Footsteps detection and identification based on distributed optical fiber sensor and double-YOLO model.

A footstep detection and recognition method based on distributed optical fiber sensor and double-YOLO method is proposed. The sound of footsteps is detected by a phase-sensitive optical time-domain reflectometry (Φ-OTDR) and the footsteps are located and identified by double-YOLO method. The Φ-OTDR can cover a much larger sensing range than traditional sensors. Based on the stride and step frequency of the gait, the double-YOLO method can determine the walker's ID. Primary field experiment results show that this method can detect, locate and identify the footsteps of three persons, and achieve about 86.0% identification accuracy, with 12.6% accuracy improvement compared to single-YOLO method. This footstep detection and recognition method may promote the development of gait-based clinical diagnosis or person identification application.

Tunable graphene oxide for the low-fouling electrochemical sensing of uric acid in human serum.

Numerous studies have been reported to improve the selectivity of uric acid (UA) by eliminating the interference from other electroactive species that coexist in biological fluids. However, two main challenges associated with the nonenzymatic electrochemical detection of UA need to be overcome to achieve practical applications in biological samples. Those are the chemical fouling of electrodes caused by the oxidation product of UA and biofouling due to the non-specific absorption of biological macromolecules. It was found that the residual oxo-functional groups and defects on graphene played a crucial part in both electrocatalysis and anti-biofouling. Here, graphene oxide (GO) was tuned by electro-oxidation and electro-reduction and was investigated in antifouling and electrocatalytic performances for the electrochemical sensing of UA by using pristine GO, BSA bound GO, electro-reduction-treated GO and electro-oxidation-treated GO. The electro-oxidation-treated GO was explored in electrochemical sensing for the first time and exhibited the highest sensitivity and low fouling properties. Holey GO might be formed on the electrode surface by the electrochemical oxidation method in a mild and green solution without the use of an acid. The different electrode interfaces as well as the interaction with BSA were investigated by Raman spectroscopy, X-ray photoelectron spectroscopy, contact angle measurements, scanning electron microscopy, electrochemistry, and electrochemical impedance spectroscopy.

2,4-DCP compromises the fertilization capacity of mouse oocytes.

2,4-DCP (2,4-dichlorophenol) is an environmental estrogen that is ubiquitously distributed in the environment and widely used to produce herbicides and pharmaceutical intermediates. Although 2,4-DCP is suspected to have endocrine disruption, the reproductive toxicity of 2,4-DCP in mammals has not been adequately assessed. In the present study, we examined the effect of 2,4-DCP on the fertility of mouse eggs. The data showed that oral administration of 2,4-DCP (180 mg/kg/day for 7 days) compromises the fertilization rate of mouse oocytes. To further analyze the mechanism by which 2,4-DCP decreases fertilization, the key regulators and events during fertilization of mouse eggs were investigated. We found that the dynamics of cortical granules (CGs) were disrupted by showing the redistribution of CG free domain in 2,4-DCP-administered oocytes. This abnormality perturbed the sperm binding site in the zona pellucida (ZP) and dramatically reduced the number of sperm binding to the ZP of 2,4-DCP-administered oocytes. In addition, the abundance of Juno, a sperm receptor on the egg membrane, was also decreased and its distribution was mislocated in 2,4-DCP-administered oocytes. Finally, we validated that the defects of fertilization participants and events caused by 2,4-DCP might be mediated by the increased level of reactive oxygen species-induced apoptosis of oocytes. Therefore, we demonstrate that 2,4-DCP compromises the fertilization ability of mouse oocytes via inducing oxidative stress.

Evaluation of N2O sources after fertilizers application in vegetable soil by dual isotopocule plots approach.

Nitrogen (N) fertilizer is the major deriver of nitrous oxide (N2O) emissions in agricultural soil. In the vegetable fields in China both inorganic and organic fertilizers are largely applied as basic sources of nitrogen. Identifying the effects of fertilizer type on soil microbial activities involved in N2O emissions would be of great help for future development of N2O reduction strategies. N2O isotopocule deltas, including δ15Nbulk, δ18O and SP (the 15N site preference in N2O), have been used to analyze microbial pathways of N2O production under different treatments, including bio-organic fertilizer treatment, half bio-organic fertilizer and half urea (mixed fertilizer) treatment, urea treatment and no fertilizer treatment. We measured environmental factors, N2O fluxes and N2O isotopocule deltas to evaluate the dynamics of N2O emissions and constructed the dual isotopocule plots (δ15Nbulk vs. SP and δ18O vs. SP) of the main N2O emission phases to assess contribution of the involved microbial processes (bacterial nitrification, bacterial denitrification, nitrifier denitrification and fungal denitrification). According to the results of the main N2O emission phases, we found that bio-organic fertilizer and mix fertilizer treatments had significantly lower N2O emissions compared to urea treatment, with average N2O fluxes of 1477 ± 204, 1243 ± 187 and 1941 ± 164 µg m-3 h-1, respectively, but there were no significant effects on mineral N and cabbage yield. In addition, the urea treatment and the mixed fertilizer treatment had close and higher nitrogen use efficiency. Furthermore, the δ18O vs. SP plot was useful for providing insight into microbial processes, showing that fungal denitrification/bacterial nitrification was the dominant microbial pathway and bio-organic fertilizer and mix fertilizer treatments had higher denitrification and N2O reduction compared to urea treatment. Those findings demonstrated that the partial replacement of urea with bio-organic fertilizer was a better choice, by means of enhancing denitrification to reduce N2O emissions and also guaranteeing the nitrogen use efficiency and the cabbage yield.

Experimental Research on Data Synchronous Acquisition Method of Subsidence Monitoring in Submarine Gas Hydrate Mining Area.

The data synchronous acquisition is crucial to the seafloor subsidence monitoring system for gas hydrate mining areas based on microelectromechanical sensors (MEMS). Because the independent and high-precision time reference sources on land cannot be used on the seafloor, especially in the deep sea, a relative time synchronization method based on input/output (I/O) and controller area network (CAN) bus was proposed to realize the internal time synchronization of the system. To demonstrate the feasibility of the proposed method, tests including the deformation test of the MEMS sensor array under high pressure, synchronous accuracy test, and landslide and collapse simulation tests were carried out. The synchronization method was performed once every 24 h, and the time drift was reduced to 0.38 ms from more than 30 ms, demonstrating that method can achieve consistent internal time of the system. The method does not require additional hardware devices and has adjustable accuracy.

Fabrication of magnetic Fe3O4 doped ß-cyclodextrin microporous organic network for the efficient extraction of endocrine disrupting chemicals from food takeaway boxes.

Endocrine disrupting chemicals (EDCs) are a typical class of natural or man-made endogenous hormone agonists or antagonists that can directly or potentially interfere with human endocrine system. However, it is still difficult to analyze trace EDCs directly from complex environment and food matrices. Therefore, the proper sample pretreatment is highly desired and the preparation of efficient adsorbents is of great challenge and importance. Herein, we report the facile one-pot solvothermal synthesis of Fe3O4 nanoparticle doped magnetic ß-cyclodextrin microporous organic network composites (MCD-MONs) for the magnetic solid phase extraction (MSPE) of four phenolic EDCs in water and food takeaway boxes prior to the high-performance liquid chromatography analysis. The sheet-like Fe3O4 doped MCD-MONs offered good magnetic property (16.5 emu g-1) and stability, and provided numerous hydrogen bonding, hydrophobic, π-π, and host-guest interaction sites for EDCs. Under the optimal experimental conditions, the established method was successfully verified with wide linear range (2.0-1000 µg L-1), low limits of detection (0.6-1.0 µg L-1), good precisions (intra-day and inter-day RSDs < 5.2 %, n = 3), large enrichment factors (88-98) and adsorption capacity (90.3-255.8 mg g-1), short extraction time (6 min), less adsorbent consumption (3 mg), and good reusability (at least 8 times) for EDCs. The proposed method was successfully applied to detect the trace EDCs in real samples with the recovery of 84.0-99.7 %. This work demonstrated the great potential of MCD-MONs for the efficient MSPE of trace EDCs from complex food takeaway boxes and water samples and uncovered the prospect of CD-based MONs in sample pretreatment.

Thiol-yne click post-synthesis of phenylboronic acid-functionalized magnetic cyclodextrin microporous organic network for selective and efficient extraction of antiepileptic drugs.

Owing to their incomplete digestion in the human body and inadequate removal by sewage treatment plants, antiepileptic drugs (AEDs) accumulate in water bodies, potentially affecting the exposed humans and aquatic organisms. Therefore, sensitive and reliable detection methods must be urgently developed for monitoring trace AEDs in environmental water samples. Herein, a novel phenylboronic acid-functionalized magnetic cyclodextrin microporous organic network (Fe3O4@CD-MON-PBA) was designed and synthesized via the thiol-yne click post-modification strategy for selective and efficient magnetic solid-phase extraction (MSPE) of trace AEDs from complex sample matrices through the specific B-N coordination, π-π, hydrogen bonding, electrostatic, and host-guest interactions. Fe3O4@CD-MON-PBA exhibited a large surface area (118.5 m2 g-1), rapid magnetic responsiveness (38.6 emu g-1, 15 s), good stability and reusability (at least 8 times), and abundant binding sites for AEDs. Under optimal extraction conditions, the proposed Fe3O4@CD-MON-PBA-MSPE-HPLC-UV method exhibited a wide linear range (0.5-1000 µg L-1), low limits of detection (0.1-0.5 µg L-1) and quantitation (0.3-2 µg L-1), good anti-interference ability, and large enrichment factors (92.2-104.3 to 92.3-98.0) for four typical AEDs. This work confirmed the feasibility of the thiol-yne click post-synthesis strategy for constructing novel and efficient multifunctional magnetic CD-MONs for sample pretreatment and elucidated the significance of B-N coordination between PBA and N-containing AEDs.

Solvent and monomer regulation synthesis of core-shelled magnetic ß-cyclodextrin microporous organic network for efficient extraction of estrogens in biological samples prior to HPLC analysis.

The abnormal estrogens levels in human body can cause many side effects and diseases, but the quantitative detection of the trace estrogens in complex biological samples still remains great challenge. Here we reported the fabrication of a novel core-shell structured magnetic cyclodextrin microporous organic network (Fe3O4@CD-MON) for rapid magnetic solid phase extraction (MSPE) of four estrogens in human serum and urine samples prior to HPLC-UV determination. The uniform spherical core-shell Fe3O4@CD-MONs was successfully regulated by altering the reactive monomers and solvents. The Fe3O4@CD-MONs owned high specific surface area, good hydrophobicity, large superparamagnetism, and abundant extraction sites for estrogens. Under optimal conditions, the proposed MSPE-HPLC-UV method provided wide linearity range (2.0-400 µg L-1), low limits of detection (0.5-1.0 µg L-1), large enrichment factors (183-198), less adsorbent consumption (3 mg), short extraction time (3 min), and good stability and reusability (at least 8 cycles). The established method had also been successfully applied to the enrichment and detection of four estrogens in serum and urine samples with a recovery of 88.4-105.1 % and a relative standard deviation of 1.0-5.9 %. This work confirmed the feasibility of solvent and monomer regulation synthesis of Fe3O4@CD-MON composites, and revealed the great prospects of magnetic CD-MONs for efficient enrichment of trace estrogens in complex biological samples.

Health status monitoring of bridge cable and telescopic compensation device based on fiber grating sensing array.

Laying power cables along the bridge is a new way of laying submarine cables across the sea. Monitoring the health status of cables and their telescopic compensation devices is necessary. In this study, fiber grating sensing technology was used to monitor the strain, temperature, and vibration of the bridge cable of the Zhoushan-Daishan Bridge in Zhoushan, Zhejiang Province, and its compensation device. Two typhoons and one invasion event happened during the monitoring period. Temperature signals, strain signals, and time domain and time-frequency domain vibration signals were analyzed. The results showed that no fire hazards or risk of external damage were found with the bridge cable, and the monitoring system filled a gap in the in situ monitoring of the bridge cable in the Zhoushan-Daishan Bridge by the State Grid.

Effects of different ecological restoration methods on the soil physicochemical properties and vegetation community characteristics of the Baotou light rare earth tailings pond in Inner Mongolia, China.

This study investigated the soil physicochemical properties and vegetation community characteristics of the Baotou light rare earth tailings pond after 10 years of aggregate spray seeding ecological restoration (S1) and ordinary soil spray seeding ecological restoration (S2), and the naturally restored dam slope area without human intervention (S3). The results showed that the vegetation community of S1 was dominated by Caragana korshinskii Kom, and its importance and abundance values were 0.40 and 38.4, respectively, while the vegetation communities of S2 and S3 mainly comprised herbaceous plants. Additionally, the vegetation biomass of S1 was significantly higher than that of S2 and S3 by 215.20% and 1345.76%, respectively, and the vegetation diversity index of S1 was the highest among the three treatment groups. The soil porosity (SP), water content (W), electrical conductivity (EC), and available K were significantly improved in S1, while soil bulk density (BD) was significantly reduced compared with that of S2 and S3. In addition, redundancy analysis revealed that SP, EC, W, and K positively correlate with the biomass, Shannon, Pielou, Simpson, and Marglef indices. Principal component analysis further showed that the comprehensive score of S1 (0.983) was higher than that of S2 (- 0.261) and S3 (- 0.648). Collectively, these findings indicate that appropriate ecological restoration can improve soil structure and vegetation community characteristics, thereby accelerating vegetation restoration, ultimately increasing the stability of the ecosystem.

An Ankle Joint Flexion and Extension Movement-Monitoring Device Based on Pressure Sensors.

Ankle joint flexion and extension movements play an important role in the rehabilitation training of patients who have been injured or bedridden for a long time before and after surgery. Accurately guiding patients to perform ankle flexion and extension movements can significantly reduce deep vein thromboembolism. Currently, most ankle rehabilitation devices focus on assisting patients with ankle flexion and extension movements, and there is a lack of devices for effectively monitoring these movements. In this study, we designed an ankle joint flexion and extension movement-monitoring device based on a pressure sensor. It was composed of an STM32 microcontroller, a pressure sensor, an HX711A/D conversion chip, and an ESP8266 WiFi communication module. The value of the force and the effective number of ankle joint flexion and extension movements were obtained. An experimental device was designed to verify the accuracy of the system. The maximum average error was 0.068 N; the maximum average relative error was 1.7%; the maximum mean-squared error was 0.00464 N. The results indicated that the monitoring device had a high accuracy and could effectively monitor the force of ankle flexion and extension movements, ultimately ensuring that the patient could effectively monitor and grasp the active ankle pump movement.

Iridium Catalysts with f-Amphbinol Ligands: Highly Stereoselective Hydrogenation of a Variety of Ketones.

A series of novel and modular ferrorence-based amino-phosphine-binol (f-amphbinol) ligands have been successfully synthesized. The f-amphbinol ligands exhibited extremely high air stability and catalytic efficiency in the Ir-catalyzed stereoselective hydrogenation of various ketones to afford corresponding stereodefined alcohols with excellent results (full conversions, cis/trans >99:1, and 83% → 99% ee, TON up to 500 000). Control experiments have shown that -OH and -NH groups played a key role in this stereoselective hydrogenation.

Simple, fast and selective detection of adenosine triphosphate at physiological pH using unmodified gold nanoparticles as colorimetric probes and metal ions as cross-linkers.

We report a simple, fast and selective colorimetric assay of adenosine triphosphate (ATP) using unmodified gold nanoparticles (AuNPs) as probes and metal ions as cross-linkers. ATP can be assembled onto the surface of AuNPs through interaction between the electron-rich nitrogen atoms and the electron-deficient surface of AuNPs. Accordingly, Cu2+ ions induce a change in the color and UV/Vis absorbance of AuNPs by coordinating to the triphosphate groups and a ring nitrogen of ATP. A detection limit of 50 nM was achieved, which is comparable to or lower than that achievable by the currently used electrochemical, spectroscopic or chromatographic methods. The theoretical simplicity and high selectivity reported herein demonstrated that AuNPs-based colorimetric assay could be applied in a wide variety of fields by rationally designing the surface chemistry of AuNPs. In addition, our results indicate that ATP-modified AuNPs are less stable in Cu2+, Cd2+ or Zn2+-containing solutions due to the formation of the corresponding dimeric metal-ATP complexes.

In Situ Electrochemical Formation of Oxo-Functionalized Graphene on Glassy Carbon Electrode with Chemical Fouling Recovery and Antibiofouling Properties for Electrochemical Sensing of Reduced Glutathione.

Electrochemical detection can be used to achieve intracellular or in vivo analysis of reduced glutathione (GSH) in tissues such as brain by using a microelectrode, which can help to better understand the complex biochemical processes of this molecule in the human body. The main challenges associated with electrochemical GSH detection are the chemical fouling of electrodes, caused by the oxidation product of GSSG, and biofouling due to the non-specific absorption of biological macromolecules. Oxo-functionalized graphene was generated in situ on the surface of a glassy carbon electrode using a green electrochemical method without using any other modifiers or materials in a mild water solution. The fabricated oxo-functionalized graphene interface was characterized by Raman spectroscopy, X-ray photoelectron spectroscopy, electrochemistry, electrochemical impedance spectroscopy, and contact angle measurements. The interface showed high electrocatalytic activity towards the oxidation of GSH, and a simple and efficient GSH sensor was developed. Interestingly, the electrode is reusable and could be recovered from the chemical fouling via electrochemical oxidation and reduction treatment. The electrode also exhibited good antibiofouling properties. The presented method could be a promising method used to treat carbon materials, especially carbon-based microelectrodes for electrochemical monitoring of intracellular glutathione or in vivo analysis.

A Micro Electrochemical Sensor for Multi-Analyte Detection Based on Oxygenated Graphene Modified Screen-Printed Electrode.

Electrode interfaces with both antibiofouling properties and electrocatalytic activity can promote the practical application of nonenzymatic electrochemical sensors in biological fluids. Compared with graphene, graphene oxide (GO) possesses unique properties such as superior solubility (hydrophilicity) in water, negative charge, and abundant oxygenated groups (oxo functionalities) in the plane and edge sites, which play an essential role in electrocatalysis and functionalization. In this work, a micro electrochemical sensor consisting of GO-modified screen-printed electrode and PDMS micro-cell was designed to achieve multi-analyte detection with excellent selectivity and anti-biofouling properties by electrochemically tuning the oxygen-containing functional species, hydrophilicity/hydrophobicity, and electrical conductivity. In particular, the presented electrodes demonstrated the potential in the analysis of biological samples in which electrodes often suffer from serious biofouling. The interaction of proteins with electrodes as well as uric acid was investigated and discussed.

Covalent Organic Frameworks-TpPa-1 as an Emerging Platform for Electrochemical Sensing.

Covalent organic frameworks (COFs) are a new type of metal-free porous architecture with a well-designed pore structure and high stability. Here an efficient electrochemical sensing platform was demonstrated based on COFs TpPa-1 constructed by 1,3,5-triformylphloroglucinol (Tp) with p-phenylenediamine (Pa-1), which possesses abundant nitrogen and oxo-functionalities. COFs TpPa-1 exhibited good water dispersibility and strong adsorption affinities for Pd2+ and thus was used as loading support to modify Pd2+. The Pd2+-modified COFs TpPa-1 electrode (Pd2+/COFs) showed high electrocatalytic activity for both hydrazine oxidation reaction and nitrophenol reduction reaction. In addition, TpPa-1-derived nitrogen-doped carbon presented high activity for the electro-oxidation of reduced glutathione (GSH), and sensitive electrochemical detection of GSH was achieved. The presented COFs TpPa-1 can be utilized as a precursor as well as support for anchoring electro-active molecules and nanoparticles, which will be useful for electrochemical sensing and electrocatalysis.

The influence of soil acidification on N2O emissions derived from fungal and bacterial denitrification using dual isotopocule mapping and acetylene inhibition.

Denitrification, as both origins and sinks of N2O, occurs extensively, and is of critical importance for regulating N2O emissions in acidified soils. However, whether soil acidification stimulates N2O emissions, and if so for what reason contributes to stimulate the emissions is uncertain and how the N2O fractions from fungal (ffD) and bacterial (fbD) denitrification change with soil pH is unclear. Thus, a pH gradient (6.2, 7.1, 8.7) was set via manipulating cropland soils (initial pH 8.7) in North China to illustrate the effect of soil acidification on fungal and bacterial denitrification after the addition of KNO3 and glucose. For source partitioning, we used and compared SP/δ18O mapping approach (SP/δ18O MAP) and acetylene inhibition technique combined isotope two endmember mixing model (AIT-IEM). The results showed significantly higher N2O emissions in the acidified soils (pH 6.2 and pH 7.1) compared with the initial soil (pH 8.7). The cumulative N2O emissions during the whole incubation period (15 days) ranged from 7.1 mg N kg-1 for pH 8.7-18.9 mg N kg-1 for pH 6.2. With the addition of glucose, relative to treatments without glucose, this emission also increased with the decrement of pH values, and were significantly stimulated. Similarly, the highest N2O emissions and N2O/(N2O + N2) ratios (rN2O) were observed in the pH 6.2 treatment. But the difference was the highest cumulative N2O + N2 emissions, which were recorded in the pH 7.1 treatment based on SP/δ18O MAP. Based on both approaches, ffD values slightly increased with the acidification of soil, and bacterial denitrification was the dominant pathway in all treatments. The SP/δ18O MAP data indicated that both the rN2O and ffD were lower compared to AIT-IEM. It has been known for long that low pH may lead to high rN2O of denitrification and ffD, but our documentation of a pervasive pH-control of rN2O and ffD by utilizing combined SP/δ18O MAP and AIT-IEM is new. The results of the evaluated N2O emissions by acidified soils are finely explained by high rN2O and enhanced ffD. We argue that soil pH management should be high on the agenda for mitigating N2O emissions in the future, particularly for regions where long-term excessive nitrogen fertilizer is likely to acidify the soils.

[Identifying the Sources of Groudwater NO3--N in Agricultural Region of Qingdao].

To increase crops yields, applying large amounts of fertilizers has become increasingly common in agricultural regions, resulting in NO3--N groundwater pollution. Agricultural non-point pollution is the main source of groundwater NO3--N pollution. To ensure drinking water safety and quality, it is crucial to clarify the sources of NO3--N pollution in agricultural regions. In this study, 35 sampling sites were randomly selected in the Qingdao agricultural area in 2009 and 2019. The spatial distribution of NO3--N concentration was analyzed by the inverse distance weighting method (IDW). The nitrogen and oxygen isotopes were used as a tool to trace sources of NO3--N and the SIAR model was used to quantify contribution proportion of pollution sources. The results showed that the concentration of NO3--N (average) in groundwater in Qingdao has been reduced from 38.49 mg·L-1 in 2009 to 22.37 mg·L-1 in 2019, but it is still higher than the maximum allowable concentration of NO3--N in drinking water set by the World Health Organization (WHO). The NO3--N concentration gradually increased from south to north both in 2009 and 2019. The cross diagram of δ15N-NO3- and δ18O-NO3- show that the main sources of NO3--N in groundwater in Qingdao are chemical fertilizers, soil nitrogen, and manure and sewage. Water isotopes indicate that precipitation was the main source of groundwater in Qingdao. The SIAR model results indicated that the contribution of each source ranked as follows:manure and sewage (47.42%) > soil nitrogen (27.80%) > chemical fertilizer (14.32%) > atmospheric nitrogen depositions (10.43%). From 2009 to 2019, the quality of groundwater in Qingdao has been improved, but NO3--N pollution still cannot be ignored. According to the results, prevention and control should be made to ensure the safety of drinking water and the sustainable development of agriculture.

Distinguishing N2O and N2 ratio and their microbial source in soil fertilized for vegetable production using a stable isotope method.

Vegetable production systems with excessive nitrogen fertilizer result in severe N2O emission. It is pivotal to identify the source of N2O for reducing N2O emission, but estimating microbial pathways of N2O production is very difficult due to the existence of N2O reduction. A promising tool can address this problem by using δ18O and δ15NSP of N2O to construct a dual isotopocule plot. For ascertaining the microbial pathways of N2O production and consumption in soil fertilized for vegetable production, four treatments were set up: urea (U), half urea and half organic fertilizer (UO), organic fertilizer (O) and no fertilizer (NF), and the experiment was carried out continuously for two years. The δ18O vs. δ15NSP plot method indicated that the nitrification/fungal denitrification was a dominant in N2O emission, and the U treatment was the highest, followed by OU, O and NF in the both years. Among the different treatments, furthermore, the N2O flux had the same trend, whereas the extent of N2O reduction showed an opposite trend. Overall, inorganic fertilizer enhances nitrification/fungal denitrification and hinders reduction of N2O to N2, resulting in a larger amount of N2O emission. However, organic fertilizer increases the contribution of denitrification and greatly improves the extent of N2O reduction, which helps to reduce N2O emission. Therefore, organic fertilizer is crucial to reducing N2O emission by enhancing N2O reduction and should be properly applied in production practice.

Expanding the family of collagen proteins: recombinant bacterial collagens of varying composition form triple-helices of similar stability.

The presence of the (Gly-Xaa-Yaa)(n) open reading frames in different bacteria predicts the existence of an expanded family of collagen-like proteins. To further explore the triple-helix motif and stabilization mechanisms in the absence of hydroxyproline (Hyp), predicted novel collagen-like proteins from Gram-positive and -negative bacteria were expressed in Escherichia coli and characterized. Soluble proteins capable of successful folding and in vitro refolding were observed for collagen proteins from Methylobacterium sp 4-46, Rhodopseudomonas palustris and Solibacter usitatus . In contrast, all protein constructs from Clostridium perfringens were found predominantly in inclusion bodies. However, attachment of a heterologous N-terminal or C-terminal noncollagenous folding domain induced the Clostridium perfringens collagen domain to fold and become soluble. The soluble constructs from different bacteria had typical collagen triple-helical features and showed surprisingly similar thermal stabilities despite diverse amino acid compositions. These collagen-like proteins provide a resource for the development of biomaterials with new properties.