{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,30]],"date-time":"2026-10-30T09:06:42Z","timestamp":1793351202305,"version":"4.5.0"},"reference-count":49,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2005,2,4]],"date-time":"2005-02-04T00:00:00Z","timestamp":1107475200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecular Microbiology"],"published-print":{"date-parts":[[2005,4]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p>Through examination of one of the fundamental <jats:italic>in vitro<\/jats:italic> characteristics of <jats:italic>Mycobacterium bovis<\/jats:italic>\u2013 its requirement for pyruvate in glycerol medium \u2013 we have revealed a lesion in central metabolism that has profound implications for <jats:italic>in vivo<\/jats:italic> growth and nutrition. Not only is <jats:italic>M. bovis<\/jats:italic> unable to use glycerol as a sole carbon source but the lack of a functioning pyruvate kinase (PK) means that carbohydrates cannot be used to generate energy. This disruption in sugar catabolism is caused by a single nucleotide polymorphism in <jats:italic>pykA<\/jats:italic>, the gene which encodes PK, that substitutes glutamic acid residue 220 with an aspartic acid residue. Substitution of this highly conserved amino acid residue renders PK inactive and thus blocks the ATP generating roles of glycolysis and the pentose phosphate pathway. This mutation was found to occur in other members of the <jats:italic>M. tuberculosis<\/jats:italic> complex, namely <jats:italic>M. microti<\/jats:italic> and <jats:italic>M. africanum<\/jats:italic>. With carbohydrates unable to act as carbon sources, the importance of lipids and gluconeogenesis for growth <jats:italic>in vivo<\/jats:italic> becomes apparent. Complementation of <jats:italic>M. bovis<\/jats:italic> with the <jats:italic>pykA<\/jats:italic> gene from <jats:italic>M. tuberculosis<\/jats:italic> H37Rv restored growth on glycerol. Additionally, the presence of a functioning PK caused the colony morphology of the complemented strain to change from the characteristic dysgonic growth of <jats:italic>M. bovis<\/jats:italic> to eugonic growth, an appearance normally associated with <jats:italic>M. tuberculosis<\/jats:italic>. We also suggest that the glycerol\u2010soaked potato slices used for the derivation of the <jats:italic>M. bovis<\/jats:italic> bacillus Calmette and Gu\u00e9rin (BCG) vaccine strain selected for an <jats:italic>M. bovis<\/jats:italic> PK<jats:sup>+<\/jats:sup> mutant, a finding that explains the alteration in colony morphology noted during the derivation of BCG. In summary, the disruption of a key step in glycolysis divides the <jats:italic>M. tuberculosis<\/jats:italic> complex into two groups with distinct carbon source utilization.<\/jats:p>","DOI":"10.1111\/j.1365-2958.2005.04524.x","type":"journal-article","created":{"date-parts":[[2005,3,9]],"date-time":"2005-03-09T12:41:36Z","timestamp":1110372096000},"page":"163-174","source":"Crossref","is-referenced-by-count":134,"title":["The pyruvate requirement of some members of the <i>Mycobacterium tuberculosis<\/i> complex is due to an inactive pyruvate kinase: implications for <i>in vivo<\/i> growth"],"prefix":"10.1111","volume":"56","author":[{"given":"Lisa A.","family":"Keating","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul R.","family":"Wheeler","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huma","family":"Mansoor","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacqueline K.","family":"Inwald","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James","family":"Dale","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R. 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