Highly efficient whole-cell biosynthesis of putrescine by recombinant Escherichia coli

Li, Guohui; Huang, Dixuan; Wang, Li; Deng, Yu

doi:10.1016/j.bej.2020.107859

Cited by 10 publications

(5 citation statements)

References 19 publications

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“…6,8 Putrescine is a polyamine found in many organisms and its biosynthesis can occur via different enzymatic pathways. 9−11 Commercially, putrescine has many industrial applications, including as a precursor for the production of polyamide polymers, 12 such as nylon-4,6, which is produced by the polycondensation of putrescine and adipic acid, 13 and is commonly used in the production of biomedical instruments and textiles as well as automobile components. 14,15 Most putrescine used today, however, is chemically produced by the hydrogenation of succinonitrile, which is neither sustainable nor environmental-friendly, due to its use of nonrenewable propene as a raw material and the in situ production of the toxic intermediate hydrogen cyanide.…”

Section: ■ Introductionmentioning

confidence: 99%

“…Putrescine is a polyamine found in many organisms and its biosynthesis can occur via different enzymatic pathways. − Commercially, putrescine has many industrial applications, including as a precursor for the production of polyamide polymers, such as nylon-4,6, which is produced by the polycondensation of putrescine and adipic acid, and is commonly used in the production of biomedical instruments and textiles as well as automobile components. , Most putrescine used today, however, is chemically produced by the hydrogenation of succinonitrile, which is neither sustainable nor environmental-friendly, due to its use of nonrenewable propene as a raw material and the in situ production of the toxic intermediate hydrogen cyanide. , In an effort to minimize the negative impact on the environment, the biosynthesis of putrescine has been extensively explored, with nearly all efforts focusing on the engineering of microorganisms, with Escherichia coli being the most studied. In many of these whole cell systems, extensive genetic engineering was performed to overexpress the enzymes associated with two key putrescine synthetic pathways: (1) the ornithine decarboxylase (ODC) pathway that utilizes ODC to directly convert l -ornithine to putrescine and (2) the arginine decarboxylase (ADC) pathway that employs ADC and agmatinase to convert l -arginine to putrescine. ,,− Additional genetic modifications, such as deletion of putrescine degradation pathways and disruption of competing pathways of precursor utilization, have also been carried out in order to enhance the production of putrescine. ,− ,, Nevertheless, the amounts produced are generally low, in part because high concentrations of putrescine are toxic to the organisms. , …”

Section: Introductionmentioning

confidence: 99%

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Genetically Encoded Multienzyme Particles for the Biosynthesis of Putrescine from l-Arginine

Xiong,

Heater,

et al. 2024

ACS Sustainable Chem. Eng.

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Immobilized enzyme biocatalysts have wide industrial applications, but the multiple steps involved in their preparation significantly increase the time, effort, and cost to produce them. Our group has previously developed a platform for the direct immobilization of enzymes based on the crystal-forming Cry3Aa protein. The application of this platform has been demonstrated for the direct immobilization of individual enzymes, with a molecular size limitation of approximately 50 kDa. However, the biosynthesis of many compounds involve cascade reactions utilizing multiple enzymes of various sizes including those larger than this limit. Herein, we report the use of a larger Cry homologue, Cry1Ab, for the in vivo coimmobilization of two enzymes involved in the biosynthesis of putrescine�human arginase I (hArg, 35 kDa) and ornithine decarboxylase (ODC, 79 kDa). We show that Cry1Ab-mediated immobilization enhanced the thermostability of these enzymes, allowing for the efficient and recyclable conversion of arginine to putrescine, yielding 30 mM putrescine per cycle at nearly 100% conversion in 30 min for the first seven cycles. Furthermore, the putrescine produced could be directly converted to the thermostable polyamide nylon-4,6.

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Section: ■ Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Genetically Encoded Multienzyme Particles for the Biosynthesis of Putrescine from l-Arginine

Xiong,

Heater,

et al. 2024

ACS Sustainable Chem. Eng.

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“…Like most of the other amino acids and their derivatives, , putrescine is most widely produced by Escherichia coli and Corynebacterium glutamicum. The pathways of ornithine decarboxylase (ODC) and arginine decarboxylase (ADC) are two different putrescine synthetic routes that are common in natural organisms . XQ52 of E.…”

Section: Introductionmentioning

confidence: 99%

“…natural organisms. 12 XQ52 of E. coli was constructed by deleting the competing pathways and overexpressing the putrescine biosynthetic genes. 13 Then, the expression level of argF (encoding ornithine carbamoyltransferase) and glnA (encoding glutamine synthetase) was repressed through the sRNA on XQ52 strain, and the final strain produced 42.3 g/L of putrescine.…”

Section: ■ Introductionmentioning

confidence: 99%

Production of Putrescine in Metabolic Engineering Corynebacterium crenatum by Mixed Sugar Fermentation

Yang

Qiu

Zhu

et al. 2022

ACS Sustainable Chem. Eng.

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Putrescine, a biogenic diamine, serves as an important component in various polyamides, medicine, and surfactants. One challenge for efficient putrescine production is to construct high-efficiency microbial cells. In this study, we designed the metabolic engineering strategies to significantly enhance the putrescine titer in Corynebacterium crenatum SYPA. First, an optimal synthetic pathway of putrescine comprising arginine decarboxylase (speA) and agmatinase (speB) genes from Escherichia coli was selected and introduced into C. crenatum. For efficient production of putrescine, the expression of agmatinase was optimized through a ribosome binding site regulation strategy. Next, the putrescine yield was furthermore increased by the knockout of snaA, speE, and cgmR to block putrescine degradation and overexpression of exporter CgmA. Additionally, the xylose utilization pathway was constructed for putrescine synthesis. Finally, the titer of putrescine was 41.5, 36.8, and 33.4 g/L from glucose, mixed sugar, and simulated wheat straw hydrolysates by fed-batch culture for 72 h, respectively. The yield was 0.18 g/g glucose, 0.15 g/g sugar, and 0.14 g/g sugar, respectively. To our knowledge, those results were the highest putrescine titers ever reported in the engineering of C. crenatum and Corynebacterium glutamicum. The engineering strains had the potential to produce putrescine from biomass hydrolysates.

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“…Qian et al ( 2009 ) overproduced putrescine through the ODC pathway in E. coli K-12, and the batch culture experiment resulted in 1.68 g/L (19.0 mM) in the 10 g/L of the glucose-enriched medium. Recently, Li et al ( 2020b ) overexpressed the ADC pathway and improved putrescine production using E. coli BL21(DE3). They produced putrescine using an SOB medium supplemented with 8 g/L of glucose and 12 g/L of arginine, resulting in 4.77 g/L of putrescine in batch culture.…”

Section: Introductionmentioning

confidence: 99%

Improvement of putrescine production through the arginine decarboxylase pathway in Escherichia coli K-12

et al. 2021

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In the bio-based polymer industry, putrescine is in the spotlight for use as a material. We constructed strains of Escherichia coli to assess its putrescine production capabilities through the arginine decarboxylase pathway in batch fermentation. N-Acetylglutamate (ArgA) synthase is subjected to feedback inhibition by arginine. Therefore, the 19th amino acid residue, Tyr, of argA was substituted with Cys to desensitize the feedback inhibition of arginine, resulting in improved putrescine production. The inefficient initiation codon GTG of argA was substituted with the effective ATG codon, but its replacement did not affect putrescine production. The essential genes for the putrescine production pathway, speA and speB, were cloned into the same plasmid with argAATG Y19C to form an operon. These genes were introduced under different promoters; lacIp, lacIqp, lacIq1p, and T5p. Among these, the T5 promoter demonstrated the best putrescine production. In addition, disruption of the puuA gene encoding enzyme of the first step of putrescine degradation pathway increased the putrescine production. Of note, putrescine production was not affected by the disruption of patA, which encodes putrescine aminotransferase, the initial enzyme of another putrescine utilization pathway. We also report that the strain KT160, which has a genomic mutation of YifEQ100TAG, had the greatest putrescine production. At 48 h of batch fermentation, strain KT160 grown in terrific broth with 0.01 mM IPTG produced 19.8 mM of putrescine.

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Highly efficient whole-cell biosynthesis of putrescine by recombinant Escherichia coli

Cited by 10 publications

References 19 publications

Genetically Encoded Multienzyme Particles for the Biosynthesis of Putrescine from l-Arginine

Genetically Encoded Multienzyme Particles for the Biosynthesis of Putrescine from l-Arginine

Production of Putrescine in Metabolic Engineering Corynebacterium crenatum by Mixed Sugar Fermentation

Improvement of putrescine production through the arginine decarboxylase pathway in Escherichia coli K-12

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