating has been analyzed. Whatever the modification, lipid adduction or glycation, barley LTP1s are highly stable proteins that resisted temperatures up to 100 degrees C. Unfolding of LTP1 occurred only when heating was conducted in the presence of a reducing agent. In the presence of sodium sulfite, the lipid-adducted barley and malt LTP1 displayed higher heat stability than the nonadducted protein. Glycation had no or weak effect on heat-induced unfolding. Finally, it was shown that unfolding occurred on wort boiling before fermentation and that the reducing conditions are provided by malt extract.  相似文献   

3.
Beer foam stability is an important characteristic. It has been suggested that isoforms of protein Z, that is, protein Z4 and protein Z7, contribute to beer foam stability. We investigated the relationship between beer foam stability and protein Z4 and protein Z7 using their deficient mutants. As a protein Z4-deficient mutant, cv. Pirkka was used. Protein Z7 deficiency was screened in 1564 barley accessions in the world collection of Okayama University, Japan. The barley samples from normal, protein Z4-deficient, protein Z7-deficient, and double-deficient were genotyped in F(2) populations and then pooled based on the DNA marker genotypes of protein Z4 and protein Z7. For a brewing trial, F(5) pooled subpopulations were used. After malting and brewing, the foam stability was determined, and the results showed that the levels of foam stability in the four samples were comparable. Two-dimensional gel electrophoresis was used to investigate the proteome in these beer samples. The results showed that low molecular weight proteins, including lipid transfer protein (LTP2), in the deficient mutants were higher than those in the normal sample. Our results suggest that the contribution of protein Z4 and protein Z7 to beer foam stability was not greater than that of other beer proteins.  相似文献   

4.
It was previously shown that ungerminated barley contains inhibitors that suppress the activities of green malt cysteine proteinases. This paper reports the purification and partial characterization of a second barley cysteine endoproteinase inhibitor, a protein called lipid transfer protein 2 (LTP2). The chromatographically purified inhibitor had a molecular mass of 7112. The amino acid composition and sequence data of the purified inhibitor indicated that it was a protein whose gene, but not the protein itself, was isolated earlier from barley aleurone tissue. The purified protein inhibited the activities of electrophoretically separated green malt cysteine proteinases but not the activities of the serine- or metalloproteinases. The purified LTP2 inhibited the same proteases as the LTP1 that was characterized previously but was present in the mature seed in much smaller amounts. Neither LTP1 nor LTP2 has been proven to transport lipids in vivo, and it seems possible that both serve to keep cysteine endoproteinases that are synthesized during barley seed development inactive until the plant needs them. The small amount of LTP2 in the seed made it impossible to determine whether it, like LTP1, is involved in beer foam formation. Because of its proteinase-inhibiting ability and its resistance to heat inactivation, some of the LTP2 may persist in beer.  相似文献   

5.
In this study, ten‐day‐old seedlings of barley {Hordeum vulgare L. cultivar Anadolu [boron (B)‐tolerant] and Hamidiye (B‐sensitive)} were used. Boron‐treated plants were grown on H3BO3 solution (final concentration of 10 mM) for five days. Control plants received no B treatment during this period. Total protein patterns were obtained by analysis of total protein extract from root and leaf tissues of control and B‐treated plants using two‐dimensional gel electrophoresis followed by silver staining. The protein profile of B‐treated seedlings of each cultivar was compared to the profile of control (no stress treatment) plants of the same cultivar. Silver‐stained gels showed that B stress caused increases or decreases in a number of proteins in root and leaf tissues. Moreover, as a result of B treatment, one newly synthesized protein with relative molecular weight (Mr) of 35.0 kDa was detected in root profile of the tolerant cultivar. This protein failed to show up in root profile of the B‐treated sensitive cultivar. Three proteins were quantitatively increased in B‐treated root profile of both cultivars. Following B treatment, three proteins were increased in root profile of the tolerant cultivar, but were not changed in the sensitive one. In leaf tissues, however, there were remarkable changes in total protein profiles after B treatment, relative to the control. Following B treatment, in leaf tissues, at least seven proteins were increased in amount in tolerant cultivar but were unchanged in the susceptible one. In tolerant and sensitive cultivars, amounts of two proteins were increased in B‐treated plants, relative to control seedlings. In addition, four proteins (Mr:29, 58, 58, and 22 kDa) were unchanged in control and B‐treated seedlings of the tolerant cultivar. In the susceptible cultivar however, among these four proteins, the first one (Mr:29) was very much reduced and the others (Mr: 58, 58, and 22 kDa) were completely lost in B‐treated seedlings. Moreover, following B treatment, a set of high‐molecular‐weight proteins was quantitatively decreased in the susceptible cultivar but was unchanged in the tolerant cultivar. These results indicate that in barley, certain proteins may be involved in tolerance to B toxicity. In this study, changes in polypeptide composition as a result of B toxic concentration in leaf tissues were more abundant than in roots. Therefore, it is suggested that these changes, especially at shoot level may form the basis of the tolerance mechanism to B toxicity.  相似文献   

6.
Beer consumers demand satisfactory and consistent foam stability; thus, it is a high priority for brewers. Beer foam is stabilized by the interaction between certain beer proteins, including lipid transfer protein 1 (LTP1), and isomerized hop alpha-acids, but destabilized by lipids. In this study it was shown that the wort boiling temperature during the brewing process was critical in determining the final beer LTP1 content and conformation. LTP1 levels during brewing were measured by an LTP1 ELISA, using antinative barley LTP1 polyclonal antibodies. It was observed that the higher wort boiling temperatures ( approximately 102 degrees C), resulting from low altitude at sea level, reduced the final beer LTP1 level to 2-3 microg/mL, whereas the lower wort boiling temperatures ( approximately 96 degrees C), resulting from higher altitudes (1800 m), produced LTP1 levels between 17 and 35 microg/mL. Low levels of LTP1 in combination with elevated levels of free fatty acids (FFA) resulted in poor foam stability, whereas beer produced with low levels of LTP1 and FFA had satisfactory foam stability. Previous studies indicated the need for LTP1 denaturing to improve its foam stabilizing properties. However, the results presented here show that LTP1 denaturation reduces its ability to act as a binding protein for foam-damaging FFA. These investigations suggest that wort boiling temperature is an important factor in determining the level and conformation of LTP1, thereby favoring satisfactory beer foam stability.  相似文献   

7.
Barley (Hordeum vulgare L.) malt contains endoproteinases belonging to all four of the commonly occurring classes, including serine proteinases. It also contains low molecular weight proteins that inhibit the activities of many of these endoproteinases, but it had never been shown that any barley or malt serine proteinases could be inhibited by any of these endogenous proteins. It is now reported that some proteins that were concentrated using an "affinity" method inhibited the activity of a malt serine endoproteinase. Two-dimensional electrophoretic and in vitro analyses showed that the inhibited enzyme was serine endoproteinase 1 (SEP-1) and that the inhibition could be quantified using a semipurified preparation of this enzyme. Amino acid sequencing and MALDI-TOF MS were used to identify the components of the partially purified inhibiting fractions. Only the "trypsin/alpha-amylase inhibitors" or chloroform/methanol (CM) proteins, most of which had truncated N and C termini, and one fragment of beta-amylase were present in the inhibitory fractions. When a CM protein fraction was prepared from barley according to traditional methods, some of its component proteins inhibited the activity of SEP-1 and some did not. This is the first report of the purification and identification of barley malt proteins that can inhibit an endogenous serine proteinase. It shows that some of the CM proteins probably play a role in controlling the activity of barley proteinases during germination, as well as possibly protecting the seed and young plant from microbes or pests.  相似文献   

8.
We examined the malting and brewing performances of a lipoxygenase‐1 (LOX‐1) null line of barley (Hordeum vulgare L.). The LOX‐normal malt and the LOX‐null malt were prepared from F4 populations derived from a single cross. We could not observe any major differences in the general malt characteristics between the two malts. A brewing trial was performed using these malts. The analysis of the wort and beer revealed that the absence of LOX‐1 had little effect on the general characteristics of the wort and beer. In contrast, beer made from the LOX‐null malt showed reduced levels of beer‐deteriorating substances, trans‐2‐nonenal (T2N), and trihydroxyoctadecenoic acid (THOD). In the sensory evaluation, well‐trained panel members recognized the significant superiority of the aged LOX‐null beer in terms of staleness. These results show that the LOX‐1 null barley line can be effectively used to improve the flavor stability of beer without changing the other important beer qualities.  相似文献   

9.
The influence of malting and brewing processes on the chemical and structural modifications occurring on LTP1 was investigated by mass spectrometry and circular dichroism. Proteins were first purified from malt, and samples were collected at various steps of beer processing performed on two barley cultivars. The levels of LTP1 found in malt were not significantly different from the amounts in barley seed. However, in malt, both LTP1b, a post-translational form of LTP1, and a third isoform named LTP1c were isolated. Moreover, both of these proteins were found to be heterogeneously glycated but still exhibited an alpha-helix structure. Both glycated LTP1 and LTP1b were recovered during mashing. It was also shown that glycated LTP1 was unfolded during heat treatment of wort boiling, which is in agreement with the denatured form previously isolated from beer.  相似文献   

10.
Ergosterol is considered to be a suitable indicator of mold infestation in barley and malt. In this study ergosterol levels in different varieties of barley and malt produced in the Czech Republic were determined. A modified high-performance liquid chromatography (HPLC) method was statistically processed, validated (Effivalidation program), and applied to 124 samples of barley and malt. Ergosterol was isolated by extraction and saponification, and the quantification was performed using HPLC with diode array detection. The content of ergosterol ranged between the limit of detection (LOD) and 36.3 mg/kg in barley and between the LOD and 131.1 mg/kg in malt. Ergosterol is presumably connected with metabolites generated when barley grain is attacked by pathogens, and such barley often shows a high overfoaming (gushing) value. However, it was found that the content of ergosterol does not correlate with the degree of beer gushing.  相似文献   

11.
Malt is known to have an impact on beer flavor stability mainly due to the presence of antioxidants. In this study, five barley varieties were malted at industrial and micro scale, and quality parameters of the resulting malts were measured (diastatic power, friability, beta-glucan content, antiradical power, reducing power, lipoxygenase activity, and nonenal potential) and correlated with the sensory data obtained for the corresponding fresh and forced aged beers. A statistical strategy using multiple linear regressions was applied to explore relationships between the malt chemical parameters and beer sensory data, showing antiradical power as the major contribution of malt to beer flavor stability. Additionally, the measured antiradical power, which is well correlated with the polyphenolic content, was found to be very similar for malt and barley, emphasizing the key role of barley endogenous polyphenols.  相似文献   

12.
The aim of the study was to determine the effect of herbicides on yields, yield components and malting quality of the grains of barley cultivars. Literature worldwide lacks works on the impact of herbicides on the brewing quality of barley. This work presents the results of field experiments and laboratory studies concerning the evaluation of the impact of herbicides, differing in terms of an active substance, on the yields, weed control and malting quality of selected cultivars of barley. The research was conducted in Poland. The herbicides affected the increase in grain yields of barley, due to their high effectiveness of weed control, resulting in an increase in the number of ears per area unit. Certain herbicides increased grain protein content and grain filling. As for malt extractivity, Kolbach index, Q index, diastatic power and beta-glucans content in wort, there were interactions between herbicides and cultivars. The impact of herbicides on parameters varied depending on cultivars. The herbicides, especially Chwastox Turbo 340 SL (MCPA?+?dicamba) and Secator 125 OD (amidosulfuron?+?iodosulfuron-methyl-sodium), worsened several (not many) malt quality parameters, depending on cultivars, but it did not contribute to disqualifying malting quality of grains. Therefore, all tested herbicides can be recommended for use by farmers.  相似文献   

13.
The formation of haze is a serious quality problem in beer production. It has been shown that the use of silica elute (SE)-ve malt (absence of molecular weight (MW) ～14000 Da) for brewing can improve haze stability in the resultant beer, and the protein was identified as a barley trypsin inhibitor of the chloroform/methanol type (BTI-CMe). The objectives of this study were to determine (1) the allelic diversity of the gene controlling BTI-CMe in cultivated and Tibetan wild barley and (2) allele-specific (AS) markers for screening SE protein type. A survey of 172 Tibetan annual wild barley accessions and 71 cultivated barley genotypes was conducted, and 104 wild accessions and 35 cultivated genotypes were identified as SE+ve and 68 wild accessions and 36 cultivated genotypes as SE-ve. The allelic diversity of the gene controlling BTI-CMe was investigated by cloning, alignment, and association analysis. It was found that there were significant differences between the SE+ve and SE-ve types in single-nucleotide polymorphisms at 234 (SNP(234)), SNP(313), and SNP(385.) Furthermore, two sets of AS markers were developed to screen SE protein type based on SNP(313). AS-PCR had results very similar to those obtained by immunoblot method. Mapping analysis showed that the gene controlling the MW～14 kDa band was located on the short arm of chromosome 3H, at the position of marker BPB-0527 (33.302 cM) in the Franklin/Yerong DH population.  相似文献   

14.
Detailed studies were carried out on the influence of corn size distribution on the values obtained for diastatic power (DP) of commercially malted barley. Malted barley was screened using a screening box, and the DP activities of the different corns retained on the different compartments of the screening box were determined. The malt samples retained on the 2.8 mm screen had the highest DP activity, whereas the small corns (相似文献   

15.
Purification and structural characterization of LTP1 polypeptides from beer   总被引：6，自引：0，他引：6  
We report on the purification of lipid transfer proteins (LTP) from barley seeds and beer with the aim of investigating the chemical modifications that occur during the brewing process. In seeds, the well-known LTP of 9 kDa (LTP1) has been found together with a second form named LTPb that displays comparable amino acid composition but was not fully sequenced. These two forms have been recovered in beer with marked chemical modifications including disulfide bond reduction and rearrangement and especially glycation by Maillard reaction. The glycation is heterogeneous with variable amounts of hexose units bound to LTPs. Circular dichroism shows that glycated LTP1 having all their disulfide bridges reduced are totally unfolded. These results provide a first basis for understanding how barley LTPs become foam-promoting agents during the malting and brewing process.  相似文献   

16.
It has been reported that germinated barley contains peptidases that are sensitive to metal-chelating agents; however, none of these enzymes have been isolated, nor have their roles in germinated barley been investigated. Anion-exchange chromatography and chromatofocusing have been used to isolate a group of peptidases from barley (Hordeum vulgare cv. Morex) green malt that are sensitive to metal-chelating agents. Their activities were studied using one- and two-dimensional polyacrylamide gel electrophoresis. When analyzed on two-dimensional PAGE gels that contained gelatin as substrate, the enzymes separated into three major and approximately six minor activity spots with acidic pI values. The enzymes were optimally active against the gelatin substrate at pH 8.0 and were completely inhibited by 1,10-phenanthroline and EDTA, indicating that they belonged to the metallopeptidase class (EC 3.4.24.x). After the enzymes were inhibited with EDTA, the activities were recovered in the presence of low concentrations of metal ions. The hydrolysis of gelatin substrate was also impaired by the presence of reducing agents. The metallopeptidases readily digested, in vitro, the barley prolamine D hordein, indicating that they may be involved in degrading storage proteins during barley germination.  相似文献   

17.
Samples of naturally contaminated hulled barley, with varying deoxynivalenol concentrations, were subjected to an abrasive type dehulling procedure. The remaining grain fractions were analyzed for weight remaining (%), deoxynivalenol (ppm), crude protein (%CP), neutral detergent fiber (%NDF), ash (%ASH), gross energy (GE; kcal/kg), and calculated digestible energy values (DE; kcal/kg). Following the initial 15 s of pearling, 85% of the grain mass remained. Additional pearling resulted in a linear decline of grain mass. Following 15 s of pearling, the grain contained 34% of the initial deoxynivalenol content, irrespective of the initial level of contamination. Further pearling resulted in continued significant (p < 0.05) reductions in the percent of deoxynivalenol remaining to a level of 7.9% after 120 s but with significant losses in grain mass. Pearling can serve as an effective means of reducing the deoxynivalenol content of barley, with improvements in nutrient levels. However, the need to reduce the deoxynivalenol content of contaminated barley to less than 1 ppm for swine will necessitate the removal of a significant amount of the grain mass for heavily contaminated samples.  相似文献   

18.
Volatile phenols have long been recognized as important flavor contributors to the aroma of various alcoholic beverages. The two main flavor-active volatile phenols in beer are 4-vinylguaiacol and 4-vinylphenol. They are the decarboxylation products of the precursors ferulic acid and p-coumaric acid, respectively, which are released during the brewing process, mainly from malt. In this study, the variability in the release of free and ester-bound hydroxycinnamic acids from nine malted barley ( Hordeum vulgare L.) varieties during wort production was investigated. A large variability between different barley malts and their corresponding worts was observed. Differences were also found between free ferulic acid levels from identical malt varieties originating from different malt houses. During mashing, free hydroxycinnamic acids in wort are both water-extracted and enzymatically released by cinnamoyl esterase activity. Esterase activities clearly differ between different barley malt varieties. Multiple linear regression analysis showed that the release of ferulic acid during mashing did not depend only on the barley malt esterase activity but also on the amount of ester-bound ferulic acid initially present in the wort and on its endoxylanase activity. The study demonstrates the importance of selecting a suitable malt variety as the first means of controlling the final volatile phenol levels in beer.  相似文献   

19.
For producing worts that are optimal for beer production, some, but not all, of the barley proteins must be degraded during malting and mashing. This protein hydrolysis is controlled by endoproteinases, and, in turn, is partially regulated by the presence of low-molecular-weight (LMW) proteinaceous inhibitors. This paper reports studies of the interactions between the proteinases and inhibitors and an "affinity" method for concentrating the inhibitors. The malt inhibitors (I) and proteinases (E) quickly formed strong (E-I) complexes when dissolved together, and all of the I was complexed. Heating at 100 degrees C, but not 70 degrees C, dissociated the complex, even though the enzyme activities were destroyed at 70 degrees C. The released I readily recomplexed with fresh E. Barley, however, contained insufficient E to complex all of its I complement. The E-I complex was treated with salts, detergents, and reducing agents to release active E molecules, but none disrupted the complex. By removing the LMW proteins from a malt E-I extract and dissociating the complex by heating, the concentration of I molecules was greatly increased. This "affinity" method can thus be used to concentrate the I molecules for further purification.  相似文献   

20.
This study examined the interaction between sorghum grain hardness and sorghum malt quality in terms of diastatic power and free amino nitrogen with endosperm modification during malting. The changes in kernel hardness during malting of four commercial sorghum cultivars of differing quality in terms of endosperm texture and potential malt quality were measured using tests for hardness and density, and endosperm modification was followed by scanning electron microscopy. The general pattern of modification during sorghum malting was confirmed to start at the endosperm–scutellum interface and then continue into the floury endosperm toward the kernel distal end. Significantly, a cultivar of intermediate hardness and low malting quality remained harder and modified more slowly than a harder cultivar of high malting quality. It appeared that intrinsic grain hardness and malt amylase and protease activity both affected malt hardness and endosperm modification, but amylase and protease activity had a greater effect because of their degradation of endosperm starch and protein. Of the hardness and density tests studied, the tangential abrasive dehulling device (TADD) gave the best measure of hardness throughout malting; maximum range was 24–100% kernel removed over five days of malting. Also, the data agreed with the observed malt modification rates. Thus, the TADD may have application as a simple and rapid test for estimating sorghum malt quality.  相似文献   

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