Biosynthesis of desferrioxamine siderophores initiated by decarboxylases: A functional investigation of two lysine/ornithine-decarboxylases from Gordonia rubripertincta CWB2 and Pimelobacter simplex 3E

Hofmann, Marika; Campo, Julia S. Martín del; Sobrado, Pablo; Tischler, Dirk

doi:10.1016/j.abb.2020.108429

Cited by 9 publications

(6 citation statements)

References 45 publications

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“…21,22 In the case of ODC in C. aceticum DSM1496, PLP served as the coenzyme. The optimal temperature for ODC activity has a proximity of ODC from Gordonia rubripertincta 23 and Clostridium thermohydrosulfuricum, 24 VkMAC-2033D. 25 The decline in ODC activity might be attributed to the dissociation of PLP coenzyme from the enzyme structure, resulting in the formation of an inactive apoprotein form, consistent with observations made for other PLP-dependent enzymes.…”

Section: Discussionsupporting

confidence: 77%

“…The optimal temperature for ODC activity has a proximity of ODC from Gordonia rubripertincta 23 and Clostridium thermohydrosulfuricum, 24 VkMAC-2033D. 25 The decline in ODC activity might be attributed to the dissociation of PLP coenzyme from the enzyme structure, resulting in the formation of an inactive apoprotein form, consistent with observations made for other PLP-dependent enzymes. 26 The optimal pH for ODC activity was determined to be approximately 6.5, similar to that of ODC from P. simplex VkMAC-2033D.…”

Section: Discussionsupporting

confidence: 77%

“…26 The optimal pH for ODC activity was determined to be approximately 6.5, similar to that of ODC from P. simplex VkMAC-2033D. 25 Under acidic conditions (pH 4.5-5.5), ODC retained around 40% of its activity, demonstrating only slight decreases. Conversely, in an alkaline environment (pH 8-9.5), ODC exhibited near-inactivity with a significant reduction in activity.…”

Section: Discussionmentioning

confidence: 87%

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Enzymatic properties of ornithine decarboxylase from Clostridium aceticum DSM1496

Tan,

Gou,

Fan

et al. 2024

Biotech and App Biochem

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Clostridium aceticum DSM1496 is an acid‐resistant strain in which ornithine decarboxylase (ODC) plays a crucial role in acid resistance. In this study, we expressed ODC derived from C. aceticum DSM1496 in Escherichia coli BL21 (DE3) and thoroughly examined its enzymatic properties. The enzyme has a molecular weight of 55.27 kDa and uses pyridoxal‐5′‐phosphate (PLP) as a coenzyme with a Km = 0.31 mM. ODC exhibits optimal activity at pH 7.5, and it maintains high stability even at pH 4.5. The peak reaction temperature for ODC is 30°C. Besides, it can be influenced by certain metal ions such as Mn2+. Although l‐ornithine serves as the preferred substrate for ODC, the enzyme also decarboxylates l‐arginine and l‐lysine simultaneously. The results indicate that ODC derived from C. aceticum DSM1496 exhibits the ability to produce putrescine, cadaverine, and agmatine through decarboxylation. These polyamines have the potential to neutralize acid in an acidic environment, facilitating the growth of microorganisms. These significant findings provide a strong basis for further investigation into the acid‐resistant mechanisms contributed by ODC.

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Section: Discussionsupporting

confidence: 77%

Section: Discussionsupporting

confidence: 77%

Section: Discussionmentioning

confidence: 87%

See 1 more Smart Citation

Enzymatic properties of ornithine decarboxylase from Clostridium aceticum DSM1496

Tan,

Gou,

Fan

et al. 2024

Biotech and App Biochem

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“…More importantly, a high concentration of putrescine not only affects the sensory characteristics of food, but also causes toxicological reactions. Putrescine can enhance the toxic effect of histamine, causing headaches, nausea, vomiting, and other adverse reactions ( Huang et al, 2015 ; Hofmann et al, 2020 ; Fu et al, 2021 ). Therefore, putrescine is an important spoilage indicator in food ( Kanjee et al, 2011 ).…”

Section: Introductionmentioning

confidence: 99%

Heterologous expression and activity verification of ornithine decarboxylase from a wild strain of Shewanella xiamenensis

et al. 2022

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Shewanella xiamenensis is widely found in spoilage fish, shrimp and other seafoods. Under suitable conditions, ornithine can be synthesized into putrescine, which may spoil food or endanger health. Our research used a wild strain of Shewanella xiamenensis isolated from “Yi Lu Xian” salted fish (a salting method for sea bass) as a research object. According to the database of National Center of Biotechnology Information (NCBI), the target ornithine decarboxylase (ODC) gene SpeF was successfully amplified using the wild strain of Shewanella xiamenensis as the template. Sequencing alignment showed that the SpeF of the wild strain had more than 98% homology compared with the standard strain. The amino acid substitution occurred in the residues of 343, 618, 705, and 708 in the wild strain. After optimizing the expression conditions, a heterologous expression system of ODC was constructed to achieve a high yield of expression. The amount of 253.38 mg of ODC per liter of LB broth was finally expressed. High performance liquid chromatography (HPLC) and subsequent ODC activity verification experiments showed that hetero-expressed ODC showed a certain enzyme activity for about 11.91 ± 0.38 U/mg. Our study gives a new way to the development of a low-cost and high-yield strategy to produce ODC, providing experimental materials for further research and elimination of putrescine in food hazards.

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“…FMOs are commonly found in natural product biosynthetic pathways, where they play a major role in the addition of functional groups essential for bioactivity. , This family has been categorized into eight classes, A–H, according to structural and mechanistic characteristics . Our group has been studying several members of class B FMOs as these enzymes perform highly specific oxidations useful for biomedical and biotechnological applications. − …”

Section: Introductionmentioning

confidence: 99%

Characterization of a Nitro-Forming Enzyme Involved in Fosfazinomycin Biosynthesis

Valentino

Sobrado

2021

Biochemistry

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N-hydroxylating monooxygenases (NMOs) are a subclass of flavindependent enzymes that hydroxylate nitrogen atoms. Recently, unique NMOs that perform multiple reactions on one substrate molecule have been identified. Fosfazinomycin M (FzmM) is one such NMO, forming nitrosuccinate from aspartate (Asp) in the fosfazinomycin biosynthetic pathway in some Streptomyces sp. This work details the biochemical and kinetic analysis of FzmM. Steady-state kinetic investigation shows that FzmM performs a coupled reaction with Asp (k cat , 3.0 ± 0.01 s −1 ) forming nitrosuccinate, which can be converted to fumarate and nitrite by the action of FzmL. FzmM displays a 70-fold higher k cat /K M value for NADPH compared to NADH and has a narrow optimal pH range (7.5−8.0). Contrary to other NMOs where the k red is rate-limiting, FzmM exhibits a very fast k red (50 ± 0.01 s −1 at 4 °C) with NADPH. NADPH binds at a K D value of ∼400 μM, and hydride transfer occurs with pro-R stereochemistry. Oxidation of FzmM in the absence of Asp exhibits a spectrum with a shoulder at ∼370 nm, consistent with the formation of a C(4a)hydroperoxyflavin intermediate, which decays into oxidized flavin and hydrogen peroxide at a rate 100-fold slower than the k cat . This reaction is enhanced in the presence of Asp with a slightly faster k ox than the k cat , suggesting that flavin dehydration or Asp oxidation is partially rate limiting. Multiple sequence analyses of FzmM to NMOs identified conserved residues involved in flavin binding but not for NADPH. Additional sequence analysis to related monooxygenases suggests that FzmM shares sequence motifs absent in other NMOs. 65 characterization of the nitro-forming NMO FzmM providing a 66 detailed report of its mechanism toward the formation of 67 nitrosuccinate.

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Biosynthesis of desferrioxamine siderophores initiated by decarboxylases: A functional investigation of two lysine/ornithine-decarboxylases from Gordonia rubripertincta CWB2 and Pimelobacter simplex 3E

Cited by 9 publications

References 45 publications

Enzymatic properties of ornithine decarboxylase from Clostridium aceticum DSM1496

Enzymatic properties of ornithine decarboxylase from Clostridium aceticum DSM1496

Heterologous expression and activity verification of ornithine decarboxylase from a wild strain of Shewanella xiamenensis

Characterization of a Nitro-Forming Enzyme Involved in Fosfazinomycin Biosynthesis

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