INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2025
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IF 8.5
Bo Zhang,Tong Wang,Siyu Ni,Yaping Zong,Nan Wang,Yunlong Si,Yong Li
Recovery of bioactive proteins from insoluble inclusion bodies (IB) is a critical challenge when purifying recombinant proteins expressed in E. coli . One practical solution is to enhance the renaturability or refolding capability of the target protein after the insoluble proteins are solubilized with denaturants such as urea. While some methods, such as using chemical additives and manipulating the physical environment, have been reported to assist in protein refolding, their effectiveness remains limited. This work evaluated the potential of a highly conserved domain found in five biotin-dependent carboxylases as a fusion tag for purifying IB proteins. We found that this tag, comprised of 67 amino acids (named P67) as eight anti-parallel β-sheets that collectively form a flattened barrel structure, significantly improved the refolding process of its IB protein partner. The yield of bioactive proteins fused with this tag recovered from the dialysis stage was markedly higher than untagged ones. We further proved that this effect relies on the structural integrity of the tag's spatial conformations. Under natural conditions, soluble proteins can be purified using the P67 tag via the biotin/streptavidin affinity system. In conclusion, this tag offers a promising methodology for purifying both insoluble and soluble recombinant proteins.
研究领域:
分子生物学
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蛋白质科学
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生物技术
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生物化学
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DOI
PubMed
高影响因子
ANALYST
·
2025
·
IF 3.3
Chenyan Li,Yilan Liang,Qincong Feng
Electrochemical biosensors are frequently employed to identify harmful microbes and disease indicators. However, their practical applicability is constrained by poor signal amplification efficiency and immobilization processes on the probe surface. To get over these restrictions, we here integrated roll-cycling amplification (RCA) and CRISPR/Cas12a gene editing tools with electrochemical biosensors. We constructed an electrochemical biosensor based on RCA to activate the cleavage activity of Cas12a. First, by double-stranded nucleic acid aptamer (ds Apt) specifically binding to E. coli-2571, Apt-b was competitively isolated to bind to T4 ligase and produce circular DNA. This in turn activates RCA, which in turn activates the accessory cleavage activity of CRISPR/Cas12a, which in turn cleaves DNA sequences loaded onto the electrode, changing electrochemical signals. With a linear range of 1 × 102–1 × 107 CFU mL−1, a detection limit of 5.28 CFU mL−1, and experimental recoveries of 93.01–101.53%, the measured electrochemical signals were positively connected with the concentration of E. coli-2571 under the optimal experimental conditions. Therefore, by combining two approaches—RCA and CRISPR/Cas12a—our electrochemical biosensor was able to detect E. coli-2571 specifically and sensitively, providing new research opportunities for the detection of other harmful bacteria.
研究领域:
分子生物学
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生物传感技术
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微生物检测
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产品关联
DOI
PubMed
高影响因子
PROCESS BIOCHEMISTRY
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2010
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IF 2.444
Runrong Ding,Zhaofeng Li,Sheng Chen,Dan Wu,Jing Wu,Jian Chen
The purpose of this study was to investigate the effect of medium additives on the secretion of recombinant α-cyclodextrin glucosyltransferase (α-CGTase) into the culture media of Escherichia coli . It is found that supplementation of the E. coli culture with SDS, glycine, Ca 2+ or Na + , individually, facilitated the secretion of α-CGTase. Orthogonal experiment showed that the optimal condition to achieve maximal secretion of α-CGTase was the supplementation with 0.03% SDS, 400 mM Na + , 0.3% glycine and 10 mM Ca 2+ together. Under this condition, extracellular enzyme activity reached 12.89 U/ml, which is 15 times higher than that of the culture without any additives. Further analysis showed that the permeability, fluidity and phosphatidylglycerol content of the E. coli cell membrane under the optimized condition were significantly increased in comparison to those under the control condition. These might be the potential mechanisms for the increased secretion of α-CGTase from the periplasmic compartment into the culture medium.
研究领域:
生物工程
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微生物学
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DOI
高影响因子
Frontiers in Bioengineering and Biotechnology
·
2022
·
IF 6.064
Tong Feifei,Qin Zongmin,Wang Hongyue,Jiang Yingying,Li Junkuan,Ming Hui,Qu Ge,Xiao Yazhong,Sun Zhoutong
Chiral amino alcohols are prevalent synthons in pharmaceuticals and synthetic bioactive compounds. The efficient synthesis of chiral amino alcohols using ammonia as the sole amino donor under mild conditions is highly desired and challenging in organic chemistry and biotechnology. Our previous work explored a panel of engineered amine dehydrogenases (AmDHs) derived from amino acid dehydrogenase (AADH), enabling the one-step synthesis of chiral amino alcohols via the asymmetric reductive amination of α-hydroxy ketones. Although the AmDH-directed asymmetric reduction is in a high stereoselective manner, the activity is yet fully excavated. Herein, an engineered AmDH derived from a leucine dehydrogenase from Sporosarcina psychrophila (SpAmDH) was recruited as the starting enzyme, and the combinatorial active-site saturation test/iterative saturation mutagenesis (CAST/ISM) strategy was applied to improve the activity. After three rounds of mutagenesis in an iterative fashion, the best variant wh84 was obtained and proved to be effective in the asymmetric reductive amination of 1-hydroxy-2-butanone with 4-fold improvements in kcat/Km and total turnover number (TTN) values compared to those of the starting enzyme, while maintaining high enantioselectivity (ee >99%) and thermostability (T5015 >53°C). In preparative-scale reaction, the conversion of 100 and 200 mM 1-hydroxy-2-butanone catalyzed by wh84 was up to 91–99%. Insights into the source of an enhanced activity were gained by the computational analysis. Our work expands the catalytic repertoire and toolbox of AmDHs.
研究领域:
生物催化
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酶工程
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有机化学
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生物技术
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DOI
PubMed
高影响因子
CHEMOSPHERE
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2020
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IF 5.778
Yuqiang Xiang,Huaduo Yan,Bingjie Zheng,Aroosha Faheem,Wenli Chen,Yonggang Hu
E. coli @UiO-67 composites were obtained using an effective and simple self-assembly method. The composites showed unique properties as a remarkable and recyclable adsorbent for the efficient removal of bisphenol A (BPA) from water with a high adsorption capacity (402.930 mg g −1 ). The increase in pore size is a key factor why E. coli @UiO-67 composites maintained high capacity. The reason might be due to that the composites with large pore sizes and defects could effectively improve mass transport and active molecular metal sites. The adsorption of BPA is a chemisorption process due to the Zr–OH groups in UiO-67 exhibit affinity toward BPA molecules, π–π interaction, and electrostatic attraction. The adsorption efficiency remained at 82.5% after 15 cycles without any remarkable changes in the PXRD patterns of E. coli @UiO-67. Moreover, the use of microorganism-loading MOFs could reduce the cost to at least 50% and minimize secondary pollution through nanoscale MOFs usage reduction. The developed composites have advantages, including low-cost, high adsorption capacity, easy to be separated and regenerated from aqueous solution, a large number of cycles, short adsorption equilibrium time, and stability, showing excellent application prospects. The presented strategy would be a potentially promising way to produce novel MOFs-based adsorbents with high-performance to control environmental pollution from wastewater.
研究领域:
环境科学
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材料科学
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生物工程
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产品关联
DOI
PubMed
高影响因子
IEEE SENSORS JOURNAL
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2025
·Vol. 26
·期 1
·41-55
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IF 4.5
Md. Abdul Khaleque,Md. Hasibul Hassan,Md. Romzan Ali,Md. Anwar Hossain,Mohamed Aly Saad Aly,Waleed Alahmad,Munawar Sultana,Md. Zaved H. Khan
Bacterial identification is challenging due to several problems, such as low sensitivity, poor selectivity, and the lack of quantitative methods. To address these obstacles, a gold nanoparticles (AuNPs)@Polyaniline (PANI)/Phage electrode was developed. The bacteriophage was immobilized on the AuNPs@PANI modified electrode, serving as an electrochemical biosensor of Escherichia coli (E. coli) DH5 $\alpha$ . In application, this biosensor can detect of E. coli DH5 $\alpha$ within a dynamic range of 10–107 CFUmL-1 with the detection limit of 1.0 CFUmL-1. Modified AuNPs@ PANI/Phage electrode exhibits a signal/noise ratio equal to 3.0, but only AuNPs@ PANI displayed 1.025 in [Fe(CN)6]3−/4− This biosensor selectively detected of E. coli DH5 $\alpha$ bacteria in the presence of comparable concentrations of different bacterial strains. In addition, the modified electrode exhibited supercapacitive behavior during electrochemical characterization and showed a high specific capacitance of 782.61 Fg-1 at 1.0 Ag-1 and 94.7% retention over 10 000 GCD cycles. Further analysis was conducted using galvanostatic charge-discharge to measure capacitance and self-discharge to assess capacity retention over time, and the Dunn method to analyze the charge storage mechanism within the electrode material. In conclusion, the developed nanostructure displayed dual electrochemical behavior as a supercapacitor and biosensor. This is the first reported PANI functionalized capacitive-controlled electrodes for bacterial detection. The fabrication and detection process for a single glass carbon electrode (GCE)/AuNPs@PANI pernigraniline salt (PANI-PS)/Phage/E. coli electrode requires ~90 min, with an estimated material cost of U.S. ${\$}$ 5.28 per electrode. Furthermore, smartphone-based testing confirmed portable, realtime detectionwith <1.5% deviation, indicating promise for multichannel clinical biosensing.
研究领域:
电化学
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生物传感器
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纳米材料
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微生物检测
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能量存储
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环境监测
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产品关联
DOI
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
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2026
·Vol. 27
·期 10
·4478
·
IF 5.6
You-Quan Ma,Lan Lin
Candida albicansis the causal agent of invasive candidiasis, which might be lethal in immunocompromised patients. Biofilm formation is considered a key virulence factor ofC. albicansand is associated with its elevated resistance to antifungals.C. albicansand bacteria likeE. coliare frequently found to form mixed biofilms on biotic or abiotic surfaces, rendering them more refractory to existing antifungals. To investigate howE. coliendogenous indole interplaying with exogenous IAA exerts modulatory effects on dual-species biofilm withC. albicans, anE. colistrain deficient in the indole biosynthetic genetnaAwas constructed, and the enzyme TnaA inhibitor was administered to block the indole production inE. colimonoculture and/orE. coli–C. albicansdual culture. Phenotypic assay revealed that indole deficiency attenuatedE. colimono-species biofilm by 12% (tnaA∆ versus WTE. coli), and the lack of indole in theE. colicell-free culture filtrate abolished the ability to promoteC. albicansbiofilms, evidenced by the fact that the treatment with WTE. coliculture supernatants exhibited a 1.7-fold promotive effect, while treatment withtnaA∆ displayed no significant difference from the broth control towardsC. albicansbiofilms. Furthermore, impairedE. coliindole production might disruptE. coli–C. albicansbiofilm, as examined by confocal laser scanning microscopy (CLSM). Moreover, indole-3-acetic acid (IAA) was found to exhibit more potent biofilm-modulatory activity than indole by CLSM imaging with dual biofilms of WTE. coli–C. albicans, in contrast to those ofE. coli tnaA∆–C. albicanspost-supplemented with exogenous IAA. This study provides evidence for indole as a signaling molecule mediating bacterial–fungal communication during mixed-biofilm formation. Indole and its derivatives, particularly in combination with existing antifungals, have potential in the development of anti-biofilm strategies to eradicate refractory fungal infections.
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DOI
PubMed
高影响因子
BioNanoScience
·
2026
·Vol. 16
·期 5
·333
·
IF 4
Zhu Mingxue,Wei Lin,Han Xiangning,Li Guoqiang,Cao Limin,Wang Xiudan,Wang Kaiqiang,Lin Hong,Sui Jianxin
The preparation of specific antibodies is of critical importance in the immunoassay of small-molecule contaminants in food. However, due to the lack of immunogenicity, these small molecules must be conjugated to a carrier to form complete antigen in order to induce the generation of antibodies, and adjuvants are typically required during the immunization when traditional proteins were used as carriers. Therefore, identifying a substance that possesses both the characteristics of carrier and adjuvant is of significant research importance for antibody preparation. Outer membrane vesicles (OMVs) are nanoscale particles with a bilayer membrane and spherical structure produced by Gram-negative bacteria through natural budding. They contain various pathogen-associated molecular patterns (PAMPs), which could be recognized by pattern recognition receptors (PRRs) on the surface of immune cells, thereby triggering a robust immune response. In this study, OMVs derived from Δ lpxl E. coli and chloramphenicol sodium succinate (CSS) were used as the carrier and hapten to synthesize the complete chloramphenicol (CAP) antigen (CSS-Δ lpxl -OMV). BALB/c mice were subsequently immunized with this antigen to generate antibodies, with the aim of evaluating the adjuvant effects of Δ lpxl -OMV and its potential as a hapten carrier. The results indicated that Δ lpxl -OMV not only served as a carrier for binding the hapten to prepare the complete antigen but also exhibited significant adjuvant properties. The synthesized complete antigen possessed significant immunogenicity and could induce the mice to produce antibodies against the target without additional adjuvants.
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DOI
高影响因子
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
·
2024
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IF 2.5
He Wang,Haidu Yuan,Jianhao Zhang,Wenjing Yan
Bacterial infections are becoming a significant threat to global human health due to the growing prevalence of biofilm-related infections and the rise in antibiotic resistance. D/ l -cysteine functionalized chiral gold nanoparticles (D/P-Au NPs or L/P-Au NPs) have demonstrated a potent antibacterial effect against E. coli , while the mechanism remains to be elucidated through additional research. Threonine deaminase (TD) is a crucial enzyme involved in branched-chain amino acid (BCAA) biosynthesis in E. coli and is involved in cysteine's antimicrobial effects. This study investigated the interaction between chiral Au NPs (D/P-Au NPs or L/P-Au NPs) and TD as well as its effect on enzyme activity. It demonstrates that chiral Au NPs interact with TD through hydrophobic forces, forming a ground state complex that induces changes in the secondary structure of TD and reduces enzyme activity in a concentration-dependent manner. We found that the exogenous supplementation of isoleucine and valine (2 mg/mL) significantly reduced the antibacterial activity of chiral Au NPs, especially for L/P-Au NPs. The proteomics results indicate that the expression of ilvA and ilvB was down-regulated after L/P-Au NPs treatment, which would interfere with the synthesis of BCAAs. These results demonstrate that chiral Au NPs cause cell death of E. coli partly due to inhibition of TD enzyme activity and the synthesis of branched-chain amino acids.
研究领域:
微生物学
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纳米生物学
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生物化学
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DOI
PubMed
高影响因子
RSC Advances
·
2020
·Vol. 10
·期 39
·22966-22971
·
IF 3.119
Ling Xiao,Dehui Chi,Guiju Sheng,Wenjuan Li,Penghui Lin,Sicheng Liang,Liangliang Zhu,Peipei Dong
UDP-glucuronosyltransferases (UGTs) and β-glucuronidase (GUS) catalyze entirely distinct metabolism reactions. UGTs are responsible for the glucuronidation of a variety of drugs, endogenous and environmental chemicals, whereas GUS hydrolyzes glucuronides and liberates the parent substrates. Information on the overlap of ligand selectivity between UGT and GUS is essential for exploring the pharmacological or toxicological effects of the inhibitors of these two metabolic enzymes. This study is conducted to test whether UGTs and GUS share common ligands, by investigating the inhibitory effects towards E. coli GUS by a series of UGT typical substrates and inhibitors. Results showed that three typical ligands of UGTs, including two specific substrates (estradiol and trifluoperazine, E2 and TFP) and one selective inhibitor (magnolol, Mag), can inhibit the activity of GUS. Kinetic assays indicated that all the three UGT specific chemicals displayed competitive inhibition, with Ki values of 31.4 (E2), 56.9 (TFP), and 16.6 μM (Mag). Docking studies further revealed that the three chemicals can enter the active sites of GUS by forming contacts with residues Glu-413, Trp-549, Asp-163, Tyr-472, Arg-562, or bound water. Our study indicates that ligand selectivity overlaps between UGTs and GUS, and some chemicals can act as co-inhibitors of these two metabolic enzymes. The pharmacological or toxicological effects of those co-inhibitors require further investigations.
研究领域:
药理学
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生物化学
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毒理学
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产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2023
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IF 8.8
Meng Liu,Liqian Guan,Yujiao Wen,Lezhu Su,Zhan Hu,Zhengjie Peng,Shikai Li,Qiyuan Tang,Zhi Zhou,Nan Zhou
The current photocatalytic bactericidal materials in the field of food pathogen control are usually consisted of metals that always suffering from poor stability and possible secondary pollution. Besides, the requirement for high energy excitation also inspires the enthusiasm on exploring non-metallic catalysts. Herein, the non-metallic composite of rice shell biochar loaded with red phosphorus (B@RP) was developed for photocatalysis and photothermal removal of bacteria. The B@RP showed effective photocatalysis performance to stimulate the generation of OH and O 2 − free radicals for the elimination of Escherichia coli ( E. coli ). At the same time, the photothermal effect of B@RP can also increase the permeability of cell membrane, which is conducive to free radicals entering the cell interior. Therefore, the non-metallic composite could achieve complete removal of E. coli within 2 h under illumination. Meanwhile, B@RP had excellent stability and the sterilization efficiency maintained 100% after 9 cycles. Hence, B@RP is expected to be a harmless and efficient bactericidal material for food industry.
研究领域:
食品科学
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环境科学
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微生物学
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产品关联
DOI
PubMed
高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
·
2023
·
IF 9.8
Wenjing Zhang,Shuxin Li,Kaichao Zhao,Juanfen Chai,Bo Wan,Yunqi Qin,Huan Huan,Simiao Sun,Yuesuo Yang,Muhammad Yousuf Jat Baloch
Because of its long survival time, high migration ability and high pathogenicity, the migration of the virus in the subsurface environment deserves in-depth exploration and research. In this study we investigated the migration behavior of E. coli phage (VI) with organic colloids (HA) or inorganic colloids (SiO 2 ) in the saturated or unsaturated bands and compared the differences in their migration mechanisms.The effects of different colloids on the surface characteristics of VI were analyzed according to particle size and zeta potential . Column experiments were conducted to simulate their migration in the subsurface environment. The results show that HA enhances the stability of VI-HA, promotes VI migration and plays a dominant role in its migration process under both saturated and unsaturated conditions. In contrast, SiO 2 puts VI-SiO 2 in an unstable state and is easily separated in the unsaturated state, thus promoting VI migration. Based on migration experiments, the extent of influence factors on VI migration was quantified and compared. The effect of colloids on VI migration is greater than that of moisture content, where the effect of HA is greater than that of SiO 2 . This study provides an experimental research idea to determine the dominant factors affecting virus migration, and provides a general direction and theoretical basis for the biological risk assessment of pathogenic microorganisms in complex underground environments , in order to enable the decision makers to make a response in time, accurately, and efficiently.
研究领域:
环境科学
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微生物学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）