Kinetics of Granular Starch Hydrolysis in Corn Dry‐Grind Process

A promising modification to the dry grind process is the use of granular starch hydrolyzing enzymes (GSHE) for corn fractionation and ethanol production. Modeling the kinetics of granular starch hydrolysis in the corn dry‐grind process is challenging given the heterogeneity of the substrate and the combined activities of α‐amylase and glucoamylase. By measuring reducing sugars, the hydrolysis progress in dry‐grind corn slurry treated with a commercial GSHE was monitored during 3 h of reaction. Progress curves consisted initially of a rapid product formation phase (<15 min), followed by a longer range kinetics characterized by relatively slower product formation. The initial product from the accelerated kinetics was proportional to the adsorbed GSHE concentration, after accounting for the contribution of dissolved starch hydrolysis. The long range kinetics was modeled by an empirical equation that yielded two parameters: a which stood for the asymptotic limit of the product concentration, and ln (b) which was the time required to reach a product concentration of a/2. In response to increasing GSHE concentration, a increased and ln (b) decreased towards a saturation limit. In response to decreasing corn particle size (geometric mean diameter), a increased and ln (b) decreased linearly. These behaviors indicated both parameters were controlled by the concentration of accessible substrates in the starch granules. The findings have implications on the optimal usage of GSHE in dry‐grind processes.     

Intestinal structure and function of broiler chickens on diets supplemented with a mannan oligosaccharide

A study was conducted to evaluate the response of broiler chickens to a commercial synthetic mannan oligosaccharide, Bio‐Mos? (BM), included in sorghum/lupin‐based diets at 0.0, 1.0, 3.0 or 5.0 g kg^?1 diet. The diets were fed between 7 and 28 days of age, and both the gross response and mechanisms involved were evaluated. The highest level of BM in the diet resulted in longer (P < 0.01) jejunal villi than those observed at other levels of supplementation. The RNA content of the ileal mucosal homogenate was significantly greater (P < 0.05) in chicks raised on diets supplemented with 3.0 and 5.0 g BM kg^?1 diet than in other groups. The protein/DNA ratio of the jejunal mucosal homogenate was also higher (P < 0.05) in chicks fed the 1.0 g BM kg^?1 diet than in chicks fed the other diets. The protein/RNA and RNA/DNA ratios in ileal homogenates were significantly (P < 0.01) influenced by the presence of BM in the diet. There were significantly greater specific activities of maltase (P < 0.01), leucine aminopeptidase (P < 0.05) and alkaline phosphatase (P < 0.001) in the jejunum as a result of supplementation with BM. Digestive enzyme activities in the ileum were unaffected. L ‐Tryptophan uptake by jejunal brush‐border membrane vesicles was significantly higher (P < 0.01) in chicks raised on the 5.0 g BM kg^?1 diet than in the other chicks. The supplement led to minor improvement in body weight but no improvement in feed conversion ratio. These effects were related to changes in digestive enzyme activities and, perhaps, amino acid transport. © 2001 Society of Chemical Industry     

Anti‐obesity effect of Liupao tea extract by modulating lipid metabolism and oxidative stress in high‐fat‐diet‐induced obese mice

Liupao tea (LPT) is traditional dark Chinese tea. The effect of LPT extract on high‐fat‐diet‐induced obese mice was investigated systematically. The results showed that LPT extract could reduce body weight and significantly alleviate liver damage and fat accumulation. LPT could also decrease the levels of alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (AKP), total cholesterol (TC), triglycerides (TG), and low‐density lipoprotein cholesterol (LDL‐C) and increase the level of high‐density lipoprotein cholesterol (HDL‐C) in the liver. It also decreased the serum levels of inflammatory cytokines, including tumor necrosis factor alpha (TNF‐α), interferon gamma (IFN‐γ), interleukin (IL)‐1β, and IL‐6 and increased the serum levels of anti‐inflammatory cytokines, including IL‐10 and IL‐4. Moreover, LPT improved the levels of total superoxide dismutase (T‐SOD), glutathione peroxidase (GSH‐Px), and catalase (CAT) and reduced the level of malondialdehyde (MDA) in the liver. Moreover, LPT could upregulate the mRNA and protein expressions of peroxisome proliferator‐activated receptor alpha (PPAR‐α), lipoprotein lipase (LPL), carnitine palmitoyltransferase 1(CPT1), and cholesterol 7 alpha‐hydroxylase (CYP7A1) and downregulate those of PPAR‐γ and CCAAT/enhancer‐binding protein alpha (C/EBP‐α) in the liver. It also increased the mRNA expression of copper/zinc superoxide dismutase (SOD1), manganese superoxide dismutase (SOD2), CAT, gamma‐glutamylcysteine synthetase 1 (GSH1), and GSH‐Px. The components of LPT extract include catechin, rutin, taxifolin, and astragalin, which possibly have a wide range of biological activities. In conclusion, our work verified that LPT extract possessed an anti‐obesity effect and alleviated obesity‐related symptoms, including lipid metabolism disorder, chronic low‐grade inflammation, and liver damage, by modulating lipid metabolism and oxidative stress.     

Dietary supplementation with polysaccharides from Semen cassiae enhances immunoglobulin production and interleukin gene expression in early‐weaned piglets

Forty piglets were used to determine the effect of 14‐day dietary supplementation with polysaccharides from Semen cassiae (PSSA) on the immunity and interleukin (IL) gene expression in early‐weaned piglets. Piglets were weaned at 21 days of age and four pigs were euthanized to provide the data at initial point. The remaining 36 pigs were fed the following corn‐ and soybean meal‐based diets: the control diet (non‐PSSA), the low‐level PSSA diet (the control diet supplemented with 0.04% of PSSA), and the high‐level PSSA diet (the control diet supplemented with 0.08% PSSA). Blood samples were collected on days 0 and 14, and lymph nodes on day 14 of the experimental period for the following measurements: IL‐1β gene expression in blood mononuclear cells and jejunal mucosa; IL‐2 gene expression in lymphatic nodes; serum levels of cytokines (IL‐1β, IL‐2, and IL‐6); serum concentrations of immunoglobulins (IgA, IgG and IgM). Weaning stress resulted in decreases in serum antibody levels and cytoimmunity. Compared with the control group, dietary supplementation with a high level of PSSA increased: (1) serum concentrations of IL‐1β, IL‐2 and IL‐6, as well as IgG, IgA and IgM; (2) expression of the IL‐1β and IL‐2 genes in blood mononuclear cells and lymphatic nodes; and (3) expression of the IL‐1β gene in the jejunal mucosa. These novel findings suggest that polysaccharides from Semen cassiae enhance both the cell‐mediated immune response and the humoral immunity in the early‐weaned piglets. Copyright © 2007 Society of Chemical Industry     

Enhanced production of α‐amylase from Bacillus subtilis subsp. spizizenii in solid state fermentation by response surface methodology and its evaluation in the hydrolysis of raw potato starch

This is an attempt to lower the cost of starch hydrolysis by the discovery of new generation α‐amylase. A natural isolate of Bacillus subtilis subsp. spizizenii was capable of producing appreciable amounts of raw potato starch digesting α‐amylase in solid state fermentation of wheat bran. The enzyme productivity has been substantially enhanced by supplementing various nutrients and statistically studying their interactions by response surface methodology. A central composite design for amylase production system elucidated a wheat bran‐based medium supplemented with soybean meal, threonine, and B‐complex vitamins predicting a yield of 521 391 U/g dry solids. The enzyme preparation could effectively digest 5–15% suspension of insoluble potato starch in 6 h revealing the dextrose equivalent of 32–44. The supplementation of a glucoamylase preparation, thereafter, brought about complete saccharification. The yield achieved in the statistically optimized amylase system may be one of the best to date and its capability in directly liquefying raw potato starch granules makes this study novel.     

Long‐term effects on the digestive tract of feeding large amounts of resistant starch: A study in pigs

The present study aimed to assess the digestive consequences of the long‐term intake of two starches providing different amounts of resistant starch. Growing pigs were used as the animal model and meal‐fed for 14 weeks on a diet containing a high amount of either raw potato starch (RPS) or corn starch (CS). Digestive adaptation was chronologically evaluated by measuring organic matter (OM), crude protein (CP), neutral detergent fibre (NDF) and starch digestibility. After 97 days, whole‐tract digestibility of OM, CP and NDF was lower for RPS‐ compared to CS‐fed pigs, whereas no differences were observed in faecal starch digestibility. In contrast, starch digestibility was reduced in the proximal compartments (ileum, caecum and proximal colon) of animals fed the RPS diet. The concentration of short‐chain fatty acids (SCFAs; P < 0.05), and purine bases (PBs; P < 0.01) was also higher in distal colon and rectum of animals fed the RPS diet. Changes in bacterial community structure (dendogram analyses) were seen in the rectum. Biodiversity tends to increase more in RPS compared to CS fed animals (34.1 vs. 28.8; P = 0.07). Among SCFAs, the proportion of butyrate was two‐fold higher in proximal colon digesta of RPS compared to CS fed pigs (0.20 vs. 0.11; P < 0.05). Increased butyrate formation in the colon reduced the number of apoptosis per crypt in the proximal colonic mucosa (0.38 vs. 0.62; P < 0.05). RPS fermentation reduced indices associated with damage to intestinal epithelial cells, such as crypt cell hyperproliferation and magnesium excretion. Long‐term ingestion of RPS induces pronounced changes of the digestive tract and their microflora, modifying mineral absorption and colonic morphology for which health benefits are likely to be associated. Copyright © 2007 Society of Chemical Industry     

Comparison of the characterisation of the fruit‐like aroma of Teucrium flavum L. subsp flavum by hydrodistillation and solid‐phase micro‐extraction

BACKGROUND: Teucrium species (Lamiaceae) are interesting for the food industry since many of them are used in the preparation of flavoured wines and beers, herbal teas, bitters and liqueurs. On this basis, the analysis of the aroma components of Teucrium flavum L. subsp. flavum growing in central Italy was carried out by means of both hydrodistillation (HD) and headspace solid‐phase micro‐extraction (HS‐SPME), coupled with GC/FID and GC/MS. RESULTS: A total of 102 components were identified in the essential oils, representing 99.0‐99.3% of the total oils. Sesquiterpenes hydrocarbons constituted the major fraction (48.5–49.4%), with the apple‐like flavour (Z,E)‐α‐farnesene being the major component. HS‐SPME allowed the analysis of the volatiles not only emitted by specific plant parts, but also by different parts within a single flower: flower calyx afforded the highest contribution, in terms of volatiles, to the aroma of the plant. CONCLUSION: The chemical profile of the volatile fraction obtained by HD and HS‐SPME, demonstrated the plant fruit‐like aroma, confirming the usefulness for flavouring wines, bitters and other kind of beverages, and also suggested other applications, as aroma and taste enhancer in food processing. In particular, SPME resulted in a very useful technique, which permits a choice between the part of the plant which has the highest concentration of a specific fragrance, and therefore establishes the best way of sampling during industrial applications of aromatic plants. Copyright © 2009 Society of Chemical Industry     

Thermal and Molecular Characterization of Aspergillus awamori Glucoamylase Catalytic and Starch‐Binding Domains

Aspergillus awamori glucoamylase catalytic domain, linker, and starch‐binding domain, the first and third expressed from yeast, have molecular masses of 56.2, 12.6 and 12.9 kDa, respectively, as determined by MALDI‐TOF mass spectroscopy, and have 10.2, 73.2 and 7.0 % (w/w) carbohydrate, respectively, showing overglycosylation by yeast. Unfolding of the starch‐binding domain monitored by circular dichroism is reversible at pH 6.0—8.0, with the unfolding T_m and ΔH increasing from 49.7 to 58.5 °C and from 284 to 351 kJ/mol, respectively, as pH decreases from 8.0 to 6.0. The catalytic domain unfolds irreversibly at pH 7.5, producing a single asymmetric endotherm by differential scanning calorimetry, with T_m and ΔH at a 1 °C/min heating rate being 60.9 °C and 1720 kJ/mol, but with both increasing as the heating rate increases. This suggests that unfolding is partially under kinetic control, while various tests show that it does not follow a simple two‐state irreversible model. Values of ΔH from calculated solvent‐accessible surface areas of unglycosylated catalytic and starch‐binding domains are about 100 kJ/mol lower than experimentally determined ΔH values of the corresponding glycosylated domains, showing the effect of glycosylation on unfolding enthalpies.     

Effect of Enzymatic Hydrolysis of Wheat Starch on Amylose‐Lipid Complexes Stability

In the process of enzymatic hydrolysis of wheat starch to glucose, the presence of amylose‐lipid complexes (AML's) decreases swelling and dissolving capacity and the water binding capacity of starch, thus impeding the access of amylolytic enzymes to the starch granules. The aim of the work was to define the relationship between the stability of AML's and the conditions of the enzymatic hydrolysis of wheat starch such as the kind of enzymatic preparation (only amylolytic or both amylolytic and lipolytic) and time of hydrolysis. Hydrolysates were produced from wheat starch liquefied with bacterial α‐amylase BAN 240L, subjected to further treatment with the enzymatic preparation Spezyme GA 300W, containing glucoamylase and lysophospholipase and, for comparison, only with a glucoamylase preparation (AMG 300L). The effect of amylolytic and lipolytic enzymes on the stability of AML's in the process of wheat starch hydrolysis was estimated. The amount of fatty acids released during hydrolysis was determined with gas‐liquid chromatography (GLC) and by differential scanning calorimetry (DSC) measuring the enthalpy of decomposition of AML's. The investigations revealed a differentiated effect of the individual enzymatic preparations on the degree of degradation of AML's. Amylose‐lipid complexes were more susceptible to the attack of preparation Spezyme GA 300W than to the digestion by α‐amylase BAN 240 L and glucoamylase AMG 300L.     

Cloning and characterization of a novel NAD+‐dependent glyceraldehyde‐3‐phosphate dehydrogenase gene from Candida glycerinogenes and use of its promoter

A 3950 bp genomic fragment from Candida glycerinogenes, WL2002‐5, containing the CgGAP gene encoding a glyceraldehyde‐3‐phosphate dehydrogenase homologous to GAP genes in other yeasts using degenerate primers, was cloned and characterized with inverse PCR. Sequence analysis revealed a 1164 bp open reading frame encoding a putative peptide of 387 deduced amino acids, with a molecular mass of 36 kDa. The CgGAP protein consisted of an N‐terminal NAD⁺‐binding domain and a central catalytic domain. Six stress‐response elements were found in the upstream region of the CgGAP gene. The influence of CgGAP on glycolysis was investigated. Functional analysis revealed that Saccharomyces cerevisiae transformed with CgGAP was restored to the wild‐type phenotype when cultured in high‐osmolarity medium, suggesting that it is a functional GAP protein. Promoter studies in S. cerevisiae using the green fluorescent protein (gfp) gene as a reporter showed that the GAP promoter (P_CgGAP) is constitutively expressed in S. cerevisiae cells grown on glucose. Copyright © 2013 John Wiley & Sons, Ltd.     

Structural Transformation of Sago Starch by Heat‐Moisture and Osmotic‐Pressure Treatment

Sago starch was modified by osmotic‐pressure treatment (OPT) and heat‐moisture treatment (HMT) and physicochemical characteristics were compared. In OPT, sago starch was suspended in saturated sodium sulfate solution and heated for 1 h at 100, 110 and 120°C, corresponding to a calculated osmotic pressure of 32,728, 33,640 and 34,552 kPa (assuming sodium sulfate dissociates completely), respectively, and in HMT, sago starch with 20% moisture content was used. Change of X‐ray diffraction pattern from C‐type to A‐type was obtained for OPT and HMT starch at 110°C and 120 °C, respectively. RVA viscograms of both OPT and HMT starch exhibited a decrease of peak and breakdown viscosity but increase of final viscosity and pasting temperature. Onset (T_o), peak (T_p), and conclusion temperature (T_c) of gelatinization of both OPT and HMT starch increased significantly with increase of treatment temperature. Biphasic broadening of T_p was observed for HMT starch indicating an inhomogeneous heat transfer during HMT. The observed narrow peaks of DSC curves indicated better homogeneity of OPT. These properties suggest that OPT starch is more suitable for large‐scale production.     

Starch sulfate as an efficient and biodegradable polymer catalyst for one‐pot,four‐component reaction of 2H‐indazolo[2,1‐b]phthalazine‐triones

A practical and green method for the synthesis of 2H‐indazolo[2,1‐b]phthalazine‐1,6,11(13H)‐trione derivatives using starch sulfate as a solid acid catalyst for the four‐component condensation reaction from hydrazinium hydroxide, phthalic anhydride, dimedone, and aromatic aldehydes under thermal solvent‐free conditions is described for the first time. Starch sulfate, a recyclable and easily handled catalyst has been demonstrated as a new catalyst for the synthesis of these classes of compounds.     

Starch but not protein digestibility is altered in pigs fed transgenic peas containing α‐amylase inhibitor

Eighteen individually housed boars were randomly allocated to one of three dietary treatments, an experimental wheat diet containing 989.4 g kg^?1 of a basal wheat diet, or this experimental wheat diet with 500 g kg^?1 of the basal wheat diet replaced with 500 g kg^?1 of either transgenic or non‐transgenic peas. The transgenic peas expressed the bean (Phaseolus vulgaris L.) α‐amylase inhibitor 1 gene. Diets contained n‐hexatriacontane (0.2 g kg^?1) as an indigestible marker to allow the determination of nutrient digestibility at the terminal ileum. Pigs were offered 1.6 kg day^?1 for 15 days, after which they were anaesthetised, the ileal and faecal digesta collected and the pigs subsequently euthanased. The ileal dry matter and starch digestibilities of the experimental wheat, non‐transgenic and transgenic pea diets were 78.3, 74.2 and 45.8% and 95.9, 95.2 and 42.4%, respectively. The apparent nutrient digestibilities of the non‐transgenic and transgenic peas were determined by difference. The ileal dry matter digestibility was significantly reduced in the transgenic peas compared with the non‐transgenic peas (12.7 and 69.9%, respectively; P = 0.006), which was largely due to a reduced starch digestibility. The apparent crude protein digestibilities of the transgenic peas were similar to the non‐transgenic, being 79.7 and 78.5%, respectively. The amino acid digestibilities of the transgenic and non‐transgenic peas were also similar. Copyright © 2006 Society of Chemical Industry     

Effect of Organic Nitrogen Sources on Raw Starch‐Digesting Glucoamylase Production of Rhizopus sp. MKU 40

In a liquid cultivation of Rhizopus sp. MKU 40, supplementation of the medium with 1.5% (w/v) organic nitrogen sources (neopeptone, casein from milk, and meat extract) had a slightly positive effect on glucoamylase (GA) (EC 3.2.1.3) activity compared with the medium lacking organic nitrogen sources. The addition of organic nitrogen sources induced production of protease. Supplementation of the medium with 1.5% (w/v) organic nitrogen sources resulted in an acid and neutral protease activity of 11 — 25 U/mL and 12 — 20 U/mL, respectively. The co‐existence of GA‐I [a highly raw starch‐digesting glucoamylase (RSDG)] and protease in the same medium leads to the production of Ga‐II (an extremely weak RSDG) from GA‐I. As a result the RSDG activity in the medium decreases. Raw starch adsorption rates of a medium without organic nitrogen sources were 100%, because the medium contained only GA‐I. In contrast, the media supplemented with organic nitrogen sources had low starch adsorption rates because the media contained both GA‐I and GA‐II. The results presented in this paper indicate that supplementation of the culture medium of Rhizopus strains with organic nitrogen sources negatively affects GA‐I production.     

Rheology of Rice Starch‐Sucrose Composites

The effect of sucrose at different concentrations (0, 10, 20 and 30%) on rheological properties of rice starch pastes (5% w/w) was investigated in steady and dynamic shear. The steady shear properties of rice starch‐sucrose composites were determined from rheological parameters for power law and Casson flow models. At 25°C all the starch‐sucrose composites exhibited a shear‐thinning flow behavior (n=0.25–0.44). The presence of sucrose resulted in the decrease in consistency index (K), apparent viscosity (η_a,100) and yield stress (σ_oc). Dynamic frequency sweeps at 25°C indicated that starch‐sucrose composites exhibited weak gel‐like behavior with storage moduli (G′) higher than loss moduli (G′′). G′ and G′′ values decreased with the increase in sucrose concentration. The dynamic (η*) and steady‐shear (η_a) viscosities at various sucrose concentrations did not follow the Cox‐Merz superposition rule. G′ values as a function of aging time (10 h) at 4°C showed a pseudoplateau region at long aging times. In general, the values of G′ and G′′ in rice starch‐sucrose composites were reduced in the presence of sucrose and depended on sucrose concentration.