Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli.docVIP

下载本文档

3
0
约4.98万字
约 11页
2017-05-07 发布于上海
举报
版权申诉

Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli.doc

1、原创力文档（book118）网站文档一经付费（服务费），不意味着购买了该文档的版权，仅供个人/单位学习、研究之用，不得用于商业用途，未经授权，严禁复制、发行、汇编、翻译或者网络传播等，侵权必究。。
2、本站所有内容均由合作方或网友上传，本站不对文档的完整性、权威性及其观点立场正确性做任何保证或承诺！文档内容仅供研究参考，付费前请自行鉴别。如您付费，意味着您自己接受本站规则且自行承担风险，本站不退款、不进行额外附加服务；查看《如何避免下载的几个坑》。如果您已付费下载过本站文档，您可以点击这里二次下载。
3、如文档侵犯商业秘密、侵犯著作权、侵犯人身权等，请点击“版权申诉”（推荐），也可以打举报电话：400-050-0827(电话支持时间：9:00-18:30)。
4、该文档为VIP文档，如果想要下载，成为VIP会员后，下载免费。
5、成为VIP后，下载本文档将扣除1次下载权益。下载后，不支持退款、换文档。如有疑问请联系我们。
6、成为VIP后，您将拥有八大权益，权益包括：VIP文档下载权益、阅读免打扰、文档格式转换、高级专利检索、专属身份标志、高级客服、多端互通、版权登记。
7、VIP文档为合作方或网友上传，每下载1次，网站将根据用户上传文档的质量评分、类型等，对文档贡献者给予高额补贴、流量扶持。如果你也想贡献VIP文档。上传文档

Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli

CombiningGenotypeImprovementandStatisticalMedia OptimizationforIsoprenoidProductioninE.coli CongqiangZhang1,XixianChen1,RuiyangZou1,KangZhou1,GregoryStephanopoulos1,3 ,Heng- PhonToo1,2 * 1ChemicalandPharmaceuticalEngineering,Singapore-MITAlliance,Singapore,Singapore,2DepartmentofBiochemistry,NationalUniversityofSingapore,Singapore, Singapore,3DepartmentofChemicalEngineering,MassachusettsInstituteofTechnology,Cambridge,Massachusetts,UnitedStatesofAmerica Abstract Isoprenoids arealargeanddiverse class ofcompounds that includesmanyhigh valuenaturalproducts andarethusin greatdemand.Tomeettheincreasingdemandforisoprenoidcompounds,metabolicengineeringofmicrobeshasbeen used to produce isoprenoids in an economical and sustainable manner. To achieve high isoprenoid yields using this technology,theavailabilityofmetabolicprecursorsfeedingthedeoxyxylulosephosphate(DXP)pathway,responsiblefor isoprenoid biosynthesis, has to be optimized. In this study, phosphoenolpyruvate, a vital DXP pathway precursor, was enriched by deleting the genes encoding the carbohydrate phosphotransferase system (PTS) in E. coli. Production of lycopene(aC40isoprenoid)wasmaximizedbyoptimizinggrowthmediumandcultureconditions.Inoptimizedconditions, thelycopeneyieldfromPTSmutantwassevenfoldhigherthanthatobtainedfromthewildtypestrain.Thisresultedinthe highestreportedspecificyieldoflycopeneproducedfromtheDXPpathwayinE.colitodate(20,000mg/gdrycellweight). Both the copy number of the plasmid encoding the lycopene biosynthetic genes and the expression were found to be increased in the optimized media. Deletion of PTS together with a similar optimization strategy was also successful in enhancing the production of amorpha-1,4-diene, a distinct C15 isoprenoid, suggesting that the approaches developed hereincanbegenerallyappliedtooptimizeproductionofotherisoprenoids. Citation:ZhangC,ChenX,ZouR,ZhouK,StephanopoulosG,etal.(2013)CombiningGenotypeImprovementandStatisticalMediaOptimizationforIsoprenoid ProductioninE