Metabolic engineering of Escherichia coli for lauryl glucoside production via a newly designed biosynthetic pathway

วิศวกรรมเมแทบอลิกของ Escherichia coli สำหรับการผลิตลอริลกลูโคไซด์ผ่านวิถีชีวสังเคราะห์ที่ออกแบบขึ้นใหม่

Name: Kasimaporn Promubon

LCSH: Glucosides

LCSH: Transgenic organisms

LCSH: Genetic engineering

Abstract: Lauryl glucoside is a mild, non-ionic surfactant widely used in cosmetic products, appreciated for its biodegradability and low toxicity. However, its conventional production process is considered unsustainable, prompting the search for alternative approaches. In this study, a newly designed biosynthetic pathway was established in Escherichia coli BL21 (DE3) to enable lauryl glucoside production using a microbial cell factory concept. The potentially toxic precursor, 1-dodecanol, showed no adverse effect on cell growth during 48 h of incubation. The production of 1-dodecanol was significantly enhanced, reaching a titre of 0.99 ± 0.02 mM (185.39 ± 3.62 mg/L) and a yield of 11.21 ± 0.28 mmol/mol glucose, by applying a solvent overlay and optimising IPTG concentrations. These optimised conditions were then used to screen for effective UDP-glycosyltransferases capable of converting 1-dodecanol into lauryl glucoside. Among six enzymes, MtH2 from Medicago truncatula exhibited the highest conversion efficiency, producing lauryl glucoside at a titre of 0.0021 ± 0.0002 mM (0.72± 0.07 mg/L) and a yield of 0.04 ± 0.0039 mmol/mol glucose. The identity of the product was confirmed through HPLC and targeted LC-MS analysis. Moreover, the study identified the limited availability of 1-dodecanol as a major bottleneck in the pathway. When cells were supplemented with 2.0 mM (372.68 mg/L) 1-dodecanol, lauryl glucoside production increased, achieving a titre of 0.04 ± 0.001 mM (13.44± 0.21 mg/L) and a yield of 0.69 ± 0.01 mmol/mol glucose. Fermentation product profiling also indicated a redirection of carbon flux from acetate toward the target product. Altogether, this study presents a successfully engineered biosynthetic route for lauryl glucoside in E. coli and highlights precursor availability as a key limiting factor, contributing to the development of more sustainable and efficient production methods.

Chiang Mai University. Library

Address: CHIANG MAI

Email: cmulibref@cmu.ac.th

Name: Pachara Sattayawat

Role: Advisor

Created: 2025

Modified: 2025-08-08

Issued: 2025-08-08

วิทยานิพนธ์/Thesis

application/pdf

eng

DegreeName: Master of Science

Level: Master's Degree

Descipline: Applied Microbiology

Grantor: Chiang Mai University

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