کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6494099 | 1418332 | 2018 | 38 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Rewiring FadR regulon for the selective production of Ï-hydroxy palmitic acid from glucose in Escherichia coli
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کلمات کلیدی
UFACyPHFASFAFASACPFFAω-hydroxylation - ω-هیدروکسیلینگFatty acid - اسید چربFree fatty acid - اسید چرب آزادsaturated fatty acid - اسید چرب اشباع شدهfatty acid synthase - اسید چرب سنتازunsaturated fatty acid - اسید چرب غیر اشباعHydroxy fatty acid - اسید چرب هیدروکسیThioesterase - تیواستروئیدTranscriptional regulator - رگولاتور رونویسیCytochrome P450 - سیتوکروم پی۴۵۰FadR - فدرالHPA - میلی بار یا هکتوپاسکالCo-culture - همکاری فرهنگیacyl carrier protein - پروتئین حامل آکیل
موضوعات مرتبط
مهندسی و علوم پایه
مهندسی شیمی
بیو مهندسی (مهندسی زیستی)
پیش نمایش صفحه اول مقاله

چکیده انگلیسی
Ï-Hydroxy palmitic acid (Ï-HPA) is a valuable compound for an ingredient of artificially synthesized ceramides and an additive for lubricants and adhesives. Production of such a fatty acid derivative is limited by chemical catalysis, but plausible by biocatalysis. However, its low productivity issue, including formations of unsaturated fatty acid (UFA) byproducts in host cells, remains as a hurdle toward industrial biological processes. In this study, to achieve selective and high-level production of Ï-HPA from glucose in Escherichia coli, FadR, a native transcriptional regulator of fatty acid metabolism, and its regulon were engineered. First, FadR was co-expressed with a thioesterase with a specificity toward palmitic acid production to enhance palmitic acid production yield, but a considerable quantity of UFAs was also produced. In order to avoid the UFA production caused by fadR overexpression, FadR regulon was rewired by i) mutating FadR consensus binding sites of fabA or fabB, ii) integrating fabZ into fabI operon, and iii) enhancing the strength of fabI promoter. This approach led to dramatic increases in both proportion (48.3-83.0%) and titer (377.8â¯mg/L to 675.8â¯mg/L) of palmitic acid, mainly due to the decrease in UFA synthesis. Introducing a fatty acid Ï-hydroxylase, CYP153A35, into the engineered strain resulted in a highly selective production of Ï-HPA (83.5â¯mg/L) accounting for 87.5% of total Ï-hydroxy fatty acids. Furthermore, strategies, such as i) enhancement in CYP153A35 activity, ii) expression of a fatty acid transporter, iii) supplementation of triton X-100, and iv) separation of the Ï-HPA synthetic pathway into two strains for a co-culture system, were applied and resulted in 401.0â¯mg/L of Ï-HPA production. For such selective productions of palmitic acid and Ï-HPA, the rewiring of FadR regulation in E. coli is a promising strategy to develop an industrial process with economical downstream processing.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Metabolic Engineering - Volume 47, May 2018, Pages 414-422
Journal: Metabolic Engineering - Volume 47, May 2018, Pages 414-422
نویسندگان
Joonwon Kim, Hee-Wang Yoo, Minsuk Kim, Eun-Jung Kim, Changmin Sung, Pyung-Gang Lee, Beom Gi Park, Byung-Gee Kim,