Showing papers on "Amylase published in 2006"

Sorghum grain as human food in Africa: relevance of content of starch and amylase activities

Abstract: Sorghum is a staple food grain in many semi-arid and tropic areas of the world, notably in Sub-Saharan Africa because of its good adaptation to hard environments and its good yield of production. Among important biochemical components for sorghum processing are levels of starch (amylose and amylopectin) and starch depolymerizing enzymes. Current research focus on identifying varieties meeting specific agricultural and food requirements from the great biodiversity of sorghums to insure food security. Results show that some sorghums are rich sources of micronutrients (minerals and vitamins) and macronutrients (carbohydrates, proteins and fat). Sorghum has a resistant starch, which makes it interesting for obese and diabetic people. In addition, sorghum may be an alternative food for people who are allergic to gluten. Malts of some sorghum varieties display α α α α-amylase and s-amylase activities comparable to those of barley, making them useful for various agro-industrial foods. The feature of sorghum as a food in developing as well as in developed countries is discussed. A particular emphasis is made on the impact of starch and starch degrading enzymes in the use of sorghum for some African foods, e.g. “to”, thin porridges for infants, granulated foods “couscous”, local beer “dolo”, as well agro-industrial foods such as lager beer and bread.

Intestinal sugar transport.

Abstract: Carbohydrates are an important component of the diet The carbohydrates that we ingest range from simple monosaccharides (glucose, fructose and galactose) to disaccharides (lactose, sucrose) to complex polysaccharides Most carbohydrates are digested by salivary and pancreatic amylases, and are further broken down into monosaccharides by enzymes in the brush border membrane (BBM) of enterocytes For example, lactase-phloridzin hydrolase and sucrase-isomaltase are two disaccharidases involved in the hydrolysis of nutritionally important disaccharides Once monosaccharides are presented to the BBM, mature enterocytes expressing nutrient transporters transport the sugars into the enterocytes This paper reviews the early studies that contributed to the development of a working model of intestinal sugar transport, and details the recent advances made in understanding the process by which sugars are absorbed in the intestine

Effects of sourdough and enzymes on staling of high-fibre wheat bread

Abstract: The effects of sourdough and enzyme mixture ( α -amylase, xylanase and lipase) on the specific volume, staling and microstructure of wheat pan bread supplemented with wheat bran were studied. Staling of bread was followed for 6 days by measuring the crumb firmness, changes in crystallization of amylopectin (DSC), increase in signal from the solid phase (NMR) and by light microscopy. The most effective treatment in improvement of quality was the combination of bran sourdough and enzyme mixture. During storage the rate of changes in crumb firmness, amylopectin crystallinity and rigidity of polymers were greatest for the white wheat bread. The most pronounced microstructural changes were swelling of starch granules and separation of amylose and amylopectin in the starch granules. Least changes in crumb firmness, amylopectin crystallinity and rigidity of polymers were observed in bran sourdough bread with enzymes. In contrast to white wheat bread, the starch granules were very much swollen in bran sourdough bread with enzyme mixture. This was hypothesized to be due to the higher water content of bran bread, and degradation of cell wall components leading to altered distribution of water among starch, gluten and bran particles during storage.

Characterization of cellulase-producing bacteria from the digestive tract of tilapia, Oreochromis mossambica (Peters) and grass carp, Ctenopharyngodon idella (Valenciennes)

Abstract: Isolation and characterization of cellulase-producing aeorobic bacterial flora in the intestine of omnivorous tilapia (Oreochromis mossambica) and phytophagous Chinese grass carp (Ctenopharyngodon idella) have been carried out using selective carboxymethylcellulose-agar (CMC-agar) medium. The cellulolytic activity was measured both qualitatively and quantitatively. It was found that the ability of different strains in degrading cellulose varies within a wide range. Among the strains isolated from the gut of each test fish, TM1 and CI3 isolated from O. mossambica and C. idella, respectively exhibited maximum cellulolytic activity (67.02 and 35.8 U mL−1 respectively). Pure cultures of these strains were selected for morphological, physiological and biochemical characterization. On the basis of these tests, the isolated strains were identified as Bacillus circulans (TM1) and Bacillus megaterium (CI3). Both the strains are rod-shaped, motile and show better temperature (15–42°C) and pH (5–11) tolerance. The selected strains were further quantitatively assayed for amylase and protease activities. Maximum amylase and protease activities were exhibited by TM1 and CI3 respectively. Information generated from the present study might contribute towards better-feed formulation incorporating plant ingredients.

Solid Culturing of Bacillus amyloliquefaciens for α-Amylase Production

Abstract: Summary Fourteen different agroresidues were screened for alpha amylase production using Bacillus amyloliquefaciens ATCC 23842. Among them, wheat bran (WB) and groundnut oil cake (GOC) in mass ratio of 1:1 was proved as the best substrate source. Supplementation with 0.01 M KH2PO4 and 1 % soluble starch enhanced the enzyme yield considerably. Maximum enzyme recovery from the solid mass was obtained when extracted with 0.1 M acetate buffer, pH=5.0. Maximum enzyme titer expressed as units per mass of dry substrate obtained was 62 470 U/g after 72 hours of fermentation at 37 °C by using the above solid substrate mixture (5 g) with the initial moisture of 85 % and inoculated with Bacillus amyloliquefaciens of 2·10 9 CFU/mL.

Paenibacillus curdlanolyticus Strain B-6 Xylanolytic-Cellulolytic Enzyme System That Degrades Insoluble Polysaccharides

Abstract: A facultatively anaerobic bacterium, Paenibacillus curdlanolyticus B-6, isolated from an anaerobic digester produces an extracellular xylanolytic-cellulolytic enzyme system containing xylanase, β-xylosidase, arabinofuranosidase, acetyl esterase, mannanase, carboxymethyl cellulase (CMCase), avicelase, cellobiohydrolase, β-glucosidase, amylase, and chitinase when grown on xylan under aerobic conditions. During growth on xylan, the bacterial cells were found to adhere to xylan from the early exponential growth phase to the late stationary growth phase. Scanning electron microscopic analysis revealed the adhesion of cells to xylan. The crude enzyme preparation was found to be capable of binding to insoluble xylan and Avicel. The xylanolytic-cellulolytic enzyme system efficiently hydrolyzed insoluble xylan, Avicel, and corn hulls to soluble sugars that were exclusively xylose and glucose. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of a crude enzyme preparation exhibited at least 17 proteins, and zymograms revealed multiple xylanases and cellulases containing 12 xylanases and 9 CMCases. The cellulose-binding proteins, which are mainly in a multienzyme complex, were isolated from the crude enzyme preparation by affinity purification on cellulose. This showed nine proteins by SDS-PAGE and eight xylanases and six CMCases on zymograms. Sephacryl S-300 gel filtration showed that the cellulose-binding proteins consisted of two multienzyme complexes with molecular masses of 1,450 and 400 kDa. The results indicated that the xylanolytic-cellulolytic enzyme system of this bacterium exists as multienzyme complexes.

Determining the binding affinities of phenolic compounds to proteins by quenching of the intrinsic tryptophan fluorescence

Abstract: The noncovalent binding of selected phenolic compounds (chlorogenic-, ferulic-, gallic acid, quercetin, rutin, and isoquercetin) to proteins (HSA, BSA, soy glycinin, and lysozyme) was studied by an indirect method applying the quenching of intrinsic tryptophan fluorescence. From the data obtained, the binding constants were calculated by nonlinear regression (one site binding; y = Bx/k + x). It has been reported that tannins inhibit human salivary amylase and that these complexes may reduce the development of cariogenic plaques. Further, amylase contains two tryptophan residues in its active site. Therefore, in a second part of the study involving 31 human subjects, evidence was sought for noncovalent interactions between the phenols of green tea and saliva proteins as measured by the fluorescence intensity. Amylase activity was determined before and after the addition of green tea to saliva of 31 subjects. Forty percent of the subjects showed an increase in amylase activity contrary to studies reporting only a decrease in activity. The interactions of tannin with amylase result in a decrease of its activity. It still remains to be elucidated why amylase does not react uniformly under conditions of applying green tea to saliva. Further, in terms of using phenols as caries inhibitors this finding should be of importance.

Roles of β‐amylase and starch breakdown during temperatures stress

Abstract: A primary role of β-amylase (BMY) is to produce maltose during hydrolytic starch degradation In photosynthetic organs, BMY activity is present in the chloroplast, where substrate starch is localized BMY activity is also found in the vacuole and cytosol, where starch is not known to be localized suggesting additional roles in glucan degradation Transcript levels and activity of different isoforms are regulated by different stimuli including cold, heat and drought stress However, little is known about the functional role of BMY during environmental stresses Recently, the functionality of one of the chloroplastic forms of Arabidopsis, ct-Bmy/BMY8/BAM3, has been shown to be needed for the protection of PSII photochemical efficiency following freezing stress by presumably catalyzing the synthesis of maltose, which acts as a cryoprotectant compound and precursor of soluble sugar metabolism Still, there remain important questions about how the mobilization of starch through maltose and glucose can lead to the accumulation of cryoprotectant soluble carbohydrates during cold acclimation

The effects of exercise in forest and urban environments on sympathetic nervous activity of normal young adults.

Abstract: In Japan, forest-air bathing and walking (shinrin-yoku) has been proposed as a health-facilitating activity in which people spend a short period of time in a forest environment. Initially, we examined the usefulness of salivary amylase activity as an indicator of an individual's stress levels in a forest environment. The circadian rhythm of salivary amylase activity was measured in healthy young male subjects under stress-free conditions. The salivary amylase activity remained relatively constant throughout the day. Salivary amylase activity was then measured before and after walking in both urban and forest environments using a hand-held monitor. Our results indicated that (i) the circadian rhythm fluctuations in salivary amylase activity were much smaller than the stressor-induced variations; (ii) salivary amylase activity was an excellent indicator of the changes in sympathetic nervous activity; and (iii) the forest was a good environment in which people could experience much less environment-derived stress.

Display of α-Amylase on the Surface of Lactobacillus casei Cells by Use of the PgsA Anchor Protein, and Production of Lactic Acid from Starch

Abstract: We developed a new cell surface engineering system based on the PgsA anchor protein from Bacillus subtilis. In this system, the N terminus of the target protein was fused to the PgsA protein and the resulting fusion protein was expressed on the cell surface. Using this new system, we constructed a novel starch-degrading strain of Lactobacillus casei by genetically displaying α-amylase from the Streptococcus bovis strain 148 with a FLAG peptide tag (AmyAF). Localization of the PgsA-AmyA-FLAG fusion protein on the cell surface was confirmed by immunofluorescence microscopy and flow cytometric analysis. The lactic acid bacteria which displayed AmyAF showed significantly elevated hydrolytic activity toward soluble starch. By fermentation using AmyAF-displaying L. casei cells, 50 g/liter of soluble starch was reduced to 13.7 g/liter, and 21.8 g/liter of lactic acid was produced within about 24 h. The yield in terms of grams of lactic acid produced per gram of carbohydrate utilized was 0.60 g per g of carbohydrate consumed at 24 h. Since AmyA was immobilized on the cells, cells were recovered after fermentation and used repeatedly. During repeated utilization of cells, the lactic acid yield was improved to 0.81 g per g of carbohydrate consumed at 72 h. These results indicate that efficient simultaneous saccharification and fermentation from soluble starch to lactic acid were carried out by recombinant L. casei cells with cell surface display of AmyA.

α-Amylase Production by Bacillus cereus MTCC 1305 Using Solid-State Fermentation

Abstract: Summary Production of a-amylase under solid-state fermentation by Bacillus cereus MTCC 1305 has been investigated using wheat bran and rice flake manufacturing waste as substrates. With wheat bran, highest enzyme production expressed as units per mass of dry substrate ((94±2) U/g) was observed. Production parameters were optimized as inoculum size 10 % (volume per mass) and substrate:moisture ratio 1:1. Among different carbon sources supplemented, glucose (0.04 g/g) showed enhanced enzyme production ((122±5) U/g). Supplementation of different nitrogen sources (0.02 g/g) showed decline in enzyme production. Optimum a-amylase enzyme activity was observed at 55 °C and pH=5. At 75 °C, enzyme showed 90 % activity compared to 55 °C.

Distinctive Sorghum Starch Granule Morphologies Appear to Improve Raw Starch Digestibility

Abstract: Various factors, including starch granule channels, have been suggested to contribute to the control of sorghum starch digestibility for animal feed. Isolated starch from two normal sorghum lines (P721N, IS6986) and one high protein digestibility (HPD) mutant line (111) that differed in starch granule morphology were selected to study the influence of these factors on starch digestibility. Scanning electron micrographs were taken of raw and digested starches. Microscopy results confirmed that in all three sorghum lines channels in starch granules are the main route of enzyme penetration and the central cavity area is the starting point of enzyme digestion. Channel density was more pronounced in the HPD sorghum mutant line than in normal lines, which may have been responsible for its relatively high digestibility. Micrographs of IS6986 showed unique starch granule morphology with a collapsed ”doughnut-shaped” structure in a portion of the granules. These unusual granules were rapidly digested and, unlike normal spherical granules, totally disappeared after 30 min of digestion. Amylases appeared to have fast access to the collapsed-appearing starch granules. Digestion profiles, following incubation with pepsin and α-amylase, showed that IS6986 and the HPD mutant (111) had the highest initial rate of starch digestion, followed by P721N. These findings provide insight as to how new sorghum cultivars might be developed with high starch digestibility for animal feed use.

Characterization of the Primary Starch Utilization Operon in the Obligate Anaerobe Bacteroides fragilis: Regulation by Carbon Source and Oxygen

Abstract: The opportunistic pathogen Bacteroides fragilis is a commensal organism in the large intestine, where it utilizes both dietary and host-derived polysaccharides as a source of carbon and energy. In this study, a four-gene operon required for starch utilization was identified. The operon also was found to be oxygen responsive and thus was designated osu for oxygen-induced starch utilization. The first three genes in the operon were predicted to encode outer membrane proteins involved in starch binding, and a fourth gene, osuD, encoded an amylase involved in starch hydrolysis. Insertional mutation of the osuA gene (ΩosuA) resulted in the inability to utilize starch or glycogen and an insertional mutation into the osuD gene (ΩosuD) was severely impaired for growth on starch media. Transcriptional studies indicated that maltose, maltooligosaccharides, and starch were inducers of osu expression and that maltose was the strongest inducer. A transcriptional activator of osuABCD, OsuR, was identified and found to mediate maltose induction. The ΩosuA and ΩosuD mutants were able to grow on maltose but not starch, whereas a mutation in osuR abolished growth on both substrates, indicating that additional genes under the control of OsuR are needed for maltose utilization. The osuABCD operon also was induced by exposure to oxygen and was shown to be part of the oxidative stress response important for aerotolerance of B. fragilis. Transcriptional analyses showed that osuA was induced 20-fold by oxygen, but OsuR was not required for this activation. Analysis of osu mutants suggested that expression of the operon was important for survival during oxygen exposure but not to hydrogen peroxide stress.

Sorghum Grain as Human Food in Africa: Relevance of Starch Content and Amylase Activities

Abstract: Sorghum is a staple food grain in many semi-arid and tropic areas of the world, notably in Sub-Saharan Africa because of its good adaptation to hard environments and its good yield of production. Among important biochemical components for sorghum processing are levels of starch (amylose and amylopectin) and starch depolymerizing enzymes. Current research focus on identifying varieties meeting specific agricultural and food requirements from the great biodiversity of sorghums to insure food security. Results show that some sorghums are rich sources of micronutrients (minerals and vitamins) and macronutrients (carbohydrates, proteins and fat). Sorghum has a resistant starch, which makes it interesting for obese and diabetic people. In addition, sorghum may be an alternative food for people who are allergic to gluten. Malts of some sorghum varieties display