Michael Perzl , Ina G. Reipen , Susanne Schmitz , Karl Poralla , Hermann Sahm , Georg A. Sprenger , Elmar L. Kannenberg
{"title":"活动单胞菌和日本慢生根瘤菌合成类藿烷脂保守基因的克隆[j]","authors":"Michael Perzl , Ina G. Reipen , Susanne Schmitz , Karl Poralla , Hermann Sahm , Georg A. Sprenger , Elmar L. Kannenberg","doi":"10.1016/S0005-2760(98)00064-2","DOIUrl":null,"url":null,"abstract":"<div><p>The squalene-hopene cyclase (SHC) is the only enzyme involved in the biosynthesis of hopanoid lipids that has been characterized on the genetic level. To investigate if additional genes involved in hopanoid biosynthesis are clustered with the <em>shc</em> gene, we cloned and analyzed the nucleotide sequences located immediately upstream of the <em>shc</em> genes from <em>Zymomonas mobilis</em> and <em>Bradyrhizobium japonicum</em>. In <em>Z. mobilis</em>, five open reading frames (ORFs, designated as <em>hpnA–E</em>) were detected in a close arrangement with the <em>shc</em> gene. In <em>B. japonicum</em>, three similarly arranged ORFs (corresponding to <em>hpnC–E</em> from <em>Z. mobilis</em>) were found. The deduced amino acid sequences of <em>hpnC–E</em> showed significant similarity (58–62%) in both bacteria. Similarities to enzymes of other terpenoid biosynthesis pathways (carotenoid and steroid biosynthesis) suggest that these ORFs encode proteins involved in the biosynthesis of hopanoids and their intermediates. Expression of <em>hpnC</em> to <em>hpnE</em> from <em>Z. mobilis</em> as well as expression of <em>hpnC</em> from <em>B. japonicum</em> in <em>Escherichia coli</em> led to the formation of the hopanoid precursor squalene. This indicates that <em>hpnC</em> encodes a squalene synthase. The two additional ORFs (<em>hpnA</em> and <em>hpnB</em>) in <em>Z. mobilis</em> showed similarities to enzymes involved in the transfer and modification of sugars, indicating that they may code for enzymes involved in the biosynthesis of the complex, sugar-containing side chains of hopanoids.</p></div>","PeriodicalId":100162,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"1998-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0005-2760(98)00064-2","citationCount":"49","resultStr":"{\"title\":\"Cloning of conserved genes from Zymomonas mobilis and Bradyrhizobium japonicum that function in the biosynthesis of hopanoid lipids1\",\"authors\":\"Michael Perzl , Ina G. Reipen , Susanne Schmitz , Karl Poralla , Hermann Sahm , Georg A. Sprenger , Elmar L. Kannenberg\",\"doi\":\"10.1016/S0005-2760(98)00064-2\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>The squalene-hopene cyclase (SHC) is the only enzyme involved in the biosynthesis of hopanoid lipids that has been characterized on the genetic level. To investigate if additional genes involved in hopanoid biosynthesis are clustered with the <em>shc</em> gene, we cloned and analyzed the nucleotide sequences located immediately upstream of the <em>shc</em> genes from <em>Zymomonas mobilis</em> and <em>Bradyrhizobium japonicum</em>. In <em>Z. mobilis</em>, five open reading frames (ORFs, designated as <em>hpnA–E</em>) were detected in a close arrangement with the <em>shc</em> gene. In <em>B. japonicum</em>, three similarly arranged ORFs (corresponding to <em>hpnC–E</em> from <em>Z. mobilis</em>) were found. The deduced amino acid sequences of <em>hpnC–E</em> showed significant similarity (58–62%) in both bacteria. Similarities to enzymes of other terpenoid biosynthesis pathways (carotenoid and steroid biosynthesis) suggest that these ORFs encode proteins involved in the biosynthesis of hopanoids and their intermediates. Expression of <em>hpnC</em> to <em>hpnE</em> from <em>Z. mobilis</em> as well as expression of <em>hpnC</em> from <em>B. japonicum</em> in <em>Escherichia coli</em> led to the formation of the hopanoid precursor squalene. This indicates that <em>hpnC</em> encodes a squalene synthase. The two additional ORFs (<em>hpnA</em> and <em>hpnB</em>) in <em>Z. mobilis</em> showed similarities to enzymes involved in the transfer and modification of sugars, indicating that they may code for enzymes involved in the biosynthesis of the complex, sugar-containing side chains of hopanoids.</p></div>\",\"PeriodicalId\":100162,\"journal\":{\"name\":\"Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1998-07-31\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/S0005-2760(98)00064-2\",\"citationCount\":\"49\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0005276098000642\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0005276098000642","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Cloning of conserved genes from Zymomonas mobilis and Bradyrhizobium japonicum that function in the biosynthesis of hopanoid lipids1
The squalene-hopene cyclase (SHC) is the only enzyme involved in the biosynthesis of hopanoid lipids that has been characterized on the genetic level. To investigate if additional genes involved in hopanoid biosynthesis are clustered with the shc gene, we cloned and analyzed the nucleotide sequences located immediately upstream of the shc genes from Zymomonas mobilis and Bradyrhizobium japonicum. In Z. mobilis, five open reading frames (ORFs, designated as hpnA–E) were detected in a close arrangement with the shc gene. In B. japonicum, three similarly arranged ORFs (corresponding to hpnC–E from Z. mobilis) were found. The deduced amino acid sequences of hpnC–E showed significant similarity (58–62%) in both bacteria. Similarities to enzymes of other terpenoid biosynthesis pathways (carotenoid and steroid biosynthesis) suggest that these ORFs encode proteins involved in the biosynthesis of hopanoids and their intermediates. Expression of hpnC to hpnE from Z. mobilis as well as expression of hpnC from B. japonicum in Escherichia coli led to the formation of the hopanoid precursor squalene. This indicates that hpnC encodes a squalene synthase. The two additional ORFs (hpnA and hpnB) in Z. mobilis showed similarities to enzymes involved in the transfer and modification of sugars, indicating that they may code for enzymes involved in the biosynthesis of the complex, sugar-containing side chains of hopanoids.