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A highly efficient poly(U)‐dependent poly(Phe) synthesis system for the extreme halophile archaebacterium Halobacterium halobium
Author(s) -
Saruyama Haruo,
Nierhaus Knud H.
Publication year - 1985
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(85)80817-6
Subject(s) - halophile , ribosome , halobacterium , yield (engineering) , biology , stereochemistry , biochemistry , chemistry , bacteria , rna , materials science , genetics , metallurgy , gene
A poly(Phe) synthesis system was optimized for the archaebacterium Halobacterium halobium . It is essential for maximal activity to isolate the ribosomes from cells in mid‐log phase. The system is characterized by the presence of tRNA from brewer's yeast, 6.4 M monovalent cations, 60 mM magnesium, 2 M sulphate anions, a pH of 7.4, and an incubation temperature of 40°C; polycations such as spermidine are not required. Under these conditions one H. halobium ribosome synthesizes statistically more than 20 Phe per h, and the synthesis is not exhausted after 2 h (40 Phe/ribosome). The yield of poly (Phe) is one to two orders of magnitude larger than corresponding yields described for other archaebacterial ribosomes. The accuracy of tRNA selection was determined, and an error fraction of about 0.5% was found. More than 30% of the ribosomes participate in poly(Phe) synthesis.