Arginine-hydrolyzing enzymes for electrochemical biosensors

Since l-Arginine (Arg) is a semi-essential amino acid for humans, its adequate amount must be consumed in the diet to prevent certain negative consequences related to insufficient synthesis of this amino acid under specific physiological conditions. Arg metabolism results in the production of a biochemically diverse range of such products as urea, some amino acids, creatine, polyamines, nitric oxide, etc. Arg, an important biomarker in clinical diagnostics, is also used for prevention/treatment of different diseases, including cancer and COVID-19. Furthermore, it serves as an indicator of food and beverages quality.A variety of optic and electrochemical methods for Arg determination have already been suggested. The biosensor systems based on the enzymes of Arg metabolism were shown to be the most promising tools for Arg assay. This review focuses on the peculiarities of electrochemical biosensors for Arg assay based on the use of Arg-degrading enzymes and on the analysis of their advantages as compared to other approaches.     

L-Arginine/nanofish bone nanocomplex enhances bone regeneration via antioxidant activities and osteoimmunomodulatory properties

In this study,a promising strategy has been developed to promote bone regeneration by combining antioxidant activities and osteoimmunomodulatory properties.Herein,an L-arginine/nanofish bone(Arg/NFB) nanocomplex has been prepared and evaluated in vitro and in vivo.The Arg/NFB nanocomplex possesses good antioxidant activities and could modulate the polarization of non-activated macrophage into different types and induce the secretion of pre-inflammato ry,anti-inflammatory,osteogenic as well as angiogenic cytokines.Additionally,the regulated immune microenvironment can enhance the osteogenic differentiation of mouse embryo osteoblast precursor cells(MC3 T3-E1) and angiogenic capacity of human umbilical vein endothelial cells(HUVECs),leading to the improved formation of mineralized nodules,alkaline phosphatase activity and angiogenic effects.In vivo results with cranial defect models reveal that the treatment of Arg/NFB nanocomplex exhibited significant improvement of new bone formation and angiogenesis.All the results demonstrate Arg/NFB nanocomplex with antioxidant activities and osteoimmunomodulatory properties could be a new idea for developing the next generation of bone regeneration biomaterials.     

Investigation of chemical activity,SCHIFF base reactions and staining effects of some amino acids by spectrophotometric and theorical methods

In this study, the staining properties of selected amino acids with Brassica oleraceae extract in alum and alum-free media were investigated. Basic, acidic and neutral amino acids (arginine, glutamic acid and glycine) were used to investigate the effect of staining. It was determined that all amino acids were stained in alum media. In the second step, the R group of amino acids found in the proteins of the cell nucleus was reacted with salicyl aldehyde. This reaction was successful only with Arginine. The staining properties of the newly formed compound were also investigated in alum and alum-free environments. Evaluation of the results was done using FT-IR and ¹H NMR methods. All compounds were optimized with the Gaussian G09 program (DFT/B3LYP/6.311 ?G(d.p) basic set. HOMO, LUMO and HOMO-LUMO gap values were determined. Chemical reaction capabilities of amino acids were discussed with the help of HOMO-LUMO gap values.     

Solubilization of organic compounds by arginine-derived polymers

Poor aqueous solubility of drugs is one of the major challenges in the pharmaceutical science. In this study, a guanidinium-containing polymer based on arginine was designed and synthesized, and was evaluated as a solubility enhancing additive for three model organic compounds (coumarin, pyrene and doxorubicin). At a guanidinium group concentration of 100 mmol/L, the polymer could significantly increase the solubility of pyrene and doxorubicin by 6-and 11-fold respectively, much more effective than arginine (2-and 3-fold, respectively). In contrast, its effect on the solubility of coumarin was less effective than arginine. The solubilizing effect may be explained by the enhanced interaction between the guanidinium group in the polymer and the aromatic compounds.     

Ketopiperazines: Conformationally Constrained Peptidomimetic of Arginine Amides

We have synthesized guanidine‐containing ketopiperazines designed to be conformational mimics of peptidomimetic arginine amides. D‐Allylglycine was converted by an efficient approach to give enantiopure ketopiperazines in which the trans stereochemistry of the C‐substituents resulted from stereospecific enolate alkylation.     

Determination of asymmetric and symmetric dimethylarginines in human plasma by HPLC with electrochemical detection

A new HPLC method was developed and validated for the determination of asymmetric and symmetric dimethylarginines and l ‐arginine in human plasma. After SPE and evaporation of the eluate, the samples were derivatised with an o‐phthaldialdehyde reagent containing 3‐mercaptopropionic acid. The derivatives formed were analysed by isocratic RP‐HPLC with electrochemical detection at +320 mV. The mobile phase consisted of 50 mM phosphate buffer (pH 6.1) containing 10% v/v acetonitrile, the flow rate was 1 mL/min. The retention times of all compounds including monomethylarginine (internal standard) were <24 min. The LODs (S/N 3:1) were 0.012 μM for both dimethylarginines and 0.013 μM for l ‐arginine; the linearity of the method was from 0.1 to 20 μM for both dimethylarginines and from 1 to 200 μM for l ‐arginine. Absolute extraction recoveries measured for all analytes ranged from 85 to 88%.     

Enzymatic Synthesis of Agmatine by Immobilized Escherichia coli Cells with Arginine Decarboxylase Activity

A new method for the enzymatic synthesis of agmatine by immobilized Escherichia coli cells with arginine decarboxylase(ADC) activity was established and a series of optimal reaction conditions was set down. The arginine decarboxylase showed the maximum activity when the pyridoxal phosphate(PLP) concentration was 50 mmol/L, pH=7 and 45 °C. The arginine decarboxylase exhibited the maximum production efficiency when the substrate concentration was 100 mmol/L and the reaction time was 15 h. It was also observed that the appropriate concentration of Mg2+, especially at 0.5 mmol/L promoted the arginine decarboxylase activity; Mn2+ had little effect on the arginine decarboxylase activity. The inhibition of Cu2+ and Zn2+ to the arginine decarboxylase activity was significant. The immobilized cells were continuously used 6 times and the average conversion rate during the six-time usage was 55.6%. The immobilized cells exhibited favourable operational stability. After optimization, the maximally cumulative amount of agmatine could be up to 20 g/L. In addition, this method can also catalyze D,L-arginine to agmatine, leaving the pure optically D-arginine simultaneously. The method has a very important guiding significance to the enzymatic preparation of agmatine.     

精氨酸侧链和核酸碱基间离子氢键作用强度分析

采用MP2/6-31+G(d,p)方法优化得到了22个由精氨酸侧链与碱基尿嘧啶、 胸腺嘧啶、 胞嘧啶、 鸟嘌呤及腺嘌呤形成的氢键复合物的气相稳定结构, 使用包含BSSE校正的MP2/aug-cc-pVTZ方法计算得到了复合物的气相结合能, 通过MP2/6-31+G(d,p)方法和PCM模型优化得到了复合物的水相稳定结构, 采用MP2/aug-cc-pVTZ方法和PCM模型计算得到了复合物的水相结合能. 研究发现, 精氨酸侧链与碱基间的离子氢键作用强度与单体间电荷转移量、 氢键临界点电子密度及二阶作用稳定化能密切相关. 与中性氢键相比, 离子氢键作用具有更显著的共价作用成分. 研究还发现, 精氨酸侧链和碱基间形成的氢键复合物的稳定性次序可以通过氢键受体碱基分子上氧原子和氮原子的质子化反应焓变进行预测, 质子化反应焓变越负, 形成的氢键复合物越稳定.     

Concept and synthetic approach for a kilogram scale synthesis of octa-d-arginine amide nonahydrochloride salt

Oligomers of arginine, such as octa-d-arginine amide, are excellent transporters for active drugs through cell membranes and tissue. The synthesis of octa-d-arginine amide, as the nonahydrochloride salt, was approached via a solution phase synthetic route involving the preparation of an octa-d-ornithine intermediate, which was then converted into the desired octa-d-arginine compound through a guanidinylation step. The multi-step synthesis was carried out at pilot scale, resulting in the preparation of 700 g of the target molecule. No chromatographic purification was needed at any step of the process.     

Evanescent wave infrared chemical sensor possessing a sulfonated sensing phase for the selective detection of arginine in biological fluids

This paper describes a new infrared (IR) sensing scheme for the determination of arginine (Arg). In this method, the surface of an IR evanescent wave sensing element was modified with sulfonic acid groups to selectively interact with Arg through specific interactions with its guanidine moiety. The sulfonated sensing phase was prepared using a two-layer modification approach. To demonstrate that this assembly could be used for selective infrared sensing, a large number of amino acids were subjected to analysis. Although the sulfonate groups on the surface of the sensing element did interact selectively with the guanidine groups of Arg species, lysine and histidine units caused some interference; this problem could be minimized because of the unique IR absorption bands of the guanidine moiety of Arg. To optimize the detection conditions, we studied the effects of both the pH and the composition of the polymer. The most intense signal was obtained at pH 9. We observed different adsorption rates for the detection of Arg at different values of pH, which we attribute to changes in the accessibility of the analytes to the pore structures of the sensing phase. The composition of the base polymer was also optimized; 60% PVBC (w/w) provided a water-stable, sensitive phase for the detection of Arg in aqueous solution. Under the optimized conditions, we obtained a linear range of detection up to 0.1 mM with a detection limit of ca. 5 μM.