Publications by authors named "Bang-Ce Ye"

225 Publications

Overexpression of Capsular Polysaccharide Biosynthesis Protein in P1 to enhance Capsular Polysaccharide production for di-n-butyl phthalate adsorption.

J Microbiol Biotechnol 2021 Apr 9. Epub 2021 Apr 9.

Lab of Biosystems and Microanalysis, Biomedical Nanotechnology Center, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.

Exopolysaccharides (EPSs) are important bioactive carbohydrate compound such as Capsular polysaccharide (CPS). EPSs are often used as bioenrichment agents and bioabsorbers to remove environmental pollutants like di-n-butyl phthalate (DBP). Among the EPSs producing bacteria, lactic acid bacteria (LAB) have gained the most attention. As generally recognized as safe (GRAS) microorganisms, LAB can produce EPSs with many different structures without any health risks. However, the EPSs production of LAB is usually not enough for large-scale application in industrial production. Here, gene (encoding CPS biosynthesis protein) was overexpressed in P1 to improve the production of EPSs and further enhance the capability of the adsorption of DBP. Compared with P1, the CPS production in overexpressed strain was increased by 11.3 mg/L, and the EPSs thickness was increased from 0.0786±0.0224μm in P1 to 0.1160±0.0480μm in P1-. These caused the adsorption ratio of DBP in P1- to be doubled. Overall, the findings in this study provided a safe solution for the adsorption and removing of DBP.
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http://dx.doi.org/10.4014/jmb.2101.01026DOI Listing
April 2021

Key role of exopolysaccharide on di-butyl phthalate adsorbing by Lactobacillus plantarum CGMCC18980.

Appl Microbiol Biotechnol 2021 Mar 5;105(6):2587-2595. Epub 2021 Mar 5.

Lab of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Meilong RD 130, Shanghai, 200237, China.

Plasticizers belong to hormone-like substances existing widely in the environment. According to the Environmental Protection Agency of China, they are considered to be the fourth class of toxic chemicals due to their harmful effects on normal endocrine system in human bodies. In the recent published work of our lab, Lactobacillus plantarum CGMCC18980 (strain P1) could reduce the toxicity of di-butyl phthalate (DBP) in rats effectively. The purpose of this study is to further explore the adsorption mechanism of di-butyl phthalate to L. plantarum CGMCC18980, based on optimizing the adsorption conditions. As a consequence, the adsorption effect of L. plantarum CGMCC18980 attributed to relationships between exopolysaccharide, membrane protein, and the cell wall. Experimental results demonstrated that exopolysaccharide and the cell wall were devoted to DBP binding. An obvious adsorption layer was observed outside of L. plantarum CGMCC18980 through scanning electron microscope (SEM) and transmission electron microscope (TEM). The Fourier transform infrared spectroscopy (FTIR) results showed that the functional groups involved in adsorption were mainly C=O, C-N, and C-O, which related to lipids and polysaccharides. Zeta potential analysis indicated that DBP adsorption had no significant relationship with surface charge. These results revealed that exopolysaccharide may be the key factor of strain CGMCC18980 in DBP adsorption. KEY POINTS: • Lactobacillus plantarum CGMCC18980 has the ability to adsorb di-butyl phthalate, reaching to 58.63%. • Exopolysaccharide is considered to play a key role in adsorption process. • Membrane protein, cell wall, and surface charge do not contribute to adsorption.
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http://dx.doi.org/10.1007/s00253-021-11145-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7954756PMC
March 2021

RegX3 Controls Glyoxylate Shunt and Mycobacteria Survival by Directly Regulating the Transcription of Isocitrate Lyase Gene in .

ACS Infect Dis 2021 04 4;7(4):927-936. Epub 2021 Mar 4.

Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou 310014, Zhejiang, China.

The glyoxylate shunt is a pathway associated with the assimilation of fatty acids and is implicated in the resistance of (). Isocitrate lyase (ICL), the first enzyme in the glyoxylate shunt, mediates infections and its survival in mice fatty acids, metabolism, and physiological functions. Here, we found that in () the two-component system SenX3-RegX3 regulated the glyoxylate shunt in response to phosphate starvation by controlling the transcription of . In response to phosphate availability, the phosphate regulator RegX3 directly bound to the upstream regulatory region of and repressed its transcription. The inactivation of increased transcription and ICL activity, causing a growth defect in with fatty acids as the sole source of carbon and energy. The growth defect was partly due to the toxicity of the excess glyoxylate produced by ICL. A decrease in glyoxylic acid levels, overexpression of , or the chemical inhibition (IA or 3-NP) of ICL restored the growth of the Regx3-deficient . Thus, we established a genetic network between the phosphate stress response and glyoxylate shunt based on the amount of intracellular ICL during mycobacterial survival on short-chain fatty acids, which contributed to its antimicrobial arsenal.
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http://dx.doi.org/10.1021/acsinfecdis.1c00067DOI Listing
April 2021

A Novel Peptide-Equipped Exosomes Platform for Delivery of Antisense Oligonucleotides.

ACS Appl Mater Interfaces 2021 Mar 23;13(9):10760-10767. Epub 2021 Feb 23.

Lab of Biosystem and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.

Exosomes are natural delivery vehicles because of their original feature such as low immunogenicity, excellent biocompatibility, and migration capability. Engineering exosomes with appropriate ligands are effective approaches to improve the low cellular uptake efficiency of exosomes. However, current strategies face considerable challenges due to the tedious and labor-intensive operational process. Here, we designed a novel peptides-equipped exosomes platform which can be assembled under convenient and mild reaction condition. Cell-penetrating peptides (CPPs) was conjugated on HepG2 cells-derived exosomes surface which can not only enhance the penetrating capacity of exosomes but also assist exosomes in loading antisense oligonucleotides (ASOs). The cellular uptake mechanism was investigated and we compared the difference between natural exosomes and modified exosomes. The resulting nanosystem demonstrated a preferential tropism for cells that are parented to their source tumor cells and could remarkably increase the cellular delivery of G3139 with efficient downregulation of antiapoptotic Bcl-2. This work developed a rapid strategy for intracellular delivery of nucleic acids, thus providing more possibilities toward personalized cancer medicine.
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http://dx.doi.org/10.1021/acsami.1c00016DOI Listing
March 2021

GlnR Regulates the Glyoxylate Cycle and the Methylcitrate Cycle on Fatty Acid Metabolism by Repressing Transcription.

Front Microbiol 2021 3;12:603835. Epub 2021 Feb 3.

Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, China.

(), along with its pathogenic counterpart (), utilizes fatty acids and cholesterol as important carbon and energy sources during the persistence within host cells. As a dual-functional enzyme in the glyoxylate cycle and the methylcitrate cycle, isocitrate lyase (ICL, encoded by or ) is indispensable for the growth of and on short-chain fatty acids. However, regulation of in mycobacteria in response to nutrient availability remains largely unknown. Here, we report that the global nitrogen metabolism regulator GlnR represses expression by binding to an atypical binding motif in the promoter region under nitrogen-limiting conditions. We further show that GlnR competes with PrpR, a transcriptional activator of , and dominantly occupies the co-binding motif in the promoter region. In the absence of GlnR or in response to the excess nitrogen condition, cells elongate and exhibit robust growth on short-chain fatty acids due to the PrpR-mediated activation of , thereby inducing enhanced apoptosis in infected macrophages. Taken together, our findings reveal the GlnR-mediated repression of on fatty acid metabolism, which might be a general strategy of nutrient sensing and environmental adaptation employed by mycobacteria.
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http://dx.doi.org/10.3389/fmicb.2021.603835DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7886694PMC
February 2021

RegX3-Mediated Regulation of Methylcitrate Cycle in .

Front Microbiol 2021 2;12:619387. Epub 2021 Feb 2.

Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, China.

is a global human pathogen that infects macrophages and can establish a latent infection. Emerging evidence has established the nutrients metabolism as a key point to study the pathogenesis of and host immunity. It was reported that fatty acids and cholesterol are the major nutrient sources of in the period of infection. However, the mechanism by which utilizes lipids for maintaining life activities in nutrient-deficiency macrophages is poorly understood. is fast-growing and generally used to study its pathogenic counterpart, . In this work, we found that the phosphate sensing regulator RegX3 of is required for its growing on propionate and surviving in macrophages. We further demonstrated that the expression of and related genes () in methylcitrate cycle could be enhanced by RegX3 in response to the phosphate-starvation condition. The binding sites of the promoter region of for RegX3 and PrpR were investigated. In addition, cell morphology assay showed that RegX3 is responsible for cell morphological elongation, thus promoting the proliferation and survival of in macrophages. Taken together, our findings revealed a novel transcriptional regulation mechanism of RegX3 on propionate metabolism, and uncovered that the nutrients-sensing regulatory system puts bacteria at metabolic steady state by altering cell morphology. More importantly, since we observed that RegX3 also binds to the operon , the RegX3-mediated regulation might be general in and other mycobacteria for nutrient sensing and environmental adaptation.
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http://dx.doi.org/10.3389/fmicb.2021.619387DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7884335PMC
February 2021

Simultaneous imaging of cancer biomarkers in live cells based on DNA-engineered exosomes.

Analyst 2021 Mar 18;146(5):1626-1632. Epub 2021 Feb 18.

Lab of Biosystem and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.

Cancer biomarkers are directly related to the development of cancers. Noninvasive identification of the location and expression levels of these biomarkers in live cancer cells offers great potential for accurate early-stage cancer diagnosis and cancer metastasis monitoring. Herein, we propose a DNA-engineered exosome (DNA-Exo) nanoplatform to image dual cancer biomarkers at the single-cell level, in which DNA probes were modified with the cholesterol group to facilely anchor on the exosomal membrane through hydrophobic interaction. Fluorophore-labeled DNA aptamer and hairpin probes targeting two kinds of cancer biomarkers of transmembrane glycoprotein mucin 1 (MUC1) and cytoplasmic microRNA-21 (miR-21), respectively, were employed for convenient dual-fluorescence imaging of cancer cells. The cellular uptake of DNA-Exos induced the specific recognition of MUC1 and miR-21, allowing the acquisition of the expression levels and spatial distributions of these two biomarkers in three tested cell lines. Our work demonstrated that the proposed DNA-Exos with designable functions have the capacity to visually discriminate different cell types based on the specific recognition of analytes.
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http://dx.doi.org/10.1039/d0an02353aDOI Listing
March 2021

Single-atom electrocatalysts templated by MOF for determination of levodopa.

Talanta 2021 Apr 27;225:122042. Epub 2020 Dec 27.

Key Laboratory for Green Processing of Chemical Engineering of Xinjiang Bingtuan, School of Chemistry and Chemical Engineering, Shihezi University, Shihezi, 832003, China; Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, Zhejiang, China. Electronic address:

To overcome the problem of incorrect levodopa (LD) dosage in the treatment of Parkinson's disease, a new analytical tool is urgently needed for accurately determining the concentration of LD in human fluids. Herein, an effective and stable sensor based on a Co-single-atomic-site catalyst (Co-SASC)-modified glassy carbon electrode (Co-SASC/GCE) was developed for the determination of LD concentration. The physicochemical characterization of Co-SASC is systematically investigated. It has excellent thermal stability, graphitization degree, and a large specific surface area. Benefiting from its porous structure for kinetically fast catalysis and component advantages for fix a single cobalt atom to improve stability, Co-SASC/GCE exhibits a superior electrochemical response. Under optimal conditions (pH 2.0, coating amount is 10 μg), an ideal linear relationship is achieved between the logarithm of the peak current of the sensor and the logarithm of LD concentration. The linear range is 0.1-200 μM, and the limit of detection (LOD) is 0.033 μM. After a simple pretreatment, LD in human serum is detected by Co-SASC/GCE with excellent stability and selectivity. As such, this work enlarges the existing electrochemical sensor toolbox by offering a reasonable design and synthesis protocol for advanced materials to accurately determine LD in human fluids for the clinical treatment of Parkinson's disease.
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http://dx.doi.org/10.1016/j.talanta.2020.122042DOI Listing
April 2021

SARS-CoV-2 ORF9b inhibits RIG-I-MAVS antiviral signaling by interrupting K63-linked ubiquitination of NEMO.

Cell Rep 2021 02 3;34(7):108761. Epub 2021 Feb 3.

Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, Zhejiang 310014, China. Electronic address:

Coronavirus disease 2019 (COVID-19) is a current global health threat caused by the novel coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Emerging evidence indicates that SARS-CoV-2 elicits a dysregulated immune response and a delayed interferon (IFN) expression in patients, which contribute largely to the viral pathogenesis and development of COVID-19. However, underlying mechanisms remain to be elucidated. Here, we report the activation and repression of the innate immune response by SARS-CoV-2. We show that SARS-CoV-2 RNA activates the RIG-I-MAVS-dependent IFN signaling pathway. We further uncover that ORF9b immediately accumulates and antagonizes the antiviral type I IFN response during SARS-CoV-2 infection on primary human pulmonary alveolar epithelial cells. ORF9b targets the nuclear factor κB (NF-κB) essential modulator NEMO and interrupts its K63-linked polyubiquitination upon viral stimulation, thereby inhibiting the canonical IκB kinase alpha (IKKα)/β/γ-NF-κB signaling and subsequent IFN production. Our findings thus unveil the innate immunosuppression by ORF9b and provide insights into the host-virus interplay during the early stage of SARS-CoV-2 infection.
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http://dx.doi.org/10.1016/j.celrep.2021.108761DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7857071PMC
February 2021

Gut Microbiota-Derived Metabolites in the Development of Diseases.

Can J Infect Dis Med Microbiol 2021 8;2021:6658674. Epub 2021 Jan 8.

Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou 310014, Zhejiang, China.

Gut microbiota is increasingly recognized as a metabolic organ essential for human health. Compelling evidences show a variety set of links between diets and gut microbial homeostasis. Changes in gut microbial flora would probably contribute to the development of certain diseases such as diabetes, heart disease, allergy, and psychiatric diseases. In addition to the composition of gut microbiota, the metabolites derived from gut microbiota have emerged as a pivotal regulator in diseases development. Since high-fat and high-protein diets substantially affect the gut microbial ecology and human health, the current review summarizes the gut microbiota-derived metabolites such as short-chain fatty acids (SCFAs), amino acids, and their derivatives and highlights the mechanisms underlying the host responses to these bioactive substances.
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http://dx.doi.org/10.1155/2021/6658674DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7815404PMC
January 2021

A Novel Wick-Like Paper-Based Microfluidic Device for 3D Cell Culture and Anti-Cancer Drugs Screening.

Biotechnol J 2021 Feb 19;16(2):e2000126. Epub 2020 Oct 19.

Lab of Biosystem and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, Shanghai, 200237, China.

Paper is increasingly recognized as a portable substrate for cell culture, due to its low-cost, flexible, and special porous property, which provides a native cellular 3D microenvironment. Therefore, paper-based microfluidics has been developed for cell culture and biomedical analysis. However, the inability of continuous medium supply limits the wide application of paper devices for cell culture. Herein, a paper-based microfluidic device is developed with novel folded paper strips as wick-like structure, which is used for medium self-driven perfusion. The paper with patterns of hydrophilic channel, culture areas, and hydrophobic barrier could be easily fabricated through wax-printing. After printing, the hydrophilic paper strip at the periphery of the lower layer is then folded at 90° and extended into the medium container for continuous automatic supply of medium to the cell culture area. Tumor cells cultured in the paper device are tested for anti-cancer drug screening. Visualized cell viability and chemical sensitivity testing can be achieved by colorimetry combined with simple smartphone imaging, effectively reducing precision instrument dependence. The wick paper-based microfluidic device for cell culture endows the method the advantages of lower cost, ease-of-operation, miniaturization, and shows a great potential for large-scale cell culture, antibody drug production, and efficient screening.
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http://dx.doi.org/10.1002/biot.202000126DOI Listing
February 2021

An aptamer biosensor based dual signal amplification system for the detection of salmonella typhimurium.

Anal Biochem 2021 02 4;615:114050. Epub 2020 Dec 4.

Lab of Biosystem and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, Shanghai, 200237, China.

Salmonella, a typical foodborne pathogen, always seriously threatens the health and even life of both humans and animals. However, highly sensitive and fast quantitative methods for its detection are remaining to be challenged. Herein, we presented an efficient method with dual signal amplification strategy by combining immune hybridization chain reaction (HCR) with surface enhanced Raman scattering (SERS) to high sensitively detect Salmonella typhimurium in food. After sample preparation, S. typhimurium were specifically captured by immunomagnetic beads (IMBs), then aptamers and hairpin-probes were added to trigger HCR to form nicked dsDNA, finally 4',6-Diamidino-2-phenylindole dihydrochloride (DAPI) was incubated with HCR products and then the whole system was mixed with AgNP colloid to detect the SERS intensity at 1610 cm. As a result, a good linear relationship was achieved between SERS intensities and corresponding concentrations of S. typhimurium ranging from 10 to 10 CFU/mL, with a limit of detection (LOD) of 6 CFU/mL in 3.5 h. The proposed method has been successfully applied to capture and detect the S. typhimurium in spiked milk samples, and the results were consistent with those of the traditional plate counting method. The method, with combination of HCR and SERS, achieves double amplification of the detection signal and significantly improves the detection sensitivity of S. typhimurium. And it also shows good application potential for the highly sensitive detection of other contaminants in food.
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http://dx.doi.org/10.1016/j.ab.2020.114050DOI Listing
February 2021

Metabolic engineering of Pseudomonas putida KT2440 for high-yield production of protocatechuic acid.

Authors:
Jin Li Bang-Ce Ye

Bioresour Technol 2021 Jan 7;319:124239. Epub 2020 Oct 7.

Laboratory of Biosystems and Microanalysis, Institute of Engineering Biology and Health, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China; Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou 310014, Zhejiang, China. Electronic address:

Protocatechuic acid (PCA) has been widely utilized in conventional pharmaceutical, cosmetic and functional food industries. Currently, chemical synthesis and solvent extraction are the main methods for commercial production, indicating several disadvantages. In this study, we developed a method for the biosynthesis of PCA in Pseudomonas putida KT2440 in high yield. First, we developed constitutive promoters with different expression intensities for fine-tuned gene expression. Second, we improved the biosynthesis of "natural" PCA in P. putida KT2440 via multilevel metabolic engineering strategies: overexpression of rate-limiting enzymes, removal of negative regulators, attenuation of pathway competition, and enhancement of precursor supply. Finally, by further bioprocess engineering efforts, the best-producing strain reached a titer of 12.5 g/L PCA from glucose at 72 h in a shake flask and 21.7 g/L in fed-batch fermentation without antibiotic pressure. This was the highest PCA titer from glucose using metabolically engineered microbial cell factories reported to date.
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http://dx.doi.org/10.1016/j.biortech.2020.124239DOI Listing
January 2021

l-arginine production in : manipulation and optimization of the metabolic process.

Crit Rev Biotechnol 2021 Mar 5;41(2):172-185. Epub 2020 Nov 5.

Jiangxi Engineering Laboratory for the Development and Utilization of Agricultural Microbial Resources, Jiangxi Agricultural University, Nanchang, China.

As an important semi-essential amino acid, l-arginine is extensively used in the food and pharmaceutical fields. At present, l-arginine production depends on cost-effective, green, and sustainable microbial fermentation by using a renewable carbon source. To enhance its fermentative production, various metabolic engineering strategies have been employed, which provide valid paths for reducing the cost of l-arginine production. This review summarizes recent advances in molecular biology strategies for the optimization of l-arginine-producing strains, including manipulating the principal metabolic pathway, modulating the carbon metabolic pathway, improving the intracellular biosynthesis of cofactors and energy usage, manipulating the assimilation of ammonia, improving the transportation and membrane permeability, and performing biosensor-assisted high throughput screening, providing useful insight into the current state of l-arginine production.
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http://dx.doi.org/10.1080/07388551.2020.1844625DOI Listing
March 2021

Acetylation of translation machinery affected protein translation in E. coli.

Appl Microbiol Biotechnol 2020 Dec 31;104(24):10697-10709. Epub 2020 Oct 31.

Laboratory of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.

Reversible lysine acetylation (RLA) of translation machinery components, such as ribosomal proteins (RPs) and translation factors (TFs), was identified in many microorganisms, while knowledge of its function and effect on translation remains limited. Herein, we show that translation machinery is regulated by acetylation. Using the cell-free translation system of E. coli, we found that AcP-driven acetylation significantly reduced the relative translation rate, and deacetylation partially restored the translation activity. Hyperacetylation caused by intracellular AcP accumulation or carbon/nitrogen fluctuation (carbon overflow or nitrogen limitation) modulated protein translation in vivo. These results uncovered a critical role of acetylation in translation regulation and indicated that carbon/nitrogen imbalance induced acetylation of ribosome in E. coli and dynamically affected translation rate via a global, uniform manner. KEY POINTS: • Acetylation of translation machinery directly regulated global translation. • K618 of EF-G, K411, and K464 of S1 are the key points influencing translation rate. • Carbon/nitrogen imbalance triggers AcP-dependent acetylation.
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http://dx.doi.org/10.1007/s00253-020-10985-2DOI Listing
December 2020

Characteristics of plant growth-promoting rhizobacteria SCPG-7 and its effect on the growth of Capsicum annuum L.

Environ Sci Pollut Res Int 2021 Mar 28;28(9):11323-11332. Epub 2020 Oct 28.

School of Chemistry and Chemical Engineering/The Key Lab. for Green Processing of Chemical Engineering of Xinjiang Bingtuan, Shihezi University, Shihezi, 832003, People's Republic of China.

The strain SCPG-7 was isolated from saline soil in a cotton field. It is confirmed that the strain SCPG-7 is Pseudomonas sp. by means of the analysis of its phenotypic features and 16S rRNA sequence. SCPG-7 was capable of dissolving mineral tri-calcium phosphate (Ca(PO)) and tri-magnesium phosphate (Mg(PO)). In contrast, no showing iron phosphate (FePO) or aluminum phosphate (AlPO) solubilizing activities were detected by this experimental approach. The ratio of the dissolved P diameter to the colony diameter was 1.86. To study the phosphate dissolving mechanisms of the strain, we analyzed the changes of the pH value, the soluble phosphate content, the concentration of alkaline phosphatase, and the production of organic acid in the insoluble phosphate liquid medium. 2-keto-D-gluconicacid, α-ketoglutaric acid, succinic acid, etc. were characterized by LC-MS/MS in NBRIP medium. The concentration of 2-keto-D-gluconicacid increased to 88.6 mg/L after being cultured for 216 h. The strain decreased the pH value of the medium from 7.4 to 4.7 and the released soluble phosphate up to 516 mg/L, which proved the production of organic acids and alkaline phosphatase to be mechanism for solubilizing P. Under low phosphorus stress, Pseudomonas global regulatory protein PhoB regulates the transcription of the alkaline phosphatase gene. IAA and siderophore were secreted by SCPG-7. After treatment with SCPG-7, the individual plant height and dry weight of pepper increased by 23.3 and 31.2%, respectively, compared to the control group. The results show that the strain SCPG-7 has the potential to convert insoluble inorganic phosphorus to plant-available phosphorus. It can enhance soil phosphorus release through biological pathways, thereby increasing crop yield, and providing germplasm resources for the development of biological fertilizers.
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http://dx.doi.org/10.1007/s11356-020-11388-6DOI Listing
March 2021

A robust electrochemical sensing based on bimetallic metal-organic framework mediated MoC for simultaneous determination of acetaminophen and isoniazid.

Anal Chim Acta 2020 Nov 27;1136:99-105. Epub 2020 Aug 27.

Key Laboratory for Green Processing of Chemical Engineering of Xinjiang Bingtuan, School of Chemistry and Chemical Engineering, Shihezi University, Shihezi, 832003, China; Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, Zhejiang, China. Electronic address:

Herein, a MoC/bimetallic zeolitic imidazolate framework-modified glassy carbon electrode (MoC@BMZIFs/GCE) was established as an electrochemical sensor for the simultaneous sensitive determination of acetaminophen (APAP) and isoniazid (INZ). The apparent morphology, structural composition, and electrochemical properties were comprehensively investigated. The outstanding electrocatalytic activity and conductivity endow the sensor desirable electrochemical performance toward APAP and INZ compared to the bare GCE, such as wide linear range, low detection limit, and high selectivity. Under the optimum conditions, a linear relationship between the oxidation peak current and the concentration of the measured object was obtained, with linear ranges from 0.1 to 300 μM for APAP and from 10 to 3500 μM for INZ. The detection limits for APAP and INZ were 0.03 μM and 1.5 μM, respectively. More importantly, the APAP and INZ oxidation peaks could be completely separated. Moreover, the highly sensitive and stable sensor was applied to detect APAP and INZ in human serum. This work can provide a viable route to rational design and construct electrochemical sensors for drug monitoring and clinical diagnosis.
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http://dx.doi.org/10.1016/j.aca.2020.08.044DOI Listing
November 2020

Ratiometric electrochemical sensing based on MoC for detection of acetaminophen.

Analyst 2020 Nov;145(23):7609-7615

Key Laboratory for Green Processing of Chemical Engineering of Xinjiang Bingtuan, School of Chemistry and Chemical Engineering, Shihezi University, Shihezi 832003, China.

In this work, MoO2 nanoparticles were synthesized and annealed to form Mo2C nanoparticles. This is the first report of a ratiometric electrochemical sensor (R-ECS) for the detection of acetaminophen (AP), in which Mo2C is used as the sensing agent and ferrocene (FC) is used as an internal reference. FC (100 μM) is added directly to the electrolyte solution for convenient operation. The synthesized materials were fully characterized with respect to composition, morphology and electrochemical performance. The oxidation peak potentials of FC (0.196 V) and AP (0.364 V) can be completely separated by the Mo2C modified glassy carbon electrode, and their ratiometric signals are used for the quantification of AP. It was found that the oxidation peak currents of AP at separated potentials on Mo2C/GCE are linear with concentration in the range of 0.5-600 μM, and the detection limit is 0.029 μM (S/N = 3). Mo2C/GCE exhibited decent repeatability, reproducibility, stability, and selectivity. The sensor was then applied to measure AP in tap water and river water.
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http://dx.doi.org/10.1039/d0an01403fDOI Listing
November 2020

Novel electrochemical sensing platform based on ion imprinted polymer with nanoporous gold for ultrasensitive and selective determination of As.

Mikrochim Acta 2020 09 17;187(10):571. Epub 2020 Sep 17.

Key Laboratory for Green Processing of Chemical Engineering of Xinjiang Production and Construction Corps, School of Chemistry and Chemical Engineering, Shihezi University, Shihezi, 832000, China.

An electrochemical sensor has been developed based on ion imprinted polymer (IIP) and nanoporous gold (NPG) modified gold electrode (IIP/NPG/GE) for determination of arsenic ion (As) in different kinds of water. NPG with high conductivity, large specific surface area, and high biocompatibility was prepared by a green electrodeposition method. Then a layer of IIP was synthesized in situ on NPG surface by electropolymerization, in which As was used as template ion and o-phenylenediamine as functional monomer. We used potassium ferricyanide and potassium ferrocyanide chelates as electrochemical probes to generate signals. The electrochemical behavior of IIP/NPG/GE (vs. Ag/AgCl) was studied by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The linear range for As was 2.0 × 10 to 9.0 × 10 M, and the lower detection limit was 7.1 × 10 M (S/N = 3). This newly developed sensor has good stability and selectivity, and has been successfully applied to the As determination of four kinds of water quality.
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http://dx.doi.org/10.1007/s00604-020-04552-9DOI Listing
September 2020

Synthesis of MOF-derived Ni@C materials for the electrochemical detection of histamine.

Talanta 2020 Nov 8;219:121360. Epub 2020 Jul 8.

Key Laboratory for Green Processing of Chemical Engineering of Xinjiang Bingtuan, School of Chemistry and Chemical Engineering, Shihezi University, Shihezi, 832003, China; Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, Zhejiang, China; State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China. Electronic address:

Histamine (HA) plays an important role in food safety supervision and is also involved in various physiological functions. Accurate and rapid detection of HA in real sample is count for much as this is the significant prerequisite for its effective monitoring. In this study, we fabricated an electrochemical sensor to detect HA via the pyrolysis of the hydrothermal Ni-MOF (metal-organic frameworks), in which the obtained Ni@C material was deployed as the sensing agent. Ni@C was comprehensively characterized in terms of its morphology, constitution, as well as its electrochemical behavior. The as-prepared sensor (Ni@C/GCE) features excellent electrocatalytic activities. It was also observed that the electrochemical property of the sensor was substantially improved because Ni@C afforded an enlarged active surface and accelerated electron transport. This sensor affords amperometric analysis in the linear range of 10-100 μM HA with a 3.2 × 10 μM low detection limit (S/N = 3). Many important features, including decent anti-interference, reproducibility, stability, and reliability, were also observed. Importantly, the sensor enabled the measurement of HA in real samples obtained from fish, thus demonstrating its practical potential as a HA analytical detector.
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http://dx.doi.org/10.1016/j.talanta.2020.121360DOI Listing
November 2020

Long-Term Rewritable Report and Recording of Environmental Stimuli in Engineered Bacterial Populations.

ACS Synth Biol 2020 09 1;9(9):2440-2449. Epub 2020 Sep 1.

Laboratory of Biosystems and Microanalysis, Institute of Engineering Biology and Health, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.

DNA writing (living sensing recorders) based whole-cell biosensors can capture transient signals and then convert them into readable genomic DNA changes. The primitive signals can be easily obtained by sequencing technology or analysis of protein activity (such as fluorescent protein). However, the functions of the current living sensing recorders still need to be expanded, and the difficulty of rewriting in complex biological environments has further limited their applications. In this study, we designed a long-term rewritable recording system using a CRISPR base editor-based synthetic genetic circuit, named CRISPR-istop. This system can convert stimuli into changes in the fluorescence intensity (reporter) and single-base mutations in genomic DNA (recording). Furthermore, we updated the biological circuit through the strategy of coupling the single-base mutation (record site) and the loss-of-function of the targeted protein (translation stopped by stop codon introduction), and we can remove edited bacteria from a population through selective sweeps upon applying a selective pressure. It successfully conducted the rewritable reporter and recording of the nutrient arabinose and pollutant arsenite with two rounds of continuous operation (10 passages/round, 12 h/passage). These observations indicated that the CRISPR-istop system can report and record stimuli over time; moreover, the recording can be manually erased and rewritten as needed. This method has great potential to be extended to more complicated recording systems to execute sophisticated tasks in inaccessible environments for synthetic biology and biomedical applications, such as monitoring disease-relevant physiological markers or other molecules.
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http://dx.doi.org/10.1021/acssynbio.0c00193DOI Listing
September 2020

A lateral flow strip combined with Cas9 nickase-triggered amplification reaction for dual food-borne pathogen detection.

Biosens Bioelectron 2020 Oct 10;165:112364. Epub 2020 Jun 10.

Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, Zhejiang, China; Lab of Biosystem and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, Shanghai, 200237, China. Electronic address:

Nucleic acid-based detection methods are accurate and rapid, which are widely-used in food-borne pathogen detection. However, traditional nucleic acid-based detection methods usually rely on special instruments, weakening their practicality for on-site tests in resource-limited locations. In this work, we developed a convenient and affordable method for food-borne pathogen detection based on a lateral flow strip combined with Cas9 nickase-triggered isothermal DNA amplification, which allows instrument-free and dual target detection. The genomic DNAs of two most common foodborne pathogens, Salmonella typhimurium and Escherichia coli, were simultaneously amplified in a one-pot reaction using specific sgRNAs and primers. The amplicons of genomic DNAs were double-labelled by digoxin/biotin and FITC/biotin tags, respectively, and directly visualized on a simple lateral flow strip. Our method exhibited a high specificity and sensitivity with a detection limit of 100 copies for genomic DNAs and 100 CFU/mL for bacteria. We believe that this method has potential to provide a convenient and low-cost point-of-care test for pathogen detection in the food quality surveillance.
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http://dx.doi.org/10.1016/j.bios.2020.112364DOI Listing
October 2020

Metabolically engineering of Yarrowia lipolytica for the biosynthesis of naringenin from a mixture of glucose and xylose.

Bioresour Technol 2020 Oct 24;314:123726. Epub 2020 Jun 24.

Laboratory of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China; Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou 310014, Zhejiang, China. Electronic address:

Xylose-inducible modules simultaneously expressing xylose utilization and naringenin biosynthesis pathways were developed in Yarrowia lipolytica to produce naringenin from a mixture of glucose and xylose. The naringenin synthetic pathway was constructed using a constitutive expression to yield 239.1 ± 5.1 mg/L naringenin. Furthermore, the introduction of an inducible pathway realized the dual function of xylose as a substrate and synthetic inducer, which coupled the xylose utilization with naringenin biosynthesis and increased production. Interestingly, the simultaneous enhancement of xylose reductase and xylose transporter expression along with that of xylitol dehydrogenase and xylulokinase can further improve the xylose utilization ability of Y. lipolytica. As expected, xylose-inducible synthesis of naringenin could achieved a titer of 715.3 ± 12.8 mg/L through the shake-flask cultivation level. Therefore, xylose-induced activation of both the xylose utilization and product biosynthesis pathway is considered to be an effective strategy for the biosynthesis of xylose-derived chemicals in yeast.
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http://dx.doi.org/10.1016/j.biortech.2020.123726DOI Listing
October 2020

A Cas12a-mediated cascade amplification method for microRNA detection.

Analyst 2020 Aug 1;145(16):5547-5552. Epub 2020 Jul 1.

Laboratory of Biosystem and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.

MicroRNAs (miRNAs) play a vital role in various biological processes and act as important biomarkers for clinical cancer diagnosis, prognosis, and therapy. Here, we took advantage of Cas12a trans-cleavage activity to develop an enzyme-assisted cascade amplification method for isothermal miRNA detection. A target miRNA-initiated ligation reaction would allow for the production of transcription templates that triggered the transcriptional amplification of RNA strands. These RNA strands were cleaved by the 8-17E DNAzyme to generate crRNAs and recycled RNAs which have the same sequence as the target miRNA. The amplified abundant crRNAs bound to Cas12a and dsDNA activators to form the complex, which trans-cleaved the ssDNA reporters to generate a fluorescence signal for miRNA quantitative analysis. The proposed method exhibits a femtomolar limit of detection and a good specificity in distinguishing the homologous sequences of miRNAs. Its practical application ability was further tested in different cell lines.
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http://dx.doi.org/10.1039/d0an00370kDOI Listing
August 2020

A novel and sensitive electrochemical sensor based on nanoporous gold for determination of As(III).

Mikrochim Acta 2020 06 20;187(7):395. Epub 2020 Jun 20.

Key Laboratory for Green Processing of Chemical Engineering of Xinjiang Bingtuan, School of Chemistry and Chemical Engineering, Shihezi University, Shihezi, 832003, China.

Three-dimensional porous gold nanoparticles (NPG) were synthesized in situ on indium-doped tin oxide (ITO) substrates by a green and convenient one-step electrodeposition method to achieve super-sensitive As(III) detection. The introduction of NPG method not only greatly improves the electron transfer capacity and surface area of sensor interface but provides more active sites for As(III) enrichment, thus boosting sensitivity and selectivity. The sensor was characterized by scanning electron microscopy, energy dispersion spectroscopy, differential pulse anode stripping voltammetry (DPASV), and electrochemical impedance to evaluate its morphology, composition, and electrochemical performance. The wall thickness of NPG was customized by optimizing the concentration of electroplating solution, dissolved electrolyte, deposition potential, and reaction time. Under optimal conditions, the electrochemical sensor showed a wide linear range from 0.1 to 50 μg/L As(III), with a detection limit (LOD) of 0.054 μg/L (S/N = 3). The LOD is far below 10 μg/L, the recommended maximum value by the world health organization for drinking water. Stability, reproducibility, and repeatability of NGP/ITO were determined to be 2.77%, 4.9%, and 4.1%, respectively. Additionally, the constructed sensor has been successfully applied to determine As(III) in three actual samples, and the results are in good agreement with that of hydride generation atomic fluorescence spectrometry (AFS). Graphical abstract.
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http://dx.doi.org/10.1007/s00604-020-04365-wDOI Listing
June 2020

Upstream ORFs Prevent MAVS Spontaneous Aggregation and Regulate Innate Immune Homeostasis.

iScience 2020 May 13;23(5):101059. Epub 2020 Apr 13.

Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, Zhejiang 310014, China. Electronic address:

The monomer-to-filament transition of MAVS is essential for the RIG-I/MDA5-mediated antiviral signaling. In quiescent cells, monomeric MAVS is under strict regulation for preventing its spontaneous aggregation, which would result in dysregulated interferon (IFN-α/β) production and autoimmune diseases like systemic lupus erythematosus. However, the detailed mechanism by which MAVS is kept from spontaneous aggregation remains largely unclear. Here, we show that upstream open reading frames (uORFs) within the MAVS transcripts exert a post-transcriptional regulation for preventing MAVS spontaneous aggregation and auto-activation. Mechanistically, we demonstrate that uORFs are cis-acting elements initiating leaky ribosome scanning of the downstream ORF codons, thereby repressing the full-length MAVS translation. We further uncover that endogenous MAVS generated from the uORF-deprived transcript spontaneously aggregates, triggering the Nix-mediated mitophagic clearance of damaged mitochondria and aggregated MAVS. Our findings reveal the uORF-mediated quantity and quality control of MAVS, which prevents aberrant protein aggregation and maintains innate immune homeostasis.
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http://dx.doi.org/10.1016/j.isci.2020.101059DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7190755PMC
May 2020

A telomerase-responsive nanoprobe with theranostic properties in tumor cells.

Talanta 2020 Aug 6;215:120898. Epub 2020 Mar 6.

Lab of Biosystem and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China; Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, Zhejiang, China; School of Chemistry and Chemical Engineering, Shihezi University, Shihezi, 832000, Xinjiang, China. Electronic address:

Multidrug resistance (MDR) is the main cause of treatment failure in clinical cancer chemotherapy due to the presence of P-glycoproteins (P-gp), which widely exist in stubborn drug-resistant tumor membranes and actively pump drugs from inside the tumor cell to the outside. In this study, we report a novel telomerase-responsive nanoprobe with theranostic properties for inhibiting P-gp expression and reversing MDR by gene silencing. This nanoprobe is composed of an AuNP assembled with telomerase primer, antisense oligonucleotide (ASO), and doxorubicin (Dox). When the designed nanoprobe is uptaken by the MDR cancer cells, the Dox and ASO are specifically released due to the extension of telomerase primer triggered by telomerase. The released ASO specifically hybridizes with multidrug resistance 1 (MDR1) mRNA sequence, which encodes the P-gp. As a result, the expression of P-gp is inhibited and the efflux of Dox is prevented with reduced MDR in cancerous cells. The results demonstrate that the nanoprobe based on telomerase switching for drug release and gene silencing, can both target cancer cells for delivering drugs and overcome the effect of efflux pumps. This work presents a novel paradigm for theranostics of MDR cancer and enhances the efficacy of chemotherapeutics.
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http://dx.doi.org/10.1016/j.talanta.2020.120898DOI Listing
August 2020

Multiple and sensitive SERS detection of cancer-related exosomes based on gold-silver bimetallic nanotrepangs.

Analyst 2020 Apr 26;145(7):2795-2804. Epub 2020 Feb 26.

Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou 310014, Zhejiang, China.

Exosomes are endogenous vesicles of cells, and can be used as important biomarkers for cancers. In this work, we developed a sensitive and reliable SERS sensor for simultaneous detection of multiple cancer-related exosomes. The SERS detection probes were made of bimetallic SERS-active nanotags, gold-silver-silver core-shell-shell nanotrepangs (GSSNTs), which were composed of bumpy surface nanorod (gold nanotrepang, GNT) cores and bilayer silver shells, and decorated with linker DNAs, which were complementary to the aptamer targeting exosomes. Three kinds of SERS detection probes were designed via the adoption of different Raman reporter molecules and linker DNAs. The capture probes were prepared by modifying specific aptamers of the target exosomes on magnetic beads (MBs). In the absence of target exosomes, SERS detection probes were coupled with MBs via specific DNA hybridization for use as aptamer-based SERS sensors. In the presence of target exosomes, the aptamer specifically recognized and captured the exosomes, and GSSNTs were subsequently released into the supernatant. Therefore, attenuated SERS signals were detected on the MBs, indicating the presence of target exosomes. The proposed aptamer-based SERS sensor is expected to be a facile and sensitive method for the multiplex detection of cancer biomarkers and has potential future applications in clinical diagnosis.
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http://dx.doi.org/10.1039/c9an02180aDOI Listing
April 2020

The ROK like protein of Myxococcus xanthus DK1622 acts as a pleiotropic transcriptional regulator for secondary metabolism.

J Biotechnol 2020 Mar 11;311:25-34. Epub 2020 Feb 11.

Department of Microbial Natural Products, Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Center for Infection Research (HZI) and Department of Pharmacy at Saarland University, Campus E8.1, 66123 Saarbrücken, Germany. Electronic address:

Myxococcus xanthus DK1622 is known as a proficient producer of different kinds of secondary metabolites (SM) with various biological activities, including myxovirescin A, myxalamide A, myxochromide A and DKxanthene. Low production of SM in the wild type bacteria makes searching for production optimization methods highly desirable. Identification and induction of endogenous key molecular feature(s) regulating the production level of the metabolites remain promising, while heterologous expression of the biosynthetic genes is not always efficient because of various complicating factors including codon usage bias. This study established proteomic and molecular approaches to elucidate the regulatory roles of the ROK regulatory protein in the modification of secondary metabolite biosynthesis. Interestingly, the results revealed that rok inactivation significantly reduced the production of the SM and also changed the motility in the bacteria. Electrophoretic mobility shift assay using purified ROK protein indicated a direct enhancement of the promoters encoding transcription of the DKxanthene, myxochelin A, and myxalamide A biosynthesis machinery. Comparative proteomic analysis by two-dimensional fluorescence difference in-gel electrophoresis (2D-DIGE) was employed to identify the protein profiles of the wild type and rok mutant strains during early and late logarithmic growth phases of the bacterial culture. Resulting data demonstrated overall 130 differently altered proteins by the effect of the rok gene mutation, including putative proteins suspected to be involved in transcriptional regulation, carbohydrate metabolism, development, spore formation, and motility. Except for a slight induction seen in the production of myxovirescin A in a rok over-expression background, no changes were found in the formation of the other SM. From the outcome of our investigation, it is possible to conclude that ROK acts as a pleiotropic regulator of secondary metabolite formation and development in M. xanthus, while its direct effects still remain speculative. More experiments are required to elucidate in detail the variable regulation effects of the protein and to explore applicable approaches for generating valuable SM in this bacterium.
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http://dx.doi.org/10.1016/j.jbiotec.2020.02.005DOI Listing
March 2020

Multimachine Communication Network That Mimics the Adaptive Immune Response.

J Am Chem Soc 2020 02 18;142(8):3851-3861. Epub 2020 Feb 18.

Laboratory of Biosystem and Microanalysis, State Key Laboratory of Bioreactor Engineering , East China University of Science and Technology , Shanghai 200237 , China.

Biological organisms capable of controlling and performing a wide variety of functions have inspired attempts to mimic biological systems with designable intelligence. Here we develop a multimachine communication network (MMCN) to mimic the operation and function of adaptive immune response (AIR) via connecting three kinds of DNA machines built from module-functionalized gold nanoparticles. These machines simulate three critical immune cells, dendritic cells, T and B lymphocytes, and their differentiation and coordinated interaction upon exposure and response to an invading pathogen. MMCN is composed of standard modules with track, movement, and fuel components that allow for the (1) integration and adaptability of a single machine, (2) convenient spatiotemporal control of the sequential activation of a single machine, and (3) rapid reaction rate and high efficiency owing to an enhanced local concentration of interacting species. We show that the proposed network can sense and clear the corresponding pathogen via consecutive activation and connection of the machines, simultaneously forming a memory to respond more rapidly and effectively upon the second invasion of the pathogen. This system may be extended to construct powerful networks to execute more sophisticated tasks and accomplish diverse functions.
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http://dx.doi.org/10.1021/jacs.9b11545DOI Listing
February 2020