Morphological and molecular characterization of L-methioninase producing Aspergillus species

Abstract

Six species of L-methioninase producing Aspergillus species, isolated from Egyptian soil, were selected for comprehensive morphotypic and molecular characterization. Based on morphological and physiological features, these isolates were identified as Aspergillus flavipes, Aspergillus carneus, Aspergillus flavus, Aspergillus tamari, Aspergillus oryzae, and Aspergillus parasiticus. Regarding to the maximum enzyme productivity by A. flavipes, it was selected as authentic strain for ribosomal ribonucleic acid (rRNA) primer design. Using primer combinations for 18S rRNA and internal transcribed spacers (ITS)1 amplification, these isolates gave the same polymerase chain reaction (PCR) amplicon size, revealing the relative molecular identity. Moreover, using ITS2 primers, among the six isolates, Aspergillus flavipes EK and A. carneus displayed PCR products on agarose gel, approving the actual morphological and biochemical similarities of these two isolates, A. flavipes group. By sequencing of ITS1-5.8S-ITS2 region, blasting and alignment from the data base, A. flavipes EK showed a typical identity to gene bank deposited A. flavipes isolates. The rRNA sequence of A. flavipes EK was deposited to genbank under accession number JF831014.

Key words: Aspergillus, morphological descriptions, 18 S rRNA, internal transcribed spacers (ITS) regions.