Characterization of a novel allele of glucose oxidase from a Korean wild type strain of Aspergillus niger.
We have identified a novel allele of the glucose oxidase (GO, EC1.1.3.4) gene (GO) from a Korean wild type strain of Aspergillus niger, ACMO4, with an increased GO activity in culture filtrate. Southern blot analyses of GO from ACMO4 (GO-ACMO4) revealed that the gene was present as a single copy in the genome of A. niger. However, its sequence differs from that of GO from A. niger ATCC 9029 (GO-ATCC9029). GO-ACMO4 appears to be a functional gene based on the fact that it is enzymatically active when heterologously expressed in yeast. Sequence comparisons of the coding region of GO-ACMO4 revealed 16 nucleotide changes that resulted in four amino acid substitutions; T432D, G517D, G530S, and Q542R. The GO proteins from both ATCC 9029 and ACMO4 were heterologously expressed, purified, and compared biochemically. The two enzymes showed no difference in their apparent Km value for glucose (30 mM), but the Vmax of GO-ACMO4 (515.6 unit/mg) was 10% higher than that of GO-ATCC 9029, resulting in a 10% higher specific activity.
Kim
M Y
MY
Institute for Molecular Biology and Genetics, Chonbuk National University, Chonju, Korea. dhkim@moak.chonbuk.ac.kr
Chung
H J
HJ
Hong
S Y
SY
Kim
H R
HR
Lee
J C
JC
Park
S M
SM
Lee
J H
JH
Yang
M S
MS
Kim
D H
DH
D000483:Alleles; D000595:Amino Acid Sequence; D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000032:analysis; D004789:Enzyme Activation / Q000235:genetics; D005949:Glucose Oxidase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D007723:Korea; D008828:Microbiological Techniques; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA
D004271:DNA, Fungal; D005949:Glucose Oxidase