NMR Fingerprinting Research Articles

NMR Studies of High Voltage Electrolyte Decomposition Reactions

Lithium-ion batteries have become an irreplaceable part of daily life, yet state-of-art LIBs could not meet the growing demand of higher energy density partially due to limited choices for the cathode and the electrolyte that can provide the desired safety and reliability. One of the solutions is to move towards higher voltage systems. To rationalize the CEI design, on the electrolyte side, fundamental studies of electrochemical decomposition behavior at high voltage of common electrolytes, typically using carbonate-based organic solvent, remain incomplete despite experimental and computational efforts. Several decomposition products of EC were identified, mostly pointing at ring-opened linear molecules such as CO, CO2, glycolic acid and oxalic acid with the presence of active/singlet oxygen or PEO-like polymer owing to Lewis acid induced polymerization. However, in some cases there were no ring opening observed with EC forming vinylene carbonate (VC) or poly-EC instead. While the mechanism of ring-opening reactions that involve deprotonation, nucleophilic and/or electrophilic reactions are straightforward and widely reported, full reaction mechanisms pathways are not yet realized. In this study we combined experimental and computational NMR to track the dynamic evolution of intermediate species to discover their chemical nature by following their NMR fingerprints. The findings will be presented to provide a clearer picture of the effects of high voltage on electrolyte systems in general and their implications on overall battery chemistry.

A multifaceted approach to understanding protein-buffer interactions in biopharmaceuticals

The excipient selection process plays a crucial role in biopharmaceutical formulation development to ensure the long-term stability of the drug product. Though there are numerous options approved by regulatory authorities, only a subset is commonly utilized. Previous research has proposed various stabilization mechanisms, including protein-excipient interactions. However, identifying these interactions remains challenging due to their weak and transient nature. In this study, we present a comprehensive approach to identify such interactions. Using the 1HT2 CPMG (Carr-Purcel-Meiboom-Gill) filter experiment we identified interactions of rituximab with certain buffers and amino acids, shedding light on its Fc fragment instability that manifested during the enzymatic cleavage of the antibody. Moreover, chemometric analyses of 2D NMR fingerprints revealed interactions of selected excipients with antibody fragments. Furthermore, molecular dynamics simulations revealed potential interacting hotspots without NMR spectra assignment. Our results highlight the importance of an orthogonal methods approach to uncovering these critical interactions, advancing our understanding of excipient stabilization mechanisms and rational formulation design in biopharmaceutics.

Authentication of apples (Malus × domesticaborkh.) according to geographical origin, variety and production method using 1H NMR spectroscopy and random forest

When buying apples, there are several characteristics that are important to the consumer. These include taxonomic variety, which influences the desired flavor and the allergenicity, and, increasingly, whether the cultivation is regional and organic. In this study, we analyzed 217 apple samples with 1H NMR spectroscopy and verified the authentication of the apples in terms of country of origin, local origin within Germany, taxonomic variety and whether they were organically produced. The obtained random forest models showed classification accuracies ranging from about 73 % for the discrimination of seven taxonomic varieties to 88.5 % for the separation of German and non-German samples. The results demonstrate that 1H NMR spectroscopy is promising for comprehensive authentication of apples. This is achieved by using different parts of the NMR fingerprint depending on the desired outcome, which was confirmed by the application of variable selection methods and the comparison of the selected variables.

NMR Fingerprinting of Conventional and Genetically Modified Soybean Plants with AtAREB1 Transcription Factors.

Drought stress impacts soybean yields and physiological processes. However, the insertion of the activated form of the AtAREB1 gene in the soybean cultivar BR16, which is sensitive to water deficit, improved the drought response of the genetically modified plants. Thus, in this study, we used 1H NMR in solution and solid-state NMR to investigate the response of genetically modified soybean overexpressing AtAREB1 under water deficiency conditions. We achieved that drought-tolerant soybean yields high content of amino acids isoleucine, leucine, threonine, valine, proline, glutamate, aspartate, asparagine, tyrosine, and phenylalanine after 12 days of drought stress conditions, as compared to drought-sensitive soybean under the same conditions. Specific target compounds, including sugars, organic acids, and phenolic compounds, were identified as involved in controlling sensitive soybean during the vegetative stage. Solid-state NMR was used to study the impact of drought stress on starch and cellulose contents in different soybean genotypes. The findings provide insights into the metabolic adjustments of soybean overexpressing AREB transcription factors in adapting to dry climates. This study presents NMR techniques for investigating the metabolome of transgenic soybean plants in response to the water deficit. The approach allowed for the identification of physiological and morphological changes in drought-resistant and drought-tolerant soybean tissues. The findings indicate that drought stress significantly alters micro- and macromolecular metabolism in soybean plants. Differential responses were observed among roots and leaves as well as drought-tolerant and drought-sensitive cultivars, highlighting the complex interplay between overexpressed transcription factors and drought stress in soybean plants.

19F NMR Probes: Molecular Logic Material Implications for the Anion Discrimination and Chemodosimetric Approach for Selective Detection of H2O2.

Detection and discrimination of similar solvation energies of bioanalytes are vital in medical and practical applications. Currently, various advanced techniques are equipped to recognize these crucial bioanalytes. Each strategy has its own benefits and limitations. One-dimensional response, lack of discrimination power for anions, and reactive oxygen species (ROS) generally limit the utilized fluorescent probe. Therefore, a cutting-edge, refined method is expected to conquer these limitations. The use of 19F NMR spectroscopy for detecting and discriminating essential analytes in practical applications is an emerging technique. As an alternative strategy, we report two fluorinated boronic acid-appended pyridinium salts 5-F-o-BBBpy (1) and 5-CF3-o-BBBpy (2). Probe (1) acts as a chemosensor for identifying and discriminating inorganic anions with similar solvation energies with strong bidirectional 19F shifts in the lower ppm range. Probe (2) turns as a chemo dosimeter for the selective detection and precise quantification of hydrogen peroxide (H2O2) among other competing ROS. To demonstrate real-life applicability, we successfully quantified H2O2 via probe (2) in different pharmaceutical, dental, and cosmetic samples. We found that tuning the -F/-CF3 moiety to the arene boronic acid enables the π-conjugation, a crucial prerequisite for the discrimination of anions and H2O2. Characteristic 19F NMR fingerprints in the presence of anions revealed a complementary implication (IMP)/not implication (NIMP) logic function. Finally, the 16 distinct binary Boolean operations on two logic values are defined for "functional completeness" using the special property of the IMP gate. Boolean logic's ability to handle information by utilizing characteristic 19F NMR fingerprints has not been seen previously in a single chemical platform for detecting and differentiating such anions.

Chemometrics analysis of camellia oil authenticity using LF NMR and fatty acid GC fingerprints

Low-field nuclear magnetic resonance (LF NMR) parameters and fatty acid contents, combined with chemometrics, were employed to detect the adulteration of camellia oil (CAO) with various oils, including rapeseed oil (RO), sunflower oil (SSO), sesame oil (SEO), corn oil (COO), rice bran oil (RBO), peanut oil (PEO), palm superolein (PAO), or high oleic-peanut oil (HO-PEO). In principal component analysis (PCA), CAO was characterized by higher levels of C18:1n9 and S22, which differentiates it from other vegetable oils. Partial least squares-discriminant analysis (PLS-DA) revealed robust clustering among CAO and other samples. Adulterated CAO samples, with an adulteration level exceeding 10 %, could be accurately classified, achieving a total discrimination accuracy of over 97.15 %. Notably, when the adulteration rate surpassed 40 %, the recognition accuracy for all samples reached 100 %. The optimized partial least squares (PLS) models, relying on fewer than 18 potential key markers identified by variable importance in projection, demonstrated efficiency and precision in predicting the adulteration levels in CAO.

Quality Evaluation of Tripterygium Glycoside Tablets Based on Quantitative Band-Selective 2D 1H-13C HSQC and 1H NMR Fingerprinting.

Tripterygium glycoside tablets (TGTs) are preparations extracted and purified from Tripterygium wilfordii Hook. F and are extensively utilized in the treatment of autoimmune diseases, such as rheumatoid arthritis (RA). However, variations in production processes among manufacturers can lead to challenges in quality control and clinical utilization of TGTs. A band-selective 2D 1H-13C HSQC quantification method was applied for the determination of 13 active ingredients in TGTs. This method was validated following the guidelines of USP-NF 2022. The results demonstrated that the quantitative method exhibited excellent signal resolution, as well as sufficient accuracy, sensitivity, and stability. In addition, the 1H NMR spectra of TGTs from three manufacturers underwent analysis using principal component analysis and orthogonal partial least-squares discriminant analysis. The results revealed significant differences among the TGTs from the three manufacturers, with manufacturer 2 and manufacturer 3 demonstrating superior product consistency compared to manufacturer 1. A quality evaluation system for TGTs was developed based on band-selective 2D 1H-13C HSQC and 1H NMR, encompassing both quality markers and fingerprinting. This system offers reliable approaches and insights for enhancing the quality control of natural products.

Breaking Cycles: Saponification-Enhanced NMR Fingerprint Matching for the Identification and Stereochemical Evaluation of Cyclic Lipodepsipeptides from Natural Sources.

We previously described NMR based fingerprint matching with peptide backbone resonances as a fast and reliable structural dereplication approach for Pseudomonas cyclic lipodepsipeptides (CLiPs). In combination with total synthesis of a small library of configurational CLiP congeners this also allows unambiguous determination of stereochemistry, facilitating structure-activity relationship studies and enabling three-dimensional structure determination. However, the on-resin macrocycle formation in the synthetic workflow brings considerable burden and limits universal applicability. This drawback is here removed altogether by also transforming the native CLiP into a linearized analogue by controlled saponification of the ester bond. This eliminates the need for macrocycle formation, limiting the synthesis effort to linear peptide analogues. NMR fingerprints of such linear peptide analogues display a sufficiently distinctive chemical shift fingerprint to act as effective discriminators. The approach is developed using viscosin group CLiPs and subsequently demonstrated on putisolvin, leading to a structural revision, and tanniamide from Pseudomonas ekonensis COR58, a newly isolated lipododecapeptide that defines a new group characterized by a ten-residue large macrocycle, the largest to date in the Pseudomonas CLiP portfolio. These examples demonstrate the effectiveness of the saponification- enhanced approach that broadens applicability of NMR fingerprint matching for the determination of the stereochemistry of CLiPs.

Mechanism of antimalarial action and mitigation of infection-mediated mitochondrial dysfunction by phyto-constituents of Andrographis paniculata ((Burm f.) Wall. ex Nees) in Plasmodium berghei-infected mice

Ethnopharmacological relevanceAndrographis paniculata (AP) ((Burm f.) Wall. ex Nees) is a medicinal plant, documented for its folkloric use in the treatment of malaria. AimThis study was designed to determine the potency of extract and fractions of A. paniculata (AP) as a curative, both for susceptible and resistant malaria and to also determine the plant's mechanism of action. This study was also designed to determine whether AP extract and its most potent fraction will mitigate infection-mediated mitochondrial dysfunction, and to assess the phytochemical constituents of the most potent fraction. Materials and methodsn-Hexane, dichloromethane, ethylacetate and methanol were used to partition the methanol extract of A. paniculata. Graded doses of these extract and fractions were used to treat mice infected with chloroquine-sensitive strain of P. berghei in a curative model. The most potent fraction was used to treat mice infected with resistant (ANKA strain) P. berghei. Inhibition of hemozoin formation, reversal of mitochondrial dysfunction and antiinflammatory potentials were determined. A combination of ultraperformance liquid chromatography-quadrupole time of flight-mass spectrometry and nuclear magnetic resonance spectroscopy were used for chemical analysis. ResultsMicroscopy revealed that the dichloromethane fraction decreased the parasite burden the most, and inhibition of the hemozoin formation is one of its mechanisms of action. The dichloromethane fraction reversed parasite-induced mitochondrial pore opening in the host, enzyme-dependent ATP hydrolysis and peroxidation of host mitochondrial membrane phospholipids as well as its antiinflammatory potentials. The UPLC-qTOF-MS report and NMR fingerprints of the dichloromethane fraction of A. paniculata yielded fourteen compounds of which sibiricinone C was identified from the plant for the first time. ConclusionFractions of A. paniculata possess antiplasmodial effects with the dichloromethane fraction having the highest potency. The potent effect of this fraction may be attributed to the phytochemicals present because it contains terpenes implicated with antimalarial and antiinflammatory activities.

Nuclear Magnetic Resonance Fingerprints and Mini DNA Markers for the Authentication of Cinnamon Species Ingredients Used in Food and Natural Health Products.

Cinnamomum verum (syn C. zeylanicum) is considered 'true' cinnamon. However, it is reported that less expensive sources of cinnamon from C. cassia (syn C. aromaticum), C. loureiroi, and C. burmannii (toxic coumarin) may be used in the place of C. verum. We lack the quality assurance tools that are required to differentiate C. verum from other cinnamon species when verifying that the correct species is sourced from ingredient suppliers. The current research on cinnamon species authentication using DNA tools is limited to a few species and the use of high-quality DNA extracted from raw leaf materials. The cinnamon bark traded in the supply chain contains much less DNA and poorer-quality DNA than leaves. Our research advances DNA methods to authenticate cinnamon, as we utilized full-length chloroplast genomes via a genome skimming approach for C. burmannii and C. cassia to facilitate the design of optimal mini DNA markers. Furthermore, we developed and validated the use of NMR fingerprints for several commercial cinnamon species, including the quantification of 16 molecules. NMR fingerprints provided additional data that were useful for quality assessment in cinnamon extract powders and product consistency. Both the new mini DNA markers and NMR fingerprints were tested on commercial cinnamon products.

Two novel ketone alkaloids from Portulaca oleraceaL. and their anti-inflammatory activities

Eleven compounds were obtained from Portulaca oleracea L., including two novel ketone alkaloids, (1, 2), 4-hydroxy-3-methoxybenzamide (3) (isolated for the first time), β-adenosine (4), oleracrylimide A and B (5, 6), oleracein H, C, D, Q and A (7–11). The two novel ketone alkaloids were identified as 5-acetyl-5-methylcyclopent-2-ene-1-carboxamide (1), named oleraciamide H, and (2 R,3S,4R,5R)-5-((R)-1,2-dihydroxyethyl)-3,4-dihydroxytetrahydrofuran-2-yl glycinate (2), named oleracone Q by spectroscopic methods, including 1D, 2D NMR and compound fingerprints. Additionally, their anti-inflammatory activities were tested via RAW 264.7 cells induced by LPS and found that they could significantly inhibit the release of IL-1β and TNF-α.

Structure Revision of Formyl Phloroglucinol Meroterpenoids: A Unified Approach Using NMR Fingerprinting and DFT NMR and ECD Analyses

NMR fingerprints are valuable tools for analyzing complex natural product mixtures and identifying incorrectly assigned structures in the literature. Our diagnostic NMR fingerprints for formyl phloroglucinol meroterpenoids revealed discrepancies in the structures reported for eucalyprobusal C (1a) and eucalypcamal K (2a). NMR fingerprinting PCA analyses identified 1a as an oxepine-diformyl phloroglucinol and 2a as an oxepine 3-acyl-1-formyl phloroglucinol, contrary to their initial assignments as pyrano-diformyl and pyrano 3-acyl-1-formyl phloroglucinols, respectively. Extensive reinterpretation of their reported one- and two-dimensional NMR data, coupled with GIAO DFT-calculated 1H and 13C NMR chemical shift and DP4+ analyses, supported the unequivocal reassignment of eucalyprobusal C to 1b and eucalypcamal K to 2b. The absolute configurations of the revised oxepine-containing phloroglucinol meroterpenoids were confirmed via the reinterpretation of their reported ROESY and NOESY NMR data, along with comparative TDDFT-calculated and experimental ECD spectra.

Metabolomics and in-vitro bioactivities studies of fermented Musa paradisiaca pulp: A potential alpha-amylase inhibitor

The in-vitro synthesis of bio-compounds via fermentation is a promising route for bioactive molecules intended for disease control and management. Therefore, this study evaluated the effect of fermentation on the antioxidants, antihyperglycemic and anti-inflammatory properties and the resultant chemometric phytochemical profiles of unripe plantain fruits. The results revealed that Escherichia coli and Propionibacterium spp. are suspected as the key fermenters. The E coli showed negative results to the pathogenicity test; Propionibacterium appeared to be opportunistic. A significant increase in the total polyphenols and protein and decreased flavonoids was recorded in the phytochemical profile of the methanolic extract of the fermented unripe plantain pulp; however, the ascorbic acid content was not significantly altered. The 1H NMR fingerprint showed that there is a closely related chemical shift among the shorter fermentation time (days 2–6) and the unfermented, while the more extended fermentation periods (days 7–12) with enhanced bioactivities were closely related based on the chemometrics analyses. Furthermore, the UPLC-QTOF-MS analysis annotated the presence of bioactive compounds in the day-9 fermented sample: polyhydroxy glucose conjugates (3-Methoxy-4-hydroxyphenyl 6-O-(3,4,5-trihydroxybenzoyl)-beta-D-glucopyranoside), short chain peptide (leucyl-glycyl-glycine), amino acid derivatives (4-Aminophenylalanine, and N-Acetylhistidine), linear and cyclic fatty acid derivatives (palmitoyl putrescine, ricinoleic acid, phytosphingosine, gabalid, rubrenoic acid, 2-aminocyclopentanecarboxylic and cystodienioc acid). The synergistic effect of these newly formed compounds and the increase in the phenolic content of the day-9 fermented unripe plantain may account for its more potent antioxidant, anti-inflammatory and antihyperglycemic activity. Therefore, the products obtained from the day 9 fermentation of unripe plantain pulp may serve as potential nutraceutical agents against gastro-enteric sugar digestion and absorption and sugar-induced oxidative stress, inflammation and metabolic disease.

Hyperpolarized 1H and 13C NMR Spectroscopy in a Single Experiment for Metabolomics.

The application of NMR spectroscopy to complex mixture analysis and, in particular, to metabolomics is limited by the low sensitivity of NMR. We recently showed that dissolution dynamic nuclear polarization (d-DNP) could enhance the sensitivity of 13C NMR for complex metabolite mixtures, leading to the detection of highly sensitive 13C NMR fingerprints of complex samples such as plant extracts or urine. While such experiments provide heteronuclear spectra, which are complementary to conventional NMR, hyperpolarized 1H NMR spectra would also be highly useful, with improved limits of detection and compatibility with the existing metabolomics workflow and databases. In this technical note, we introduce an approach capable of recording both 1H and 13C hyperpolarized spectra of metabolite mixtures in a single experiment and on the same hyperpolarized sample. We investigate the analytical performance of this method in terms of sensitivity and repeatability, and then we show that it can be efficiently applied to a plant extract. Significant sensitivity enhancements in 1H NMR are reported with a repeatability suitable for metabolomics studies without compromising on the performance of hyperpolarized 13C NMR. This approach provides a way to perform both 1H and 13C hyperpolarized NMR metabolomics with unprecedented sensitivity and throughput.

Stereomeric Lipopeptides from a Single Non-Ribosomal Peptide Synthetase as an Additional Source of Structural and Functional Diversification in Pseudomonas Lipopeptide Biosynthesis.

In Pseudomonas lipopeptides, the D-configuration of amino acids is generated by dedicated, dual-function epimerization/condensation (E/C) domains. The increasing attention to stereochemistry in lipopeptide structure elucidation efforts has revealed multiple examples where epimerization does not occur, even though an E/C-type domain is present. While the origin of the idle epimerization in those E/C-domains remains elusive, epimerization activity has so far shown a binary profile: it is either 'on' (active) or 'off' (inactive). Here, we report the unprecedented observation of an E/C-domain that acts 'on and off', giving rise to the production of two diastereoisomeric lipopeptides by a single non-ribosomal peptide synthetase system. Using dereplication based on solid-phase peptide synthesis and NMR fingerprinting, we first show that the two cyclic lipopeptides produced by Pseudomonas entomophila COR5 correspond to entolysin A and B originally described for P. entomophila L48. Next, we prove that both are diastereoisomeric homologues differing only in the configuration of a single amino acid. This configurational variability is maintained in multiple Pseudomonas strains and typically occurs in a 3:2 ratio. Bioinformatic analysis reveals a possible correlation with the composition of the flanking sequence of the N-terminal secondary histidine motif characteristic for dual-function E/C-type domains. In permeabilization assays, using propidium iodide entolysin B has a higher antifungal activity compared to entolysin A against Botrytis cinerea and Pyricularia oryzae spores. The fact that configurational homologues are produced by the same NRPS system in a Pseudomonas strain adds a new level of structural and functional diversification to those already known from substrate flexibility during the recruitment of the amino acids and fatty acids and underscores the importance of complete stereochemical elucidation of non-ribosomal lipopeptide structures.

The origin of suspended particulate matter in the Great Barrier Reef

River run-off has long been regarded as the largest source of organic-rich suspended particulate matter (SPM) in the Great Barrier Reef (GBR), contributing to high turbidity, pollutant exposure and increasing vulnerability of coral reef to climate change. However, the terrestrial versus marine origin of the SPM in the GBR is uncertain. Here we provide multiple lines of evidence (13C NMR, isotopic and genetic fingerprints) to unravel that a considerable proportion of the terrestrially-derived SPM is degraded in the riverine and estuarine mixing zones before it is transported further offshore. The fingerprints of SPM in the marine environment were completely different from those of terrestrial origin but more consistent with that formed by marine phytoplankton. This result indicates that the SPM in the GBR may not have terrestrial origin but produced locally in the marine environment, which has significant implications on developing better-targeted management practices for improving water quality in the GBR.

1H NMR-based approach to determine the geographical origin and cultivation method of roasted coffee

A comprehensive study of 603 roasted arabica coffee samples using NMR fingerprinting and multivariate data analysis was performed to differentiate coffee samples according to their geographical origin and cultivation method. Both lipophilic and hydrophilic coffee metabolites were recorded using 1H NMR spectroscopy, and principal component analysis followed by linear discriminant analysis (PCA-LDA) was applied. Coffee samples were fist differentiated according to their continents of origin followed by discrimination of coffee samples from Brazil, Ethiopia, and Colombia from coffee samples originating from another continent. Discrimination of coffee samples according to their continent of origin and additional assignment to the countries Brazil and Ethiopia were successful. However, an unambiguous separation of Colombian coffee samples from coffee samples of another continent (other than South America) was not possible. Also, differentiation of organically and conventionally produced coffee samples by using 1H NMR and PCA-LDA was not achieved.

A multi-block data approach to assessing beef quality: ComDim analysis of hyperspectral imaging, 1H NMR, electronic nose and quality parameters data

Several factors affect the quality of beef. In the field of chemometrics, multi-block data analysis methods are useful for examining multiple sources of information from a sample. This study focuses on the application of ComDim, a multi-block data analysis method, to evaluate beef from different parts of hyperspectral spectrum and image texture information, 1H NMR fingerprints, quality parameters and electronic nose. Compared to principal component analysis (PCA) methods based on low-level data fusion, ComDim is more efficient and powerful, because it reveals the relationships between the methods and techniques studied, as well as the variability of beef quality across multiple metrics. The quality and metabolite composition of beef tenderloin and hindquarters were differentiated, with low L* value and high shear tenderloin distinguished from hindquarters with opposite characteristics. The proposed strategy demonstrates that ComDim approach can be used to characterize samples when different techniques describe the same set of samples.

NMR Fingerprints of Formyl Phloroglucinol Meroterpenoids and Their Application to the Investigation of Eucalyptus gittinsii subsp. gittinsii.

NMR fingerprints provide powerful tools to identify natural products in complex mixtures. Principal component analysis and machine learning using 1H and 13C NMR data, alongside structural information from 180 published formyl phloroglucinols, have generated diagnostic NMR fingerprints to categorize subclasses within this group. This resulted in the reassignment of 167 NMR chemical shifts ascribed to 44 compounds. Three pyrano-diformyl phloroglucinols, euglobal In-1 and psiguadiols E and G, contained 1H and 13C NMR data inconsistent with their predicted phloroglucinol subclass. Subsequent reinterpretation of their 2D NMR data combined with DFT 13C NMR chemical shift and ECD calculations led to their structure revisions. Direct covariance processing of HMBC data permitted 1H resonances for individual compounds in mixtures to be associated, and analysis of their 1H/13C HMBC correlations using the fingerprint tool further classified components into phloroglucinol subclasses. NMR fingerprinting HMBC data obtained for six eucalypt flower extracts identified three subclasses of pyrano-acyl-formyl phloroglucinols from Eucalyptus gittinsii subsp. gittinsii. New, eucalteretial F and (+)-eucalteretial B, and known, (-)-euglobal VII and eucalrobusone C, compounds, each belonging to predicted subclasses, were isolated and characterized. Staphylococcus aureus and Plasmodium falciparum screening revealed eucalrobusone C as the most potent antiplasmodial formyl phloroglucinol to date.

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 5.79mg/L and is stable under various stresses. Moreover, viscosin was produced at 0.42g/L at 48h 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 (0.04mg/L), 2.21mg/L and 1.27mg/L fluoranthene was recovered from 100mg/L and 200mg/L viscosin treatment, respectively. However, only 200mg/L viscosin has significantly enhanced the solubilization of phenanthrene (0.75mg/L) and benzo[a]pyrene (0.51mg/L) compared to each control (0.23mg/L for phenanthrene and 0.09mg/L for benzo[a]pyrene). Viscosin treatment of oily sludge (recovering of 0.58g oil) has shown a significant oil recovery compared to that of control (recovering of 0.42g oil). This study shows the great potential of viscosin-type biosurfactant on oily sludge treatment.