EVALUATION OF LIPASE ACTIVITY FROM Burkholderia lata LBBIO-BL02 in vitro USING COMPLEX SUBSTRATES AND SIMULATED INTESTINAL FLUID

• Grant number: 23/03334-9
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: June 01, 2023
• End date: December 31, 2024
• Field of knowledge: Biological Sciences - Biochemistry - Enzymology
• Principal Investigator: Valéria Marta Gomes de Lima
• Grantee: Pedro Castro Baltazar
• Host Institution: Faculdade de Ciências e Letras (FCL-ASSIS). Universidade Estadual Paulista (UNESP). Campus de Assis. Assis , SP, Brazil

Abstract

Lipases are promising tools for enzymatic replacement therapy. For this purpose, an ideal lipase replacement therapy should (i) maintain adequate levels of lipolytic activity at acidic and relatively neutral pH levels and in the presence of normal and low physiological concentrations of intra-intestinal bile salts, (ii) resist proteolytic digestion by pepsin and trypsin, and (iii) actively hydrolyze high diversity of dietary triacylglycerols (TAG). However, most microbial lipases have alkaline properties, instable or even inactivated in the digestive environment, due to acidity, physiological concentrations of bile salts, and digestive proteases (pepsin, trypsin and chymotrypsin) in intestine and stomach. This study explores the characterization of the lipase from Burkholderia lata LBBIO-BL02 (BLL) and its use in hydrolysis in digestive environments. This strain was chosen because, in previous studies by our research group, conditions for producing the enzyme in a low-cost culture medium (chicken meat processing industry waste as a source of carbon and inorganic nitrogen) and purification by an unprecedented and low-cost alternative method have already been established (protected by patent). Specially, All the found results clearly show that BLL is an enzyme as potent as the gastric and pancreatic lipases, while combining properties of these two lipases, acting on a large pH range covering pH variations in the human gastrointestinal tract, acting in a wide variety of substrates and resisting the proteases action and bile salts. Even so, the lipase activity of the LBBIO-BL02 strain needs to be studied in different reaction conditions that simulate human digestive conditions, and thus confirm the potential of this enzyme as a digestive supplement and justify more complex assays, using animal models, for example. This study seeks to evaluate the activity of the lipase produced by B. lata LBBIO-BL02 in a simulated intestinal environment, characterizing the hydrolytic capacity on varied substrates consistent with lipids consumed daily, such as vegetable oils and animal fats, isolated or associated with different foods. The objective of this evaluation is to analyze the efficiency of the lipase in different conditions aiming at a possible application in organisms with pancreatic dysfunctions, being an alternative to the current commercial lipases.