Identification of new galactose oxidase genes in Fusarium spp.

Abstract

Galactose oxidase (GO) converts galactose to an aldehyde and has several biotechnological applications, including cancer diagnosis. It is mainly produced by Fusarium austroamericanum but is also produced by Fusarium acuminatum and by isolates of the Fusarium graminearum and Gibberella fujikuroi complexes. The F. austroamericanum GO gaoA gene has been cloned, but the GO genes from other secreting species have not been characterized. Problems associated with the F. austroamericanum GO such as high pI and low catalytic efficiency and thermostability, and the difficult purification process makes the search for homologous genes attractive. In this work, the GO genes from Fusarium verticillioides and Fusarium subglutinans, two species of the G. fujikuroi complex, were cloned, sequenced, and analyzed. New GO genes were found in databases and were used to construct a phylogenetic tree, which revealed the existence of three orthologous lineages of GO genes in Fusarium spp. In addition, RT-PCR analyses revealed that the new GO cloned gene may be endogenously expressed in F. subglutinans but not in F. verticillioides, in the used culture conditions.

Galactose Oxidase from Fusarium oxysporum - Expression in E. coli and P. pastoris and Biochemical Characterization

“…Furthermore, the gao gene has 96%, 62%, and 67% similarity with the gaoA (BK007071), gaoB (BK007067) and gaoC (BK007074) gene from F. oxysporum f. sp. lycopersici , respectively [8]. Hence the cloned gao gene from F. oxysporum belongs to the gaoA cluster of GalOx genes.…”

Isolation of genes related to abscisic acid production in Botrytis cinerea TB‐3‐H8 by cDNA‐AFLP

“…Trizol (Invitrogen, USA) was used to extract the total RNA of those samples following procedures as described in the manufacturer's instructions. Contaminated DNA was removed by digestion with DNase I (Takara) and verified by PCR analysis with the RNA as the template . mRNA was prepared from total RNA using EZ Spin Column mRNA Purification Kit (Bio Basic, Inc., China).…”

New PCR Assays for the Identification of Fusarium verticillioides, Fusarium subglutinans, and Other Species of the Gibberella fujikuroi Complex

“…Initially, the gaoB genes from F. verticillioides and F. subglutinans [24] (GenBank AN HM069186 and HM069185, respectively) and the gaoA gene from F. austroamericanum [20] (GenBank AN M86819) were aligned with the ClustalW program. Low similarity regions were chosen for primer design.…”

“…Galactose oxidase is a copper enzyme that catalyzes the oxidation of primary alcohols to aldehydes with the concomitant reduction of O 2 to H 2 O 2 [23]. Three ortholog lineages of this gene have been identified in Fusarium and the lineage gaoB has been cloned in our laboratory from F. verticillioides and from F. subglutinans [24]. The gaoB lineage lacks introns and is 2040 bp in length [24].…”

“…Three ortholog lineages of this gene have been identified in Fusarium and the lineage gaoB has been cloned in our laboratory from F. verticillioides and from F. subglutinans [24]. The gaoB lineage lacks introns and is 2040 bp in length [24]. …”