Extracellular expression of natural cytosolic arginine deiminase from Pseudomonas putida and its application in the production of l-citrulline

Su, Lingqia; Ma, Yue; Wu, Jang‐Yen

doi:10.1016/j.biortech.2015.07.081

Cited by 20 publications

(12 citation statements)

References 30 publications

(45 reference statements)

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“…(c)]. This is likely the result of rapid synthesis and misfolding of recombinant protein at 37 °C . The best results were achieved at 30 °C.…”

Section: Resultsmentioning

confidence: 99%

“…In order to obtain the maximum level of target protein through a balance of cell growth and protein expression, which should both be considered, as reported, enzyme induction was performed using lactose feeding rates of 0.1, 0.2 and 0.4 g L −1 h −1 . The results are shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…4(c)]. This is likely the result of rapid synthesis and misfolding of recombinant protein at 37 ∘ C. 34,35 The best results were achieved at 30 ∘ C. Half of the total SP Lc activity was released into the culture medium, leading to the highest extracellular activity (949.7 U mL −1 ) among the three induction temperatures [ Fig. 4(b)].…”

Section: Effect Of Induction Temperature On Cell Growth and Protein Pmentioning

confidence: 99%

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Highly efficient extracellular expression of naturally cytoplasmic Leuconostoc mesenteroides sucrose phosphorylase

Tian

et al. 2018

J of Chemical Tech & Biotech

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BACKGROUND We previously showed that extracellular production of naturally cytoplasmic proteins could be realized through co‐expression with phospholipase C (PLC), which enhances cell membrane permeability through limited hydrolysis of membrane phospholipids. Leuconostoc mesenteroides sucrose phosphorylase (SPLc), a naturally cytoplasmic enzyme, exhibits excellent properties for synthesis of α‐arbutin, but no systemic investigation of its preparation has thus far been reported. RESULTS Three PLCs were selected and expressed, without signal peptide, in Escherichia coli. More than 90% of the PLCs from Listeria monocytogenes and Bacillus cereus were present in the culture medium. When SPLc was co‐expressed with these PLCs, and the location of the PLC and SPLc genes in the co‐expression vector was optimized, the highest production was obtained when SPLc gene was inserted upstream of the B. cereus PLC gene, as in BL21(DE3)/pETDuet‐sp‐bcplc. SPLc production was then scaled up to a 3 L fermenter and fermentation conditions were optimized. When the initial medium contained 0.6 mmol L−1 ZnCl2 and protein expression was induced at a dry cell weight of 15 g L−1 by the addition of lactose at 0.2 g L−1 h−1 at 30 °C, the extracellular SPLc activity reached its highest level of 1445.1 U mL−1 This constituted 81.6% of total activity; the remainder of the enzyme (18.4%) remained inside the cells. CONCLUSION This is the first report describing extracellular expression of sucrose phosphorylase, and the highest yield ever reported was obtained. This provides the basis for industrial‐scale production and application of sucrose phosphorylase. It also provides a potential method for improving the production of other proteins and fine chemicals. © 2018 Society of Chemical Industry

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“…(c)]. This is likely the result of rapid synthesis and misfolding of recombinant protein at 37 °C . The best results were achieved at 30 °C.…”

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Effect Of Induction Temperature On Cell Growth and Protein Pmentioning

confidence: 99%

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Highly efficient extracellular expression of naturally cytoplasmic Leuconostoc mesenteroides sucrose phosphorylase

Tian

et al. 2018

J of Chemical Tech & Biotech

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“…All these results revealed that the gene codon optimization could obviously enhance the enzyme activity, which was similar to the previous study (Su et al . ). In addition, these results also provided a theoretical basis for further study.…”

Section: Discussionmentioning

confidence: 97%

“…It has been reported that gene codon optimization could significantly increase the expression level of the target enzyme (Su et al . ,b). Therefore, codon optimization of the ksdd gene based on the codon preference of B. subtilis is necessary for KSDD enzyme activity improvement.…”

Section: Introductionmentioning

confidence: 99%

Efficient androst-1,4-diene-3,17-dione production by co-expressing 3-ketosteroid-Δ¹-dehydrogenase and catalase inBacillus subtilis

et al. 2016

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Characterization of a recombinant arginine deiminase from Halothermothrix orenii and its application in citrulline production

Wang

Miao

et al. 2022

Biotech and App Biochem

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In recent years, arginine deiminase (ADI, EC 3.5.3.6) has attracted much attention as a biocatalyst that produces the functional amino acid l‐citrulline from l‐arginine and also as an anticancer enzyme. Here, we identified and characterized a putative ADI from the thermophilic bacterium Halothermothrix orenii. The H. orenii ADI (H‐ADI) protein was expressed in Escherichia coli BL21(DE3) with a specific activity of 91.8 U/mg protein at 55°C and pH 6.5. The enzyme remained at 74% relative activity after incubation at 45°C for 180 min, only 25% at 50°C. The melting temperature was 56°C. H‐ADI is not a metal‐requiring enzyme; Ni2+ slightly improved the catalytic activity. The Km and Vmax for l‐arginine were 55.5 mM and 156.8 μmol/min/mg protein, respectively. Moreover, three residues (Arg183, Arg237, and His273) were key to the formation of l‐citrulline, as analyzed by alanine‐scanning mutagenesis. Finally, the enzymatic synthesis of l‐citrulline was carried out at 50°C with a conversion ratio reaching 99.03%. Together, these findings show that H‐ADI is a promising biocatalyst for the production of l‐citrulline.

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Extracellular expression of natural cytosolic arginine deiminase from Pseudomonas putida and its application in the production of l-citrulline

Cited by 20 publications

References 30 publications

Highly efficient extracellular expression of naturally cytoplasmic Leuconostoc mesenteroides sucrose phosphorylase

Highly efficient extracellular expression of naturally cytoplasmic Leuconostoc mesenteroides sucrose phosphorylase

Efficient androst-1,4-diene-3,17-dione production by co-expressing 3-ketosteroid-Δ¹-dehydrogenase and catalase inBacillus subtilis

Characterization of a recombinant arginine deiminase from Halothermothrix orenii and its application in citrulline production

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Extracellular expression of natural cytosolic arginine deiminase from Pseudomonas putida and its application in the production of l-citrulline

Cited by 20 publications

References 30 publications

Highly efficient extracellular expression of naturally cytoplasmic Leuconostoc mesenteroides sucrose phosphorylase

Highly efficient extracellular expression of naturally cytoplasmic Leuconostoc mesenteroides sucrose phosphorylase

Efficient androst-1,4-diene-3,17-dione production by co-expressing 3-ketosteroid-Δ1-dehydrogenase and catalase inBacillus subtilis

Characterization of a recombinant arginine deiminase from Halothermothrix orenii and its application in citrulline production

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Efficient androst-1,4-diene-3,17-dione production by co-expressing 3-ketosteroid-Δ¹-dehydrogenase and catalase inBacillus subtilis