TY - JOUR
T1 - Effect of deglycosylation on the properties of thermophilic invertase purified from the yeast Candida guilliermondii MpIIIa
AU - Plascencia-Espinosa, Miguel
AU - Santiago-Hernández, Alejandro
AU - Pavón-Orozco, Patricia
AU - Vallejo-Becerra, Vanessa
AU - Trejo-Estrada, Sergio
AU - Sosa-Peinado, Alejandro
AU - Benitez-Cardoza, Claudia G.
AU - Hidalgo-Lara, María Eugenia
N1 - Funding Information:
This work was supported by CINVESTAV-IPN and by doctoral fellowship (M.P-E) from the Consejo Nacional de Ciencia y Tecnología, México .
PY - 2014/9
Y1 - 2014/9
N2 - Invertase from Candida guilliermondii MpIIIa was purified and biochemically characterized. The purified enzyme (INV3a-N) is a glycoprotein with a carbohydrate composition comprising nearly 74% of its total molecular weight (MW) and specific activity of 82,027 U/mg of protein. The enzyme displayed optimal activity at pH 5.0 and 65 °C. The Km and Vmax values for INV3a-N were 0.104 mM and 10.9 μmol/min/mg of protein, respectively, using sucrose as the substrate. The enzyme retained 50% and 20% of its maximal activity after 168 h and 30 days, respectively, at 50 °C. INV3a-N was fully active at sucrose concentrations of 400 mM and the activity of the enzyme dropped slowly at higher substrate concentration. Interestingly, the deglycosylated form of INV3a-N (INV3a-D) displayed 76-92% lower thermostability than that of INV3a-N at all temperatures assayed (50-70 °C), and was inhibited at sucrose concentrations of 200 mM. Findings here indicate glycosylation plays an important role, not only in the thermostability of INV3a-N, but also in the inhibition of the enzyme by sucrose. Since the enzyme is active at high sucrose concentrations, INV3a-N may be considered a suitable candidate for numerous industrial applications involving substrates with high sugar content or for improvement of ethanol production from cane molasses.
AB - Invertase from Candida guilliermondii MpIIIa was purified and biochemically characterized. The purified enzyme (INV3a-N) is a glycoprotein with a carbohydrate composition comprising nearly 74% of its total molecular weight (MW) and specific activity of 82,027 U/mg of protein. The enzyme displayed optimal activity at pH 5.0 and 65 °C. The Km and Vmax values for INV3a-N were 0.104 mM and 10.9 μmol/min/mg of protein, respectively, using sucrose as the substrate. The enzyme retained 50% and 20% of its maximal activity after 168 h and 30 days, respectively, at 50 °C. INV3a-N was fully active at sucrose concentrations of 400 mM and the activity of the enzyme dropped slowly at higher substrate concentration. Interestingly, the deglycosylated form of INV3a-N (INV3a-D) displayed 76-92% lower thermostability than that of INV3a-N at all temperatures assayed (50-70 °C), and was inhibited at sucrose concentrations of 200 mM. Findings here indicate glycosylation plays an important role, not only in the thermostability of INV3a-N, but also in the inhibition of the enzyme by sucrose. Since the enzyme is active at high sucrose concentrations, INV3a-N may be considered a suitable candidate for numerous industrial applications involving substrates with high sugar content or for improvement of ethanol production from cane molasses.
KW - Candida guilliermondii
KW - Glycosylation
KW - Invertase
KW - Substrate inhibition
KW - Thermostability
UR - http://www.scopus.com/inward/record.url?scp=84906783611&partnerID=8YFLogxK
U2 - 10.1016/j.procbio.2014.05.022
DO - 10.1016/j.procbio.2014.05.022
M3 - Artículo
SN - 1359-5113
VL - 49
SP - 1480
EP - 1487
JO - Process Biochemistry
JF - Process Biochemistry
IS - 9
ER -