Screening of microorganisms producing a novel protein-asparaginase and characterization of the enzyme derived from Luteimicrobium album
| Autor: | Takayuki Kajiura, Noriko Miwa, Mayu Mitsuhashi |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
0301 basic medicine Asparaginase Microbacterium Bioengineering 01 natural sciences Applied Microbiology and Biotechnology Enrichment culture 03 medical and health sciences chemistry.chemical_compound 010608 biotechnology Casein Asparagine Deamidation chemistry.chemical_classification biology Temperature Hydrogen-Ion Concentration biology.organism_classification Agromyces Actinobacteria 030104 developmental biology Enzyme chemistry Biochemistry Biocatalysis Peptides Biotechnology |
| Zdroj: | Journal of bioscience and bioengineering. 127(3) |
| ISSN: | 1347-4421 |
| Popis: | A screening system using enrichment culture has been established with the aim of obtaining a novel enzyme for protein modification that has not been previously reported. This enzyme catalyzes deamidation of the side-chain amide group of asparagine in proteins. Enrichment culture of 390 soil samples was carried out with Z-Asn-Gly as the sole source of nitrogen, and the reaction product, Z-Asp-Gly, was detected in the culture supernatant of 102 strains. Strains with particularly high activity were Leifsonia sp., Luteimicrobium sp., Microbacterium sp., and Agromyces sp., all belonging to the class Actinobacteria. Of these, a protein-asparaginase (PA) was obtained from the culture supernatant of Luteimicrobium album 333B-h1, and its reactivity with different substrates and its basic enzymatic characteristics were investigated. Addition of the enzyme solution resulted in specific deamidation of only the asparagine residue in insulin chain B. The enzyme showed no reactivity with free asparagine or asparagine in low molecular weight peptides. It was demonstrated that the enzyme reacts with various protein substrates. In particular, proteins that have open structures, such as casein or gelatin, were good substrates. The activity and stability of PA at different temperatures and pH values were investigated. It was found that a temperature of 37°C and a roughly neutral pH are optimal conditions for the enzyme. |
| Databáze: | OpenAIRE |
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