کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6494198 | 44798 | 2016 | 13 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
CRISPR interference-guided balancing of a biosynthetic mevalonate pathway increases terpenoid production
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کلمات کلیدی
UTR1-deoxy-D-xylulose 5-phosphateHMG-CoAdCas9CRISPRiIPPRT-PCRMEPsfGFPPAMDXPDMAPPIPTGCRISPR interferenceRBSPBSCATT7RNAPCRISPRISPA3-hydroxy-3-methyl-glutaryl-CoAMevalonateReal-time PCR - PCR به موقعsingle guide RNA - RNA راهنما تنهاsgRNA - scRNAT7 RNA polymerase - T7 RNA پلیمرازflame ionization detector - آشکارساز یونیزاسیون شعلهEscherichia coli - اشریشیا کُلیisopropyl β-D-1-thiogalactopyranoside - ایزوپروپیل β-D-1-thiogalactopyranosideisopentenyl diphosphate - ایسپنتنیل دی فسفاتclustered regularly interspaced short palindromic repeats - به طور منظم تکرار پینگندرومی کوتاه مدت میان دو طرف تقسیم می شودTerpenoid - ترپنوئیدFID - درdimethylallyl diphosphate - دی متیللید دی فسفاتribosome binding site - سایت اتصال ریبوزومterrific broth - سوپ فوق العادهfarnesyl diphosphate synthase - فارسیل دی فسفات سنتازPhosphate buffered saline - فسفات بافر شورLuria-Bertani - لواریا بارتانیMVA - مالیات بر ارزش افزودهmethylerythritol 4-phosphate - متیلیرتریتریل 4-فسفاتMevalonate pathway - مسیر مولکولیuntranslated region - منطقه غیر ترجمهprotospacer adjacent motif - موتیف مجاور protospacerpolymerase chain reaction - واکنش زنجیره ای پلیمرازPCR - واکنش زنجیرهٔ پلیمرازGas chromatography - کروماتوگرافی گازیchloramphenicol acetyl transferase - کلرامفنیکول استیل ترانسفراز
موضوعات مرتبط
مهندسی و علوم پایه
مهندسی شیمی
بیو مهندسی (مهندسی زیستی)
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: CRISPR interference-guided balancing of a biosynthetic mevalonate pathway increases terpenoid production CRISPR interference-guided balancing of a biosynthetic mevalonate pathway increases terpenoid production](/preview/png/6494198.png)
چکیده انگلیسی
Methods for simple and efficient regulation of metabolic pathway genes are essential for maximizing product titers and conversion yields, and for minimizing the metabolic burden caused by heterologous expression of multiple genes often in the operon context. Clustered regularly interspaced short palindromic repeats (CRISPR) interference (CRISPRi) is emerging as a promising tool for transcriptional modulation. In this study, we developed a regulatable CRISPRi system for fine-tuning biosynthetic pathways and thus directing carbon flux toward target product synthesis. By exploiting engineered Escherichia coli harboring a biosynthetic mevalonate (MVA) pathway and plant-derived terpenoid synthases, the CRISPRi system successfully modulated the expression of all the MVA pathway genes in the context of operon and blocked the transcription of the acetoacetyl-CoA thiolase enzyme that catalyzes the first step in the MVA pathway. This CRISPRi-guided balancing of expression of MVA pathway genes led to enhanced production of (-)-α-bisabolol (C15) and lycopene (C40) and alleviation of cell growth inhibition that may be caused by expression of multiple enzymes or production of toxic intermediate metabolites in the MVA pathway. Coupling CRISPRi to cell growth by regulating an endogenous essential gene (ispA) increased isoprene (C5) production. The regulatable CRISPRi system proved to be a robust platform for systematic modulation of biosynthetic and endogenous gene expression, and can be used to tune biosynthetic metabolic pathways. Its application can enable the development of microbial 'smart cell' factories that can produce other industrially valuable products in the future.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Metabolic Engineering - Volume 38, November 2016, Pages 228-240
Journal: Metabolic Engineering - Volume 38, November 2016, Pages 228-240
نویسندگان
Seong Keun Kim, Gui Hwan Han, Wonjae Seong, Haseong Kim, Seon-Won Kim, Dae-Hee Lee, Seung-Goo Lee,