Acuna, J.

Greiner, Ralf

Jorquera, M.

Menezes-Blackburn, D.

Sajidan, A.

English

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BIOAVAILABILITY 2014UNDERSTANDING THE BIOAVAILABILITY OF MICRONUTRIENTS AND BIOACTIVE COMPOUNDS SO AS TO IMPROVE PUBLIC HEALTHMAY 12-14, 2014 - HOTEL RECANTO - IGUASSU FALLS - BRAZIL32* Names, titles and abstracts are reproduced exactly as it was entered by the authors, therefore, replications and errors are their responsibility.EP-40 - INFLUENCE OF DIET, MENSTRUATION, AND GENETIC FACTORS ON IRON STATUS IN SPANISH WOMEN OF CHILDBEARING AGE Authors: RUTH BLANCO-ROJO; LAURA TOXQUI; ANA M LÓPEZ-PARRA; CARLOS BAEZA-RICHER; ANA M PÉREZ-GRANADOS; EDUARDO ARROYO-PARDO; M PILAR VAQUERO Institution: ICTAN-CSIC Country: SPAIN Session: TV4 - Date: 12/05/2014 - Room:  FOYER - Time: 15:24-15:29:00 BACKGROUNDPrevalence of  iron deficiency in women at fertile age is high, and may have substantial health and economic costs.OBJECTIVESThe aim of  this study was to investigate the combined influence of  diet, menstruation and genetic factors on iron status in Spanish menstruating women.METHODSA group of  142 healthy menstruating women participated in the study. Dietary intake was assessed by a 72-h detailed dietary report and menstrual blood loss by a questionnaire designed by the research group, to determine a Menstrual Blood Loss Coefficient (MBLC). Five selected SNPs (1,2) were genotyped by minisequencing assay. Iron biomarkers (haemoglobin, ferritin, soluble transferrin receptor, transferrin, serum iron and red blood cell distribution width) were determined and a cluster analysis using the k-means algorithm was performed. A categorical regression with optimal scaling was performed to identify the factors associated with cluster belonging.RESULTSThree clusters were identified: women with poor iron status (cluster 1, n=26); women with mild iron deficiency (cluster 2, n=59) and women with sufficient iron status (cluster 3, n=57). Three independent factors were included in the model that better explained cluster belonging (R2=0.142 and p<0.001). The most influential factor was red meat consumption (p=0.001 and importance coefficient=0.677); higher red meat intake predicts better iron status. MBLC was inversely related to clusters belonging, therefore, the higher MBLC the lower iron status (p=0.085 and importance coefficient=0.170). Thirdly, presence of  the minor allele of  SNP rs1800562 (mutation C282Y) was related to higher iron status (p=0.095 and importance coefficient=0.153).CONCLUSIONSWe found, by a cluster approach method, that three factors: red meat consumption, menstrual blood loss and the C282Y mutation of  the HFE gene, play a significant role in the iron status of  menstruating women. These findings could help choose adequate strategies to improve iron status and prevent the development of  iron deficiency anaemia in population at risk.Supported by Project AGL2009 11437. Blanco-Rojo R. and Toxqui L. were supported by JAE-predoc grants from European Social Fund.1-Baeza-Richer C, Blanco-Rojo R, Lopez-Parra AM et al Dis Markers 2013 34:121-1292-Blanco-Rojo R, Baeza-Richer C, Lopez-Parra AM et al Nutr Metab (Lond) 2011 8:69EP-39 - SCREENING FOR PHYTASES USING A METAGENOMIC APPROACH Authors: RALF GREINER; ADJID SAJIDAN; DANIEL BLACKBURN-MENEZ; JACQUELINNE ACUÑA; MILKO JORQUERA Institution: MAX RUBNER-INSTITUT Country: GERMANY Session: TV4 - Date: 12/05/2014 - Room:  FOYER - Time: 15:18-15:29:00 BACKGROUNDPhytase [myo-inositol(1,2,3,4,5,6)hexakisphosphate phosphohydrolase], a phytate-specific phosphatase, is already used as a supplement in diets for simple-stomached animals to improve phosphate utilisation from phytate [myo-inositol(1,2,3,4,5,6)hexakisphosphate], the major storage form of  phosphate in plant seeds and grains. In recent years, this class of  enzymes has also found increasing interest to be used in food processing and manufacturing, particularly because reduction in dietary phytate is seen as a possibility to combat zinc and iron deficiencies by enhancing their bioavailability in plant-based foods. Several phytase have been purified and characterized in respect to their biotechnological application and some are commercially available in the meantime. However, especially for food applications a higher temperature stability of  the enzyme would be of  advantage.OBJECTIVESTherefore, screening soil microorganisms for the production of  a thermostable phytase by a metagenomic approach was the aim of  the studies.METHODSExpression of  environmental DNA in bacteria (metagenomic screening) has proven highly useful for identification of  novel enzymes or enzymes with improved properties. Genomic DNA was obtained from agricultural soil using a method based on direct lysis and purified by gel filtration. The recovered DNA was of  high molecular mass and sufficiently pure for subsequent cloning. Following enzymatic digestion, the DNA was cloned into the expression vector pBluescript SK+. The resulting expression plasmid library comprised a total of  25000 clones.RESULTSThe library was screened for phosphatase activity using X-phosphate as a substrate. 27 clones with improved phosphatase activities were identified. Using sodium phytate as a test substrate revealed that 15 clones showed also an improved phytase activity. Compared to many phytases described in the scientific literature so far, 4 of  the phytases identified by metagenomic screening exhibited an improved temperature stability. Residual activity after exposure to 80°C for 10 min was determined to be higher than 70%.CONCLUSIONSIn summary, is was shown that direct cloning of  environmental DNA is a suitable strategy to utilize the metabolic diversity in a given habitat for biotechnical innovations.back to index

Screening for phytases using a metagenomic approach

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Screening for phytases using a metagenomic approach

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