Advancements in biocatalysis: From computational to metabolic engineering

Through several waves of technological research and un-matched innovation strategies, bio-catalysis has been widely used at the industrial level. Because of the value of enzymes, methods for producing value-added compounds and industrially-relevant fine chemicals through biological methods have been developed. A broad spectrum of numerous biochemical pathways is catalyzed by enzymes, including enzymes that have not been identified. However, low catalytic efficacy, low stability, inhibition by non-cognate substrates, and intolerance to the harsh reaction conditions required for some chemical processes are considered as major limitations in applied bio-catalysis. Thus, the development of green catalysts with multi-catalytic features along with higher efficacy and induced stability are important for bio-catalysis. Implementation of computational science with metabolic engineering, synthetic biology, and machine learning routes offers novel alternatives for engineering novel catalysts. Here, we describe the role of synthetic biology and metabolic engineering in catalysis. Machine learning algorithms for catalysis and the choice of an algorithm for predicting protein-ligand interactions are discussed. The importance of molecular docking in predicting binding and catalytic functions is reviewed. Finally, we describe future challenges and perspectives.     

Synthesis and characterization of polyaniline–zirconium dioxide and polyaniline–cerium dioxide composites with enhanced photocatalytic degradation of rhodamine B dye

For the first time, chloroform and 2-butanol were used as solvent systems for the preparation of ZrO₂–PANI and CeO₂–PANI composites. Solubility of the synthesized composites was studied in chloroform, N-methyl-2-pyrrolidinone (NMP), and in mixture of toluene?+?2-propanol (2:1). XRD and cyclic voltammetry data showed that the ZrO₂–PANI and CeO₂–PANI composites possess both crystalline and amorphous domains indicating some sort of conductivity. TGA results showed that ZrO₂–PANI composite have a better thermal stability than pure PANI; however, CeO₂–PANI composite has lower thermal stability than pure PANI. The conjugated unsaturated structure of PANI is responsible for the enhanced photocatalytic properties of ZrO₂–PANI and CeO₂–PANI. Photocatalytic results showed that, at photolysis time of 60 min, rhodamine B (RhB) dye was degraded up to 34 and 35% by ZrO₂–PANI and CeO₂–PANI, respectively. The degradation products of RhB were quantified by LC–MS and GC–MS, and accordingly, a detailed pathway was proposed.     

Photoaddition reactions of acetylene and butadiyne derivatives to benzodithiophene

The photochemical [2pi +2pi] cycloaddition of dimethyl acetylenedicarboxylate to benzo[1,2-b:4,5-b']dithiophene has been used to synthesize substituted cyclobuta[b]thieno[2,3-f][1]benzothiophene. The first [2pi + 2pi] photocycloaddition reaction of a series of butadiynes to benzodithiophene is reported to yield regioselective and acetylene-substituted cyclobutene derivatives containing an aromatic thiophene moiety.     

Ion products of water in 1m NaCl under hydrothermal conditions

The apparent ion product of water Q _w has been determined potentiometrically at pressure up to 96.7 MPa and temperatures up to 246°C in solutions of ionic strength 1.019m (NaCl). A concentration cell employing hydrogen electrodes was used. It was found that-logQ _w varies from 13.64 at 4.1 MPa and 20°C to 9.98 at 96.7 MPa and 230°C.     

Quantitation of the enantiomers of ofloxacin by capillary electrophoresis in the parts per billion concentration range for in vitro drug absorption studies

Ofloxacin, a chiral fluoroquinolone, possesses two optical isomers. The antibacterial activity of S-(-)-ofloxacin is reported to be 8-128 times higher than that of R-(+)-ofloxacin. A capillary zone electrophoresis method has been developed to quantify the enantiomers of ofloxacin in high diluted samples (20-700 ng/ml for each enantiomer). After fluid-fluid extraction of ofloxacin from physiological solution electrokinetic injection was employed to improve the sensitivity. The method was optimised using a central composite design. Four experimental factors were investigated: the background electrolyte concentration, the methyl-beta-cyclodextrin concentration, the buffer pH and the temperature. The amount migrated into the capillary, determined by the peak area, the resolution between the ofloxacin enantiomers, the migration time and the generated current were evaluated as responses. The quantification limit is 11.4 ng/ml for S-ofloxacin and 10.8 ng/ml for R-ofloxacin. The method has shown good validation data in terms of precision and recovery rate.     

Quantitation of talinolol and other beta-blockers by capillary electrophoresis for in vitro drug absorption studies

A capillary zone electrophoresis method is described for the enantioseparation of talinolol using heptakis(2,3-diacetyl-6-sulfo)-beta-cyclodextrin (HDAS-beta-CD) as a chiral selector. After liquid-liquid extraction of talinolol from physiological solution, electrokinetic injection was employed to improve the sensitivity. The use of a coated capillary was necessary to achieve stable and reproducible enantioseparations. A baseline separation of the talinolol enantiomers was achieved in less than 10 min using 100 mM phosphate solution as background electrolyte and pH 3.5, at the presence of 3.0 mM HDAS-beta-CD and at 20 degrees C. In addition, this analytical condition proved to be useful for the enantioseparation of a number of other beta-blocking agents such as alprenolol, atenolol, bisoprolol, celiprolol, metipranolol, oxprenolol, and sotalol. For determining talinolol, the method could be validated in terms of precision, accuracy and linearity, and was found to be suitable in determination of talinolol enantiomers in highly diluted samples obtained from in vitro experiments.     

Development and Validation of an HPLC Method for the Determination of Dexamethasone, Dexamethasone Sodium Phosphate and Chloramphenicol in Presence of Each Other in Pharmaceutical Preparations

An HPLC method for the determination of dexamethasone, dexamethasone sodium phosphate and chloramphenicol in presence of each other in pharmaceutical preparations has been developed using a Shim-Pack CLC-ODS column (6.0 × 150 mm²). These analytes were separated under isocratic conditions. Various chromatographic parameters including linearity, precision and accuracy have been evaluated. The method was found to be suitable for analysis of these drug substances in presence of each other. The run time was less than 15 min. This method is suitable for application to various dosage forms.     

Crystallization and Organic Field-Effect Transistor Performance of a Hydrogen-Bonded Quaterthiophene

Crystalline thin films of π-conjugated molecules are relevant as the active layers in organic electronic devices. Therefore, materials with enhanced control over the supramolecular arrangement, crystallinity, and thin-film morphology are desirable. Herein, it is reported that hydrogen-bonded substituents serve as additional structure-directing elements that positively affect crystallization, thin-film morphology, and device performance of p-type organic semiconductors. It is observed that a quaterthiophene diacetamide exhibits a denser packing than that of other quaterthiophenes in the single-crystal structure and, as a result, displays enhanced intermolecular electronic interactions. This feature was preserved in crystalline thin films that exhibited a layer-by-layer morphology, with large domain sizes and high internal order. As a result, organic field-effect transistors of these polycrystalline thin films showed mobilities in the range of the best mobility values reported for single-crystalline quaterthiophenes. The use of hydrogen-bonded groups may, thus, provide an avenue for organic semiconducting materials with improved morphology and performance.     

Monodisperse CuPt alloy nanoparticles assembled on reduced graphene oxide as catalysts in the transfer hydrogenation of various functional organic groups

We present herein a new nanocatalyst, namely binary CuPt alloy nanoparticles (NPs) supported on reduced graphene oxide (CuPt‐rGO), as a highly active heterogeneous catalyst for the transfer hydrogenation (TH) protocol that is demonstrated to be applicable over the reduction of various unsaturated organic compounds (olefins, aldehydes/ketones and nitroarenes) in aqueous solutions at room temperature. CuPt alloy NPs were synthesized by the co‐reduction of metal (II) acetylacetonates by borane‐tert‐butylamine (BTB) complex in hot oleylamine (OAm) solution and then assembled on reduced graphene oxide (rGO) via ultrasonic‐assisted liquid phase self‐assembly method. The structure of yielded CuPt NPs and CuPt‐rGO nanocatalyst were characterized by TEM, XRD and ICP‐MS. The activity of Cu₇Pt₃‐rGO nanocatalysts were then tested for the THs that were conducted in a commercially available high‐pressure tube using water as sole solvent and ammonia borane as a hydrogen donor at room temperature. The presented catalytic TH protocol was successfully applied over nitroarenes, olefines and aldehydes/ketones, and all the tested compounds were converted to corresponding reduction products with the yields reaching up to 99% under ambient conditions. Moreover, the Cu₇Pt₃‐rGO nanocatalyst was also reusable in the TH by providing 99% yield after five consecutive runs in TH of nitrobenzene as an example.     

从野生与突变株云芝IBL-04提纯锰过氧化物酶及其表征

由野生及突变株云芝IBL-04制得细胞外锰过氧化物酶(MnPs)，并经过硫酸铵沉淀、透析、离子交换和凝胶渗透层析法等步骤提纯.纯化的酶在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)上于43 kDa区域呈现单一谱带，它适宜的pH值和温度分别为5.0和40°C.突变株MnPs表现出比野生株MnPs更宽的活性pH值范围和更高的热稳定性.从所选突变株所得纯化的MnPs表现出与野生株MnP几乎相同的电泳性质、稳态动力学、金属离子和EDCs降解效率.该生物酶与Mn2+一起催化的反应速率最快，但最高的亲和性对应于ABTS、甲氧基羟苯基乙二醇、4-氨基苯酚和活性染料. Mn2+和Cu2+可显著提高MnPs的活性，但Zn2+， Fe2+， EDTA和半胱氨酸则会不同程度地抑制其活性， Hg2+是最强的活性抑制剂.所有来源的MnPs均可有效催化EDCs、壬基苯酚和二氯苯氧氯酚降解，处理3 h可除去80%以上，在MnPs-介质体系中可进一步提高到90%.综上，云芝MnPs生物酶具有较高的pH适用性和热稳定性、独特的Michaelis-Menten动力学参数和高的EDCs去除效率等特点，因而有望工业化应用.