{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,7]],"date-time":"2025-11-07T13:15:30Z","timestamp":1762521330475},"reference-count":33,"publisher":"Springer Science and Business Media LLC","issue":"1","content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Microb Cell Fact"],"published-print":{"date-parts":[[2010,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:sec>\n            <jats:title>Background<\/jats:title>\n            <jats:p>Nikkomycins are a group of peptidyl nucleoside antibiotics produced by <jats:italic>Streptomyces ansochromogenes<\/jats:italic>. They are competitive inhibitors of chitin synthase and show potent fungicidal, insecticidal, and acaricidal activities. Nikkomycin X and Z are the main components produced by <jats:italic>S. ansochromogenes<\/jats:italic>. Generation of a high-producing strain is crucial to scale up nikkomycins production for further clinical trials.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>To increase the yields of nikkomycins, an additional copy of nikkomycin biosynthetic gene cluster (35 kb) was introduced into nikkomycin producing strain, <jats:italic>S. ansochromogenes<\/jats:italic> 7100. The gene cluster was first reassembled into an integrative plasmid by Red\/ET technology combining with classic cloning methods and then the resulting plasmid(pNIK)was introduced into <jats:italic>S. ansochromogenes<\/jats:italic> by conjugal transfer. Introduction of pNIK led to enhanced production of nikkomycins (880 mg L<jats:sup>-1<\/jats:sup>, 4 -fold nikkomycin X and 210 mg L<jats:sup>-1<\/jats:sup>, 1.8-fold nikkomycin Z) in the resulting exconjugants comparing with the parent strain (220 mg L<jats:sup>-1<\/jats:sup> nikkomycin X and 120 mg L<jats:sup>-1<\/jats:sup> nikkomycin Z). The exconjugants are genetically stable in the absence of antibiotic resistance selection pressure.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusion<\/jats:title>\n            <jats:p>A high nikkomycins producing strain (1100 mg L<jats:sup>-1<\/jats:sup> nikkomycins) was obtained by introduction of an extra nikkomycin biosynthetic gene cluster into the genome of <jats:italic>S. ansochromogenes<\/jats:italic>. The strategies presented here could be applicable to other bacteria to improve the yields of secondary metabolites.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/1475-2859-9-6","type":"journal-article","created":{"date-parts":[[2010,1,23]],"date-time":"2010-01-23T07:14:39Z","timestamp":1264230879000},"update-policy":"http:\/\/dx.doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":43,"title":["Cloning, reassembling and integration of the entire nikkomycin biosynthetic gene cluster into Streptomyces ansochromogenes lead to an improved nikkomycin production"],"prefix":"10.1186","volume":"9","author":[{"given":"Guojian","family":"Liao","sequence":"first","affiliation":[]},{"given":"Jine","family":"Li","sequence":"additional","affiliation":[]},{"given":"Lei","family":"Li","sequence":"additional","affiliation":[]},{"given":"Haihua","family":"Yang","sequence":"additional","affiliation":[]},{"given":"Yuqing","family":"Tian","sequence":"additional","affiliation":[]},{"given":"Huarong","family":"Tan","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2010,1,23]]},"reference":[{"issue":"5","key":"396_CR1","doi-asserted-by":"publisher","first-page":"281","DOI":"10.1016\/j.ymben.2008.07.001","volume":"10","author":"C Olano","year":"2008","unstructured":"Olano C, Lombo F, Mendez C, Salas JA: Improving production of bioactive secondary metabolites in actinomycetes by metabolic engineering. Metab Eng. 2008, 10 (5): 281-292. 10.1016\/j.ymben.2008.07.001.","journal-title":"Metab Eng"},{"issue":"2","key":"396_CR2","doi-asserted-by":"publisher","first-page":"168","DOI":"10.1016\/j.cbpa.2008.01.017","volume":"12","author":"CN Santos","year":"2008","unstructured":"Santos CN, Stephanopoulos G: Combinatorial engineering of microbes for optimizing cellular phenotype. Curr Opin Chem Biol. 2008, 12 (2): 168-176. 10.1016\/j.cbpa.2008.01.017.","journal-title":"Curr Opin Chem Biol"},{"issue":"13","key":"396_CR3","doi-asserted-by":"publisher","first-page":"6200","DOI":"10.1073\/pnas.92.13.6200","volume":"92","author":"F Fierro","year":"1995","unstructured":"Fierro F, Barredo JL, Diez B, Gutierrez S, Fernandez FJ, Martin JF: The penicillin gene cluster is amplified in tandem repeats linked by conserved hexanucleotide sequences. Proc Natl Acad Sci USA. 1995, 92 (13): 6200-6204. 10.1073\/pnas.92.13.6200.","journal-title":"Proc Natl Acad Sci USA"},{"issue":"6","key":"396_CR4","doi-asserted-by":"publisher","first-page":"1137","DOI":"10.1111\/j.1365-2958.1995.tb02338.x","volume":"16","author":"U Peschke","year":"1995","unstructured":"Peschke U, Schmidt H, Zhang HZ, Piepersberg W: Molecular characterization of the lincomycin-production gene cluster of Streptomyces lincolnensis 78-11. Mol Microbiol. 1995, 16 (6): 1137-1156. 10.1111\/j.1365-2958.1995.tb02338.x.","journal-title":"Mol Microbiol"},{"issue":"25","key":"396_CR5","doi-asserted-by":"publisher","first-page":"9661","DOI":"10.1073\/pnas.0603251103","volume":"103","author":"K Yanai","year":"2006","unstructured":"Yanai K, Murakami T, Bibb M: Amplification of the entire kanamycin biosynthetic gene cluster during empirical strain improvement of Streptomyces kanamyceticus. Proc Natl Acad Sci USA. 2006, 103 (25): 9661-9666. 10.1073\/pnas.0603251103.","journal-title":"Proc Natl Acad Sci USA"},{"issue":"2","key":"396_CR6","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1038\/2417","volume":"20","author":"Y Zhang","year":"1998","unstructured":"Zhang Y, Buchholz F, Muyrers JP, Stewart AF: A new logic for DNA engineering using recombination in Escherichia coli. Nat Genet. 1998, 20 (2): 123-128. 10.1038\/2417.","journal-title":"Nat Genet"},{"issue":"12","key":"396_CR7","doi-asserted-by":"publisher","first-page":"1314","DOI":"10.1038\/82449","volume":"18","author":"Y Zhang","year":"2000","unstructured":"Zhang Y, Muyrers JP, Testa G, Stewart AF: DNA cloning by homologous recombination in Escherichia coli. Nat Biotechnol. 2000, 18 (12): 1314-1317. 10.1038\/82449.","journal-title":"Nat Biotechnol"},{"issue":"3","key":"396_CR8","doi-asserted-by":"publisher","first-page":"349","DOI":"10.1016\/j.chembiol.2004.12.012","volume":"12","author":"SC Wenzel","year":"2005","unstructured":"Wenzel SC, Gross F, Zhang Y, Fu J, Stewart AF, Muller R: Heterologous expression of a myxobacterial natural products assembly line in pseudomonads via red\/ET recombineering. Chem Biol. 2005, 12 (3): 349-356. 10.1016\/j.chembiol.2004.12.012.","journal-title":"Chem Biol"},{"issue":"12","key":"396_CR9","doi-asserted-by":"publisher","first-page":"7485","DOI":"10.1128\/AEM.01503-06","volume":"72","author":"O Perlova","year":"2006","unstructured":"Perlova O, Fu J, Kuhlmann S, Krug D, Stewart AF, Zhang Y, Muller R: Reconstitution of the myxothiazol biosynthetic gene cluster by Red\/ET recombination and heterologous expression in Myxococcus xanthus. Appl Environ Microbiol. 2006, 72 (12): 7485-7494. 10.1128\/AEM.01503-06.","journal-title":"Appl Environ Microbiol"},{"issue":"4","key":"396_CR10","doi-asserted-by":"publisher","first-page":"603","DOI":"10.1002\/cbic.200700483","volume":"9","author":"M Wolpert","year":"2008","unstructured":"Wolpert M, Heide L, Kammerer B, Gust B: Assembly and heterologous expression of the coumermycin A1 gene cluster and production of new derivatives by genetic engineering. Chembiochem. 2008, 9 (4): 603-612. 10.1002\/cbic.200700483.","journal-title":"Chembiochem"},{"issue":"3","key":"396_CR11","doi-asserted-by":"publisher","first-page":"447","DOI":"10.1002\/cbic.200700549","volume":"9","author":"TM Binz","year":"2008","unstructured":"Binz TM, Wenzel SC, Schnell HJ, Bechthold A, Muller R: Heterologous expression and genetic engineering of the phenalinolactone biosynthetic gene cluster by using red\/ET recombineering. Chembiochem. 2008, 9 (3): 447-454. 10.1002\/cbic.200700549.","journal-title":"Chembiochem"},{"issue":"10","key":"396_CR12","doi-asserted-by":"publisher","first-page":"1603","DOI":"10.1002\/cbic.200800029","volume":"9","author":"Y Hu","year":"2008","unstructured":"Hu Y, Phelan VV, Farnet CM, Zazopoulos E, Bachmann BO: Reassembly of anthramycin biosynthetic gene cluster by using recombinogenic cassettes. Chembiochem. 2008, 9 (10): 1603-1608. 10.1002\/cbic.200800029.","journal-title":"Chembiochem"},{"issue":"17","key":"396_CR13","doi-asserted-by":"publisher","first-page":"e113","DOI":"10.1093\/nar\/gkn499","volume":"36","author":"J Fu","year":"2008","unstructured":"Fu J, Wenzel SC, Perlova O, Wang J, Gross F, Tang Z, Yin Y, Stewart AF, Muller R, Zhang Y: Efficient transfer of two large secondary metabolite pathway gene clusters into heterologous hosts by transposition. Nucleic Acids Res. 2008, 36 (17): e113-10.1093\/nar\/gkn499.","journal-title":"Nucleic Acids Res"},{"issue":"1","key":"396_CR14","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1128\/CMR.6.1.1","volume":"6","author":"RF Hector","year":"1993","unstructured":"Hector RF: Compounds active against cell walls of medically important fungi. Clin Microbiol Rev. 1993, 6 (1): 1-21.","journal-title":"Clin Microbiol Rev"},{"issue":"5","key":"396_CR15","doi-asserted-by":"publisher","first-page":"662","DOI":"10.1007\/s004380000352","volume":"264","author":"B Lauer","year":"2001","unstructured":"Lauer B, Russwurm R, Schwarz W, Kalmanczhelyi A, Bruntner C, Rosemeier A, Bormann C: Molecular characterization of co-transcribed genes from Streptomyces tendae Tu901 involved in the biosynthesis of the peptidyl moiety and assembly of the peptidyl nucleoside antibiotic nikkomycin. Mol Gen Genet. 2001, 264 (5): 662-673. 10.1007\/s004380000352.","journal-title":"Mol Gen Genet"},{"issue":"1","key":"396_CR16","first-page":"102","volume":"262","author":"C Bruntner","year":"1999","unstructured":"Bruntner C, Lauer B, Schwarz W, Mohrle V, Bormann C: Molecular characterization of co-transcribed genes from Streptomyces tendae Tu901 involved in the biosynthesis of the peptidyl moiety of the peptidyl nucleoside antibiotic nikkomycin. Mol Gen Genet. 1999, 262 (1): 102-114.","journal-title":"Mol Gen Genet"},{"issue":"6","key":"396_CR17","doi-asserted-by":"publisher","first-page":"1855","DOI":"10.1111\/j.1365-2958.2005.04512.x","volume":"55","author":"G Liu","year":"2005","unstructured":"Liu G, Tian Y, Yang H, Tan H: A pathway-specific transcriptional regulatory gene for nikkomycin biosynthesis in Streptomyces ansochromogenes that also influences colony development. Mol Microbiol. 2005, 55 (6): 1855-1866. 10.1111\/j.1365-2958.2005.04512.x.","journal-title":"Mol Microbiol"},{"key":"396_CR18","volume-title":"Structure and Function of Gene Involved in Nikkomycin Biosynthesis and Differentiation in Streptomyces ansochromogenes. PhD Dissertation","author":"W Li","year":"2003","unstructured":"Li W: Structure and Function of Gene Involved in Nikkomycin Biosynthesis and Differentiation in Streptomyces ansochromogenes. PhD Dissertation. 2003, Beijing: Institute of microbiology, CAS"},{"issue":"4","key":"396_CR19","doi-asserted-by":"publisher","first-page":"379","DOI":"10.1002\/1097-0290(20000220)72:4<379::AID-BIT1000>3.0.CO;2-5","volume":"72","author":"H Theilgaard","year":"2001","unstructured":"Theilgaard H, Berg van Den M, Mulder C, Bovenberg R, Nielsen J: Quantitative analysis of Penicillium chrysogenum Wis54-1255 transformants overexpressing the penicillin biosynthetic genes. Biotechnol Bioeng. 2001, 72 (4): 379-388. 10.1002\/1097-0290(20000220)72:4<379::AID-BIT1000>3.0.CO;2-5.","journal-title":"Biotechnol Bioeng"},{"issue":"4","key":"396_CR20","doi-asserted-by":"publisher","first-page":"697","DOI":"10.1007\/s00253-009-1933-8","volume":"83","author":"S Baba","year":"2009","unstructured":"Baba S, Abe Y, Suzuki T, Ono C, Iwamoto K, Nihira T, Hosobuchi M: Improvement of compactin (ML-236B) production by genetic engineering in compactin high-producing Penicillium citrinum. Appl Microbiol Biotechnol. 2009, 83 (4): 697-704. 10.1007\/s00253-009-1933-8.","journal-title":"Appl Microbiol Biotechnol"},{"key":"396_CR21","doi-asserted-by":"publisher","first-page":"1222","DOI":"10.1038\/nbt1088-1222","volume":"6","author":"CW Chen","year":"1988","unstructured":"Chen CW, Lin H-F, Kuo CL, Tsai H-L, Tsai JF-Y: Cloning and Expression of a DNA Sequence Conferring Cephamycin C Production. Bio\/Technology. 1988, 6: 1222-1224. 10.1038\/nbt1088-1222.","journal-title":"Bio\/Technology"},{"issue":"1","key":"396_CR22","doi-asserted-by":"publisher","first-page":"43","DOI":"10.1016\/0378-1119(92)90627-2","volume":"116","author":"M Bierman","year":"1992","unstructured":"Bierman M, Logan R, O'Brien K, Seno ET, Rao RN, Schoner BE: Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. Gene. 1992, 116 (1): 43-49. 10.1016\/0378-1119(92)90627-2.","journal-title":"Gene"},{"issue":"3","key":"396_CR23","doi-asserted-by":"publisher","first-page":"171","DOI":"10.1016\/j.plasmid.2008.12.002","volume":"61","author":"B Ostash","year":"2009","unstructured":"Ostash B, Makitrinskyy R, Walker S, Fedorenko V: Identification and characterization of Streptomyces ghanaensis ATCC14672 integration sites for three actinophage-based plasmids. Plasmid. 2009, 61 (3): 171-175. 10.1016\/j.plasmid.2008.12.002.","journal-title":"Plasmid"},{"issue":"8","key":"396_CR24","doi-asserted-by":"publisher","first-page":"760","DOI":"10.1038\/nbt.1555","volume":"27","author":"KE Tyo","year":"2009","unstructured":"Tyo KE, Ajikumar PK, Stephanopoulos G: Stabilized gene duplication enables long-term selection-free heterologous pathway expression. Nat Biotechnol. 2009, 27 (8): 760-765. 10.1038\/nbt.1555.","journal-title":"Nat Biotechnol"},{"key":"396_CR25","first-page":"325","volume-title":"Cutaneous antifungal agents:selected compounds in clinical practice and development","author":"H-P Fiedler","year":"1993","unstructured":"Fiedler H-P, Schuz T, Decker H: An overview of nikkomycins: history, biochemistry and applications. Cutaneous antifungal agents:selected compounds in clinical practice and development. Edited by: Rippon JW, Fromtling RA. 1993, New York: Marcel Dekker, Inc, 325-352."},{"issue":"1","key":"396_CR26","doi-asserted-by":"publisher","first-page":"103","DOI":"10.1016\/S1074-5521(02)00090-X","volume":"9","author":"H Chen","year":"2002","unstructured":"Chen H, Hubbard BK, O'Connor SE, Walsh CT: Formation of beta-hydroxy histidine in the biosynthesis of nikkomycin antibiotics. Chem Biol. 2002, 9 (1): 103-112. 10.1016\/S1074-5521(02)00090-X.","journal-title":"Chem Biol"},{"issue":"3","key":"396_CR27","doi-asserted-by":"publisher","first-page":"229","DOI":"10.1023\/B:BILE.0000013720.14278.07","volume":"26","author":"G Wang","year":"2004","unstructured":"Wang G, Tan H: Enhanced production of nikkomycin X by over-expression of SanO, a non-ribosomal peptide synthetase in Streptomyces ansochromogenes. Biotechnol Lett. 2004, 26 (3): 229-233. 10.1023\/B:BILE.0000013720.14278.07.","journal-title":"Biotechnol Lett"},{"key":"396_CR28","volume-title":"Practical StreptomycesGenetics","author":"T Kieser","year":"2000","unstructured":"Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA: Practical Streptomyces Genetics. 2000, the John Innes Foundation, Norwich, England"},{"issue":"20","key":"396_CR29","doi-asserted-by":"publisher","first-page":"5746","DOI":"10.1128\/JB.184.20.5746-5752.2002","volume":"184","author":"P Combes","year":"2002","unstructured":"Combes P, Till R, Bee S, Smith MC: The streptomyces genome contains multiple pseudo-attB sites for the (phi)C31-encoded site-specific recombination system. J Bacteriol. 2002, 184 (20): 5746-5752. 10.1128\/JB.184.20.5746-5752.2002.","journal-title":"J Bacteriol"},{"key":"396_CR30","doi-asserted-by":"publisher","first-page":"487","DOI":"10.1016\/S0021-9673(00)96176-4","volume":"316","author":"HP Fiedler","year":"1984","unstructured":"Fiedler HP: Screening for new microbial products by high-performance liquid chromatography using a photodiode array detector. J Chromatogr. 1984, 316: 487-494. 10.1016\/S0021-9673(00)96176-4.","journal-title":"J Chromatogr"},{"issue":"3","key":"396_CR31","doi-asserted-by":"publisher","first-page":"175","DOI":"10.1007\/s00284-001-0115-4","volume":"45","author":"H Zeng","year":"2002","unstructured":"Zeng H, Tan H, Li J: Cloning and function of sanQ: a gene involved in nikkomycin biosynthesis of Streptomyces ansochromogenes. Curr Microbiol. 2002, 45 (3): 175-179. 10.1007\/s00284-001-0115-4.","journal-title":"Curr Microbiol"},{"issue":"1","key":"396_CR32","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1046\/j.1365-2958.1999.01452.x","volume":"33","author":"MS Paget","year":"1999","unstructured":"Paget MS, Leibovitz E, Buttner MJ: A putative two-component signal transduction system regulates sigmaE, a sigma factor required for normal cell wall integrity in Streptomyces coelicolor A3(2). Mol Microbiol. 1999, 33 (1): 97-107. 10.1046\/j.1365-2958.1999.01452.x.","journal-title":"Mol Microbiol"},{"issue":"4","key":"396_CR33","doi-asserted-by":"publisher","first-page":"1541","DOI":"10.1073\/pnas.0337542100","volume":"100","author":"B Gust","year":"2003","unstructured":"Gust B, Challis GL, Fowler K, Kieser T, Chater KF: PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin. Proc Natl Acad Sci USA. 2003, 100 (4): 1541-1546. 10.1073\/pnas.0337542100.","journal-title":"Proc Natl Acad Sci USA"}],"container-title":["Microbial Cell Factories"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/1475-2859-9-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,9,1]],"date-time":"2021-09-01T11:53:48Z","timestamp":1630497228000},"score":1,"resource":{"primary":{"URL":"https:\/\/microbialcellfactories.biomedcentral.com\/articles\/10.1186\/1475-2859-9-6"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2010,1,23]]},"references-count":33,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2010,12]]}},"alternative-id":["396"],"URL":"https:\/\/doi.org\/10.1186\/1475-2859-9-6","relation":{},"ISSN":["1475-2859"],"issn-type":[{"value":"1475-2859","type":"electronic"}],"subject":[],"published":{"date-parts":[[2010,1,23]]},"assertion":[{"value":"14 November 2009","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"23 January 2010","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"23 January 2010","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}}],"article-number":"6"}}