Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.1.3.8 (phytase)
1,997 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Twenty-four Hansenula polymorpha transformants were passaged and stabilised in glucose medium and screened in glycerol medium for recombinant phytase in shaken test tubes. The cultivations were performed under either limited or non-limited oxygen supply. Maximum oxygen transfer capacities of test tubes were assessed by sulfite oxidation. Oxygen-limited glucose cultures resulted in a partially anaerobic metabolism and formation of 4.1 g ethanol l(-1), which was subsequently aerobically metabolised. Non-limited oxygen supply led to overflow metabolism and to accumulation of 2.1 g acetic acid l(-1), reducing the biomass yield. The use of glycerol in the screening main cultures prevented by-product formation irrespective of oxygen supply. Preculturing in glucose medium under non-limited oxygen supply resulted in a 20-h lag phase of the screening main culture. This lag phase was not observed when preculturing was performed under oxygen limitation. Phytase activity was on average 25% higher in cultures passaged, stabilised and screened under limited oxygen supply than in cultures under non-limited oxygen supply.
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PMID:Effect of oxygen supply on passaging, stabilising and screening of recombinant Hansenula polymorpha production strains in test tube cultures. 1461 84

A phytase from Candida krusei WZ-001 isolated from soil was purified to electrophoretic homogeneity by ion-exchange chromatography, hydrophobic interaction chromatography, and gel filtration. The phytase is composed of two different subunits with molecular masses of 116 kDa and 31 kDa on SDS-PAGE (or 120 kDa and 30 kDa on gel chromatography), with the larger subunit having a glycosylation rate of around 35%. The phytase has an optimum pH of 4.6, an optimum temperature of 40 degrees C and a pI value of 5.5. The phytase activity was stimulated by 2-mercapto-ethanol and dithiothreitol (DTT), and inhibited by Zn2+, Mg2+, iodoacetate, pI value of 5.5. The phytase activity was stimulated by 2-mercapto-ethanol ethanol and dithiothreitol (DTT), and inhibited by Zn2+, Mg2+, iodoacetate, p-chroloromercuribenzoate (pCMB) and phenylmethylsulfonyl fluoride (PMSF). The phytase displayed a broad substrate specificity and the K(m) for phytate was 0.03 mM. Phytate was sequentially hydrolyzed by the phytase. Furthermore, 1D and 2D NMR analyses and bioassay of myoinositol indicated that the end hydrolysis product of phytate was myoinositol 2-monophosphate.
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PMID:Purification and properties of a phytase from Candida krusei WZ-001. 1623 28

The metabolism of myo-inositol-2-(14)C, d-glucuronate-1-(14)C, d-glucuronate-6-(14)C, and l-methionine-methyl-(14)C to cell wall polysaccharides was investigated in excised root-tips of 3 day old Zea mays seedlings. From myo-inositol, about one-half of incorporated label was recovered in ethanol insoluble residues. Of this label, about 90% was solubilized by treatment, first with a preparation of pectinase-EDTA, then with dilute hydrochloric acid. The only labeled constituents in these hydrolyzates were d-galacturonic acid, d-glucuronic acid, 4-O-methyl-d-glucuronic acid, d-xylose, and l-arabinose, or larger oligosaccharide fragments containing these units. Medium external to excised root-tips grown under sterile conditions in myo-inositol-2-(14)C contained labeled polysaccharide.When label was supplied in the form of d-glucuronate, the pattern of labeled uronic acid and pentose units in cell wall polysaccharides resembled that obtained from labeled myo-inositol, indicating that both substances were metabolized along a common path during polysaccharide formation, and that methylation occurred at a step subsequent to uronic acid formation. When label was supplied in the form of l-methionine-methyl-(14)C, 4-O-methyl-d-glucuronic acid was the only labeled monosaccharide component that survived enzymatic or acid hydrolysis.Zea mays endosperm, a known source of phytin, developed maximal phytase activity after the third day of germination. Results obtained here suggest that myo-inositol released by hydrolysis of phytin represents the initial precursor of a normal, possibly predominant pathway for the formation of uronic acids in plants.
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PMID:Inositol Metabolism in Plants. V. Conversion of Myo-inositol to Uronic Acid and Pentose Units of Acidic Polysaccharides in Root-tips of Zea mays. 1665 71

When Wolffiella floridana, an aquatic angiosperm in the family, Lemnaceae, was grown in axenic culture under continuous light in E medium containing 1.0% sucrose and a micromolar amount of (14)C-labeled myo-inositol (MI), MI was taken up by the growing plants and converted to phytic acid. After 13 weeks in labeled medium during which time there was a 1000-fold increase in fresh weight, 30% of the (14)C was recovered in ethanol insoluble residue. Extraction of this residue with EDTA released 70% of the label into solution. Phytic acid, identified by paper electrophoresis, ion exchange chromatography, and hydrolysis with phytase, accounted for most of this radioactivity although some label was also found in pentaphosphate and lower phosphate esters of MI. Very little MI was converted to cell wall polysaccharides under the conditions used. Results of this study indicate that Wolffiella floridana is a convenient tissue for the study of phytic acid biosynthesis under laboratory conditions.Lemna gibba G3, grown under short day conditions in medium of the same composition as that used for W. floridana, also formed labeled phytic acid as well as other labeled lower phosphate esters of MI.
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PMID:Inositol Metabolism in Plants. VI. Conversion of Myo-Inositol to Phytic Acid in Wolffiella floridana. 1665 59

Distillers' dried grains with solubles (DDGS) and corn gluten feed (CGF) are major coproducts of ethanol production from corn dry grind and wet milling facilities, respectively. These coproducts contain important nutrients and high levels of phytates. The phytates in these products cannot be digested by nonruminant animals; consequently, large quantities of phytate phosphorus (P) are deposited into the soil with the animal wastes which potentially could cause P pollution in soil and underground water resources. To reduce phytates in DDGS and CGF, a phytase from Aspergillus niger, PhyA, was investigated regarding its capability to catalyze the hydrolysis of phytates in light steep water (LSW) and whole stillage (WS). LSW and WS streams are the intermediate streams in the production of CGF and DDGS, respectively, and contribute to most of the P in these streams. Enzyme loadings with activity of 0.1, 1, 2, and 4 FTU/g substrate and temperatures of 35 and 45 degrees C were investigated regarding their influences on the degree of hydrolysis. The analysis of the hydrolyzate suggested to a sequentially degradation of phytates to lower order myo-inositol phosphate isomers. Approximately 90% phytate P of LSW and 66% phytate P of WS were released, suggesting myo-inositol monophosphate as the end product. The maximum amount of released P was 4.52 +/- 0.03 mg/g LSW and 0.86 +/- 0.01 mg/g WS.
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PMID:Degradation of phytates in distillers' grains and corn gluten feed by Aspergillus niger phytase. 1881 3

Combination of physical and chemical mutagenesis was used to isolate hyper secretory strains of Aspergillus niger NCIM 563 for phytase production. Phytase activity of mutant N-1 and N-79 was about 17 and 47% higher than the parent strain. In shake flask the productivity of phytase in parent, mutant N-1 and N-79 was 6,181, 7,619 and 9,523 IU/L per day, respectively. Up scaling of the fermentation from shake flask to 3 and 14 L New Brunswick fermenter was studied. After optimizing various fermentation parameters like aeration, agitation and carbon source in fermentation medium the fermentation time to achieve highest phytase activity was reduced considerably from 14 days in shake flask to 8 days in 14 L fermenter. Highest phytase activity of 80 IU/ml was obtained in 1% rice bran-3.5% glucose containing medium with aeration 0.2 vvm and agitation 550 rpm at room temperature on 8th day of fermentation. Addition of either bavistin (0.1%), penicillin (0.1%), formalin (0.2%) and sodium chloride (10%) in fermented broth were effective in retaining 100% phytase activity for 8 days at room temperature while these reagents along with methanol (50%) and ethanol (50%) confer 100% stability of phytase activity at 4 degrees C till 20 days. Among various carriers used for application of phytase in feed, wheat bran and rice bran were superior to silica and calcium carbonate. Thermo stabilization studies indicate 100% protection of phytase activity in presence of 12% skim milk at 70 degrees C, which will be useful for its spray drying.
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PMID:Strain improvement and up scaling of phytase production by Aspergillus niger NCIM 563 under submerged fermentation conditions. 1908 44

For determining variation in mineral composition and phosphorus (P) profile among streams of dry-grind ethanol production, samples of ground corn, intermediate streams, and distillers dried grains with solubles (DDGS) were obtained from three commercial plants. Most attributes (dry matter concentrations) increased significantly from corn to cooked slurry but fermentation caused most significant increase in all attributes. During centrifugation, more minerals went into thin stillage than wet grains, making minerals most concentrated in the former. Mineral increase in DDGS over corn was about 3 fold, except for Na, S, Ca, and Fe. The first three had much higher fold of increase, presumably due to exogenous addition. During fermentation, phytate P and inorganic P had 2.54 and 10.37 fold of increase over corn, respectively, while relative to total P, % phytate P decreased and % inorganic P increased significantly. These observations suggest that phytate underwent some degradation, presumably due to activity of yeast phytase.
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PMID:Changes in mineral concentrations and phosphorus profile during dry-grind processing of corn into ethanol. 2105 25

In recent years, increasing demand for ethanol as a fuel additive and decreasing dependency on fossil fuels have resulted in a dramatic increase in the amount of grains used for ethanol production. Dry-grind is the major process, resulting in distillers dried grains with solubles (DDGS) as a major coproduct. Like fuel ethanol, DDGS has quickly become a global commodity. However, high compositional variation has been the main problem hindering its use as a feed ingredient. This review provides updated information on the chemical composition of distillers grains in terms of nutrient levels, changes during dry-grind processing, and causes for large variation. The occurrence in grain feedstock and the fate of mycotoxins during processing are also covered. During processing, starch is converted to glucose and then to ethanol and carbon dioxide. Most other components are relatively unchanged but concentrated in DDGS about 3-fold over the original feedstock. Mycotoxins, if present in the original feedstock, are also concentrated. Higher fold of increases in S, Na, and Ca are mostly due to exogenous addition during processing, whereas unusual changes in inorganic phosphorus (P) and phytate P indicate phytate hydrolysis by yeast phytase. Fermentation causes major changes, but other processing steps are also responsible. The causes for varying DDGS composition are multiple, including differences in feedstock species and composition, process methods and parameters, the amount of condensed solubles added to distiller wet grains, the effect of fermentation yeast, and analytical methodology. Most of them can be attributed to the complexity of the dry-grind process itself. It is hoped that information provided in this review will improve the understanding of the dry-grind process and aid in the development of strategies to control the compositional variation in DDGS.
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PMID:Chemical composition of distillers grains, a review. 2129 15

Both the high phosphorus (P) content and P bioavailability of the animal feed coproducts of the corn-ethanol industry could potentially contribute to increased manure and soil P levels and associated environmental issues (e.g., eutrophication). Therefore, a detailed modeling of total P mass flow to the coproducts (i.e., dry distillers grains with solubles, DDGS) was performed. Distribution of P between inorganic P and phytase-hydrolyzable P forms was quantified for selected coproducts (thin stillage, DDGS, modified DDGS [mDDGS]). The P mass balance indicated that although corn is the major P contributor to the coproducts (80.2%), a substantial portion (19.4%) comes from yeast addition. Of the two components constituting DDGS, wet distillers grains and condensed solubles, the latter contributes to only one-third of the mass but, importantly, yields 70.9% of P. The phytase enzyme used, , was very effective in hydrolyzing the nonorthophosphate P components of thin stillage, DDGS and mDDGS. Our results would help track P movement during various dry-grind processing steps and formulate strategies for phytase enzyme supplementation to various postfermentation coproducts from corn-ethanol plants.
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PMID:Phosphorus flow and characterization in dry-grind corn ethanol plants. 2309 62

The yeast Arxula adeninivorans is considered to be a promising producer of recombinant proteins. However, growth characteristics are poorly investigated and no industrial process has been established yet. Though of vital interest for strain screening and production processes, rationally defined culture conditions remain to be developed. A cultivation system was evolved based on targeted sampling and mathematical analysis of rationally designed small-scale cultivations in shake flasks. The oxygen and carbon dioxide transfer rates were analyzed as conclusive online parameters. Oxygen limitation extended cultivation and led to ethanol formation in cultures supplied with glucose. Cultures were inhibited at pH-values below 2.8. The phosphorus demand was determined as 1.55 g phosphorus per 100 g cell dry weight. Synthetic SYN6 medium with 20 g glucose l(-1) was optimized for cultivation in shake flasks by buffering at pH 6.4 with 140 mmol MES l(-1). Optimized SYN6 medium and operating conditions provided non-limited cultivations without by-product formation. A maximal specific growth rate of 0.32 h(-1) and short fermentations of 15 h were achieved. A pH optimum curve was derived from the oxygen transfer rates of differently buffered cultures, showing maximal growth between pH 2.8 and 6.5. Furthermore, it was shown that the applied medium and cultivation conditions were also suitable for non-limiting growth and product formation of a genetically modified A. adeninivorans strain expressing a heterologous phytase.
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PMID:Definition of culture conditions for Arxula adeninivorans, a rational basis for studying heterologous gene expression in this dimorphic yeast. 2466 17


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