Open AccessPhysiological regulation, purification and properties of urease from Methylophilus methylotrophus
Author(s) -
Greenwood Jaqueline A,
Mills James,
Tyler Paul D,
Jones Colin W
Publication year - 1998
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1998.tb12902.x
Subject(s) - fast protein liquid chromatography , urea , urease , formamide , chemistry , acetamide , chromatography , ammonium , size exclusion chromatography , biochemistry , ammonium sulfate precipitation , ammonia , enzyme , organic chemistry
The methylotrophic bacterium Methylophilus methylotrophus hydrolyses urea to ammonia using a cytoplasmic urease (EC 3.5.1.5). During growth in continuous culture under various nutrient limitations, urease was induced by urea and short‐chain amides (formamide and urea≫acetamide), and repressed by excess ammonia. The enzyme was purified using ammonium sulfate fractionation and fast protein liquid chromatography (FPLC), and exhibited a narrow substrate specificity (urea≫formamide and acetamide; no activity with other amides) with a K m for urea of 3.8 mM. Gel filtration FPLC and SDS‐PAGE showed that the enzyme had a native M r of approximately 190 000 and was composed of α ( M r 64 000), β ( M r 15 500) and γ ( M r 15 000) subunits in the probable ratio α 2 β 2 γ 2 . The physiological regulation and biochemical properties of the M. methylotrophus urease are compared with those of other bacterial ureases.