Cloning and sequencing the genomic encoding region of copper–zinc superoxide dismutase enzyme from several marine strains of the genus Debaryomyces (Lodder & Kreger‐van Rij)

Abstract

Copper-zinc superoxide dismutase (SODC) is a cytosolic enzyme which catalyses the dismutation of the superoxide radical. Due to its physiological importance, the encoding genes have been cloned from several species of higher eukaryotes. However, genes from moulds and yeast have not been studied extensively. In this paper, the encoding region of this gene (sod1) has been cloned from several strains of marine yeast belonging to the genus Debaryomyces (dvv sod1, dvy sod1 and dh sod1-61) through genomic DNA-PCR amplification. Fragments of 480-486 nucleotides were obtained, which contain information for products of 153-156 amino acids with calculated molecular masses of 15.8-16.6 kDa.The deduced amino acid sequence shows that D. vanrijiae enzymes present three additional amino acids not closely related to the active site conformation. In addition, in D. vanrijiae var. vanrijiae (strain 020), one histidine residue is apparently replaced by a proline; the incidence and function of other aromatic or heterocyclic amino acids is discussed. Homology and phylogenetic trees were constructed from amino-acid sequence multialignment analyses; the interrelationships among fungi are discussed.

Debaryomyces hansenii — an extremophilic yeast with biotechnological potential

“…SOD production using D. hansenii is very competitive compared to current methods, due to its ability to grow, as halotolerant yeast, either in sea‐ or freshwater‐formulated media, with a wide range of cultivation parameters137. Hernandez‐Saavedra and Romero‐Geraldo83 first described the cloning of the SOD gene of D. hansenii .…”

Oxidative stress sensitivity inDebaryomyces hansenii

“…For example, we have identified in D. hansenii genes similar to SOD 1 , SOD 2 , GPX 1 , TRX 1 and GSH 1 coding for putative superoxide dismutases (SOD), a peroxidase, a thioredoxin and a glutathione synthetase, respectively. However, information has been published only about the existence of SOD activity (Ramírez‐Orozco et al , 1998) and a putative SOD 1 gene (Hernández‐Saavedra & Romero‐Geraldo, 2001).…”

Cu,Zn superoxide dismutase in Rhodotorula and Udeniomyces spp. isolated from sea water: cloning and sequencing the encoding region

“…With this framework, we found, after induction by copper addition, that U. piniceus, but not Rhodotorula spp., responded as reported (Gralla et al, 1991;Ramírez-Orozco et al, 1999). To discard an inactive form of Cu,Zn-SOD in the Rhodotorula spp., we made a Western blot analysis, finding no recognition by polyclonal antibodies raised against D. hansenii Cu,Zn-SOD enzyme (Hernández-Saavedra and Ochoa, 1999), contrary to different recognition levels observed for other species (data not shown; Hernández-Saavedra and Romero, 2001). From these results, we assumed absence of the Cu,Zn-SOD enzyme in the studied Rhodotorula spp.…”

“…neoformans AAK31914-17/AF2448045-48 (Chaturvedi et al, 2001); Filobasidiella neoformans var. bacillispora AF248049-51/AAK31918-20 (unpublished data); and the marine yeasts Debaryomyces hansenii strain C11 O42724/ AFO-16383 (Hernández-Saavedra et al, 1998) (Hernández-Saavedra and Romero, 2001).…”