{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,20]],"date-time":"2026-09-20T04:06:44Z","timestamp":1789877204901,"version":"4.0.1"},"reference-count":43,"publisher":"National Academy of Sciences","issue":"21","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2008,5,27]]},"abstract":"\n To identify the genes for biosynthesis of the off-flavor terpenoid alcohol, 2-methylisoborneol (2-MIB), the key genes encoding monoterpene cyclase were located in bacterial genome databases by using a combination of hidden Markov models, protein\u2013family search, and the sequence alignment of their gene products. Predicted terpene cyclases were classified into three groups: sesquiterpene, diterpene, and other terpene cyclases. Genes of the terpene cyclase group that form an operon with a gene encoding\n S<\/jats:italic>\n -adenosyl-\n l<\/jats:sc>\n -methionine (SAM)-dependent methyltransferase were found in genome data of seven microorganisms belonging to actinomycetes,\n Streptomyces ambofaciens<\/jats:italic>\n ISP5053,\n Streptomyces coelicolor<\/jats:italic>\n A3(\n 2<\/jats:xref>\n ),\n Streptomyces griseus<\/jats:italic>\n IFO13350,\n Streptomyces lasaliensis<\/jats:italic>\n NRRL3382R,\n Streptomyces scabies<\/jats:italic>\n 87.22,\n Saccharopolyspora erythraea<\/jats:italic>\n NRRL2338, and\n Micromonospora olivasterospora<\/jats:italic>\n KY11048. Among six microorganisms tested,\n S. ambofaciens<\/jats:italic>\n ,\n S. coelicolor<\/jats:italic>\n A3(\n 2<\/jats:xref>\n ),\n S. griseus<\/jats:italic>\n , and\n S. lasaliensis<\/jats:italic>\n produced 2-MIB but\n M. olivasterospora<\/jats:italic>\n produced 2-methylenebornane (2-MB) instead. The regions containing monoterpene cyclase and methyltransferase genes were amplified by PCR from\n S. ambofaciens<\/jats:italic>\n ,\n S. lasaliensis<\/jats:italic>\n , and\n Saccharopolyspora erythraea<\/jats:italic>\n , respectively, and their genes were heterologously expressed in\n Streptomyces avermitilis<\/jats:italic>\n , which was naturally deficient of 2-MIB biosynthesis by insertion and deletion. All exoconjugants of\n S. avermitilis<\/jats:italic>\n produced 2-MIB. Full-length recombinant proteins, monoterpene cyclase and methyltransferase of\n S. lasaliensis<\/jats:italic>\n were expressed at high level in\n Escherichia coli<\/jats:italic>\n . The recombinant methyltransferase catalyzed methylation at the C2 position of geranyl diphosphate (GPP) in the presence of SAM. 2-MIB was generated by incubation with GPP, SAM, recombinant methyltransferase, and terpene cyclase. We concluded that the biosynthetic pathway involves the methylation of GPP by GPP methyltransferase and its subsequent cyclization by monoterpene cyclase to 2-MIB.\n <\/jats:p>","DOI":"10.1073\/pnas.0802312105","type":"journal-article","created":{"date-parts":[[2008,5,20]],"date-time":"2008-05-20T23:33:39Z","timestamp":1211326419000},"page":"7422-7427","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":235,"title":["Identification and functional analysis of genes controlling biosynthesis of 2-methylisoborneol"],"prefix":"10.1073","volume":"105","author":[{"given":"Mamoru","family":"Komatsu","sequence":"first","affiliation":[{"name":"*Kitasato Institute for Life Sciences, Kitasato University, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555, Japan;"},{"name":"The Kitasato Institute, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8642, Japan; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muneya","family":"Tsuda","sequence":"additional","affiliation":[{"name":"Division of Chemistry, Graduate School of Science, Hokkaido University, Sapporo 060-0810, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Satoshi","family":"\u014cmura","sequence":"additional","affiliation":[{"name":"The Kitasato Institute, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8642, Japan; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hideaki","family":"Oikawa","sequence":"additional","affiliation":[{"name":"Division of Chemistry, Graduate School of Science, Hokkaido University, Sapporo 060-0810, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haruo","family":"Ikeda","sequence":"additional","affiliation":[{"name":"*Kitasato Institute for Life Sciences, Kitasato University, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555, Japan;"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2008,5,27]]},"reference":[{"key":"e_1_3_3_1_2","volume-title":"Encyclopedia of the Terpenoids","author":"Glasby JS","year":"1982","unstructured":"JS Glasby Encyclopedia of the Terpenoids (John Wiley, Chichester, U.K., 1982)."},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.277.5333.1788"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1021\/jf0116754"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.62.4.1295-1299.1996"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.3109\/10408417909082014"},{"key":"e_1_3_3_6_2","first-page":"453","volume-title":"The Cyanobacteria","author":"J\u00fcttner F","year":"1987","unstructured":"F J\u00fcttner, Volatile organic substances. The Cyanobacteria, eds B Fay, CV Baalen (Elsevier, Amsterdam), pp. 453\u2013469 (1987)."},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.43.3.708-714.1982"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF00408253"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1002\/anie.200702496"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1021\/jf00035a053"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.58.9.3170-3172.1992"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1139\/f76-004"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.2166\/wst.1983.0125"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.2166\/wst.1983.0155"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1021\/jf020806d"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1021\/jf00063a005"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.7164\/antibiotics.47.434"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0040-4039(00)89625-2"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.7164\/antibiotics.22.508"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1021\/es60028a008"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0337625100"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1038\/ja.2006.66"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja061292s"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1093\/genetics\/158.2.811"},{"key":"e_1_3_3_25_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja015747j"},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-08-091283-7.00040-0"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(81)80723-5"},{"key":"e_1_3_3_28_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/14.9.755"},{"key":"e_1_3_3_29_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/30.1.276"},{"key":"e_1_3_3_30_2","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio803"},{"key":"e_1_3_3_31_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.183.20.6085-6094.2001"},{"key":"e_1_3_3_32_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.277.5333.1820"},{"key":"e_1_3_3_33_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja026058q"},{"key":"e_1_3_3_34_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.277.5333.1815"},{"key":"e_1_3_3_35_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M000433200"},{"key":"e_1_3_3_36_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.231313098"},{"key":"e_1_3_3_37_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.022627099"},{"key":"e_1_3_3_38_2","doi-asserted-by":"publisher","DOI":"10.1093\/molbev\/msl108"},{"key":"e_1_3_3_39_2","doi-asserted-by":"publisher","DOI":"10.7164\/antibiotics.35.507"},{"key":"e_1_3_3_40_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00204-08"},{"key":"e_1_3_3_41_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkf436"},{"key":"e_1_3_3_42_2","first-page":"4673","volume":"22","author":"Thompson JD","year":"1994","unstructured":"JD Thompson, DG Higgins, TJ Gibson CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice Nucleic Acids Res 22, 4673\u20134680 (1994).","journal-title":"CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice Nucleic Acids Res"},{"key":"e_1_3_3_43_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.169.12.5615-5621.1987"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.0802312105","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,12]],"date-time":"2022-04-12T16:46:55Z","timestamp":1649782015000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.0802312105"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2008,5,27]]},"references-count":43,"journal-issue":{"issue":"21","published-print":{"date-parts":[[2008,5,27]]}},"alternative-id":["10.1073\/pnas.0802312105"],"URL":"https:\/\/doi.org\/10.1073\/pnas.0802312105","relation":{"has-review":[{"id-type":"doi","id":"10.3410\/f.1115030.570998","asserted-by":"object"}]},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2008,5,27]]},"assertion":[{"value":"2007-12-12","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2008-05-27","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}