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Bruno Tilocca, Maren Witzig, Markus Rodehutscord, Jana Seifert
The chicken gastrointestinal tract (GIT) harbours a complex microbial community, involved in several physiological processes such as host immunomodulation and feed digestion. For the first time, the present study analysed dietary effects on the protein inventory of the microbiome in crop and ceca of broilers. We performed quantitative label-free metaproteomics by using 1-D-gel electrophoresis coupled with LC-MS/MS to identify the structural and functional changes triggered by diets supplied with varying amount of mineral phosphorous (P) and microbial phytase (MP)...
2016: PloS One
M Brnić, R F Hurrell, L T Songré-Ouattara, B Diawara, A Kalmogho-Zan, C Tapsoba, C Zeder, R Wegmüller
BACKGROUND/OBJECTIVES: Fortifying cereal staples with zinc is a strategy for increasing zinc intake in young children in developing countries. However, phytic acid (PA) naturally present in cereals strongly decreases zinc absorption. A stable-isotope zinc absorption study was conducted in young children to investigate the ability of the PA-degrading enzyme phytase to improve zinc absorption, when added to a cereal porridge immediately before consumption. SUBJECTS/METHODS: Fractional absorption of zinc (FAZ) was estimated in 35 young healthy Burkinabe children using the double-isotopic tracer ratio method with (67)Zn as oral tracer and (70)Zn as intravenous tracer, in a crossover design...
October 19, 2016: European Journal of Clinical Nutrition
Jianhua Yang, Anna Joëlle Ruff, Stefanie Nicole Hamer, Feng Cheng, Ulrich Schwaneberg
Escherichia coli is a common host for recombinant protein production in which production titers are highly dependent on the employed expression system. Promoters are thereby a key element to control gene expression levels. In this study, a novel PLICable promoter toolbox was developed which enables in a single cloning step and after a screening experiment to identify out of ten IPTG-inducible promoters (T7, A3, lpp, tac, pac, Sp6, lac, npr, trc and syn) the most suitable one for high level protein production...
October 18, 2016: Biotechnology Journal
Ivo M Rodrigues, M Graça V S Carvalho, Jorge M S Rocha
BACKGROUND: Rapeseed meal is a good source of high-quality vegetal protein but contains antinutritional compounds which limit its use for human and animal feed. The aim of this study was to develop a methodology to enhance alkaline protein extraction of rapeseed meal and to produce protein rich products with low levels of phytic acid. Different phytase dosages and operating conditions were used for rapeseed meal pretreatment followed by alkaline extraction at different temperatures, time, pH and solid/liquid ratios (S/L)...
October 14, 2016: Journal of the Science of Food and Agriculture
Patricia M Valdespino-Castillo, Rocío J Alcántara-Hernández, Martín Merino-Ibarra, Javier Alcocer, Miroslav Macek, Octavio A Moreno-Guillén, Luisa I Falcón
Microbes can modulate ecosystem function since they harbor a vast genetic potential for biogeochemical cycling. The spatial and temporal dynamics of this genetic diversity should be acknowledged to establish a link between ecosystem function and community structure. In this study, we analyzed the genetic diversity of bacterial phosphorus utilization genes in two microbial assemblages, microbialites and bacterioplankton of Lake Alchichica, a semiclosed (i.e., endorheic) system with marked seasonality that varies in nutrient conditions, temperature, dissolved oxygen, and water column stability...
October 10, 2016: Microbial Ecology
Jorge A M Delezuk, Adriana Pavinatto, Marli L Moraes, Flávio M Shimizu, Valquíria C Rodrigues, Sérgio P Campana-Filho, Sidney J L Ribeiro, Osvaldo N Oliveira
In this paper, we show that chitosan may induce conformation changes in silk fibroin (SF) in layer-by-layer (LbL) films, which were used as matrix for immobilization of the enzyme phytase to detect phytic acid. Three chitosan (CH) samples possessing distinct molecular weights were used to build CH/SF LbL films, and a larger change in conformation from random coils to β-sheets for SF was observed for high molecular weight chitosan (CHH). The CHH/SF LbL films deposited onto interdigitated gold electrodes were coated with a layer of phytase, with which phytic acid could be detected down to 10(-9)M using impedance spectroscopy as the principle of detection and treating the data with a multidimensional projection technique...
January 2, 2017: Carbohydrate Polymers
V G Schramm, J F Durau, L N E Barrilli, J O B Sorbara, A J Cowieson, A P Félix, A Maiorka
An experiment was carried out to evaluate the digestibility and metabolizability of corn and a corn/soy-based diet with the inclusion or not of xylanase and/or phytase in broilers. In the trial, 1,120 broiler chicks were distributed according to a completely randomized experimental design, consisting of 16 treatments, with 10 replicates of 7 birds each. Treatments were evaluated following a factorial arrangement (4 × 2 × 2), with 4 xylanase levels (zero, 50, 100, or 150 fungal β-xylanase units/kg; FXU), 2 phytase levels (zero or 1,000 phytase units/kg; FTU), and 2 diets (corn/soy or pure corn)...
October 2, 2016: Poultry Science
Anay Kumar Maurya, Deepak Parashar, T Satyanarayana
Thermophilc mold Sporotrichum thermophile secretes an acidstable and thermostable phytase, which finds application as a food and feed additive because of its adequate thermostability, acid stability, protease insensitivity and broad substrate spectrum. Low extracellular phytase production by the mold is a major bottleneck for its application on a commercial scale. We have successfully overcome this problem by constitutive secretary expression of codon optimized rStPhy under glyceraldehyde phosphate dehydrogenase (GAP) promoter in Pichia pastoris...
September 30, 2016: International Journal of Biological Macromolecules
M S Oliveira, H H Stein
Three experiments were conducted to determine standardized ileal digestibility (SID) of CP and AA, DE and ME, and apparent total tract digestibility (ATTD) and standardized total tract digestibility (STTD) of P in a new source of soy protein concentrate (SPC) and in soybean meal (SBM). In Exp. 1, 9 barrows (initial BW: 13.08 ± 1.98 kg) were prepared with a T-cannula in the distal ileum and allotted to a triplicated 3 × 3 Latin square design with 3 diets and 3 periods. A nitrogen-free diet and 2 diets that contained corn starch and SPC or SBM as the sole source of CP and AA were formulated...
August 2016: Journal of Animal Science
Nabeela Abid, Asia Khatoon, Asma Maqbool, Muhammad Irfan, Aftab Bashir, Irsa Asif, Muhammad Shahid, Asma Saeed, Henrik Brinch-Pedersen, Kauser A Malik
Phytate is a major constituent of wheat seeds and chelates metal ions, thus reducing their bioavailability and so the nutritional value of grains. Transgenic plants expressing heterologous phytase are expected to enhance degradation of phytic acid stored in seeds and are proposed to increase the in vitro bioavailability of mineral nutrients. Wheat transgenic plants expressing Aspergillus japonicus phytase gene (phyA) in wheat endosperm were developed till T3 generation. The transgenic lines exhibited 18-99 % increase in phytase activity and 12-76 % reduction of phytic acid content in seeds...
September 29, 2016: Transgenic Research
N P Balaban, A D Suleimanova, L R Valeeva, E V Shakirov, M R Sharipova
β-Propeller phytases of Bacillus are unique highly conservative and highly specific enzymes capable of cleaving insoluble phytate compounds. In this review, we analyzed data on the properties of these enzymes, their differences from other phytases, and their unique spatial structures and substrate specificities. We considered influences of different factors on the catalytic activity and thermostability of these enzymes. There are few data on the hydrolysis mechanism of these enzymes, which makes it difficult to analyze their mechanism of action and their final products...
August 2016: Biochemistry. Biokhimii︠a︡
Z Z Ren, D E Bütz, A N Wahhab, A J Piepenburg, M E Cook
Phytase hydrolyzes phytate rendering phosphorus available for intestinal absorption, while systemic neutralization of fibroblast growth factor 23 (FGF-23), using anti-FGF-23 antibody, has been shown to increase phosphate retention. Hence, neutralization of FGF-23 should be additive with phytase in reducing dietary non-phytate phosphorus (nPP) needs in chickens fed plant-based diets rich in phytic acid. This study was designed to test the additive effects of maternally derived anti-FGF-23 antibody and dietary phytase on the performance of chicks fed a low nPP diet from one to 14 d...
September 24, 2016: Poultry Science
Lan Shen, Xiao-Qin Wu, Qing-Wei Zeng, Hong-Bin Liu
Phytate-mineralizing rhizobacteria (PMR) play an important role in providing phosphorus for the sustainable plant growth. It is important to investigate the ability of PMR to produce phytase under different phosphate levels for its application. The effects of different concentrations of soluble phosphate on the ability of phytate mineralization of Pseudomonas fluorescens JZ-DZ1, a phytate-mineralizing rhizobacterium, were investigated in both solid and liquid media. The results on solid media showed that halo zone width gradually reduced with concentrations of soluble phosphate increasing from 0...
December 2016: Current Microbiology
Inger Baeksted Holme, Giuseppe Dionisio, Claus Krogh Madsen, Henrik Brinch-Pedersen
The phytase purple acid phosphatase (HvPAPhy_a) expressed during barley seed development was evaluated as transgene for overexpression in barley. The phytase was expressed constitutively driven by the cauliflower mosaic virus 35S-promoter and the phytase activity was measured in the mature grains, the green leaves and in the dry mature vegetative plant parts left after harvest of the grains. The T2-generation of HvPAPhy_a transformed barley showed phytase activity increases up to 19 fold (29000 phytase units (FTU) per kg in mature grains...
September 16, 2016: Plant Biotechnology Journal
Alexandre Maller, Thays Cristina Oliveira de Quadros, Otto M Junqueira, Alfredo Lora Graña, Ana Paula de Lima Montaldi, Ricardo Fernandes Alarcon, João Atílio Jorge, Maria de Lourdes T M Polizeli
Phytases are enzymes that hydrolyze the ester linkage of phytic acid, releasing inositol and inorganic phosphate. The phytic acid (phytate) is a major form of phosphorus in plant foods. Knowing that diet for animal of production has the cereal base (corn and soybean), primarily, broilers need for an alternative to use of the phosphate present in these ingredients, since it does not naturally produce the enzyme phytase, which makes it available. The aims of this work was studding the safe supplementation of Aspergillus japonicus var...
2016: SpringerPlus
Vikas Sharma, Ajit Kumar, G Archana, G Naresh Kumar
The Escherichia coli phytase gene appA encoding enzyme AppA was cloned in a broad host range plasmid pBBR1MCS2 (lac promoter), termed pVA1, and transformed into the Ensifer meliloti 1020. Transformation of pVA1 in Ensifer meliloti {E. m (pVA1)} increased its phosphatase and phytase activity by ∼9- and ∼50-fold, respectively, compared to the transformants containing empty plasmid as control {E. m (pBBR1MCS2)}. The western blot experiments using rabbit anti-AppA antibody showed that AppA is translocated into the periplasm of the host after its expression...
October 2016: Die Naturwissenschaften
A M Amerah, L F Romero, A Awati, V Ravindran
Two trials (a 42-d performance and a 21-d cohort digestibility) were conducted to evaluate the performance and nutrient digestibility of broilers fed corn diets supplemented with exogenous xylanase, amylase, and protease as single or combined activities. A nutritionally adequate, positive control (PC) diet was formulated. The negative control (NC) diet was formulated to be lower in metabolizable energy (∼86 kcal/kg diet) and digestible amino acids (1 to 2%) compared to PC. The other 4 treatments were based on the NC and they were either supplemented with xylanase (X), amylase (A), protease (P), or a combination of X, A, and P (XAP; to provide 2,000 U of X, 200 U of A, and 4,000 U of P/kg diet)...
September 2, 2016: Poultry Science
Mannanthendil Kumaran Asha, Debnath Debraj, Shekhar Dethe, Anirban Bhaskar, Nithyanantham Muruganantham, Mundkinajeddu Deepak
Flavonoid-rich extract prepared from Glycyrrhiza glabra has been found to be beneficial in patients with functional dyspepsia and was reported to possess some gut health-promoting properties such as antioxidant, anti-inflammatory and anti-Helicobacter pylori activities. In the present study, the flavonoid-rich extract of Glycyrrhiza glabra was evaluated for its compatibility with probiotic strains (Lactobacillus casei, Lactobacillus fermentum, Lactobacillus plantarum, and Streptococcus thermophilus), commercial probiotic drinks, and digestive enzymes (pancreatic α-amylase, α-glucosidase, phytase, xylanase, and pancreatic lipase)...
September 2, 2016: Journal of Dietary Supplements
C Schoene, S P Bennett, M Howarth
Enzymes often have marginal stability, with unfolding typically leading to irreversible denaturation. This sensitivity is a major barrier, both for de novo enzyme development and for expanding enzyme impact beyond the laboratory. Seeking an approach to enhance resilience to denaturation that could be applied to a range of different enzymes, we developed SpyRing cyclization. SpyRings contain genetically encoded SpyTag (13 amino acids) on the N-terminus and SpyCatcher (12kDa) on the C-terminus of the enzyme, so that the Spy partners spontaneously react together through an irreversible isopeptide bond...
2016: Methods in Enzymology
Yanhua Tan, Xiaoping Yi, Limin Wang, Cunzhi Peng, Yong Sun, Dan Wang, Jiaming Zhang, Anping Guo, Xuchu Wang
To investigate unintended effects in genetically modified crops (GMCs), a comparative proteomic analysis between the leaves of the phytase-transgenic maize and the non-transgenic plants was performed using two-dimensional gel electrophoresis and mass spectrometry. A total of 57 differentially expressed proteins (DEPs) were successfully identified, which represents 44 unique proteins. Functional classification of the identified proteins showed that these DEPs were predominantly involved in carbohydrate transport and metabolism category, followed by post-translational modification...
2016: Frontiers in Plant Science
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