TY - JOUR
T1 - Response surface methodology (RSM) for analysing culture conditions of acidocella facilis strain USBA-GBX-505 and partial purification and biochemical characterization of lipase 505 LIP
AU - Bernal, Luisa Fernanda
AU - López, Gina
AU - Ruiz, Mónica
AU - Vera-Bravo, Ricardo
AU - Reyes, Alejandro
AU - Baena, Sandra
PY - 2017
Y1 - 2017
N2 - Using Response Surface Methodology (RSM) we evaluated the culture conditions (nitrogen source, carbon source, pH and agitation rate) that increase the biomass of Aádocella facilis strain USBA-GBX-505 and therefore enhance the production of its lipolytic enzyme, 505 LIP. RSM results revealed that yeast extract and agitation were key culture factors that increased the growth-associated lipolytic activity by 4.5-fold (from 0.13 U.mg-1 to 0.6 U.mg-1). The 505 LIP lipase was partially purified using size-exclusion chromatography and ion-exchange chromatography. Its molecular weight was >77 kDa. The enzyme shows its optimum catalytic activity at 55 °C and pH 7.5. EDTA, PMSF, 1-butanol and DMSO inhibited enzymatic activity, whereas Tween 20, acetone, glycerol and methanol increased it. Metallic ions are not required for the activity of 505 LIP, and even have an inhibitory effect on the enzyme. This study shows the potential use of A. facilis strain USBA-GBX-505 for the production of a newly identified lipolytic enzyme, 505 LIP, which is stable at moderate temperatures and in the presence of organic solvents. These are important characteristics for the synthesis of many useful products.
AB - Using Response Surface Methodology (RSM) we evaluated the culture conditions (nitrogen source, carbon source, pH and agitation rate) that increase the biomass of Aádocella facilis strain USBA-GBX-505 and therefore enhance the production of its lipolytic enzyme, 505 LIP. RSM results revealed that yeast extract and agitation were key culture factors that increased the growth-associated lipolytic activity by 4.5-fold (from 0.13 U.mg-1 to 0.6 U.mg-1). The 505 LIP lipase was partially purified using size-exclusion chromatography and ion-exchange chromatography. Its molecular weight was >77 kDa. The enzyme shows its optimum catalytic activity at 55 °C and pH 7.5. EDTA, PMSF, 1-butanol and DMSO inhibited enzymatic activity, whereas Tween 20, acetone, glycerol and methanol increased it. Metallic ions are not required for the activity of 505 LIP, and even have an inhibitory effect on the enzyme. This study shows the potential use of A. facilis strain USBA-GBX-505 for the production of a newly identified lipolytic enzyme, 505 LIP, which is stable at moderate temperatures and in the presence of organic solvents. These are important characteristics for the synthesis of many useful products.
KW - Acidocella facilis
KW - Acidophiles
KW - Central composite design
KW - Lipases
KW - Plackett-Burman design
KW - Stability in solvents
UR - http://www.scopus.com/inward/record.url?scp=85015096288&partnerID=8YFLogxK
U2 - 10.11144/Javenana.SC22-1.rsmr
DO - 10.11144/Javenana.SC22-1.rsmr
M3 - Article
AN - SCOPUS:85015096288
SN - 0122-7483
VL - 22
SP - 45
EP - 70
JO - Universitas Scientiarum
JF - Universitas Scientiarum
IS - 1
ER -