Pilot-scale field studies on activated microbial remediation of petroleum-contaminated soil.

Soil contamination by petroleum, including crude oil from various sources, is increasingly becoming a pressing global environmental concern, necessitating the exploration of innovative and sustainable remediation strategies. The present field-scale study developed a simple, cost-effective microbial remediation process for treating petroleum-contaminated soil. The soil treatment involves adding microbial activators to stimulate indigenous petroleum-degrading microorganisms, thereby enhancing the total petroleum hydrocarbons (TPH) degradation rate. The formulated microbial activator provided a growth-enhancing complex of nitrogen and phosphorus, trace elements, growth factors, biosurfactants, and soil pH regulators. The field trials, involving two 500 m3 soil samples with the initial TPH content of 5.01% and 2.15%, were reduced to 0.41% and 0.02% in 50 days, respectively, reaching the national standard for cultivated land category II. The treatment period was notably shorter than the commonly used composting and bioaugmentation methods (typically from 8 to 12 weeks). The results indicated that the activator could stimulate the functional microorganisms in the soil and reduce the phytotoxicity of the contaminated soil. After 40 days of treatment, the germination rate of rye seeds increased from 20 to 90%, indicating that the microbial activator could be effectively used for rapid on-site remediation of oil-contaminated soils.

In-situ noncovalent interaction of ammonium ion enabled C-H bond functionalization of polyethylene glycols.

The noncovalent interactions of ammonium ion with multidentate oxygen-based host has never been reported as a reacting center in catalytic reactions. In this work, we report a reactivity enhancement process enabled by non-covalent interaction of ammonium ion, achieving the C-H functionalization of polyethylene glycols with acrylates by utilizing photoinduced co-catalysis of iridium and quinuclidine. A broad scope of alkenes can be tolerated without observing significant degradation. Moreover, this cyano-free condition respectively allows the incorporation of bioactive molecules and the PEGylation of dithiothreitol-treated bovine serum albumin, showing great potentials in drug delivery and protein modification. DFT calculations disclose that the formed α-carbon radical adjacent to oxygen-atom is reduced directly by iridium before acrylate addition. And preliminary mechanistic experiments reveal that the noncovalent interaction of PEG chain with the formed quinuclidinium species plays a unique role as a catalytic site by facilitating the proton transfer and ultimately enabling the transformation efficiently.

Photoinduced [3 + 2] Cycloadditions of Aryl Cyclopropyl Ketones with Alkynes and Alkenes.

The five-membered ring skeleton is one of the most pivotal in the area of pharmaceutical and natural products. [3 + 2] cycloadditions of cyclopropyl and unsaturated compounds are a highly efficient and atom-economical way to build a five-member compound. The previous works about the kind of [3 + 2] cycloadditions usually utilized metal or organic small molecule catalysts. However, an ideal [3 + 2] cycloaddition reaction that smoothly happens without any additives and catalysts under mild conditions is underdeveloped. Hence, we report [3 + 2] cycloadditions of aryl cyclopropyl without any additives and catalysts under purple LED. In this method, a broad scope of cyclopropyl, alkyne, and alkene was very compatible, especially drug derivatives ibuprofen and Ioxoprofen, to obtain the corresponding cycloaddition product with a good yield up to 93%.

Iron-Catalyzed Csp2-Csp3 Cross-Coupling via Double Decarboxylation: One Step Synthesis of Remote Polar Alkenes.

Herein, we report an efficient photoinduced iron-catalyzed strategy for cross-couplings of alkyl carboxylic and acrylic acids, which provides a powerful tool for the synthesis of a variety of alkenes with polar functional groups. This novel synthetic methodology can also be applied to the preparation of ketones by using α-keto acids. Mechanistic experiments revealed preliminary mechanistic details. Diverse functionalization could be achieved, which may help streamline the synthesis of complex analogues for drug discovery.

Microbial enhanced oil recovery (MEOR): recent development and future perspectives.

After conventional oil recovery operations, more than half of the crude oil still remains in a form, which is difficult to extract. Therefore, exploring and developing new enhanced oil recovery (EOR) technologies have always been priority research in oilfield development. Microbial enhanced oil recovery (MEOR) is a promising tertiary oil recovery technology that has received widespread attention from the global oil industry in recent years due to its environmental friendliness, simplicity of operation, and cost-effectiveness. This review presents the: principle, characteristics, classification, recent development, and applications of MEOR technology. Based on hundreds of field trials conducted worldwide, the microbial strains, nutrient systems, and actual effects used in these technologies are summarized, with an emphasis on the achievements made in the development and application of MEOR in China in recent years. These technical classifications involve: microbial huff and puff recovery (MHPR), microbial flooding recovery (MFR), microbial selective plugging recovery (MSPR), and microbial wax removal and control (MWRC). Most of them have achieved good results, with a success rate of approximately 80%. These successful cases have accumulated into rich experiential indications for the popularization and application of MEOR technology, but there are still important yet uncertain factors that hinder the industrialization of this technology. Finally, based on the extensive research and development of MEOR by the authors, especially in both laboratory and industrial large scales, the main challenges and future perspectives of the industrial application for MEOR are presented.

Characterization and Production of a Biosurfactant Viscosin from Pseudomonas sp. HN11 and its Application on Enhanced oil Recovery During oily Sludge Cleaning.

Biosurfactants are renewable resources with versatile applications on environmental bioremediation and industrial processes. Pseudomonas species are one of the promising biosurfactant producers. However, besides rhamnolipids, little is known about Pseudomonas-derived biosurfactants on solubilization of polycyclic aromatic hydrocarbons (PAHs) and oily sludge treatment. In this study, Pseudomonas sp. HN11-derived biosurfactant was purified by chromatographic methods and was characterized as viscosin via bioinformatic analysis, spectrometric and spectroscopic analyses, Marfey's method and (C-H)α NMR fingerprint matching approach. Viscosin is a potent biosurfactant with critical micelle concentration of 5.79 mg/L and is stable under various stresses. Moreover, viscosin was produced at 0.42 g/L at 48 h of liquid fermentation. Further data have shown that emulsifying agent viscosin is capable of promoting the solubilization of PAHs and displays enhanced oil recovery during oily sludge treatment. More specifically, viscosin has shown significantly enhanced solubilization on fluoranthene compared with control (0.04 mg/L), 2.21 mg/L and 1.27 mg/L fluoranthene was recovered from 100 mg/L and 200 mg/L viscosin treatment, respectively. However, only 200 mg/L viscosin has significantly enhanced the solubilization of phenanthrene (0.75 mg/L) and benzo[a]pyrene (0.51 mg/L) compared to each control (0.23 mg/L for phenanthrene and 0.09 mg/L for benzo[a]pyrene). Viscosin treatment of oily sludge (recovering of 0.58 g oil) has shown a significant oil recovery compared to that of control (recovering of 0.42 g oil). This study shows the great potential of viscosin-type biosurfactant on oily sludge treatment.

Bioremediation of oily sludge by solid complex bacterial agent with a combined two-step process.

In the present research, a bioremediation process was developed using solid complex bacterial agents (SCBA) through a combined two-step biodegradation process. Four isolated strains showed high efficiency for the degradation of total petroleum hydrocarbons (TPH) and the reduction of COD of the oily sludge, at 96.6% and 92.6%, respectively. The mixed strains together with bran prepared in form of SCBA exhibited improved performance compared to individual strains, all of which had an optimal temperature of around 35 °C. The use of SCBA provided advantages over commonly used liquid media for storage and transportation. The two-step process, consisting of firstly biosurfactant-assisted oil recovery and secondly biodegradation of the remaining TPH with SCBA, demonstrated the capability for treating oily sludge with high TPH content (>10 wt%) and short process period (60 days). The large-scale (5 tons oily sludge) field test, achieving a TPH removal efficiency of 93.8% and COD reduction of 91.5%, respectively, confirmed the feasibility and superiority of the technology for industrial applications.

Ligand-free copper-catalyzed C(sp3)-H imidation of aromatic and aliphatic methyl sulfides with N-fluorobenzenesulfonimide.

A novel and efficient process has been developed for copper-catalyzed C(sp3)-H direct imidation of methyl sulfides with N-fluorobenzenesulfonimide(NFSI). Without using any ligands, various methyl sulfides including aromatic and aliphatic methyl sulfides, can be transformed to the corresponding N-((phenylthio)methyl)-benzenesulfonamide derivatives in good to excellent yields.

A 3D Cu(ii) tetrazolate coordination polymer based on pentanuclear units with a large coercive field.

A novel 3D coordination polymer {[Cu4.5 (BTZE)1.5 (µ3-OH)3(µ-OH)(SO4)(H2O)1.5·4H2O]}n (1) was synthesized by a solvothermal reaction of 1,2-bis(tetrazol-5-yl) ethane (BTZE) with copper sulfate. Compound (1) contained triangular [Cu3(µ3-OH)] cluster based magnetic Δ-chains linked with in situ generated µ2-BTZE ligands to form a 2D cyclic annular layer. This 2D layer structure was further modified with sulfate and symmetry-related µ3-OH groups, extending to a 3D coordination framework structure. The magnetic performance of (1) was characterized in the temperature range of 2-300 K in terms of direct-current and alternating-current magnetic susceptibilities, revealing that (1) was a canted ferromagnet with a critical temperature (Tc) of 9.5 K. Notably, (1) behaved as a hard magnet with a coercive field of 2.3 kOe at 2 K, showing significant unique characteristics compared to those of the reported spin canting systems based on pure Cu(ii) ions.

Biodegradation of crude oil by Chelatococcus daeguensis HB-4 and its potential for microbial enhanced oil recovery (MEOR) in heavy oil reservoirs.

Biodegradation of crude heavy oil was investigated with Chelatococcus daeguensis HB-4 that was isolated from the produced fluid of Baolige Oilfield in China. Batch growth characterization and crude oil degradation tests confirmed HB-4 to be facultative anaerobic and able to degrade heavy oil. The oil degradation was found to occur through degrading long hydrocarbons chains to shorter ones, resulting in oil viscosity reduction. By mixing crude oil with glucose, or using sole crude oil as carbon source, the content of light fractions (C8-C22) increased by 4.97% while heavy fractions (C23-C37) decreased by 7.98%. It was also found that bioemulsifiers were produced rather than commonly observed biosurfactants in the fermentation process, which was attributed to the extracellular degradation of hydrocarbons. Core flooding tests demonstrated 20.5% oil recovery by microbial enhancement, and 59.8% viscosity reduction, showing potential of strain HB-4 for application in the oil industry, especially in enhanced heavy oil recovery.

High efficiency and fast separation of active proteins by HIC chromatographic pie with sub-2 µm polymer packings.

This paper reports the development of hydrophobic interaction chromatography (HIC) by synthesizing sub-2 µm polymer packings which was packed into a chromatographic pie for fast separation of native proteins at low pressures demonstrating high efficiency. Using styrene as monomer and ethylene dimethacrylate (EDMA)as swelling agent, the polystyrene seeds with an average particle size of 0.8 µm and monodisperse polymeric microspheres with a particle size of 1.5-5.0 µm were synthesized through dispersion polymerization and one-step swelling method, respectively. In order to separate active proteins, the microspheres were modified to hydrophobic chromatographic packings through covalent bonding with benzene methanol. Compared with the traditional column chromatography, the sub-2 µm polymer packings in chromatographic pie exhibited higher column efficiency for protein separation at lower column pressures, even at higher flow rates. The van Deemter curve showed that the flow rate had insignificant effect on column efficiency of chromatographic pie. Seven example proteins were clearly separated within 3 min at a flow rate of 10 mL/min. The applicability of this method was further demonstrated by the separation of human serum samples. The results indicated that this chromatographic mode can be potentially applied for the fast separation of complex active proteins, such as protein drugs from natural products.

Luteimonas huabeiensis sp. nov., isolated from stratum water.

A yellow-coloured bacterial strain, designated HB2(T), isolated from stratum water was investigated using a polyphasic taxonomic approach. Cells were Gram-stain-negative, aerobic, non-spore-forming, non-flagellated and rod-shaped. Phylogenetic analysis based on 16S rRNA gene sequences showed that the strain was a member of the genus Luteimonas, its three closest neighbours being Luteimonas aquatica BCRC 17731(T) (97.5% similarity), Luteimonas marina JCM 12488(T) (97.3%) and Luteimonas aestuarii DSM 19680(T) (96.9%). Strain HB2(T) could clearly be distinguished from these type strains based on phylogenetic analysis, DNA-DNA hybridization, fatty acid composition and a range of physiological and biochemical characteristics. It is evident from the genotypic and phenotypic data that strain HB2(T) represents a novel species of the genus Luteimonas, for which the name Luteimonas huabeiensis sp. nov. is proposed. The type strain is HB2(T) ( =DSM 26429(T) =CICC 11005s(T)).

A new approach for characterizing the intermediate feature of α-chymotrypsin refolding by hydrophobic interaction chromatography.

A new approach for characterizing the intermediate of urea-denatured alpha-chymotrypsin (alpha-Chy) by hydrophobic interaction chromatography (HIC) is presented. The contact surface region (Z, S), affinity (logI), and the character of interaction force (j) of the alpha-Chy to the stationary phase of HIC (STHIC) between the intermediate (M) and native (N) states were found to be quite different as urea concentration (C(urea)) changes. With the changes in C(urea), a linear relationship between logI and Z was found to exist only for its N state, not for M state, indicating the interaction force between alpha-Chy in N state to the STHIC to be non-selective, but selective one for its M state. Also, the measured magnitude of both logI and Z in M state is only a fifth of that in N state. All three parameters were employed to distinguish protein in the N state from that in the M state. It would be expected that this result could be employed to distinguish any kind of non-functional protein having correct three-, or four-dimensional molecular structure from their stable M state of any kinds of proteins, and/or other proteins in proteome investigation, separation process of protein, and intensively understanding the intrinsic rule of protein folding in molecular biology.

On-line separation of native proteins by two-dimensional liquid chromatography using a single column.

This paper reports the on-line separation of native (N) proteins by two-dimensional liquid chromatography (2D-LC) using a single column with one phase (called 2D column). The 2D column exhibits excellent resolution, selectivity, and retention of proteins in the N state and functions in two retention modes--hydrophobic interaction chromatography (HIC) and weak-cation exchange chromatography (WCX). We describe a new approach to on-line buffer exchange and collection of fractions from the first retention mode and their quantitative re-injection into the same column, followed by re-separation in the second retention mode. Thus, liquid chromatography in a closed system and in an on-line manner could be successfully carried out. This method was termed on-line protein separation by 2D-LC using only a single column (on-line 2D-LC-1C). The applicability of this method was experimentally demonstrated using standard proteins and a human serum sample. The total hypothetical maximum possible peak capacity n(c,total) and total sample peak capacity n(c,total)(*) of the 2D column were 329 and 199, respectively. By comparison against several popular commercially available columns, it was found that the 2D column had not only comparable resolution and better selectivity but also some unique characteristics. This 2D-LC-1C method could be applied to the fast purification of intact proteins in the N state, such protein drugs from natural products, and recombinant proteins and also for the fast pre-fractionation of intact proteins in the "top-down" MS strategy in proteomics.

A fast method for preparation of cytochrome-C of high purity from pig heart by hydrophobic interaction chromatography.

A fast method for the preparation of cytochrome C (Cyt-C) of high purity from pig heart is presented. A simple-type column of hydrophobic interaction chromatography packed with big particles was employed for the fast purification in laboratory scale. The purity of the purified Cyt-C was verified by reversed-phase liquid chromatography, UV spectroscopy, and iron content in it. Compared with the common method, the new method has advantages of higher speed, simpler purification processes, higher mass recoveries and higher purity.

Design and resolution of a simple-type column for renaturation with simultaneous purification of proteins and its application to lysozyme refolding.

A new kind of column called simple-type chromatographic column using the packings of large particles, which resembles columns used in traditional low pressure liquid chromatography in both shape and operation but has resolution of biopolymers similar to high performance liquid chromatography, was designed and investigated. In addition, so long as a suitable stationary phase, such as hydrophobic interaction chromatographic stationary phase, is packed, the column can be employed for the purification with simultaneous renaturation of proteins. The structure, operation and characteristics of the designed column including pressure drop, column life, and resolution were investigated. Lysozyme was selected as the model protein for investigating the renaturation. The recoveries of mass and bioactivity were found to be (96.6 +/- 1.3)% and (101.1 +/- 6.0)% respectively when the original concentration of lysozyme was 50.0 g/L. The simple-type column is very cheap and easy to use, and can be applied to the purification of proteins under the condition of low pressure. Thus, it has great potential applications for the purification and/or renaturation of proteins in large scale, down-stream processing of therapeutic proteins in biotechnology.

Preparation of weak cation exchange packings based on monodisperse poly(glycidyl methacrylate- co-ethylene dimethacrylate) beads and their chromatographic properties.

The monodisperse, macroporous poly(glycidyl methacrylate- co-ethylene dimethacrylate) beads were synthesized by a single-step swelling and polymerization method. Based on this media, a weak cation exchange (WCX) stationary phase for HPLC was synthesized by a new chemically modified method. The prepared resin has advantages for biopolymer separation, high column efficiency, low column backpressure, high protein mass recovery, and good resolution for proteins. The measured bioactivity recovery for lysozyme was 98+/-5%. The dynamic protein loading capacity of the WCX packings was 17.3 mg g(-1). The experimental results show that the synthesized WCX resin has very weak hydrophobicity.