Intestinal alkaline phosphatase activity of LPS-desfosforilase from broilers

• Grant number: 13/14773-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: October 01, 2013
• End date: November 30, 2016
• Field of knowledge: Biological Sciences - Biochemistry - Enzymology
• Principal Investigator: João Martins Pizauro Junior
• Grantee: Gabriella Cavazzini Pavarina
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil

Abstract

Phosphatases are phosphomonohydrolases widely distributed among living organisms, being found in microorganisms, animals and plants. They are divided into three major groups according to the pH of activity, in acid and alkaline and protein phosphatase. Alkaline phosphatase is normally membrane-associated and distributed in almost all tissues, but its highest activity is found in the intestinal epithelium, tubular renal cells, osteoblasts, liver and placenta. The animals' gastrointestinal tract is colonized by primordial microorganisms that along with a matrix of cells, a complete immune system and colonizer microorganisms, protect the intestinal mucosa. There are also indigenous bacteria present in the microbiota that are gram-negative, presenting an outer membrane composed by molecules of lipopolysaccharide (LPS). These indigenous bacteria are present in the intestine and are responsible for the production of high levels of LPS, which have the potential to trigger inflammatory and bacterial infections. Studies suggest that there is a function of the intestinal alkaline phosphatase related to the destruction of lipopolysaccharides toxicity. The destruction of the toxicity responsible for the virulence of intestinal bacteria is an important object for studies, suggesting the possibility of a new physiological role of the enzyme by the dephosphorylation of glucophospholipid, participating of mechanisms that prevent the pathogenicity of gram-negative bacteria. This function is supported also by the fact that LPS, in its dephosphorylated form, does not present toxicity. Samples will be obtained by approximately 20 centimeters from the jejunum (portion corresponding to the small intestine after the duodenum and anterior to Meckel's diverticulum). Then they will be opened longitudinally, clamped by the extremities on the cardboard and the mucosa will be washed with buffered saline (pH 7.4) at 4°C. Then the mucosa will be scraped with the aid of a sterile metal blade and the obtained material will be wrapped in aluminum paper, frozen in liquid nitrogen and stored in a freezer at -70°C for subsequent laboratory processing. The project aims to analyze the activity of intestinal alkaline LPS-dephosphorilase in broilers (Cobb-500).