2024-03-29T12:22:16Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/1596512018-09-11T10:24:54Zcom_10261_15com_10261_6col_10261_268
González Grau, Juan Miguel
Kato, C.
Horikoshi, Kobi
2018-01-25T11:24:31Z
2018-01-25T11:24:31Z
1996-01
Journal of Marine Biotechnology 4(3):159-164 (1996)
0941-2905
http://hdl.handle.net/10261/159651
Sodium dodecyl sulfate (SDS) extracts from intact Thermococcus peplonophilus cells resulted in high proteolytic activity. After 24 h of incubation in the presence of 0.5% SDS at 90°C, the extracts still exhibited 12.5% of the initial protease activity. Treated extracts showed the presence of a single band in SDS-PAGE gels. Molecular weight, assessed by SDS-PAGE, was about 14 kDa. The treated extract showed high thermostable proteolytic activity; measured half-life times were 10 h at 90°C, 8 h at 100°C, and 19 min at 120°C. This proteolytic activity was resistant to denaturation by SDS, dithiothreitol, deoxycholate, urea, and β-mercaptoethanol. The proteolytic activity of the treated extract was inhibited by phenylmethylsulfonyl fluoride, diisopropyl fluorophosphate and, partially, by iodoacetic acid; no inhibition was observed by EDTA or ethylene glycolbis(β-aminoethyl ether)-N, N, N′, N′-tetraacetic acid. These results indicate that the extracts contain a serine protease. The hydrolysis of several synthetic substrates was assayed; proteolytic activity of treated extracts was hydrolytically active towards N-α-benzoyl-L-arginine-4-nitroanilide (with high affinity), L-leucine-p-nitroanilide (with low affinity), and L-lycine-p-nitroanilide (with low affinity). Extracts can be stored at -80°C for at least one month with minimum activity losses.
eng
closedAccess
Membrane-bound, sodium dodecyl sulfate-resistant proteolytic activity from the extremely thermophilic archaeon Thermococcus peptonophilus
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