RESUMEN
The use of starch in food systems in many instances relies on its thickening and gelling capacity. When native starches fail to match process and/or product-specific requirements, starches are physically and/or chemically modified to meet end-use demands. Evidently, differences between starches of varying botanical origin have to be considered when selecting or modifying starches for particular applications. Potato starch (PS) ranks third in world production after maize and wheat starches. Its unique properties differ from those of cereal and pulse starches and are directly related to its molecular structure and organization. This review summarizes the differences between PS and cereal and pulse starches and how they set it apart in terms of gelatinization, pasting, gelation, and retrogradation. Recent advances in improving PS pasting and gelation using enzyme technology and mineral ions are also described.
Asunto(s)
Tubérculos de la Planta/química , Solanum tuberosum/química , Almidón/química , Tecnología de Alimentos , Geles/química , ViscosidadRESUMEN
Potato chips are a major product in the savory snack market and are consumed worldwide because of their enjoyable and distinctive organoleptic properties. They are conventionally produced by deep-frying thin slices of fresh potato. In contrast, potato crisps are manufactured from dried potato derivatives such as potato flakes (PFs). Their production is reviewed in this manuscript and requires the formation of dough based on hydrated PFs. Expansion of the dough during deep-frying provides the crisps with their desired crunchy texture. As part of an overall trend, the consumer search for calorie-reduced food products has also stimulated research to lower oil uptake during crisp production. However, minimizing oil absorption without losing the characteristic palatability of deep-fried products is challenging and requires fundamental knowledge on factors determining product texture and oil absorption. The transformations and functional role of starch, potato's main constituent, during crisp making are key in this respect and are reviewed here.
Asunto(s)
Culinaria , Bocadillos , Solanum tuberosum/química , Almidón/química , Absorción Fisicoquímica , Aceites de Plantas/análisis , Propiedades de SuperficieRESUMEN
Gradientless baking by means of ohmic heating was used for the first time in gluten-free (GF) bread making. Combination thereof with in-line measurements of batter height, viscosity and carbon dioxide (CO2) release proved to be powerful for studying structure formation in GF breads. GF breads studied here were based on (i) a mixture of potato and cassava starches and egg white powder (C/P-S+EW), (ii) rice flour (RF) or (iii) a mixture of RF and egg white powder (RF+EW). The work revealed that bread volume and crumb structure rely heavily on the balance between the moment of CO2 release from batter during baking and that of crumb setting. At the moment of CO2 release, C/P-S+EW bread crumb had already (partly) set, while this was not the case for RF bread crumb, resulting in a collapse and thus low volume of the latter. When a part of RF was replaced by egg white powder, the moment of CO2 release was postponed and the batter collapse was less pronounced, leading to a higher volume and a finer crumb. The presence of egg white proteins in C/P-S+EW or RF+EW batters improved gas cell stabilization. Thus, increasing batter stability or altering the moment of crumb setting results in GF breads with higher volume and a finer crumb structure.
Asunto(s)
Pan/análisis , Culinaria/instrumentación , Dieta Sin Gluten , Impedancia Eléctrica , Dióxido de Carbono/análisis , Proteínas Dietéticas del Huevo/análisis , Diseño de Equipo , Fermentación , Calor , Manihot , Oryza , Solanum tuberosum , Almidón/análisis , Viscosidad , Agua/análisisRESUMEN
The impact of different hydration levels, on gelatinization of potato starch (PS), rice starch (RS) and a 1:1 blend thereof, was investigated by differential scanning calorimetry and related to nuclear magnetic resonance proton distributions of hydrated samples, before and after heating. At 20% or 30% hydration, the visual appearance of all samples was that of a wet powder, and limited, if any, gelatinization occurred upon heating. At 30% hydration, changes in proton distributions were observed and related to plasticization of amorphous regions in the granules. At 50% hydration, the PS-RS blend appeared more liquid-like than other hydrated samples and showed more pronounced gelatinization than expected based on additive behavior of pure starches. This was due to an additional mobile water fraction in the unheated PS-RS blend, originating from differences in water distribution due to altered stacking of granules and/or altered hydration of PS due to presence of cations in RS.
Asunto(s)
Rastreo Diferencial de Calorimetría/métodos , Imagen por Resonancia Magnética/métodos , Oryza/química , Solanum tuberosum/química , Almidón/química , Agua/química , CalefacciónRESUMEN
Granular normal maize starch was reacted with sodium trimetaphosphate in deionized water ( [Formula: see text] ), aqueous sodium sulfate solution ( [Formula: see text] ), aqueous ethanol (MSethanol) or aqueous acetone (MSacetone) under otherwise identical reaction conditions. Analysis of the resultant starches by Rapid Visco Analysis (RVA) showed that the starch was cross-linked to a higher degree in aqueous ethanol or aqueous acetone than in water or sodium sulfate solution, and with minimal starch leaching. While MSacetone and MSethanol had incorporated similar levels of phosphorous, RVA analysis and microscopic analysis showed that MSacetone granules were more effectively stabilized by cross-linking than MSethanol granules. Cross-linking in aqueous acetone is believed to either contain the greater numbers of distarch monophosphate (versus monostarch monophosphate), or occur more intensively at the granule outer layers than that in aqueous ethanol and, at the same time, to account for the greater granular strength of MSethanol than that of MSacetone.
Asunto(s)
Almidón/química , Zea mays/química , Rastreo Diferencial de Calorimetría , Etanol/química , Solanum tuberosum/química , Sulfatos/química , Temperatura , Viscosidad , Agua/química , Zea mays/metabolismoRESUMEN
Wheat bran extract (WBE), containing arabinoxylan-oligosaccharides that are potential prebiotic substrates, has been shown to modify bacterial colonic fermentation in human subjects and to beneficially affect the development of colorectal cancer (CRC) in rats. However, it is unclear whether these changes in fermentation are able to reduce the risk of developing CRC in humans. The aim of the present study was to evaluate the effects of WBE on the markers of CRC risk in healthy volunteers, and to correlate these effects with colonic fermentation. A total of twenty healthy subjects were enrolled in a double-blind, cross-over, randomised, controlled trial in which the subjects ingested WBE (10 g/d) or placebo (maltodextrin, 10 g/d) for 3 weeks, separated by a 3-week washout period. At the end of each study period, colonic handling of NH3 was evaluated using the biomarker lactose[15N, 15N']ureide, colonic fermentation was characterised through a metabolomics approach, and the predominant microbial composition was analysed using denaturing gradient gel electrophoresis. As markers of CRC risk, faecal water genotoxicity was determined using the comet assay and faecal water cytotoxicity using a colorimetric cell viability assay. Intake of WBE induced a shift from urinary to faecal 15N excretion, indicating a stimulation of colonic bacterial activity and/or growth. Microbial analysis revealed a selective stimulation of Bifidobacterium adolescentis. In addition, WBE altered the colonic fermentation pattern and significantly reduced colonic protein fermentation compared with the run-in period. However, faecal water cytotoxicity and genotoxicity were not affected. Although intake of WBE clearly affected colonic fermentation and changed the composition of the microbiota, these changes were not associated with the changes in the markers of CRC risk.
Asunto(s)
Fibras de la Dieta/análisis , Disbiosis/prevención & control , Microbioma Gastrointestinal , Extractos Vegetales/uso terapéutico , Prebióticos , Semillas/química , Triticum/química , Adulto , Anticarcinógenos/efectos adversos , Anticarcinógenos/uso terapéutico , Bélgica/epidemiología , Biomarcadores/análisis , Neoplasias Colorrectales/epidemiología , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/microbiología , Neoplasias Colorrectales/prevención & control , Estudios Cruzados , Método Doble Ciego , Disbiosis/metabolismo , Disbiosis/microbiología , Heces/química , Heces/microbiología , Femenino , Fermentación , Fármacos Gastrointestinales/efectos adversos , Fármacos Gastrointestinales/uso terapéutico , Humanos , Masculino , Extractos Vegetales/efectos adversos , Prebióticos/efectos adversos , Riesgo , Adulto JovenRESUMEN
OBJECTIVES: We assessed whether wheat bran extract (WBE) containing arabinoxylan-oligosaccharides (AXOS) elicited a prebiotic effect and modulated gastrointestinal (GI) parameters in healthy preadolescent children upon consumption in a beverage. METHODS: This double-blind randomized placebo-controlled crossover trial evaluated the effects of consuming WBE at 0 (control) or 5.0 g/day for 3 weeks in 29 healthy children (8-12 years). Fecal levels of microbiota, short-chain fatty acids, branched-chain fatty acids, ammonia, moisture, and fecal pH were assessed at the end of each treatment and at the end of a 1-week run-in (RI) period. In addition, the subjects completed questionnaires scoring distress severity of 3 surveyed GI symptoms. Finally, subjects recorded defecation frequency and stool consistency. RESULTS: Nominal fecal bifidobacteria levels tended to increase after 5 g/day WBE consumption (P = 0.069), whereas bifidobacteria expressed as percentage of total fecal microbiota was significantly higher upon 5 g/day WBE intake (P = 0.002). Additionally, 5 g/day WBE intake induced a significant decrease in fecal content of isobutyric acid and isovaleric acid (P < 0.01), markers of protein fermentation. WBE intake did not cause a change in distress severity of the 3 surveyed GI symptoms (flatulence, abdominal pain/cramps, and urge to vomit) (P > 0.1). CONCLUSIONS: WBE is well tolerated at doses up to 5 g/day in healthy preadolescent children. In addition, the intake of 5 g/day exerts beneficial effects on gut parameters, in particular an increase in fecal bifidobacteria levels relative to total fecal microbiota, and reduction of colonic protein fermentation.
Asunto(s)
Fibras de la Dieta , Tracto Gastrointestinal/microbiología , Microbiota/efectos de los fármacos , Oligosacáridos/administración & dosificación , Extractos Vegetales/administración & dosificación , Xilanos/administración & dosificación , Dolor Abdominal/etiología , Amoníaco/análisis , Bifidobacterium/aislamiento & purificación , Niño , Estudios Cruzados , Fibras de la Dieta/análisis , Método Doble Ciego , Ácidos Grasos/análisis , Ácidos Grasos Volátiles/análisis , Heces/química , Heces/microbiología , Femenino , Flatulencia/inducido químicamente , Tracto Gastrointestinal/efectos de los fármacos , Humanos , Concentración de Iones de Hidrógeno , Masculino , Oligosacáridos/análisis , Cooperación del Paciente , Placebos , Extractos Vegetales/efectos adversos , Prebióticos , Xilanos/análisisRESUMEN
A large and diverse material collection of whole grain wheat samples (n=129) was analysed for total dietary fibre (TDF) content and composition, including fructan (11.5-15.5%). Correlations between the dietary fibre components, associated bioactive components (e.g. tocols, sterols, phenolic acids and folates) and agronomic properties previously determined on the same samples were found with multivariate analysis (PCA). Samples from the same countries had similar characteristics. The first PC described variation in components concentrated in the starchy endosperm (e.g. starch, ß-glucan and fructan) and the dietary fibre components concentrated in the bran (e.g. TDF, arabinoxylan and cellulose). The second PC described the variation in kernel weight and other bran components such as alkylresorcinols, tocols and sterols. Interestingly, there was no correlation among these different groups of bran components, which reflected their concentration in different bran tissues. The results are of importance for plant breeders who wish to develop varieties with health-promoting effects.
Asunto(s)
Biodiversidad , Fibras de la Dieta/análisis , Extractos Vegetales/análisis , Triticum/química , Cruzamiento , Celulosa/análisis , Ácido Fólico/análisis , Genotipo , Humanos , Esteroles/análisis , Triticum/genéticaRESUMEN
A unique series of potato (mutant) starches with highly different amylopectin/amylose (AP/AM) ratios was annealed in excess water at stepwise increasing temperatures to increase the starch melting (or gelatinization) temperatures in aqueous suspensions. Small-angle X-ray scattering (SAXS) experiments revealed that the lamellar starch crystals gain stability upon annealing via thickening for high-AM starch, whereas the crystal surface energy decreases for AM-free starch. In starches with intermediate AP/AM ratio, both mechanisms occur, but the surface energy reduction mechanism prevails. Crystal thickening seems to be associated with the cocrystallization of AM with AP, leading to very disordered nanomorphologies for which a new SAXS data interpretation scheme needed to be developed. Annealing affects neither the crystal internal structure nor the spherulitic morphology on a micrometer length scale.
Asunto(s)
Almidón/química , Amilopectina/química , Amilosa/química , Cristalización , Solanum tuberosum/química , TemperaturaRESUMEN
Wheat bran extract (WBE) is a food-grade soluble fibre preparation that is highly enriched in arabinoxylan oligosaccharides. In this placebo-controlled cross-over human intervention trial, tolerance and effects on colonic protein and carbohydrate fermentation were studied. After a 1-week run-in period, sixty-three healthy adult volunteers consumed 3, 10 and 0 g WBE/d for 3 weeks in a random order, with 2 weeks' washout between each treatment period. Fasting blood samples were collected at the end of the run-in period and at the end of each treatment period for analysis of haematological and clinical chemistry parameters. Additionally, subjects collected a stool sample for analysis of microbiota, SCFA and pH. A urine sample, collected over 48 h, was used for analysis of p-cresol and phenol content. Finally, the subjects completed questionnaires scoring occurrence frequency and distress severity of eighteen gastrointestinal symptoms. Urinary p-cresol excretion was significantly decreased after WBE consumption at 10 g/d. Faecal bifidobacteria levels were significantly increased after daily intake of 10 g WBE. Additionally, WBE intake at 10 g/d increased faecal SCFA concentrations and lowered faecal pH, indicating increased colonic fermentation of WBE into desired metabolites. At 10 g/d, WBE caused a mild increase in flatulence occurrence frequency and distress severity and a tendency for a mild decrease in constipation occurrence frequency. In conclusion, WBE is well tolerated at doses up to 10 g/d in healthy adults volunteers. Intake of 10 g WBE/d exerts beneficial effects on gut health parameters.
Asunto(s)
Fibras de la Dieta/análisis , Tracto Gastrointestinal/efectos de los fármacos , Promoción de la Salud , Oligosacáridos/administración & dosificación , Extractos Vegetales/administración & dosificación , Xilanos/administración & dosificación , Adulto , Bifidobacterium/crecimiento & desarrollo , Cresoles/orina , Estudios Cruzados , Método Doble Ciego , Ácidos Grasos Volátiles/análisis , Heces/química , Heces/microbiología , Femenino , Fermentación , Enfermedades Gastrointestinales/inducido químicamente , Humanos , Concentración de Iones de Hidrógeno , Masculino , Persona de Mediana Edad , Oligosacáridos/metabolismo , Placebos , Extractos Vegetales/efectos adversos , Extractos Vegetales/química , Xilanos/metabolismoRESUMEN
Ball milling was used for producing complex arabinoxylan oligosaccharides (AXOS) and low molecular mass arabinoxylans (AX) from wheat bran, pericarp-enriched wheat bran, and psyllium seed husk. The arabinose to xylose ratio of the samples produced varied between 0.14 and 0.92, and their average degree of polymerization (avDP) ranged between 42 and 300. Their fermentation for 48 h in an in vitro system using human colon suspensions was compared to enzymatically produced wheat bran AXOS with an arabinose to xylose ratio of 0.22 and 0.34 and an avDP of 4 and 40, respectively. Degrees of AXOS fermentation ranged from 28% to 50% and were lower for the higher arabinose to xylose ratio and/or higher avDP materials. Arabinose to xylose ratios of the unfermented fractions exceeded those of their fermented counterparts, indicating that molecules less substituted with arabinose were preferably fermented. Xylanase, arabinofuranosidase, and xylosidase activities increased with incubation time. Enzyme activities in the samples containing psyllium seed husk AX or psyllium seed husk AXOS were generally higher than those in the wheat bran AXOS preparations. Fermentation gave rise to unbranched short-chain fatty acids. Concentrations of acetic, propionic, and butyric acids increased to 1.9-2.6, 1.9-2.8, and 1.3-2.0 times their initial values, respectively, after 24 h incubation. Results show that the human intestinal microbiota can at least partially use complex AXOS and low molecular mass AX. The tested materials are thus interesting physiologically active carbohydrates.
Asunto(s)
Fibras de la Dieta/análisis , Fermentación , Oligosacáridos/metabolismo , Psyllium/química , Xilanos/metabolismo , Glicósido Hidrolasas/metabolismo , Humanos , Peso Molecular , Semillas/química , Xilanos/químicaRESUMEN
Starch, an essential component of an equilibrated diet, is present in cereals such as common and durum wheat, maize, rice, and rye, in roots and tubers such as potato and cassava, and in legumes such as peas. During food processing, starch mainly undergoes nonchemical transformations. Here, we focus on the occurrence of starch in food raw materials, its composition and properties, and its transformations from raw material to final products. We therefore describe a number of predominant food processes and identify research needs. Nonchemical transformations that are dealt with include physical damage to starch, gelatinization, amylose-lipid complex formation, amylose crystallization, and amylopectin retrogradation. A main focus is on wheat-based processes. (Bio)chemical modifications of starch by amylolytic enzymes are dealt with only in the context of understanding the starch component in bread making.
Asunto(s)
Manipulación de Alimentos , Almidón/química , Pan/análisis , Fenómenos Químicos , Grano Comestible/química , Comida Rápida/análisis , Harina/análisis , Tubérculos de la Planta/química , Solanum tuberosum/química , Almidón/metabolismoRESUMEN
Psyllium (Plantago ovata Forsk) seed husk (PSH) is very rich in arabinoxylan (AX). However, its high gelling capacity and the complex nature of the AX make it difficult to process. In this study, ball milling was investigated as a tool for enhancing PSH AX water extractability and molecular mass (MM). A 48 h laboratory-scale ball mill treatment under standardized optimal conditions reduced the PSH average particle size from 161 microm for the untreated sample to 6 microm. Concurrently, it increased the water-extractable AX (WE-AX) level from 13 (untreated PSH) to 90% of the total PSH AX. While the WE-AX of the untreated PSH had a peak MM of 216 kDa and an arabinose to xylose (A/X) ratio of 0.20, WE-AX fragments from ball mill-pretreated PSH had a peak MM of 22 kDa and an A/X ratio of 0.31. Ball milling further drastically reduced the intrinsic viscosity of PSH extracts and their water-holding capacity. Prolonged treatment brought almost all AX (98%) in solution and yielded WE-AX fragments with an even higher A/X ratio (0.42) and a lower peak MM (11 kDa). While impact and jet milling of PSH equally led to significant reductions in particle size, these technologies only marginally affected the water extractability of PSH AX. This implies that ball milling affects PSH particles and their constituent molecules differently than impact and jet milling.
Asunto(s)
Manipulación de Alimentos , Extractos Vegetales/química , Psyllium/química , Semillas/química , Xilanos/química , Peso Molecular , Tamaño de la Partícula , ViscosidadRESUMEN
OBJECTIVE: Arabinoxylooligosaccharides (AXOS) are non-digestible in the upper gastrointestinal tract and have been shown to exert prebiotic effects in animals. The aim of this study was to characterize the influence of AXOS with an average degree of polymerization of 15 and an average degree of arabinose substitution of 0.26 (AXOS-15-0.26) on gastrointestinal motility and colonic bacterial metabolism in healthy human volunteers. METHODS: Twelve healthy volunteers received five test meals, containing different amounts of AXOS-15-0.26, with one week intervals between each test meal. Breath tests were used to measure gastric emptying rate, oro-cecal transit time (OCTT) and hydrogen excretion. Colonic bacterial metabolism was estimated using the biomarkers lactose-[(15)N, (15)N']-ureide ((15)N-LU) and p-cresol. RESULTS: Gastric emptying and OCTT were not influenced by addition of varying amounts of AXOS-15-0.26. Administration of 2.2g or 4.9 g AXOS-15-0.26 significantly decreased the urinary (15)N-excretion (respectively p = 0.008 and p = 0.035) as compared to the baseline, whereas fecal (15)N-excretion was significantly increased (respectively p = 0.034 and p = 0.019). This shift from urinary to fecal (15)N-excretion suggests a higher uptake or incorporation by bacteria due to the stimulation of colonic bacterial growth and/or metabolic activity. Furthermore, a significant increase in hydrogen excretion after administration of 2.2g (p = 0.002) and 4.9 g (p = 0.004) AXOS-15-0.26 was observed. No influence on urinary p-cresol excretion was observed. CONCLUSION: These findings suggest that a minimal dose of 2.2g AXOS-15-0.26 favorably modulates the colonic bacterial metabolism in healthy humans. However, long term studies are required to confirm a possible prebiotic effect.
Asunto(s)
Colon/metabolismo , Colon/microbiología , Motilidad Gastrointestinal/efectos de los fármacos , Oligosacáridos/farmacología , Extractos Vegetales/farmacología , Probióticos/farmacología , Adulto , Biomarcadores/análisis , Relación Dosis-Respuesta a Droga , Grano Comestible , Femenino , Alimentos Fortificados , Tránsito Gastrointestinal , Humanos , Isótopos/análisis , Masculino , Oligosacáridos/administración & dosificación , Adulto JovenRESUMEN
The action pattern of several amylases was studied at 35, 50, and 70 degrees C using potato amylose, a soluble (Red Starch) and insoluble (cross-linked amylose) chromophoric substrate. With potato amylose as substrate, Bacillus stearothermophilus alpha-amylase (BStA) and porcine pancreatic alpha-amylase displayed a high degree of multiple attack (DMA, i.e., the number of bonds broken during the lifetime of an enzyme-substrate complex minus one), the fungal alpha-amylase from Aspergillus oryzae a low DMA, and the alpha-amylases from B. licheniformis, Thermoactinomyces vulgaris, B. amyloliquifaciens, and B. subtilis an intermediate DMA. These data are discussed in relation to structural properties of the enzymes. The level of multiple attack (LMA), based on the relation between the drop in iodine binding of amylose and the increase in total reducing value, proved to be a good alternative for DMA measurements. The LMA of the endo-amylases increased with temperature to a degree depending on the amylase. In contrast, BStA showed a decreased LMA when temperature was raised. Furthermore, different enzymes had different activities on Red Starch and cross-linked amylose. Hence, next to the temperature, the action pattern of alpha-amylases is influenced by structural parameters of the starch substrate.
Asunto(s)
Amilasas/química , Bioquímica/métodos , Amilosa/química , Animales , Aspergillus oryzae/enzimología , Geobacillus stearothermophilus/enzimología , Concentración de Iones de Hidrógeno , Modelos Químicos , Modelos Estadísticos , Páncreas/enzimología , Solanum tuberosum/enzimología , Porcinos , Temperatura , alfa-Amilasas/químicaRESUMEN
The search for functional foods or functional food ingredients, i.e. foods or food ingredients that can enhance health, is beyond any doubt one of the leading trends in today's food industry. In this context, probiotics, i.e. living microbial food supplements, and prebiotics, i.e. non-digestible food ingredients, receive much attention. Both popular concepts target the gastrointestinal microbiota. While in the Western world, intake of probiotics has been recommended for long, prebiotics in general, and non-digestible oligosaccharides in particular, have only recently received attention. This review deals with production and characterization of non-digestible oligosaccharides and focuses on their role in promoting health and treating diseases. Attention is paid to the effects of non-digestible oligosaccharides on constipation, mineral absorption, lipid metabolism, cancer prevention, hepatic encephalopathy, glycemia/insulinemia, and immunomodulation.
Asunto(s)
Digestión , Oligosacáridos/administración & dosificación , Probióticos/administración & dosificación , Bacterias/efectos de los fármacos , Bacterias/metabolismo , Cromatografía , Fibras de la Dieta/administración & dosificación , Enzimas , Manipulación de Alimentos/métodos , Tracto Gastrointestinal/microbiología , Promoción de la Salud , Humanos , Oligosacáridos/efectos adversos , Oligosacáridos/química , Oligosacáridos/uso terapéutico , Probióticos/uso terapéutico , Análisis EspectralRESUMEN
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.
Asunto(s)
Endo-1,4-beta Xilanasas/análisis , Semillas/enzimología , Triticum/enzimología , Triticum/microbiología , Endo-1,4-beta Xilanasas/antagonistas & inhibidores , Extractos VegetalesRESUMEN
On-line size-exclusion chromatography monitoring of potato phosphorylase catalyzed amylose synthesis--starting from alpha-D-glucose-1-P and maltohexaose--revealed rather monodisperse amylose populations. In the presence of lipids, amylose-lipid complexes spontaneously formed and precipitated. They were recovered by centrifugation, freeze-dried, and characterized by wide-angle X-ray diffraction and differential scanning calorimetry. The presence of lipids during amylose synthesis led to lower amylose degrees of polymerization (DP). Lipid chain length defined amylose DP, which increased in the order myristic acid (C14), glyceryl monostearate (GMS), stearic acid (C18), and docosanoic acid (C22). The thermal stability of the complexes increased in the same manner, with the C22 complexes having the highest dissociation temperature. In addition, we hypothesized that these results provide additional evidence for the fringed micellar organization of (semi-enzymically synthesized) amylose-lipid complexes.
Asunto(s)
Amilosa/química , Sustancias Macromoleculares/química , Fosforilasas/química , Solanum tuberosum/enzimología , Rastreo Diferencial de Calorimetría , Conformación de Carbohidratos , Secuencia de Carbohidratos , Catálisis , Cromatografía , Ácidos Grasos/química , Glicéridos/química , Calor , Lípidos/química , Micelas , Modelos Químicos , Datos de Secuencia Molecular , Peso Molecular , Ácido Mirístico/química , Polímeros/química , Almidón/química , Ácidos Esteáricos/química , Temperatura , Factores de Tiempo , Difracción de Rayos XRESUMEN
Wheat grains contain Triticum aestivum xylanase inhibitor (TAXI) proteins which inhibit microbial xylanases, some of which are used in cereal based food industries. These inhibitors may play a role in plant defence. Among the TAXI isoforms described so far, TAXI-II displays a deviating inhibition specificity pattern. Here, we report on the molecular identity of TAXI-II and the basis of its inhibition specificity. Three candidate TAXI-II encoding sequences were isolated and recombinantly expressed in Pichia pastoris. To identify TAXI-II, the resulting proteins were tested against glycoside hydrolase family (GHF) 11 xylanases of Aspergillus niger (ANX) and Bacillus subtilis (BSX). One of these proteins (rTAXI-IB) inhibited both enzymes, like natural TAXI-I. The other candidates (rTAXI-IIA and rTAXI-IIB) showed an inhibition pattern typical for natural TAXI-II, only clearly inhibiting BSX. Comparative analysis of these highly similar sequences with distinct inhibition activity patterns, combined with information on the structural basis for ANX inhibition by TAXI-I [S. Sansen, C.J. De Ranter, K. Gebruers, K. Brijs, C.M. Courtin, J.A. Delcour, A. Rabijns, Structural basis for inhibition of Aspergillus niger xylanase by Triticum aestivum xylanase inhibitor-I, J. Biol. Chem. 279 (2004) 36022-36028], indicated a crucial role for Pro294 of TAXI-IIA and Gln376 of TAXI-IIB in determining the reduced inhibition activity towards ANX. Consequently, single point mutants rTAXI-IIA[P294L] and rTAXI-IIB[Q376H], both displaying the Leu/His combination corresponding to TAXI-I, were able to inhibit ANX. These results show that TAXI-II inhibition specificity bears on the identity of two key residues at positions 294 and 376, which are involved in the interaction at the -2 glycon subsite and the active site of GHF 11, respectively.