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NCS orthologs in available transcriptome databases were screened for variants that might improve the low yield of BIAs in engineered microorganisms. Databases for 21 BIA-producing species from four plant families yielded 33 assembled contigs with homology to characterized <jats:italic>NCS<\/jats:italic> genes. Predicted translation products generated from nine contigs consisted of two to five sequential repeats, each containing most of the sequence found in single-domain enzymes. Assembled contigs containing tandem domain repeats were detected only in members of the Papaveraceae family, including opium poppy (<jats:italic>Papaver somniferum<\/jats:italic>). Fourteen cDNAs were generated from 10 species, five of which encoded NCS orthologs with repeated domains. Functional analysis of corresponding recombinant proteins yielded six active NCS enzymes, including four containing either two, three or four repeated catalytic domains. Truncation of the first 25 N-terminal amino acids from the remaining polypeptides revealed two additional enzymes. Multiple catalytic domains correlated with a proportional increase in catalytic efficiency. Expression of <jats:italic>NCS<\/jats:italic> genes in <jats:italic>Saccharomyces cereviseae<\/jats:italic> also produced active enzymes. The metabolic conversion capacity of engineered yeast positively correlated with the number of repeated domains.<\/jats:p>","DOI":"10.1038\/srep39256","type":"journal-article","created":{"date-parts":[[2016,12,19]],"date-time":"2016-12-19T10:41:06Z","timestamp":1482144066000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":40,"title":["Genes encoding norcoclaurine synthase occur as tandem fusions in the Papaveraceae"],"prefix":"10.1038","volume":"6","author":[{"given":"Jing","family":"Li","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eun-Jeong","family":"Lee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Limei","family":"Chang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter J.","family":"Facchini","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2016,12,19]]},"reference":[{"key":"BFsrep39256_CR1","doi-asserted-by":"crossref","first-page":"1083","DOI":"10.1016\/0031-9422(89)80187-6","volume":"28","author":"R Stadler","year":"1989","unstructured":"Stadler, R., Kutchan, T. M. & Zenk, M. H. (S)-Norcoclaurine is the central intermediate in benzylisoquinoline alkaloid biosynthesis. Phytochemistry 28, 1083\u20131086 (1989).","journal-title":"Phytochemistry"},{"key":"BFsrep39256_CR2","doi-asserted-by":"crossref","unstructured":"Stadler, R. & Zenk, M. H. A revision of the generally accepted pathway for the biosynthesis of the benzyltetrahydroisoquinoline reticuline. Liebigs Ann. Chem. 555\u2013562 (1990).","DOI":"10.1002\/jlac.1990199001104"},{"key":"BFsrep39256_CR3","doi-asserted-by":"crossref","first-page":"3242","DOI":"10.1039\/c0cc05282e","volume":"47","author":"T Pesnot","year":"2011","unstructured":"Pesnot, T., Gershater, M. C., Ward, J. M. & Hailes, H. C. Phosphate mediated biomimetic synthesis of tetrahydroisoquinoline alkaloids. Chem. Commun. 47, 3242\u20133244 (2011).","journal-title":"Chem. 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P.J.F. receives research funding from, owns stock in, serves on the Board of Directors of, and is provided compensation by Epimeron. J.L. and L.C. also own stock or stock options in Epimeron.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"39256"}}