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Hippocampal deficits and metabolic dysregulations such as dyslipidemia have been frequently reported in schizophrenia and are suggested to contribute to the pathophysiology of schizophrenia. Hippocampus is particularly susceptible to environmental challenges including metabolism and inflammation. However, evidence linking hippocampal alterations and metabolic dysregulations are quite sparse in drug-naïve schizophrenia. A total of 166 drug-naïve patients with first-episode schizophrenia (FES) and 78 healthy controls (HC) underwent measures for several serum metabolic markers, structural and resting-state functional magnetic resonance imaging (rs-fMRI), as well as diffusion tensor imaging (DTI). Seed-to-voxel functional connectivity (FC) and probabilistic tractography were performed to assess the functional and microstructural connectivity of the bilateral hippocampi. Clinical symptoms were evaluated with Positive and Negative Syndrome Scale (PANSS). Patients with FES showed significantly decreased total cholesterol (Chol) level. Patients showed elevated FC between the left hippocampus and bilateral thalami while showing decreased microstructural connectivity between the left hippocampus and bilateral thalami. Multiple regression analyses showed that FC from the left hippocampus to the right superior frontal gyrus (SFG), bilateral frontal pole (FP), and right thalamus were negatively associated with the Chol level, while no association was observed in the HC group. Our study validated alterations in both functional and microstructural thalamo-hippocampal connectivities, and abnormal cholesterol level in FES. Moreover, decreased cholesterol level is associated with elevated thalamo-hippocampal functional connectivity in patients with FES, suggesting that dyslipidemia may interact with the hippocampal dysfunction in FES.

With the rapid development of green chemistry, the design and application of the related methods and requisite solvents have received increasing attention in recent years. Deep eutectic solvents (DESs) are mixtures formed from a hydrogen bond acceptor (HBA) and a hydrogen bond donor (HBD). Generally, ionic liquids (ILs) and DESs have similar physical and chemical properties, and hence, find application in the same fields. However, DESs have many advantages over ILs, such as non-toxicity, environmental friendliness, low cost, and biodegradability. Thus, there are many areas where DESs play a key role and act as new, efficient green extraction solvents. DESs can aid the extraction and separation of different target compounds from a variety of samples, thus promoting the rapid development of sample pretreatment technology. As extraction solvents, DESs offer unique advantages. In dispersive liquid-liquid microextraction (DLLME), DESs show incredible ability to extract residual drugs, metal ions, and bioactive components from complex matrices, which would require complicated sample preparation steps when using traditional organic extraction solvents. Compared with traditional organic extraction solvents, DESs have considerable merits of greenness, hypotoxicity, higher extraction efficiency, etc. Moreover, as a dispersant, a DES can accelerate the diffusion of the extractant in the sample solution during DLLME, owing to its benefits of miniaturization and low cost. Traditional dispersants such as methanol and acetonitrile have many disadvantages, including high volatility, flammability, and toxicity, while DESs are environmentally friendly. Therefore, the combination of DES and DLLME has recently gained prominence in the field of sample preparation. Additionally, the combination of DES and solid-phase extraction (SPE) has broad application prospects. By virtue of their diverse functions, DESs have been used as eluents, in combination with a solid-phase extraction column and a stir bar, to elute analytes from the sorbent surface. The molar ratio of the HBA and HBD is one of the important factors influencing the elution efficiency. DESs can be combined with magnetic multiwalled carbon nanotubes, magnetic graphene oxide, and other nanocomposites to specifically adsorb target analytes through hydrogen bonding, π-π forces, and electrostatic forces. In addition, the DES can be used in the synthesis of magnetic nanocomposites and molecularly imprinted polymers when combined with magnetic materials. Magnetic nanocomposites functionalized with DES show excellent performance and high efficiency in the extraction process. The combination of DES and magnetic materials would promote the development of magnetic materials for green chemistry and expand the application of DES to several other fields. However, to the best of our knowledge, research on the microstructure, physical and chemical properties, and extraction mechanism of DESs is still in its nascent stage. Therefore, exploring the theoretical mechanism and applications of new DESs with special functions would be an essential future research direction. This article integrates the research progress of DESs in extraction separation technology; introduces the preparation, properties, and classification of DESs; and summarizes the applications of DESs in DLLME and SPE.

The efficiency and accuracy of the synthesis of structural lipids are closely related to the regiospecificity of lipases. Understanding the structural mechanism of their regiospecificity contributes to the regiospecific redesign of lipases for meeting the technological innovation needs. Here, we used a thermostable lipase from Streptomyces sp. W007 (MAS1), which has been recently reported to show great potential in industry, to gain an insight into the structural basis of its regiospecificity by molecular modelling and mutagenesis experiments. The results indicated that increasing the steric hindrance of the site for binding a non-reactive carbonyl group of TAGs could transform the non-specific MAS1 to a α-specific lipase, such as the mutants G40E, G40F, G40Q, G40R, G40W, G40Y, N45Y, H108W and T237Y (PSI > 80). In addition, altering the local polarity of the site as well as the conformational stability of its composing residues could also impact the regiospecificity. Our present study could not only aid the rational design of the regiospecificity of lipases, but open avenues of exploration for further industrial applications of lipases.

This study reports the properties of immobilized MAS1-H108A lipase from marine Streptomyces sp. strain W007 on XAD1180 resin and its application in the synthesis of n-3 polyunsaturated fatty acids (PUFA)-rich triacylglycerols (TAG) for the first time. It was found that the optimal temperature and pH for both immobilized MAS1-H108A lipase and free lipase MAS1-H108A were 70 °C and 7.0, respectively. However, immobilized MAS1-H108A lipase exhibited higher thermostability when compared with free lipase MAS1-H108A. It was also interesting that both immobilized MAS1-H108A lipase and free lipase MAS1-H108A showed no regiospecificity in the hydrolysis of triolein. Subsequently, immobilized MAS1-H108A lipase and free lipase MAS1-H108A were employed to catalyze glycerolysis of n-3 PUFA-rich ethyl esters (EE) and esterification of n-3 PUFA with glycerol under vacuum in the solvent-free system. The results showed that n-3 PUFA-rich TAG were synthesized efficiently by non-regiospecific immobilized MAS1-H108A lipase and TAG contents separately reached 92.07% and 76.13% during the esterification and glycerolysis reactions, which were significantly higher than those (71.82% and 39.62%, respectively) obtained by free lipase MAS1-H108A. Besides, TAG exhibited similar n-3 PUFA composition to the substrate. These findings indicated that non-regiospecific immobilized MAS1-H108A lipase is a promising and efficient biocatalyst for the industrial synthesis of n-3 PUFA-rich TAG.

Enzymatic degumming is an essential refining process to improve oil quality. In this study, a monoacylglycerol lipase GMGL was derived from marine Geobacillus sp., and was found that not only took monoacylglycerol (MAG) as substrate, but also had activity toward lysophosphatidylcholine (LPC), lysophosphatidylethanolamine (LPE) and glycerolphosphatidylcholine (GPC). Binding free energy showed LPC and LPE could bind with enzyme stably as MAG. It presented great potential in the field of enzymatic degumming. The phosphorus content in crude soybean oil decreased from 680.50 to 2.01 mg/kg and the yield of oil reached to 98.80 % after treating with phospholipase A1 (Lecitase Ultra) combined with lipase GMGL. An ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was developed to identify 21 differential phospholipids between crude soybean oil and enzymatic treatment. This work might shed some light on understanding the catalytic mechanism of monoacylglycerol lipase and provide an effective strategy for enzymatic degumming.

Traditionally, it is believed that the substrate and products of a monoacylglycerol lipase (MGL) share the same path to enter and exit the catalytic site. Glycerol (a product of MGL), however, was recently hypothesized to be released through a different path. In order to improve the catalytic efficacy and thermo-stability of MGL, it is important to articulate the pathways of a MGL products releasing. In this study, with structure biological approaches, biochemical experiments, and in silico methods, we prove that glycerol is released from a different path in the catalytic site indeed. The fatty acid (another product of MGL) does share the same binding path with the substrate. This discovery paves a new road to design MGL inhibitors or optimize MGL catalytic efficacy.

Microservice architecture (MSA) is a new software architecture, which divides a large single application and service into dozens of supporting microservices. With the increasingly popularity of MSA, the security issues of MSA get a lot of attention. In this paper, we propose an algorithm for mining causality and the root cause. Our algorithm consists of two parts: invocation chain anomaly analysis based on robust principal component analysis (RPCA) and a single indicator anomaly detection algorithm. The single indicator anomaly detection algorithm is composed of Isolation Forest (IF) algorithm, One-Class Support Vector Machine (SVM) algorithm, Local Outlier Factor (LOF) algorithm, and $3\sigma $ principle. For general and network time-consuming anomaly in the process of the MSA, we formulate different anomaly time-consuming detection strategies. We select a batch of sample data and three batches of test data of the 2020 International AIOps Challenge to debug our algorithm. According to the scoring criteria of the competition organizers, our algorithm has an average score of 0.8304 (The full score is 1) in the four batches of data. Our proposed algorithm has higher accuracy than some traditional machine learning algorithms in anomaly detection.

n-3 polyunsaturated fatty acid (PUFA)-rich lysophosphatidylcholine (LPC) with many beneficial effects was effectively synthesized by immobilized MAS1 lipase-catalyzed esterification of n-3 PUFA with sn-glycero-3-phosphatidylcholine (GPC) under vacuum in a solvent-free system. Immobilized MAS1 lipase was found to be a more suitable catalyst for the production of n-3 PUFA-rich LPC when compared with Novozym 435. The maximal GPC conversion and LPC content (93.12% and 90.77 mol %) were obtained under the optimized conditions (enzyme loading of 300 U/g substrate, temperature of 55 °C, and n-3 PUFA/GPC molar ratio of 20:1). Moreover, it was observed that 1-acyl-sn-glycero-3-lysophosphatidylcholine (sn-1 acyl LPC) was the main reaction product, as demonstrated by molecular docking. These results showed that immobilized MAS1 lipase had high phospholipase activity with a predominant specificity for the sn-1 hydroxyl position of GPC to efficiently synthesize highly pure n-3 PUFA-rich LPC from GPC for industrial application.

Unspecific peroxygenases (UPOs, EC 1.11.2.1) are promising oxyfunctionalization catalysts because of their unique stereoselectivity. However, so far only a few UPOs have been reported. In this study, gene mining was used to identify a gene from Galerina marginata that coded for a novel UPO (GmaUPO). GmaUPO was expressed in Pichia pastoris X-33 by scale-up fermentation (the UPO activity of the culture supernatant was 118 U/L). GmaUPO exhibited a molecular weight of 40 kDa and exhibited highest activity at 35°C and pH 9, respectively. Furthermore, GmaUPO was demonstrated to catalyze the epoxidation, sulfoxidation, and hydroxylation of common substrates, particularly fatty acids such as tridecanoic acid. The molecular basis for GmaUPO regioselectivity for fatty acid hydroxylation was explored by molecular modelling. The regioselectivity was mostly governed by the architecture of the enzyme's active site.

With the rapid development of green chemistry, the design and application of the related methods and requisite solvents have received increasing attention in recent years. Deep eutectic solvents (DESs) are mixtures formed from a hydrogen bond acceptor (HBA) and a hydrogen bond donor (HBD). Generally, ionic liquids (ILs) and DESs have similar physical and chemical properties, and hence, find application in the same fields. However, DESs have many advantages over ILs, such as non-toxicity, environmental friendliness, low cost, and biodegradability. Thus, there are many areas where DESs play a key role and act as new, efficient green extraction solvents. DESs can aid the extraction and separation of different target compounds from a variety of samples, thus promoting the rapid development of sample pretreatment technology. As extraction solvents, DESs offer unique advantages. In dispersive liquid-liquid microextraction (DLLME), DESs show incredible ability to extract residual drugs, metal ions, and bioactive components from complex matrices, which would require complicated sample preparation steps when using traditional organic extraction solvents. Compared with traditional organic extraction solvents, DESs have considerable merits of greenness, hypotoxicity, higher extraction efficiency, etc. Moreover, as a dispersant, a DES can accelerate the diffusion of the extractant in the sample solution during DLLME, owing to its benefits of miniaturization and low cost. Traditional dispersants such as methanol and acetonitrile have many disadvantages, including high volatility, flammability, and toxicity, while DESs are environmentally friendly. Therefore, the combination of DES and DLLME has recently gained prominence in the field of sample preparation. Additionally, the combination of DES and solid-phase extraction (SPE) has broad application prospects. By virtue of their diverse functions, DESs have been used as eluents, in combination with a solid-phase extraction column and a stir bar, to elute analytes from the sorbent surface. The molar ratio of the HBA and HBD is one of the important factors influencing the elution efficiency. DESs can be combined with magnetic multiwalled carbon nanotubes, magnetic graphene oxide, and other nanocomposites to specifically adsorb target analytes through hydrogen bonding

n-3 polyunsaturated fatty acids (PUFA)-rich triacylglycerols (TAG) with many beneficial effects are still difficult to be synthesized efficiently and rapidly by current synthetic techniques. This study reports the fatty acid specificity of immobilized MAS1 lipase and its efficient synthesis of n-3 PUFA-rich TAG by esterification of glycerol with n-3 PUFA in natural deep eutectic solvents (NADES) systems. Immobilized MAS1 lipase showed the highest preference for capric acid [C10:0, the highest specificity constant (1/α)=1] whereas it discriminated strongly against docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) due to their lowest specificity constants (1/α=0.19 and 0.2). Moreover, the highest n-3 PUFA-rich TAG content (55.8%) with similar n-3 PUFA composition to the substrate was obtained in choline chloride/glycerol (CG) system. There was a 1.38-fold increase of TAG content in CG system compared with that in the solvent-free system. Interestingly, immobilized MAS1 lipase exhibited no regiospecificity in the solvent-free and various NADES systems. Besides, the potential reaction mechanism of immobilized MAS1 lipase-catalyzed esterification of glycerol with n-3 PUFA in NADES systems was described. It was found that the use of NADES as solvents could greatly enhance TAG content, and make it easy to separate the product. These results indicated that immobilized MAS1 lipase is a promising biocatalyst for the efficient synthesis of n-3 PUFA-rich TAG by esterification of glycerol with n-3 PUFA in NADES systems.

This study reports the properties of immobilized MAS1-H108A lipase from marine Streptomyces sp. strain W007 on XAD1180 resin and its application in the synthesis of n-3 polyunsaturated fatty acids (PUFA)-rich triacylglycerols (TAG) for the first time. It was found that the optimal temperature and pH for both immobilized MAS1-H108A lipase and free lipase MAS1-H108A were 70 °C and 7.0, respectively. However, immobilized MAS1-H108A lipase exhibited higher thermostability when compared with free lipase MAS1-H108A. It was also interesting that both immobilized MAS1-H108A lipase and free lipase MAS1-H108A showed no regiospecificity in the hydrolysis of triolein. Subsequently, immobilized MAS1-H108A lipase and free lipase MAS1-H108A were employed to catalyze glycerolysis of n-3 PUFA-rich ethyl esters (EE) and esterification of n-3 PUFA with glycerol under vacuum in the solvent-free system. The results showed that n-3 PUFA-rich TAG were synthesized efficiently by non-regiospecific immobilized MAS1-H108A lipase and TAG contents separately reached 92.07% and 76.13% during the esterification and glycerolysis reactions, which were significantly higher than those (71.82% and 39.62%, respectively) obtained by free lipase MAS1-H108A. Besides, TAG exhibited similar n-3 PUFA composition to the substrate. These findings indicated that non-regiospecific immobilized MAS1-H108A lipase is a promising and efficient biocatalyst for the industrial synthesis of n-3 PUFA-rich TAG.

An effective and reliable method, magnetic solid phase extraction based on magnetic silica aerogels, has been developed and used for the enrichment of 3 pyrethroid insecticides (including fenpropathrin, lambda-cyhalothrin and cypermethrin) from juices and tea beverages. The magnetic silica aerogels were successfully synthesized by an acid–base two-step catalytic sol–gel method using tetraethylorthosilicate (TEOS) as a precursor solution, and characterized. Several influencing factors that could affect magnetic solid-phase extraction (MSPE) performance including adsorbent dosage, adsorption time, the pH of the solution, ionic strength, elution solvent type, the volume of elution solvent and the elution time were systematically investigated and optimized. Under the optimal conditions, an excellent linearity was shown for fenpropathrin and lambda-cyhalothrin in the 0.04–4 ng mL−1 range and cypermethrin in the 0.08–8 ng mL−1 range, and the correlation coefficients were in the range of 0.9985–0.9996. The recoveries of fenpropathrin, lambda-cyhalothrin and cypermethrin ranged from 79.5% to 102.7% with relative standard deviation (RSD) less than 9.7%. The detection limit of the method was from 0.008 to 0.024 ng mL−1. The MSPE method, based on magnetic silica aerogels, possessed high sensitivity, efficiency, and convenience, and was suitable to determine the trace pyrethroid insecticides in juices and tea beverages.

Penicillium camembertii (PCL), a mono- and di-acylglycerol lipase (DGL), has the vital potential in the oil chemistry for food industry. However, known DGLs are mesophilic enzymes which restricts its application in the industry. To improve thermostability of PCL, we used amino acid substitution by comparison of amino acids compositions of PCL and protein sequences from typical thermophilic bacteria. Then, some conservative residues around active center were avoided to mutate according to homologous alignment analyses. Furthermore, the list was narrowed down to 28 candidate mutational sites of PCL by analyzing the hydrophobic interaction of amino acids in the structure. And among them only the mutant PCL-D25R had formed an additional salt bridge between R25-D32 and increased more hydrogen bonds interaction. Therefore, mutant PCL-D25R were constructed and expressed. Thermal inactivation assay showed that the half-life of mutant PCL-D25R at 45 °C increased 4-fold compared to that of PCL-WT. Melting temperature of mutant PCL-D25R increased to 49.5 °C from 46.5 °C by fluorescence-based thermal stability assay. This study provides a valuable strategy for engineering DGL thermostability.

MAS1 from marine Streptomyces sp. strain W007 belongs to the bacterial lipase I.7 subfamily and is characterized as a thermostable and nonregiospecific lipase. To shed light on the catalytic mechanism of MAS1, we determined its crystal structure with closed conformation in two crystal forms at 2.3 A resolution. MAS1 adopts the canonical α/β hydrolase core fold with its catalytic triad being formed by S109, D200 and H232. Structural analysis and biochemical assays revealed that disulfide bonds and salt bridges play a vital role in the thermostability of MAS1. In addition, we discovered that the replacement of H108 with a tryptophan converts MAS1 from a nonregiospecific to an sn-1,3-specific lipase, suggesting the functional importance of the second position from the conserved pentapeptide motif in defining the regiospecificity of MAS1. Our present study provides insights into the molecular basis for the thermostability and regiospecificity of MAS1, and it may aid in the rational design of thermostable or regiospecific lipases for potential industrial applications. Database Structural data are available in the Protein Data Bank database under the accession numbers 5H6B and 5H6G.

The mechanism of lipase binding to the lipid-water interface is crucial for substrate specificity and kinetic properties. In this study, the chain-length specificity, regiospecificity and substrate specificity of Phospholipase A1 (PLA1) and its parent enzyme Thermomyces lanuginosus lipase (TLL) have been investigated using a classical emulsion system. The results show that both PLA1 and TLL are 1,3-regioselective lipases. Additionally, the hydrolytic activity of PLA1 is comparatively lower on short-chain triacylglyceride (TAG) and higher on phosphatidylcholine (PC) than the hydrolytic activity of TLL. Further, the results obtained with monolayer film techniques demonstrate that the C-terminal region regulates the binding of PLA1 to PC. A hypothesis is presented according to which the α9 helix of C-terminal region in PLA1 not only controls the opening of lid but also serves as a membrane anchor that assists in binding to PC. These findings bring new insight into rational design of novel lipases with intriguing functionalities.

Lipases with unique substrate specificity are highly desired in biotechnological applications. In this study, a putative marine Geobacillus sp. monoacylglycerol lipase (GMGL) encoded gene was identified by a genomic mining strategy. The gene was expressed in Escherichia coli as a His-tag fusion protein and purified by affinity chromatography with a yield of 264 mg per liter fermentation broth. The recombinant GMGL shows the highest hydrolysis activity at 60 &deg, C and pH 8.0, and the half-life was 60 min at 70 &deg, C. The GMGL is active on monoacylglycerol (MAG) substrate but not diacylglycerol (DAG) or triacylglycerol (TAG), and produces MAG as the single product in the esterification reaction. Modeling structure analysis showed that the catalytic triad is formed by Ser97, Asp196 and His226, and the flexible cap region is constituted by residues from Ala120 to Thr160. A mutagenesis study on Leu142, Ile145 and Ile170 located in the substrate binding tunnel revealed that these residues were related with its substrate specificity. The kcat/Km value toward the pNP-C6 substrate in mutants Leu142Ala, Ile145Ala and Ile170Phe increased to 2.3-, 1.4- and 2.2-fold as compared to that of the wild type, respectively.

A rapid, reliable and eco-friendly method for the determination of three sex hormones in five kinds of milk was developed and validated by combining vortex-assisted liquid-liquid microextraction (VALLME) and magnetic solid-phase extraction (MSPE). Deep eutectic solvents (DESs) such as choline chloride/urea were considered as the extraction solvent in VALLME and multi-walled carbon nanotubes (MMWCNTs) were used as the adsorbent which could adsorb DESs on the surface. The optimum experimental conditions were as follows: amount of MMWCNTs for 10 mg, volume of acetone for 4 mL, no sodium chloride and extraction pH at 7. After the optimization of several main variables, satisfactory sensitivity levels were achieved as low as 1.0–1.3 ng mL−1 and 2.5–4.5 ng mL−1 for the limit of method detections and the limit of method quantitation, respectively. The recoveries of the three hormones in different milk samples were in the range of 80.1%-116.4%. Consequently, this method is suitable for monitoring the trace amount of sex hormones in milk matrices.

Diacylglycerol (DAG)-enriched oil has drawn considerable attention for the prevention of obesity and other lifestyle-related diseases. In this study, a mutant lipase of SMG1 (SMG1-F278D) from Malassezia globosa was studied for the production of DAG. The SMG1-F278D exhibited 4-fold increased esterification activity as well as superior fatty acid (FA) specificity for medium chain FAs. Molecular docking study suggested that the caprylic acid (CA) was strongly bound to the catalytic residue Ser171 present in the SMG1-F278D as compared to SMG1. Molecular Dynamics (MD) simulations were employed in order to understand the structural conformations of SMG1 and SMG1-F278D. The structure of SMG1-F278D was found to be more compact as well as less deviated from its native conformation. There was an increase in β-sheet as well as α-helix in the SMG1-F278D that may stabilize the protein structure due to point mutation. Finally, the capability of SMG1-F278D in synthesis of DAG was evaluated. Effects of reaction parameters such as substrate molar ratio of glycerol to FAs, enzyme loading and reaction temperature on the esterification were investigated. The optimal reaction conditions were achieved as 4:1 molar ratio of glycerol to FAs, enzyme loading of 100 U/g (U/w, with respect to the total substrates) and temperature of 25 °C. The present study provides important information about SMG1 mutant for better utilization in the oils and fats industries.

A simple rapid and efficient deep eutectic solvent-based magnetic colloidal gel (DES-MCG) assisted magnetic solid-phase extraction (MSPE) method followed by high performance liquid chromatography with a diode array detector (HPLC-DAD) was established for determination of four sex hormones (including ethinylestradiol, norgestrel, megestrol acetate and medroxyprogesterone acetate) in cosmetic skin care toners. The DES-MCG with the desirable advantages of high adsorbing ability was prepared by combining choline chloride/urea deep eutectic solvent and magnetic multiwalled carbon nanotubes (MMWCNTs). The synthesized DES-MCG was characterized using fourier transform infrared spectrometry (FTIR), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and vibrating sample magnetometry (VSM). The cosmetic skin care toners were concentrated by a rotary evaporator and the obtained solutions were further purified by DES-MCG assisted magnetic solid-phase extraction. Response surface methodology (RSM) was applied for efficient optimization of the main variables in the extraction procedure. Under the optimized conditions, method detection limits and method quantitation limits were in the range of 1.2–6.6 ng mL−1 and 4.4–26.6 ng mL−1, respectively. The recoveries of the four sex hormones in different cosmetic skin care toners ranged from 80.1% to 118.8% and the precisions were no more than 0.35%. The developed method was successfully applied for the determination of sex hormones in cosmetic skin care toners.

A green and highly efficient ultrasound-assisted deep eutectic solvent extraction combined with functionalized magnetic multi-walled carbon nanotubes solid-phase extraction method for determination of seven pesticide residues in food products was developed. Various types of deep eutectic solvents (DESs) were screened for high extraction efficiencies and DES composing of proline and propylene glycol at 1:3 M ratio was selected as it exhibited highest yields. Extraction conditions were statistically optimized through response surface methodology using a Box-Behnken design. The established method was linear, precise, and accurate over the range of 0.1–50 μg mL−1. Limit of detection and limit of quantification were in the range of 0.02–0.05 μg mL−1 and 0.05–0.10 μg mL−1, respectively. The mean recoveries for pesticides ranged from 76.09 to 97.96% with relative standard deviation of 0.13–10.05%. The proposed method was successfully applied to analysis of the pesticides in real samples, which is a potentially promising technique for food matrix.

Fungal lipases are efficient and environment-friendly biocatalysts for many industrially relevant processes. One of the most widely applied lipases in the manufacturing industry is Candida antarctica lipase B (CALB). Here, we report the biochemical and structural characterization of a novel CALB-like lipase from an important human pathogen-Aspergillus fumigatus (AFLB), which has high sn-1,3-specificity toward triolein. AFLB crystal structure displays a CALB-like catalytic domain and hosts a unique tightly closed 'lid' domain that contains a disulfide bridge, as well as an extra N-terminal subdomain composed of residues 1-128 (including the helix α1-α5 located above the active site). To gain insight into the function of this novel lid and N-terminal subdomain, we constructed and characterized a series of mutants in these two domains. Deleting the protruding bulk lid's residues, replacing the bulk and tight lid with a small and loose lid from CALB, or breaking the disulfide bridge increased the affinity of CALB for glyceride substrates and improved its catalytic activity, along with the loss of enzyme fold stability and thermostability. N-terminal truncation mutants revealed that the N-terminal peptide (residues 1-59) is a strong inhibitor of AFLB binding to lipid films. This peptide thus limits AFLB's penetration power and specific activity, revealing a unique enzyme activity regulatory mechanism. Our findings on the functional and structural properties of AFLB provide a better understanding of the functions of the CALB-like lipases and pave the way for its future protein engineering. DATABASE: Structural data are available in the Protein Data Bank under the accession numbers 6IDY.

In Gas-Insulated Substation (GIS), the Transient Enclosure Voltage (TEV) would threaten the reliability of secondary devices. Studying the propagation law of TEV can provide reference for suppression method of TEV. In this paper, field measurement and theoretical simulation were taken to study the TEV of a 330kV GIS. At first, the TEV caused by operation of disconnecting switch was measured and analyzed. Then, this paper built a simulating model based on electrical main connection to obtain the TEV under the maximum residual voltage. Finally, a method was proposed to reduce the TEV. The simulation results showed that reducing the grounding inductance can reduce the TEV effectively. The study shows that: (1) The maximum voltage of TEV can reach to 4.9kV, the transient time is among 100ms to 160ms, and the main frequency ranged from 11MHz to 35MHz. the maximum value of TEV does not always appear at the enclosure under out-line bushing in an operation, which is related to the TEV propagation path. (2) The TEV generated by the disconnecting switch near out-line bushing is much more serious than the TEV generated near the bus. The TEV generated during opening operation is more serious than the TEV generated during closing. (3) Reducing the equivalent inductance of grounding can reduce the value of TEV. When the equivalent inductance decrease 40%, the maximum voltage can be reduced by 21%. So reducing the equivalent inductance may be a possible effective way to reduce the value of TEV.

In this paper, the AC and DC test platforms are set up to study the change regularity of MOA characteristic parameters with various factors and typical faults by online and offline detection methods. The sensitivity and uniqueness of the characteristic parameters are summarized and analyzed to determine the state of MOA or varistors. The test results show that the resistive fundamental current is the most sensitive to the environment temperature and it increases about 2.27 times when the temperature changes from 20 C to 80 C. The resistive fundamental current is also the most sensitive to the change of MOA surface state and it increases about 1.52 times when the equivalent salt deposit density of surface is 0.06mg/cm2, but the DC leakage current is also very sensitive and it increases to 72.0μA. The characteristic parameters change a little after the deterioration test of MOA. The DC reference voltage is the most sensitive to the overvoltage deterioration of MOA and it decreases 6.4% when part of MOA is in the state of severe deterioration. Based on the test results, a qualitative state detection method is put forward.

Using the classical emulsified system and the monomolecular film technique, the substrate specificity of recombinant Gibberella zeae lipase (rGZEL) that originates from Gibberella zeae was characterized in detail. Under the emulsified reaction system, both phospholipase and glycolipid hydrolytic activities were observed, except for the predominant lipase activity. The optimum conditions for different activity exhibition were also determined. Compared with its lipase activity, a little higher ratio of glycolipid hydrolytic activity (0.06) than phospholipase activity (0.02) was found. rGZEL preferred medium chain-length triglycerides, while lower activity was found for the longer-chain triglyceride. Using the monomolecular film technique, we found that the preference order of rGZEL to different phospholipids was 1,2-diacyl-sn-glycero-3-phospho-l-serine (PS) > 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (PG) > 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) > l-α-phosphatidylinositol (PI) > cardiolipin (CL) > 3-sn-phosphatidic acid sodium salt (PA) > l-α-phosphatidylethanolamine (PE), while no hydrolytic activity was detected for sphingomyelin (SM). Moreover, rGZEL showed higher galactolipase activity on 1,2-distearoyimonoglactosylglyceride (MGDG). A kinetic study on the stereo- and regioselectivity of rGZEL was also performed by using three pairs of pseudodiglyceride enantiomers (DDGs). rGZEL presented higher preference for distal DDG enantiomers than adjacent ester groups, however, no hydrolytic activity to the sn-2 position of diglyceride analogs was found. Furthermore, rGZEL preferred the R configuration of DDG enantiomers. Molecular docking results were in concordance with in vitro tests.

Two effective molecularly imprinted polymers for the adsorption of alpha-lipoic acid (ALA) were synthesized by the cross-linking of chitosan with epichlorohydrin (ECH) and glutaraldehyde (GLU), respectively, in the presence of ALA as template molecules. Investigations on the molar ratios of ALA and chitosan (&ndash, NH2) in the preparation of chitosan molecularly imprinted polymers (MIPs) were carried out with a factor of ALA rebinding capabilities. The surface morphology and chemical properties of the polymers were characterized. The optimized MIPs crosslinked by ECH (MIPs&ndash, ECH) and MIPs crosslinked by GLU (MIPs&ndash, GLU) had adsorption capabilities of 12.09 mg/g and 19.72 mg/g for ALA, respectively. The adsorption behaviors of two kinds of chitosan MIPs including adsorption kinetics and isotherms were investigated in detail. Adsorption and kinetic binding experiments showed that the prepared MIPs&ndash, ECH and MIPs&ndash, GLU had selective adsorption and excellent affinity for ALA. In addition, the possible binding models between ALA and chitosan oligosaccharide were predicted by molecular dynamics simulation.

Lipases are important industrial enzymes. Most of the lipases operate at lipid–water interfaces enabled by a mobile lid domain located over the active site. Lid protects the active site and hence responsible for catalytic activity. In pure aqueous media, the lid is predominantly closed, whereas in the presence of a hydrophobic layer, it is partially opened. Hence, the lid controls the enzyme activity. In the present review, we have classified lipases into different groups based on the structure of lid domains. It has been observed that thermostable lipases contain larger lid domains with two or more helices, whereas mesophilic lipases tend to have smaller lids in the form of a loop or a helix. Recent developments in lipase engineering addressing the lid regions are critically reviewed here. After on, the dramatic changes in substrate selectivity, activity, and thermostability have been reported. Furthermore, improved computational models can now rationalize these observations by relating it to the mobility of the lid domain. In this contribution, we summarized and critically evaluated the most recent developments in experimental and computational research on lipase lids.

The mechanism of lipase binding to the lipid-water interface is crucial for substrate specificity and kinetic properties. In this study, the chain-length specificity, regiospecificity and substrate specificity of Phospholipase A