Structure characterization of the central repetitive domain of high molecular weight gluten proteins. I. Model studies using cyclic and linear peptides.

The high molecular weight (HMW) proteins from wheat contain a repetitive domain that forms 60-80% of their sequence. The consensus peptides PGQGQQ and GYYPTSPQQ form more than 90% of the domain; both are predicted to adopt beta-turn structure. This paper describes the structural characterization of these consensus peptides and forms the basis for the structural characterization of the repetitive HMW domain, described in the companion paper. The cyclic peptides cyclo-[PGQGQQPGQGQQ] (peptide 1), cyclo-[GYYPTSPQQGA] (peptide 2), and cyclo-[PGQGQQGYYPTSPQQ] (peptide 3) were prepared using a novel synthesis route. In addition, the linear peptides (PGQGQQ)n (n = 1, 3, 5) were prepared. CD, FTIR, and NMR data demonstrated a type II beta-turn structure at QPGQ in the cyclic peptide 1 that was also observed in the linear peptides 9PGQGQQ)n. A type I beta-turn was observed at YPTS and SPQQ in peptides 2 and 3, with additional beta-turns of either type I or II at GAGY (peptide 2) and QQGY (peptide 3). The proline in YPTS showed considerable cis/trans isomerization, with up to 50% of the population in the cis-conformation; the other prolines were more than 90% in the trans conformation. The conversion from trans to cis destroys the type I beta-turn at YPTS, but leads to an increase in turn character at SPQQ and GAGY (peptide 2) or QQGY (peptide 3).     

Structure characterization of the central repetitive domain of high molecular weight gluten proteins. II. Characterization in solution and in the dry state.

The structure of the central repetitive domain of high molecular weight HMW) wheat gluten proteins was characterized in solution and in the dry state using HMW proteins Bx6 and Bx7 and a subcloned, bacterially expressed part of the repetitive domain of HMW Dx5. Model studies of the HMW consensus peptides PGQGQQ and GYYPTSPQQ formed the basis for the data analysis (van Dijk AA et al., 1997, Protein Sci 6:637-648). In solution, the repetitive domain contained a continuous nonoverlapping series of both type I and type II II beta-turns at positions predicted from the model studies; type II beta-turns occurred at QPGQ and QQGY sequences and type I beta-turns at YPTS and SPQQ. The subcloned part of the HMW Dx5 repetitive domain sometimes migrated as two bands on SDS-PAGE; we present evidence that this may be caused by a single amino acid insertion that disturbs the regular structure of beta-turns. The type I beta-turns are lost when the protein is dried on a solid surface, probably by conversion to type II beta-turns. The homogeneous type II beta-turn distribution is compatible with the formation of a beta-spiral structure, which provides the protein with elastic properties. The beta-turns and thus the beta-spiral are stabilized by hydrogen bonds within and between turns. Reformation of this hydrogen bonding network after, e.g., mechanical disruption may be important for the elastic properties of gluten proteins.     

Effects of relaxation of gluten network on rehydration kinetics of pasta

The aim of this study was to investigate the effects of the relaxation of the gluten network on pasta rehydration kinetics. The moisture content of pasta, under conditions where the effects of the diffusion of water on the moisture content were negligible, was estimated by extrapolating the average moisture content of pasta of various diameters to 0?mm. The moisture content of imaginary, infinitely thin pasta did not reach equilibrium even after 1?h of rehydration. The rehydration of pasta made of only gluten was also measured. The rate constants estimated by the Long and Richman equation for both the pasta indicated that the rehydration kinetics of infinitely thin pasta were similar to those of gluten pasta. These results suggest that the swelling of starch by fast gelatinization was restricted by the honeycomb structural network of gluten and the relaxation of the gluten network controlled pasta rehydration kinetics.     

Visualization of Gluten and Starch Distributions in Dough by Fluorescence Fingerprint Imaging

A novel method combining imaging techniques and fluorescence fingerprint (FF) data measurement was developed to visualize the distributions of gluten and starch in dough without any preprocessing. Fluorescence images of thin sections of gluten, starch, and dough were acquired under 63 different combinations of excitation and emission wavelengths, resulting in a set of data consisting of the FF data for each pixel. Cosine similarity values between the FF of each pixel in the dough and those of gluten and starch were calculated. Each pixel was colored according to the cosine similarity value to obtain a pseudo-color image showing the distributions of gluten and starch. The dough sample was then fluorescently stained for gluten and starch. The stained image showed patterns similar to the pseudo-color FF image, validating the effectiveness of the FF imaging method. The method proved to be a powerful visualization tool, applicable in fields other than food technology.     

Effect of diet composition and lysine supply on growth and body composition in juvenile turbot (Psetta maxima)

A 10-week feeding trial was conducted to study the effect of feeding level and dietary lysine concentration on growth, protein and lysine retention, and body composition in juvenile turbot. Maintenance requirement for lysine and the efficiency of lysine utilisation were determined as well. Two experimental diets were formulated based on fishmeal or wheat gluten as main protein sources, containing 6.4 g (Diet A, control) and 4.5 g lysine per 100 g CP (Diet B), respectively. Diets were fed once daily at six feeding levels (per day 0.3%, 0.6%, 0.9%, 1.2%, and 1.5% of body weight [BW] and ad libitum) to a total of 432 fish of 48 g initial BW. No differences in the growth parameters were observed between diets at the same feeding level, except a lower feed to gain ratio (p < 0.05) at the highest feeding level at Diet B. Whole-body composition was not affected by diet, whereas muscle protein concentration was significantly lower for fish fed Diet B. Amino acid concentration in whole-body protein was affected by dietary treatment and fish fed Diet B showed lower concentrations of all essential amino acids. In fish muscle protein, lysine, methionine, leucine, isoleucine, and valine concentrations were significantly lower in Diet B. Efficiency of lysine utilisation for growth (k_lys) was determined by linear regression analysis and amounted for 0.69 for Diet B. The maintenance lysine requirement defined at zero lysine retention was 6.5 mg · kg^?0.8 · d^?1. Lysine intakes at zero protein retention were 13.0 mg and 12.9 mg · kg^?0.8 · d^?1 for Diet A and B, respectively. Growth and nutrient retention were similar for both diets and, therefore, a lysine deficiency in Diet B did not occur. In conclusion, a proportion of 330 g wheat gluten per kg feed did not influence growth performance and maintenance requirement for lysine in juvenile turbot. However, the effect of diet composition on the amino acid profile of body protein might be relevant for the derivation of the amino acid requirement from protein retention.     

Molecular characterization and expression analysis of BmNOX in two strains of Bombyx mori with contrasting viral resistance phenotype

We recently documented the identification of a 26.5 kDa protein named BmNox in the gut fluid of Nistari strain of Bombyx mori, which possessed antiviral activity against BmNPV in vitro. In this report, we report the characterization of the full‐length gene encoding BmNOX and the levels of expression of this gene in select tissues of silkworm larvae from a BmNPV‐susceptible and a BmNPV‐resistant strain to the defense capability in Bombyx mori larvae challenged with BmNPV. We also evaluated the BmNox expression in various stages of larval life of a resistant and a susceptible strain of Bombyx mori selected from among a panel of strains of silkworm. Nistari, a multivoltine strain of silkworm, expressed BmNOX during all five larval stages, and were highly resistant to BmNPV infection. In sharp contrast, CSR₂, a bivoltine strain, showed weaker expression of BmNOX in the anterior midgut in larval life and was highly susceptible to BmNPV infection. BmNOX is a secretory protein with dual expression in gut fluid and mid gut tissue. BmNOX is expressed heavily in the posterior mid gut, with weaker expression in the fore‐ and mid‐gut regions. © 2010 Wiley Periodicals, Inc.     

施钾对强筋小麦生理特性及产量的效应

以强筋面包小麦临优145为试验材料,于大田生产条件下研究了施用钾肥对强筋小麦叶片光合、膜脂氧化及产量特性的影响.结果显示,适量施钾能显著降低开花14d以后旗叶中超氧阴离子的产生强度和MDA含量,并显著提高旗叶的叶绿素含量、光合速率和籽粒灌浆速率,显著增加千粒重;在施钾(K2O)37.5~150.0kg/hm2范围内,随施钾量增加,各处理产量呈先上升后下降趋势且均比对照极显著增产14.5%~30.7%,各处理间产量差异显著,以施钾(K2O)112.5kg/hm2时籽粒产量最高.结果表明,中产条件下施用钾肥可显著提高强筋小麦籽粒产量,且以施钾(K2O)103.0~112.5kg/hm2为宜.     

改性谷朊粉在新型水性涂料中的应用研究

研究了谷朊粉的酸法改性及其在新型水性涂料中的应用.改性谷朊粉除具有优良的粘结性、流动性和流平性外,还提高了与涂料乳液的相容性,解决了谷朊粉原胶在涂料使用过程中的开裂严重,使得涂料具有更优的流平性和涂膜外观.