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Minimizing endogenous cryptic plasmids to construct antibiotic-free expression systems for Escherichia coli Nissle 1917

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Synthetic and Systems biotechnology 2024年第1期9卷 165-175页

作者： Siyan Zhou Linlin Zhao Wenjie Zuo Yilin Zheng Ping Zhang Yanan Sun Yang Wang Guocheng Du Zhen Kang The science Center for Future Foods Jiangnan UniversityWuxi214122China The Key laboratory of Carbohydrate Chemistry and biotechnology Ministry of EducationSchool of BiotechnologyJiangnan UniversityWuxi214122China The Key laboratory of Industrial biotechnology Ministry of EducationSchool of BiotechnologyJiangnan UniversityWuxi214122China

The probiotic bacterium Escherichia coli Nissle 1917(EcN)holds significant promise for use in clinical and biological ***,the reliance on antibiotics to maintain plasmid-borne genes has overshadowed its *** this study,we addressed this issue by engineering the endogenous cryptic plasmids pMUT1 and *** non-essential elements were removed to create more stable derivatives pMUT1NR△and pMUT2HBC△.Synthetic promoters by integrating binding motifs on sigma factors were further constructed and applied for expression of Bacteroides thetaiotaomicron heparinaseⅢand the biosynthesis of *** to traditional antibiotic-dependent expression systems,our newly constructed antibiotic-free expression systems offer considerable advantages for clinical and synthetic biology applications.

关键词： Cryptic plasmids Antibiotic-free Gene expression Plasmid stability E.coli Nissle 1917

Enhanced axonal regeneration and functional recovery of the injured sciatic nerve in a rat model by lithium-loaded electrospun nanofibrous scaffolds

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Bio-Design and Manufacturing 2024年第5期7卷 701-720页

作者： Banafsheh Dolatyar Bahman Zeynali Iman Shabani Azita Parvaneh Tafreshi Reza Karimi-Soflou Developmental Biology laboratory School of BiologyCollege of ScienceUniversity of TehranTehran ***Iran Department of Biomedical Engineering Amirkabir University of Technology(Tehran Polytechnic)Tehran ***Iran Department of Molecular Medicine Faculty of Medical BiotechnologyNational Institute of Genetic Engineering and BiotechnologyTehran ***Iran 123

Increasing evidence indicates that engineered nerve grafts have great potential for the regeneration of peripheral nerve injuries(PNIs).While most studies have focused only on the topographical features of the grafts,we have considered both the biophysical and biochemical manipulations in our applied *** achieve this,we fabricated an electrospun nanofibrous scaffold(ENS)containing polylactide nanofibers loaded with lithium(Li)ions,a Wnt/β-catenin signaling *** addition,we seeded human adipose-derived mesenchymal stem cells(hADMSCs)onto this engineered scaffold to examine if their differentiation toward Schwann-like cells was *** further examined the efficacy of the scaffolds for nerve regeneration in vivo via grafting in a PNI rat *** results showed that Li-loaded ENSs gradually released Li within 11 d,at concentrations ranging from 0.02 to(3.64±0.10)mmol/L,and upregulated the expression of Wnt/β-catenin target genes(cyclinD1 and c-Myc)as well as those of Schwann cell markers(growth-associated protein 43(GAP43),S100 calcium binding protein B(S100B),glial fibrillary acidic protein(GFAP),and SRY-box transcription factor 10(SOX10))in differentiated *** the PNI rat model,implantation of Li-loaded ENSs with/without cells improved behavioral features such as sensory and motor functions as well as the electrophysiological characteristics of the injured *** improved function was further validated by histological analysis of sciatic nerves grafted with Li-loaded ENSs,which showed no fibrous connective tissue but enhanced organized myelinated *** potential of Li-loaded ENSs in promoting Schwann cell differentiation of hADMSCs and axonal regeneration of injured sciatic nerves suggests their potential for application in peripheral nerve tissue engineering.

关键词： Stem cell Schwann cell differentiation Electrospun nanofibrous scaffold Lithium ion Nerve regeneration

Supramolecular complex glycoconjugate vaccine generates self-enhancement effects for carbohydrate antigen delivery

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Chinese Chemical Letters 2024年第2期35卷 317-322页

作者： Han Lin Yanchun Li Kun Zhou Dan Li Hangyan Yu Jie Zhao Haofei Hong Zhifang Zhou Zhimeng Wu Key laboratory of Carbohydrate Chemistry & biotechnology Ministry of EducationSchool of BiotechnologyJiangnan UniversityWuxi 214122China

Targeting delivery of tumor-associated carbohydrate antigen(TACA)-based vaccine to antigen-presenting cells(APCs)mediated by endogenous antibodies can improve the immunogenicity of ***,an essential requirement of this approach is to generate high titers of endogenous antibodies in vivo through pre-immunization,which complicates the immunization procedure and may cause side ***,we report a new generation of APC-targeting TACA-based supramolecular complex vaccine,assembled by sialyl Thomsen-nouveau-bovine serum albumin-adamantine(sTn-BSA-Ada)and heptavalent rhamnose(Rha)-modifiedβ-cyclodextrin(β-CD)via host-guest *** complex vaccine retained anti-Rha antibodies recruiting capability and facilitated the APCs uptake of the vaccine via the interaction of the Fc-domain with the Fc receptors on *** demonstrate that direct immunization of complex vaccine elicited anti-Rha and anti-sTn specific immune response synchronously,generating a novel self-enhancement effect that can improve the antigen delivery to APCs in high *** structure-activity relationship(SAR)study proved that complex vaccine 4 with polyethylene glycol 6(PEG 6)linker in host molecule provoked a robust and specific sTn immune response comparable to the pre-immunization *** antisera induced by complex vaccine,either through direct immunization or pre-immunization,exhibited equal potency of cytotoxicity against the sTn expression cancer *** study provides a general platform for TACA-based vaccines with self-enhancement effects without the need for pre-immunization.

关键词： Glycoconjugate vaccine Endogenous antibody APC-targeting Supramolecular complex sTn antigen

An engineered Cas12i nuclease that is an efficient genome editing tool in animals and plants

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The Innovation 2024年第2期5卷 61-69页

作者： Zhiqiang Duan Yafeng Liang Jialei Sun Hongjin Zheng Tong Lin Pengyu Luo Mengge Wang Ruiheng Liu Ying Chen Shuhua Guo Nannan Jia Hongtao Xie Meili Zhou Minghui Xia Kaijun Zhao Shuhui Wang Na Liu Yongling Jia Wei Si Qitong Chen Yechun Hong Ruilin Tian Jian-Kang Zhu Institute of Advanced biotechnology and school of Medicine Southern University of Science and TechnologyShenzhen 518055China Bellagen biotechnology Jinan 250000China Department of medical Neuroscience School of MedicineKey University Laboratory of Metabolism and Health of GuangdongSouthern University of Science and TechnologyShenzhenGuangdong Province 518055China

The type V-I CRISPR-Cas system is becoming increasingly more attractive for genome ***,natural nucleases of this system often exhibit low efficiency,limiting their ***,we used structure-guided rational design and protein engineering to optimize an uncharacterized Cas12i nuclease,*** a result,we developed Cas-SF01,a Cas12i3 variant that exhibits significantly improved gene editing activity in mammalian ***-SF01 shows comparable or superior editing performance compared to SpCas9 and other Cas12 *** to natural Cas12i3,Cas-SF01 has an expanded PAM range and effectively recognizes NTTN and noncanonical NATN and TTVN *** addition,we identified an amino acid substitution,D876R,that markedly reduced the off-target effect while maintaining high on-target activity,leading to the development of CasSF01^(HiFi)(high-fidelity Cas-SF01).Finally,we show that Cas-SF01 has high gene editing activities in mice and *** results suggest that CasSF01 can serve as a robust gene editing platform with high efficiency and specificity for genome editing applications in various organisms.

关键词： system attractive rational

The interaction mechanism between zein and folic acid in alkaline aqueous solutions:an experimental and molecular simulation study

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Food Quality and Safety 2024年第1期8卷 166-178页

作者： Jun Zhu Yiling Yao Yuying Liao Lala Cai Wenqi Zhang Nuo Hong Wenlin Sun Chunhua Xiong Lanying Yao school of Food science and biotechnology Zhejiang Gongshang UniversityHangzhouChina

Objectives:Folic acid is a vitamin that is not highly soluble in water and is sensitive to the ***,it is important to find suitable *** study aimed to exemplify the interaction of folic acid with zein in alkaline aqueous solutions and shed light on how zein can be used as a carrier for folic *** and Methods:Zein and folic acid were separately dissolved in NaOH solutions with a PH of *** solution and folic acid solution were combined in specific *** methods including multi-spectroscopy,dynamic light scattering,and electron microscopy combining with molecular dynamics simulations were used to study the interaction mechanism between zein and folic acid in alkaline aqueous ***:Fluorescence spectroscopy showed that the quenching of zein by folic acid was mainly static,and the main driving force behind this interaction was van der Waals forces and hydrogen *** formation of zein–folic acid complexes was confirmed by ultraviolet–visible spectroscopy,Fourier transform infrared spectroscopy,and circular dichroism *** results also showed that the structure of zein changed when it interacted with folic *** light scattering analysis revealed that the addition of folic acid caused proteins to *** aggregates of the complexes had an irregular shape and were large,as observed by scanning electron microscopy and transmission electron *** simulation was used to further investigate the interaction *** to these findings,the folic acid molecule interacted with zein in a shallow recess near the protein *** dominant forces at play in the zein–folic acid interaction were van der Waals forces and electrostatic forces,including hydrogen ***:The zein alkali-soluble system is very suitable for folic acid delivery.

关键词： Zein folic acid alkaline aqueous solutions molecular simulation interaction mechanism.

RebL1 is required for macronuclear structure stability and gametogenesis in Tetrahymena thermophila

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Marine Life science & Technology 2024年第2期6卷 183-197页

作者： Huijuan Hao Yinjie Lian Chenhui Ren Sitong Yang Min Zhao Tao Bo Jing Xu Wei Wang Key laboratory of Chemical Biology and Molecular Engineering of Ministry of Education Institute of BiotechnologyShanxi UniversityTaiyuan 030006China school of Life science Shanxi UniversityTaiyuan 030006China Shanxi Key laboratory of biotechnology Taiyuan 030006China

Histone modification and nucleosome assembly play important roles in chromatin-related *** chaperones form different complexes and coordinate histone transportation and *** histone chaperone complexes have been identified in different *** ciliate protozoa(ciliates)have various chromatin structures and different nuclear ***,histone chaperone components and functions of different subunits remain unclear in ***-rahymema thermophila contains a transcriptionally active macronucleus(MAC)and a transcriptionally inactive micronu-cleus(MIC)which exhibit multiple replication and various chromatin remodeling progresses during vegetative growth and sexual developmental ***,we found histone chaperone RebL1 not only localized evenly in the transcriptionally active MAC but also dynamically changed in the MIC during vegetative growth and sexual developmental ***1 knockdown inhibited cellular *** macronuclear morphology became bigger in growing *** abnormal macronuclear structure also occurred in the starvation ***,micronuclear meiosis was disturbed during sexual development,leading to a failure to generate new gametic ***1 potentially interacted with various factors involved in histone-modifying complexes and chromatin remodeling complexes in different developmental ***1 knockdown affected expression levels of the genes involved in chromatin organization and *** together,RebL1 plays a vital role in maintaining macronuclear structure stability and gametogenesis in ***.

关键词： Gametogenesis Histone chaperone Macronuclear structure RebL1 Tetrahymena thermophilaIntroduction

Rational design of a single-component mRNA vaccine against orthopoxvirus and SARS-CoV-2

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science China(Life sciences) 2024年第6期67卷 1311-1313页

作者： Yilong Yang Xiaofan Zhao Yu Li Xiaodong Zai Xinyi Wang Yue Zhang Xiaolin Wang Peng Lv Jun Zhang Lihua Hou Junjie Xu Wei Chen laboratory of Advanced biotechnology Beijing Institute of BiotechnologyAcademy of Military Medical SciencesBeijing 100071China

Dear Editor,In the era of COVID-19,the prevalence of viruses from other families poses new threats to *** the eradication of smallpox in 1980,the global outbreak of monkeypox in 2022(Lum et al.,2022;Nuzzo et al.,2022)alerted that the re-emerging orthopoxvirus(OPXV)remained a persistent threat to global *** includes a number of viruses with a broad host range,including variola virus(VARV),monkeypox virus(MPXV),vaccinia virus(VACV).

关键词： vaccine Rational

Photo-controllable antifouling hydrogels embedded with AgNPs coated spiropyran functionalized mesoporous silica for long-term antibacterial activity

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Nano Research 2024年第9期17卷 8337-8349页

作者： Wei Su Fei Wang Bing Chen Ruoke Li Jindian Liu Junjie Xu Jianhua Fan Yueling Liu State Key laboratory of Bioreactor Engineering East China University of Science and TechnologyShanghai 200237China Department of Applied Biology School of BiotechnologyEast China University of Science and TechnologyShanghai 200237China

Silver nanoparticles(AgNPs)are widely used in antimicrobial ***,its easy aggregation and rapid loss hinder the effective *** address this issue,a novel stimuli-responsive antibacterial nanocomposite(Ag@SP-MSN)was developed based on spiropyran covalently conjugated mesoporous silica nanoparticles(284.6 nm)and AgNPs(27.1 nm)via strong electrostatic *** transmission electron microscopy(TEM)and atomic force microscopy(AFM)images proved the successful modification of AgNPs onto *** light-induced maximum loading amount towards AgNPs was calculated to be 95.0 wt.%after ultraviolet irradiation,while the amount of AgNPs released from Ag@SP-MSN was 94.4 wt.%under visible light in the aid of ammonia(0.1%,v/v).Upon cycled light irradiation,Ag@SP-MSN could recover 84.5 wt.%of AgNPs even after four *** proposed Ag@SP-MSN exhibited better antibacterial activity against both *** and *** than Ag@MSN under visible light illumination,indicating the efficient photo-responsive isomerization of ***,the Ag@SP-MSN embedded gel demonstrated outstanding antifouling ability even after 21 days when compared to AgNPs *** long-term photo-controllable antifouling property proved the excellent reversible absorption and release of Ag@SP-MSN towards *** work provides new insights into the safe utilization of nanomaterials,offering promising advancements to meet clinical antibacterial requirements.

关键词： light-responsive silver nanoparticles spiropyran mesoporous silica nanoparticles antibacterial activity

Sinensetin protects against periodontitis through binding to Bach1 enhancing its ubiquitination degradation and improving oxidative stress

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International Journal of Oral science 2024年第3期16卷 553-562页

作者： Zhiyao Yuan Junjie Li Fuyu Xiao Yu Wu Zhiting Zhang Jiahong Shi Jun Qian Xudong Wu Fuhua Yan Department of Periodontology Nanjing Stomatological HospitalAffiliated Hospital of Medical SchoolInstitute of StomatologyState Key Laboratory of Pharmaceutical BiotechnologySchool of Life SciencesNanjing UniversityNanjingChina

Periodontitis is a chronic inflammatory and immune reactive disease induced by the subgingival *** therapeutic effect for susceptible patients is often unsatisfactory due to excessive inflammatory response and oxidative ***(Sin)is a nature polymethoxylated flavonoid with anti-inflammatory and antioxidant *** study aimed to explore the beneficial effect of Sin on periodontitis and the specific molecular *** found that Sin attenuated oxidative stress and inflammatory levels of periodontal ligament cells(PDLCs)under inflammatory *** Sin to rats with ligation-induced periodontitis models exhibited a protective effect against periodontitis in *** molecular docking,we identified Bach1 as a strong binding target of Sin,and this binding was further verified by cellular thermal displacement assay and immunofluorescence *** immunoprecipitation-quantitative polymerase chain reaction results also revealed that Sin obstructed the binding of Bach1 to the HMOX1 promoter,subsequently upregulating the expression of the key antioxidant factor *** functional experiments with Bach1 knocked down and overexpressed verified Bach1 as a key target for Sin to exert its antioxidant ***,we demonstrated that Sin prompted the reduction of Bach1 by potentiating the ubiquitination degradation of Bach1,thereby inducing HO-1 expression and inhibiting oxidative ***,Sin could be a promising drug candidate for the treatment of periodontitis by targeting binding to Bach1.

关键词： Bach1 period prompt