Engineering <i>Escherichia coli</i> for efficient coproduction of polyhydroxyalkanoates and 5-aminolevulinic acid

Zhang, Xue; Zhang, Jian; Xu, Jiasheng; Zhao, Qian; Wang, Qian; Qi, Qingsheng

doi:10.1007/s10295-017-1990-4

Cited by 19 publications

(10 citation statements)

References 40 publications

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“…The parent strain BW24 produced 5.3 g/L ALA and the highest OD 600 reached 30.5 (Figure b,c). In previous studies, acetate accumulation was observed for the ALA‐producing E. coli strain (X. Zhang et al, ). In our case, acetate was also the main by‐product, which was accumulated at 6.1 g/L at the end of fermentation (Figure d).…”

Section: Resultsmentioning

confidence: 82%

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Enhancing 5‐aminolevulinic acid tolerance and production by engineering the antioxidant defense system of Escherichia coli

Zhu

Chen

Wang

et al. 2019

Biotech & Bioengineering

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5-Aminolevulinic acid (ALA) is a value-added compound with potential applications in the fields of agriculture and medicine. Although massive efforts have recently been devoted to building microbial producers of ALA through metabolic engineering, few studies focused on the cellular response and tolerance to ALA. In this study, we demonstrated that ALA caused severe cell damage and morphology change of Escherichia coli via generating reactive oxygen species (ROS), which were further determined to be mainly hydrogen peroxide and superoxide anion radical. ALA treatment activated the native antioxidant defense system by upregulating catalase (CAT) and superoxide dismutase (SOD) expression to combat ROS. Further overexpressing CAT (encoded by katG and katE) and SOD (encoded by sodA, sodB, and sodC) not only improved ALA tolerance but also its production level. Notably, coexpression of katE and sodB in an ALA synthase expressing strain enhanced the biomass and final ALA titer by 81% and 117% (11.5 g/L) in a 5 L bioreactor, respectively. This study demonstrates the importance of tolerance engineering in strain development. Reinforcing the antioxidant defense system holds promise to improve the bioproduction of chemicals that cause oxidative stress.

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Section: Resultsmentioning

confidence: 82%

“…Cell integrity was monitored by double staining of cells by fluorescent probes fluorescein diacetate (FDA) and propidium iodide (PI) as described previously (M. Zhang et al, ). Bacterial cells with intact membranes will exclude PI but be stained by FDA and emit green fluorescence.…”

Section: Methodsmentioning

confidence: 99%

Enhancing 5‐aminolevulinic acid tolerance and production by engineering the antioxidant defense system of Escherichia coli

Zhu

Chen

Wang

et al. 2019

Biotech & Bioengineering

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“…To explore the feasibility of T4 DNA ligase-mediated microbial mutagenesis, the polyhydroxybutyrate (PHB) producing E. coli XLPHB, which harbors chromosomal integrated PHB biosynthesis genes phbCAB from Ralstonia eutropha , was subjected to mutagenesis and screening with T4 DNA ligase expressed in vivo [12] . Generally, there was nearly no colony on the plate after the cells were treated by ARTP for 60-s (Additional file 1: Fig.…”

Section: Resultsmentioning

confidence: 99%

“…E. coli DH5α strain was used for molecular cloning and plasmids propagation. Engineered PHB producing E. coli XLPHB strain was subjected to ARTP treatment [12, 31].…”

Section: Methodsmentioning

confidence: 99%

“…With the development of molecular biology, system biology and synthetic biology, rational modification of microbes to obtain desired properties became fashionable and efficient [10–12]. Endogenous genes were up or down regulated to redirect the metabolic flow to desired routes, while exogenous genes were adopted to obtain the properties that was not existed in the host [13–16].…”

Section: Introductionmentioning

confidence: 99%

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The phage T4 DNA ligase in vivo improves the survival-coupled bacterial mutagenesis

Wang

Liu

et al. 2019

Microb Cell Fact

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Background Microbial mutagenesis is an important avenue to acquire microbial strains with desirable traits for industry application. However, mutagens either chemical or physical used often leads narrow library pool due to high lethal rate. The T4 DNA ligase is one of the most widely utilized enzymes in modern molecular biology. Its contribution to repair chromosomal DNA damages, therefore cell survival during mutagenesis will be discussed. Results Expression of T4 DNA ligase in vivo could substantially increase cell survival to ionizing radiation in multiple species. A T4 mediated survival-coupled mutagenesis approach was proposed. When polyhydroxybutyrate (PHB)-producing E . coli with T4 DNA ligase expressed in vivo was subjected to ionizing radiation, mutants with improved PHB production were acquired quickly owing to a large viable mutant library generated. Draft genome sequence analysis showed that the mutants obtained possess not only single nucleotide variation (SNV) but also DNA fragment deletion, indicating that T4 DNA ligase in vivo may contribute to the repair of DNA double strand breaks. Conclusions Expression of T4 DNA ligase in vivo could notably enhance microbial survival to excess chromosomal damages caused by various mutagens. Potential application of T4 DNA ligase in microbial mutagenesis was explored by mutating and screening PHB producing E. coli XLPHB strain. When applied to atmospheric and room temperature plasma (ARTP) microbial mutagenesis, large survival pool was obtained. Mutants available for subsequent screening for desirable features. The use of T4 DNA ligase we were able to quickly improve the PHB production by generating a larger viable mutants pool. This method is a universal strategy can be employed in wide range of bacteria. It indicated that traditional random mutagenesis became more powerful in combine with modern genetic molecular biology and has exciting prospect. Electronic supplementary material The online version of this article (10.1186/s12934-019-1160-7) contains supplementary material, which is available to authorized users.

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Integrated Biorefinery Design with Techno‐Economic and Life Cycle Assessment Tools in Polyhydroxyalkanoates Processing

Andhalkar,

Foong,

Kee

et al. 2023

Macro Materials & Eng

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To support and move toward a sustainable bioeconomy, the production of polyhydroxyalkanoates (PHAs) using renewable biomass has acquired more attention. However, expensive biomass pretreatment and low yield of PHAs pose significant disadvantages in its large‐scale production. To overcome such limitations, the most recent advances in metabolic engineering strategies used to develop high‐performance strains that are leading to a new manufacturing concept converting biomass to PHAs with co‐products such as amino acids, proteins, biohydrogen, biosurfactants, and various fine chemicals are critically summarized. This review article presents a comprehensive roadmap that highlights the integrated biorefinery strategies, lifecycle analysis, and techno‐economic assessment for sustainable and economic PHAs production. Finally, current and future challenges that must be addressed to transfer this technology to real‐world applications are reviewed.

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Engineering Escherichia coli for efficient coproduction of polyhydroxyalkanoates and 5-aminolevulinic acid

Cited by 19 publications

References 40 publications

Enhancing 5‐aminolevulinic acid tolerance and production by engineering the antioxidant defense system of Escherichia coli

Enhancing 5‐aminolevulinic acid tolerance and production by engineering the antioxidant defense system of Escherichia coli

The phage T4 DNA ligase in vivo improves the survival-coupled bacterial mutagenesis

Integrated Biorefinery Design with Techno‐Economic and Life Cycle Assessment Tools in Polyhydroxyalkanoates Processing

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