Improving the tolerance of Escherichia coli to medium-chain fatty acid production

Sherkhanov, Saken; Korman, Tyler P.; Bowie, James U.

doi:10.1016/j.ymben.2014.06.003

Cited by 68 publications

(58 citation statements)

References 32 publications

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“…In an attempt to produce shorter fatty acids, we expressed heterologous acyl-ACP thioesterase enzymes from five different species previously shown to have specificity toward acyl-ACP chains eight carbons in length (McMahon and Prather 2014; Sherkhanov et al 2014). We evaluated Y. lipolytica PO1f strains with acyl-ACPthioesterases from A. tetradius, Cuphea hookeriana, Cuphea palustris, Clostridium perfringens, and U. californica randomly integrated into the genome (referred to as PO1f-AtACPT, PO1f-AChACPT, PO1f-CpaACPT, PO1f-CpeACPT, and PO1f-UcACPT, respectively) for their ability to produce medium-chain fatty acids (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…These results demonstrate that an acyl-ACP thioesterase with specificity for shorter acyl-ACP molecules is capable of associating with the FAS complex and catalyzing the termination of fatty acid elongation to produce medium-chain fatty acids. It was previously shown that all enzymes tested catalyzed formation of octanoic acid in an E. coli system (McMahon and Prather 2014;Sherkhanov et al 2014), yet decanoic acid was the product in most cases when they were expressed in Y. lipolytica. It is unknown how a plant or bacterial acyl-ACP thioesterase is capable of associating with the fungal type I FAS, whose structure is known to orient most of the pertinent catalytic sites within a capsule-like structure (Lomakin et al 2007;Leibundgut et al 2008).…”

Section: Discussionmentioning

confidence: 98%

“…Enzymes previously shown to produce fatty acids as short as octanoic acid in E. coli (McMahon and Prather 2014;Sherkhanov et al 2014) were able to produce either octanoic or decanoic acid in Y. lipolytica. The production of novel medium-chain fatty acids accompanied reduction in native long-chain fatty acid production, which resulted in a reduction of cell growth and an increase in lipid yields.…”

Section: Discussionmentioning

confidence: 99%

“…Many acyl-ACP thioesterases have been identified that use mediumchain acyl-ACP molecules as their substrate (McMahon and Prather 2014;Voelker et al 1997;Pollard et al 1991;Dörmann et al 1993). Numerous studies have shown that expression of a medium-chain acyl-ACP-specific acyl-ACP thioesterase leads to production of shorter fatty acids in Escherichia coli as well as several plant species (Eccleston et al 1996;Sherkhanov et al 2014;Liu et al 2010;Lu et al 2008;Steen et al 2010;Voelker et al 1992).…”

Section: Introductionmentioning

confidence: 99%

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Engineering Yarrowia lipolytica for production of medium-chain fatty acids

Rutter

Zhang

Rao

2015

Appl Microbiol Biotechnol

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Lipids are naturally derived products that offer an attractive, renewable alternative to petroleum-based hydrocarbons. While naturally produced long-chain fatty acids can replace some petroleum analogs, medium-chain fatty acid would more closely match the desired physical and chemical properties of currently employed petroleum products. In this study, we engineered Yarrowia lipolytica, an oleaginous yeast that naturally produces lipids at high titers, to produce medium-chain fatty acids. Five different acyl-acyl carrier protein (ACP) thioesterases with specificity for medium-chain acyl-ACP molecules were expressed in Y. lipolytica, resulting in formation of either decanoic or octanoic acid. These novel fatty acid products were found to comprise up to 40 % of the total cell lipids. Furthermore, the reduction in chain length resulted in a twofold increase in specific lipid productivity in these engineered strains. The medium-chain fatty acids were found to be incorporated into all lipid classes.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Engineering Yarrowia lipolytica for production of medium-chain fatty acids

Rutter

Zhang

Rao

2015

Appl Microbiol Biotechnol

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“…Avoiding costly gene expression during growth phases of a culture should help ensure that the final population has not lost those functions when they are eventually expressed. For example, delayed selection can be achieved in cell culture by adding a chemical that activates a heterologous metabolic pathway late in the growth phase [30]. This method has obvious parallels to the traditional approach of limiting an intervention to late ages to weaken selection [3], but many new methods for engineering cells to implement this type of spatiotemporal control are now available.…”

Section: Engineering Fitness Landscapes To Suppress Evolutionmentioning

confidence: 99%

Arresting Evolution

Bull

Barrick

2017

Trends in Genetics

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Evolution in the form of selective breeding has long been harnessed as a useful tool by humans. However, rapid evolution can also be a danger to our health and a stumbling block for biotechnology. Unwanted evolution can underlie the emergence of drug and pesticide resistance, cancer, and weeds. It makes live vaccines and engineered cells inherently unreliable and unpredictable, and therefore potentially unsafe. Yet, there are strategies that have been and can possibly be used to stop or slow many types of evolution. We review and classify existing population genetics-inspired methods for arresting evolution. Then, we discuss how genome editing techniques enable a radically new set of approaches to limit evolution.

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Engineering Robustness of Microbial Cell Factories

Gong

Nielsen

Zhou

2017

Biotechnology Journal

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Metabolic engineering and synthetic biology offer great prospects in developing microbial cell factories capable of converting renewable feedstocks into fuels, chemicals, food ingredients, and pharmaceuticals. However, prohibitively low production rate and mass concentration remain the major hurdles in industrial processes even though the biosynthetic pathways are comprehensively optimized. These limitations are caused by a variety of factors unamenable for host cell survival, such as harsh industrial conditions, fermentation inhibitors from biomass hydrolysates, and toxic compounds including metabolic intermediates and valuable target products. Therefore, engineered microbes with robust phenotypes is essential for achieving higher yield and productivity. In this review, the recent advances in engineering robustness and tolerance of cell factories is described to cope with these issues and briefly introduce novel strategies with great potential to enhance the robustness of cell factories, including metabolic pathway balancing, transporter engineering, and adaptive laboratory evolution. This review also highlights the integration of advanced systems and synthetic biology principles toward engineering the harmony of overall cell function, more than the specific pathways or enzymes.

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Improving the tolerance of Escherichia coli to medium-chain fatty acid production

Cited by 68 publications

References 32 publications

Engineering Yarrowia lipolytica for production of medium-chain fatty acids

Engineering Yarrowia lipolytica for production of medium-chain fatty acids

Arresting Evolution

Engineering Robustness of Microbial Cell Factories

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