l-Methioninase from some Streptomyces isolates I: Isolation identification of best producers and some properties of the crude enzyme produced

摘要

Among 60 isolates of Streptomyces tested; only 40 isolates were capable to utilize l-methionine as the only source of nitrogen in medium. In addition, 24 of these isolates could grow in medium amended with l-methionine as a source of nitrogen and carbon. Qualitative rapid plate assay test shows the ability of 18 of these isolates to grow with a pink color surrounding their colonial growth, while 6 of these isolates could grow and utilize l-methionine without any pink color around their colonial growth. Quantitative assay test shows the rate of l-methioninase production by all isolates tested. Permeabilization treatment including chemical and physical methods proved that l-methioninase was found to be extracellularly produced. The results also indicate that l-methioninase production was not correlated with growth rate or l-methionine consumption in medium. On the other hand, quantitative assay test shows that these six isolates were l-methioninase negative and failed to produce any amount of l-methioninase. In addition, results also show that isolates No. 4 and No. 60 were the most suitable for l-methioninase production, these two isolates were characterized and identified as Streptomyces sp. DMMMH 4 and Streptomyces sp. MDMMH 60 using 16S rRNA with accession No. in gene bank. Furthermore, optimal conditions for enzyme activity produced by the two isolates were established in relation to temperature, pH, reaction time and type of buffer used and its molarities.