TY - JOUR
T1 - Production and characterization of extracellular α-amylase produced by wickerhamia sp. X-Fep
AU - Hernández-Montañez, Zahuiti
AU - Juárez-Montiel, Margarita
AU - Velázquez-Ávila, Martha
AU - Cristiani-Urbina, Eliseo
AU - Hernández-Rodríguez, César
AU - Villa-Tanaca, Lourdes
AU - Chávez-Camarillo, Griselda
PY - 2012/8
Y1 - 2012/8
N2 - A yeast isolate able to produce high levels of extracellular a-amylase was selected from a collection of 385 yeasts and identified as Wickerhamia sp. by the sequence of the D1/D2 domain of the 26 S rDNA gene. Part of the nucleotide sequence of the amy1-Wgene was cloned, and a sequence of 191 amino acids deduced from this gene was analyzed. The peptide contains three characteristic well-conserved regions in the active sites of a-amylases (EC 3.2.1.1). The enzyme was purified and in situ activity showed only one band with amylolytic activity. The molecular mass of the a-amylase was estimated at 54 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Enzymatic activity on soluble starch as substrate was optimal at pH 5-6 and 50 °C. This thermostable enzyme was inhibited by EDTA-Na2 and 1,10-phenanthroline; the activity of the dialyzed enzyme was reactivated with Ca 2+ + and Mg 2+ + cations, which indicates that the a-amylase is ametalloenzyme. a-Amylase productionwas induced by starch andmaltose and repressed by glucose. The high yield and productivity found in this work makes this Wickerhamia sp. strain a promising candidate for the biotechnological production of α-amylase.
AB - A yeast isolate able to produce high levels of extracellular a-amylase was selected from a collection of 385 yeasts and identified as Wickerhamia sp. by the sequence of the D1/D2 domain of the 26 S rDNA gene. Part of the nucleotide sequence of the amy1-Wgene was cloned, and a sequence of 191 amino acids deduced from this gene was analyzed. The peptide contains three characteristic well-conserved regions in the active sites of a-amylases (EC 3.2.1.1). The enzyme was purified and in situ activity showed only one band with amylolytic activity. The molecular mass of the a-amylase was estimated at 54 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Enzymatic activity on soluble starch as substrate was optimal at pH 5-6 and 50 °C. This thermostable enzyme was inhibited by EDTA-Na2 and 1,10-phenanthroline; the activity of the dialyzed enzyme was reactivated with Ca 2+ + and Mg 2+ + cations, which indicates that the a-amylase is ametalloenzyme. a-Amylase productionwas induced by starch andmaltose and repressed by glucose. The high yield and productivity found in this work makes this Wickerhamia sp. strain a promising candidate for the biotechnological production of α-amylase.
KW - Amylase
KW - Enzyme production
KW - Metalloenzyme
KW - Starch
KW - Wickerhamia sp.
KW - Yeast
UR - http://www.scopus.com/inward/record.url?scp=84866431329&partnerID=8YFLogxK
U2 - 10.1007/s12010-012-9736-2
DO - 10.1007/s12010-012-9736-2
M3 - Artículo
SN - 0273-2289
VL - 167
SP - 2117
EP - 2129
JO - Applied Biochemistry and Biotechnology
JF - Applied Biochemistry and Biotechnology
IS - 7
ER -