{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,19]],"date-time":"2026-03-19T00:57:59Z","timestamp":1773881879830,"version":"3.50.1"},"reference-count":62,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2005,3,3]],"date-time":"2005-03-03T00:00:00Z","timestamp":1109808000000},"content-version":"vor","delay-in-days":4536,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["European Journal of Biochemistry"],"published-print":{"date-parts":[[1992,10]]},"abstract":"<jats:p>From a genomic library of <jats:italic>Chromatium vinosum<\/jats:italic> strain D in \u03bbL47, a 16.5\u2010kbp <jats:italic>Eco<\/jats:italic>RI\u2010restriction fragment was identified by hybridization with a DNA fragment harboring the operon for <jats:italic>Alcaligenes eutrophus<\/jats:italic> poly(3\u2010hydroxyalkanoate) (PHA) synthesis. This fragment and subfragments thereof restored the ability to synthesize and accumulate PHA in PHA\u2010negative mutants of <jats:italic>A. eutrophus<\/jats:italic>. A region of 6977 bp was sequenced; seven open reading frames (ORFs) were identified which probably represent coding regions; six of these are most probably relevant for PHA biosynthesis in <jats:italic>C. vinosum<\/jats:italic>. The structural genes for biosynthetic acetyl\u2010CoA acyltransferase (\u03b2\u2010ketothiolase; <jats:italic>phbA<\/jats:italic><jats:sub>Cv<\/jats:sub>, 1188 bp) and NADH\u2010dependent acetoacetyl\u2010CoA reductase (<jats:italic>phbB<\/jats:italic><jats:sub>Cv<\/jats:sub>, 741 bp) were separated by ORF4 (462 bp) and ORF5 (369 bp). Downstream of <jats:italic>phbB<\/jats:italic><jats:sub>Cv<\/jats:sub> ORF7 (471 pb) was identified which was not completed at the 3\u2032 terminus. The functions of ORF4, ORF5, and ORF7 are not known. The amino acid sequences of \u03b2\u2010ketothiolase and acetoacetyl\u2010CoA reductase deduced from <jats:italic>phbA<\/jats:italic><jats:sub>Cv<\/jats:sub> and <jats:italic>phbB<\/jats:italic><jats:sub>Cv<\/jats:sub>, exhibited a similarity of 68.2% and 56.4% identical amino acids, respectively, to the corresponding enzymes of <jats:italic>A. eutrophus<\/jats:italic>. Antilinear to and upstream of the genes mentioned above, two genes were identified which were transcribed from a \u03c3<jats:sup>70<\/jats:sup>\u2010dependent promoter. This promoter overlapped with and was divergent to the <jats:italic>phbA<\/jats:italic><jats:sub>Cv<\/jats:sub> promoter; the transcriptional start sites were mapped by S1 nuclease protection assays. These genes were ORF2 (1074 bp), whose function is not known but whose presence in <jats:italic>Escherichia coli<\/jats:italic> is essential for expression of PHA synthase activity, and the structural gene for a PHA synthase of low <jats:italic>M<\/jats:italic><jats:sub>r<\/jats:sub> (<jats:italic>phbC<\/jats:italic><jats:sub>Cv<\/jats:sub>, 1068 bp). The gene products of ORF2 and <jats:italic>phbC<\/jats:italic><jats:sub>Cv<\/jats:sub>, with <jats:italic>M<\/jats:italic><jats:sub>r<\/jats:sub> of 40525 and 39730, respectively, were expressed in <jats:italic>E. coli<\/jats:italic> applying the T7 RNA polymerase\/promoter system. Although the amino acid sequence of PHA synthase deduced from <jats:italic>phbC<\/jats:italic><jats:sub>Cv<\/jats:sub> exhibited only 24.7% overall similarity with the PHA synthase of <jats:italic>A. eutrophus<\/jats:italic>, highly conserved regions were identified.<\/jats:p>","DOI":"10.1111\/j.1432-1033.1992.tb17270.x","type":"journal-article","created":{"date-parts":[[2005,3,4]],"date-time":"2005-03-04T07:59:14Z","timestamp":1109923154000},"page":"135-150","source":"Crossref","is-referenced-by-count":114,"title":["Cloning and nucleotide sequences of genes relevant for biosynthesis of poly(3\u2010hydroxybutyric acid) in <i>Chromatium vinosum<\/i> strain D"],"prefix":"10.1111","volume":"209","author":[{"given":"Matthias","family":"LIEBERGESELL","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexander","family":"STEINB\u00dcCHEL","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2005,3,3]]},"reference":[{"key":"e_1_2_3_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(88)90421-0"},{"key":"e_1_2_3_3_1","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1128\/mr.54.4.450-472.1990","article-title":"Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxy\u2010alkanoates","volume":"54","author":"Anderson A. 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