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BstB7SI (R↓CCGGY), a thermostable isoschizomer of Cfr10I, from a strain of Bacillus stearothermophilus isolated from oil-contaminated soil in Kuwait

Isolates of thermophilic bacteria from desert soil in Kuwait, heavily contaminated with crude oil, have been screened for the presence of restriction endonuclease activity. One of the isolates (B7S), identified as Bacillus stearothermophilus, showed a high level of restriction endonuclease activity...

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Bibliographic Details
Published in:FEMS microbiology letters 1998-03, Vol.160 (2), p.205-208
Main Authors: Al-Awadhi, Sameera, Welch, Simon G., Smith, Kelvin E., Williams, Ralph A.D.
Format: Article
Language:English
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Summary:Isolates of thermophilic bacteria from desert soil in Kuwait, heavily contaminated with crude oil, have been screened for the presence of restriction endonuclease activity. One of the isolates (B7S), identified as Bacillus stearothermophilus, showed a high level of restriction endonuclease activity when a cell-free extract was incubated with λ bacteriophage DNA at 65°C. A type II restriction endonuclease ( BstB7SI) has been partially purified from this isolate. BstB7SI recognises the six-base sequence RCCGGY (R=A or G; Y=T or C) and hydrolyses the phosphodiester bond in both strands of the DNA substrate between the first and second bases of the recognition sequence 5′-R↓CCGGY-3′ producing four-base 5′ overhangs. BstB7SI is therefore an isoschizomer of the mesophilic prototype restriction endonuclease Cfr10I. BstB7SI has a pH optimum of 9.7, requires 10 mM MgCl 2 and 75 mM NaCl for maximum activity, and retains full enzyme activity when incubated for 5 min at temperatures up to 70°C.
ISSN:0378-1097
1574-6968
DOI:10.1016/S0378-1097(98)00032-9