Efficient agmatine production using an arginine decarboxylase with substrate‐specific activity

Sun, Anran; Song, Wei; Qiao, Weihua; Chen, Xiulai; Liu, Jia; Luo, Qiuling; Li, Liming

doi:10.1002/jctb.5245

Cited by 7 publications

(4 citation statements)

References 23 publications

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“…Phospholipases have been used in scientific and medical research, such as inhibitors for generating anti-inflammatory agents and as diagnostic markers for microbial infections [ 30 , 61 ]. Arginine decarboxylase catalyses conversion of L-arginine into agmatine, a valuable pharmaceutical intermediate with various potential therapeutic functions in neurotransmitter systems, nitric oxide synthesis, and polyamine metabolism [ 60 ].…”

Section: Discussionmentioning

confidence: 99%

Biotechnologically potential genes in a polysaccharide-degrading epibiont of the Indonesian brown algae Hydroclathrus sp.

Ethica

Zilda²,

Oedjijono

et al. 2023

Journal of Genetic Engineering and Biotechnology

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Background Marine bacteria have recently attracted increasing attention to be harnessed for the production of valuable enzymes, vitamins, and bioactive compounds. Bacteria associated with the surfaces of marine macroalgae, called epibionts, are particularly interesting from ecological and biotechnological points of view, as they often exhibit antimicrobial activities to compete with pathogenic bacteria for nutrients and spaces. In search for biotechnologically potential genes from marine bacteria, we sequenced and analysed the genome of the epibiont HI03-3b, a polysaccharide-degrading bacterium associated with the surface of the Indonesian brown algae Hydroclathrus sp. Results The algal epibiont HI03-3b has a genome of approximately 4,860,704 bp in size with 42.02 mol% G + C content, consisting of 5655 open reading frames (ORFs), 4409 genes coding for proteins (CDSs), 94 genes for tRNAs, and 32 genes for rRNAs. The genome sequence of HI03-3b was most closely related to that of Cytobacillus firmus NCTC10335 with the average amino acid identity (AAI) of 95.0 %, average nucleotide identity (ANI) of 94.1 %, and a recommended DNA-DNA hybridization (DDH) of 57.60 %. These scores are lower than the most frequently used standard for species demarcation (95% ANI cutoff) and the new species threshold (DDH > 70.0% for the same bacterial species). Some differences in genome features and gene composition were observed between HI03-3b and NCTC10335, such as genes encoding carbohydrate active enzymes. These suggest that HI03-3b is unique and likely a novel species within Cytobacillus genus, and we therefore proposed its name as Cytobacillus wakatobiense HI03-3b. Genome sequence analyses indicated the presence of genes involved not only in polysaccharide and protein degradation but also in vitamin and secondary metabolite biosynthesis. Some of them encode enzymes and compounds with biotechnological interest, such as protease, chitinase, subtilisin, pullulanase, and bacillolysin, which are often associated with antimicrobial or antibiofilm activities. This antimicrobial potential is supported by our finding that the extracellular protein fraction of this epibiont inhibited the growth of the bacterial pathogen Staphylococcus aureus. Conclusion The epibiont Cytobacillus HI03-3b harbours genes for polysaccharide and protein degradation as well as for natural product biosynthesis, suggesting its potential ecological roles in outcompeting other bacteria during biofilm formation as well as in protecting its algal host from predation. Due to the presence of genes for vitamin biosynthesis, it might also provide the algal host with vitamins for growth and development. Some of these metabolic genes are biotechnologically important, as they could become a platform for bioengineering to generate various seaweed-derived substances sustainably, such as antibiofilm agents and vitamins, which are beneficial for human health.

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Section: Discussionmentioning

confidence: 99%

Biotechnologically potential genes in a polysaccharide-degrading epibiont of the Indonesian brown algae Hydroclathrus sp.

Ethica

Zilda²,

Oedjijono

et al. 2023

Journal of Genetic Engineering and Biotechnology

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“…The supernatant of induced cells was added to the mixed reaction to determine the enzyme activity. The activity of ADC was measured by observing the production of agmatine . The amount of ADC that generated 1 μmol agmatine per minute was defined as one unit of enzyme activity.…”

Section: Methodsmentioning

confidence: 99%

“…The activity of ADC was measured by observing the production of agmatine. 32 The amount of ADC that generated 1 μmol agmatine per minute was defined as one unit of enzyme activity. The reaction was incubated at 37 °C for 10 min with a mixture of 50 mM L-arginine, 0.6 mM PLP, 2.5 mM MgSO 4 , and a certain amount of crude enzyme extracts in 50 mM Tris-HCl buffer (pH 8.0).…”

Section: ■ Materials and Methodsmentioning

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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“…The enzyme activity of ADC was determined according to the amount of agmatine produced at 37 °C for 10 min [ 35 ]. The reaction mixture contained 50 mM Tris-HCl buffer (pH 8.0), 50 mM l -arginine, 2.5 mM MgSO 4 , 0.6 mM pyridoxal phosphate (PLP) and an appropriate amount of purified enzyme.…”

Section: Methodsmentioning

confidence: 99%

High-level production of the agmatine in engineered Corynebacterium crenatum with the inhibition-releasing arginine decarboxylase

Yang

Zhu

et al. 2022

Microb Cell Fact

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Background Agmatine is a member of biogenic amines and is an important medicine which is widely used to regulate body balance and neuroprotective effects. At present, the industrial production of agmatine mainly depends on the chemical method, but it is often accompanied by problems including cumbersome processes, harsh reaction conditions, toxic substances production and heavy environmental pollution. Therefore, to tackle the above issues, arginine decarboxylase was overexpressed heterologously and rationally designed in Corynebacterium crenatum to produce agmatine from glucose by one-step fermentation. Results In this study, we report the development in the Generally Regarded as Safe (GRAS) l-arginine-overproducing C. crenatum for high-titer agmatine biosynthesis through overexpressing arginine decarboxylase based on metabolic engineering. Then, arginine decarboxylase was mutated to release feedback inhibition and improve catalytic activity. Subsequently, the specific enzyme activity and half-inhibitory concentration of I534D mutant were increased 35.7% and 48.1%, respectively. The agmatine production of the whole-cell bioconversion with AGM3 was increased by 19.3% than the AGM2. Finally, 45.26 g/L agmatine with the yield of 0.31 g/g glucose was achieved by one-step fermentation of the engineered C. crenatum with overexpression of speAI534D. Conclusions The engineered C. crenatum strain AGM3 in this work was proved as an efficient microbial cell factory for the industrial fermentative production of agmatine. Based on the insights from this work, further producing other valuable biochemicals derived from l-arginine by Corynebacterium crenatum is feasible.

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Efficient agmatine production using an arginine decarboxylase with substrate‐specific activity

Cited by 7 publications

References 23 publications

Biotechnologically potential genes in a polysaccharide-degrading epibiont of the Indonesian brown algae Hydroclathrus sp.

Biotechnologically potential genes in a polysaccharide-degrading epibiont of the Indonesian brown algae Hydroclathrus sp.

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

High-level production of the agmatine in engineered Corynebacterium crenatum with the inhibition-releasing arginine decarboxylase

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