Substrate affinity patterns showing by lipase and esterase activities from Aspergillus niger MYA 135

ROMERO CM, PERA LM, RODRIGUEZ, E. BAIGORÍ MD.

Pinamar.Buenos Aires

Congreso; X Congreso de PABMB, XLI Reunión Anual, Sociedad Argentina de Investigación en Bioquímica y Biología Molecular (SAIB); 2005

SAIB

SUBSTRATE AFFINITY PATTERNS SHOWING BY LIPASE AND ESTERASE ACTIVITIES FROM Aspergillus niger MYA 135 Romero CM, Pera LM, Rodriguez E, Baigorí MD. PROIMI-CONICET Av. Belgrano y Pje. Caseros, 4000 Tucumán. Tel: 4344888, E-mail: baigori@proimi.org.ar Introduction. Lipases (EC 3.1.1.3) are produced by various microorganisms either alone or together with esterases (EC 3.1.1.1). Lipases and esterases are biotechnologically important enzymes that catalyze the hydrolysis and synthesis of a wide array of esters. The explosive growth of biocatalytics process has led to an increasing demand for simple substrate affinity detection. In this work we analyzed substrate affinity patterns showing by extracellular lipase and esterase activities from Aspergillus niger MYA 135. Methods. Proteins were separated by native polyacrylamide gel electrophoresis. Lipases and esterases activities were detected using 1.3 mM of a-naphtyl derivates of acetate, propionate, butyrate, caproate, caprylate, caprate, laurate, myristate, palmitate or estereate as substrate. Released naphthol was coupled with 1mM Fast Blue to give a colored product. Reactions were carried out at 37°C in shaken plates containing 0.1M phosphate buffer (pH7). Both the substrate affinity and the number of band were analyzed. Results and conclusions. Supernatant of A. niger developed in mineral medium plus olive oil, exhibited the following substrate affinity patterns: 4 bands with acetate, propionate, butyrate or caproate; 2 bands with caprylate; and only 1 band with caprate, laurate, myristate, palmitate or estereate. This work were supported by grant PIP 2052/00 CONICET and CABBIO 2000 cod 012.