{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,13]],"date-time":"2026-06-13T05:49:07Z","timestamp":1781329747201,"version":"3.54.1"},"reference-count":36,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2003,8,1]],"date-time":"2003-08-01T00:00:00Z","timestamp":1059696000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2003,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The cyanide dihydratase in\n            <jats:italic>Bacillus pumilus<\/jats:italic>\n            was shown to be an 18-subunit spiral structure by three-dimensional reconstruction of electron micrographs of negatively stained material at its optimum pH, 8.0. At pH 5.4, the subunits rearrange to form an extended left-handed helix. Gel electrophoresis of glutaraldehyde cross-linked enzyme suggests that the fundamental component of the spiral is a dimer of the 37-kDa subunit. The gene was cloned, and the recombinant enzyme was readily expressed at high levels in\n            <jats:italic>Escherichia coli<\/jats:italic>\n            . Purification of the recombinant enzyme was facilitated by the addition of a C-terminal six-histidine affinity purification tag. The tagged recombinant enzyme has\n            <jats:italic>\n              K\n              <jats:sub>m<\/jats:sub>\n            <\/jats:italic>\n            and\n            <jats:italic>V<\/jats:italic>\n            <jats:sub>max<\/jats:sub>\n            values similar to those published for the native enzyme. This is the first cyanide dihydratase from a gram-positive bacterium to be sequenced, and it is the first description of the structure of any member of this enzyme class. The putative amino acid sequence shares over 80% identity to the only other sequenced cyanide dihydratase, that of the gram-negative\n            <jats:italic>Pseudomonas stutzeri<\/jats:italic>\n            strain AK61, and is similar to a number of other bacterial and fungal nitrilases. This sequence similarity suggests that the novel short spiral structure may be typical of these enzymes. In addition, an active cyanide dihydratase from a non-cyanide-degrading isolate of\n            <jats:italic>B<\/jats:italic>\n            .\n            <jats:italic>pumilus<\/jats:italic>\n            (strain 8A3) was cloned and expressed. This suggests that\n            <jats:italic>cynD<\/jats:italic>\n            , the gene coding for the cyanide dihydratase, is not unique to the C1 strain of\n            <jats:italic>B<\/jats:italic>\n            .\n            <jats:italic>pumilus<\/jats:italic>\n            and is not a reflection of its origin at a mining waste site.\n          <\/jats:p>","DOI":"10.1128\/aem.69.8.4794-4805.2003","type":"journal-article","created":{"date-parts":[[2003,8,5]],"date-time":"2003-08-05T22:09:55Z","timestamp":1060121395000},"page":"4794-4805","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":64,"title":["CynD, the Cyanide Dihydratase from\n            <i>Bacillus pumilus<\/i>\n            : Gene Cloning and Structural Studies"],"prefix":"10.1128","volume":"69","author":[{"given":"Dakshina","family":"Jandhyala","sequence":"first","affiliation":[{"name":"Department of Biology and Biochemistry"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark","family":"Berman","sequence":"additional","affiliation":[{"name":"Electron Microscopy Unit"},{"name":"Department of Molecular and Cell Biology, University of Cape Town, Cape Town, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul R.","family":"Meyers","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cell Biology, University of Cape Town, Cape Town, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"B. Trevor","family":"Sewell","sequence":"additional","affiliation":[{"name":"Electron Microscopy Unit"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard C.","family":"Willson","sequence":"additional","affiliation":[{"name":"Department of Biology and Biochemistry"},{"name":"Department of Chemical Engineering, University of Houston, Houston, Texas 77204"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael J.","family":"Benedik","sequence":"additional","affiliation":[{"name":"Department of Biology and Biochemistry"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/7.6.1513"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(76)90527-3"},{"key":"e_1_3_2_4_2","unstructured":"Bron S. 1990. Plasmids p. 15-174. In C. R. Harwood and S. M. Cutting (ed.) Molecular biological methods for Bacillus . John Wiley & Sons Inc. 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