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Minimizing endogenous cryptic plasmids to construct antibiotic-free expression systems for Escherichia coli Nissle 1.1.

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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 {1. of Carbohydrate Chemistry and biotechnology Ministry of EducationSchool of BiotechnologyJiangnan UniversityWuxi214122China The key {1. of industrial biotechnology Ministry of EducationSchool of BiotechnologyJiangnan UniversityWuxi214122China

The probiotic bacterium Escherichia coli Nissle 1.1.(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 pMUT1.R△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 1.1.

Regulatory RNAs in Bacillus subtilis:A review on regulatory mechanism and applications in synthetic biology

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Synthetic and Systems biotechnology 2024年第2期9卷 223-233页

作者： Anqi Peng Guobin Yin Wenjie Zuo Luyao Zhang Guocheng Du Jian Chen Yang Wang Zhen Kang The Science Center for Future Foods Jiangnan UniversityWuxi214122China The key {1. of Carbohydrate Chemistry and biotechnology Ministry of EducationSchool of BiotechnologyJiangnan UniversityWuxi214122China The key {1. of industrial biotechnology Ministry of EducationSchool of BiotechnologyJiangnan UniversityWuxi214122China

Bacteria exhibit a rich repertoire of RNA molecules that intricately regulate gene expression at multiple hierarchical levels,including small RNAs(sRNAs),riboswitches,and antisense ***,the majority of these regulatory RNAs lack or have limited protein-coding capacity but play pivotal roles in orchestrating gene expression by modulating transcription,post-transcription or translation *** and redesigning these regulatory RNA elements have emerged as pivotal strategies in the domains of metabolic engineering and synthetic *** previous investigations predominantly focused on delineating the roles of regulatory RNA in Gram-negative bacterial models such as Escherichia coli and Salmonella enterica,this review aims to summarize the mechanisms and functionalities of endogenous regulatory RNAs inherent to typical Gram-positive bacteria,notably Bacillus ***,we explore the engineering and practical applications of these regulatory RNA elements in the arena of synthetic biology,employing *** as a foundational chassis.

关键词： Regulatory RNA devices Small RNAs Riboswitches Synthetic biology Bacillus subtilis

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 {1. 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

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Design-build-test of recombinant Bacillus subtilis chassis cell by lifespan engineering for robust bioprocesses

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Synthetic and Systems biotechnology 2024年第3期9卷 470-480页

作者： Kexin Ren Qiang Wang Jianghua Chen Hengwei Zhang Zhoule Guo Meijuan Xu Zhiming Rao Xian Zhang key {1. of industrial biotechnology of the ministry of education School of BiotechnologyJiangnan UniversityWuxiJiangsu214122China Yixing Institute of Food and biotechnology Co. LtdYixing214200China

Microbial cell factories utilize renewable raw materials for industrial chemical production,providing a promising path for sustainable *** subtilis is widely used in industry for its food safety properties,but challenges remain in the limitations of microbial *** study proposes a novel strategy based on lifespan engineering to design robust *** chassis cells to supplement traditional metabolic modification strategies that can alleviate cell autolysis,tolerate toxic substrates,and get a higher mass transfer *** modified chassis cells could produce high levels of L-glutaminase,and tolerate hydroquinone to produceα-arbutin *** a 5 L bioreactor,the L-glutaminase enzyme activity of the final strain CRE1.TG was increased to 281..4±21.7 U m L^(-1.,about 1.98-fold compared with that of the wild ***α-arbutin yield of strain CRE1.A was increased to 1.4.7 g L^(-1.,about 1.34-fold compared with that of the *** our knowledge,both of the products in this study performed the highest yields reported so *** chassis modification strategy described in this study can Improve the utilization efficiency of chassis cells,mitigate the possible adverse effects caused by excessive metabolic modification of engineered strains,and provide a new idea for the future design of microbial cell factories.

关键词： Bacillus subtilis Chassis cell Lifespan engineering Robustness industrial production

Cell-free biocatalysis coupled with photo-catalysis and electro-catalysis: Efficient CO2-to-chemical conversion

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Green Energy & Environment 2024年第9期9卷 1366-1383页

作者： Junzhu Yang Chi-Kit Sou Yuan Lu key {1. of industrial Biocatalysis Ministry of Education Tsinghua University Department of Chemical Engineering Tsinghua University

The increasing atmospheric carbon dioxide （CO2） concentration has exposed a series of crises in the earth's ecological *** to effectively fix and convert carbon dioxide into products with added value has attracted the attention of many ***-free enzyme catalytic system coupled with electrical and light have been a promising attempt in the field of biological carbon fixation in recent *** this review,the research progresses of photoenzyme catalysis,electroenzyme catalysis and photo-electroenzyme catalysis for converting carbon dioxide into chemical products in cell-free systems are systematically *** focus on reviewing and comparing various coupling methods and principles of photoenzyme catalysis and electroenzyme catalysis in cell-free systems,especially the materials used in the construction of the coupling system,and analyze and point out the characteristics and possible problems of different coupling ***,we discuss the major challenges and prospects of coupling physical signals and cell-free enzymatic catalytic systems in the field of CO2fixation,suggesting possible strategies to improve the carbon sequestration capacity of such systems.

关键词： CO 2 fixation Cell-free system Enzyme Photoenzyme catalysis Electroenzyme catalysis

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Cross-linkedα-amylase aggregates on Fe3O4 magnetic nanoparticles modified with polydopamine/polyethyleneimine for efficient hydrolysis of starch

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Particuology 2024年第7期90卷 98-105页

作者： Jiandong Cui Xiuming Tang Qingqing Ma Yuyan Chang Qunli Zhang Shiru Jia State key {1. of Food Nutrition and Safety Laboratory of Industrial Fermentation MicrobiologyMinistry of EducationTianjin University of Science and TechnologyTianjin 300457China key {1. of Biomass Chemical Engineering of ministry of education Zhejiang UniversityHangzhou 310027China

In this workα-amylase was immobilized on magnetic Fe3O4 nanoparticles with polyethylenimine(PEI)/polydopamine(PDA)coating or 3-aminopropyl triethoxysilane(APTES)for the first time via adsorption–precipitation–*** with the freeα-amylase,the resultant magnetic cross-linkedα-amylase aggregates(PEI/PDA-M-CLEAs and N-M-CLEAs)exhibited excellent thermal and storage stability as well as pH *** storage at 25°C for 60 days,freeα-amylase only retained 60%of its initial activity,while PEI/PDA-M-CLEAs and N-M-CLEAs retained 80%and 78%of their initial activities,***,N-M-CLEAs and PEI/PDA-M-CLEAs showed good *** 6 repeated uses,PEI/PDA-M-CLEAs and N-M-CLEAs still maintained 65%and 62%of their initial activities,***,PEI/PDA-M-CLEAs and N-M-CLEAs exhibited higher starch hydrolysis efficiency than freeα-*** maximum dextrose equivalent(DE)values of starch hydrolysis by PEI/PDA-M-CLEAs and N-M-CLEAs reached 29.24%and 28.79%within 90 min,***,the maximum DE values of starch hydrolysis by the freeα-amylase was only 27.89%even in 1.0 *** magnetic cross-linkedα-amylase aggregates could be introduced as effective biocatalyst for industrial applications in production of maltose syrups.

关键词： α-amylase Magnetic nanoparticles Functional modification Magnetic cross-linked enzyme aggregates Hydrolysis of starch

Conductive and stable polyphenylene/CNT composite membrane for electrically enhanced membrane fouling mitigation

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Frontiers of Environmental Science & Engineering 2024年第1期18卷 35-47页

作者： Huijuan Xie Haiguang Zhang Xu Wang Gaoliang Wei Shuo Chen Xie Quan key {1. of industrial Ecology and Environmental Engineering(ministry of education) School of Environmental Science and TechnologyDalian University of TechnologyDalian 116024China

Nanocarbon-based conductive membranes, especially carbon nanotube (CNT)-based membranes, have tremendous potential for wastewater treatment and water purification because of their excellent water permeability and selectivity, as well as their electrochemically enhanced performance (e.g., improved antifouling ability). However, it remains challenging to prepare CNT membranes with high structural stability and high electrical conductivity. In this study, a highly electroconductive and structurally stable polyphenylene/CNT (PP/CNT) composite membrane was prepared by electropolymerizing biphenyl on a CNT hollow fiber membrane. The PP/CNT membrane showed 3.4 and 5.0 times higher electrical conductivity than pure CNT and poly(vinyl alcohol)/CNT (PVA/CNT) membranes, respectively. The structural stability of the membrane was superior to that of the pure CNT membrane and comparable to that of the PVA/CNT membrane. The membrane fouling was significantly alleviated under an electrical assistance of −2.0 V, with a flux loss of only 1..7% after 5 h filtration of humic acid, which is significantly lower than those of PP/CNT membranes without electro-assistance (56.8%) and commercial polyvinylidene fluoride (PVDF) membranes (64.1.). Additionally, the rejection of negatively charged pollutants (humic acid and sodium alginate) was improved by the enhanced electrostatic repulsion. After four consecutive filtration-cleaning cycle tests, the flux recovery rate after backwashing reached 97.2%, which was much higher than those of electricity-free PP/CNT membranes (67.0%) and commercial PVDF membranes (61.1.). This study offers insights into the preparation of stable conductive membranes for membrane fouling control in potential water treatment applications.

关键词： Polyphenylene CNTs Membrane Electro-assistance Membrane fouling mitigation

Characterization,expression dynamics,and potential function of OPA1.for regulation of mitochondrial morphology during spermiogenesis in Phascolosoma esculenta

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Journal of Oceanology and Limnology 2024年第1期42卷 187-200页

作者： Xinming GAO Binbin FENG Chen DU Congcong HOU Shan JIN Daojun TANG Junquan ZHU key {1. of Applied Marine biotechnology by the ministry of education and key {1. of Marine biotechnology of Zhejiang Province School of Marine SciencesNingbo UniversityNingbo 315211China College of Ecology Lishui UniversityLishui 323000China

Mitochondria undergo morphological changes during spermatogenesis in some *** mechanism and role of mitochondrial morphology regulation,however,remain somewhat *** this study,we analyzed the molecular characteristics,expression dynamics and subcellular localization of optic atrophy protein 1.OPA1.,a mitochondrial fusion and cristae maintenance-related protein,to reveal the possible regulatory mechanisms underlying mitochondrial morphology in Phascolosoma esculenta *** full-length cDNA of the *** opa1.gene(Pe-opa1.is 3743 bp in length and encodes 975 amino *** Pe-OPA1.protein is highly conservative and includes a transmembrane domain,a GTPase domain,two helical bundle domains,and a lipid-interacting *** and protein expression was higher in the coelomic fluid(a site of spermatid development)of male *** and increased first and then decreased from March to ***,their expression during the breeding stage was significantly higher than during the non-breeding stage,suggesting that Pe-OPA1.is involved in *** *** Pe-OPA1.protein was more abundant in components consisting of many spermatids than in components without,indicating that Pe-OPA1.mainly plays a role in the spermatid in coelomic ***,Pe-OPA1.was mainly detected in the spermatid *** experiments showed that the Pe-OPA1.are constitutively expressed and co-localized with mitochondria during spermiogenesis,suggesting its involvement in *** *** results provide evidence for Pe-OPA1.s involvement in the regulation of mitochondrial morphology during spermiogenesis.

关键词： optic atrophy protein 1.OPA1. spermiogenesis Phascolosoma esculenta mitochondrial dynamics spermatid

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Characterization of the effects of hyaluronan oligosaccharides on human gut microbiota using a simulated batch fermentation system

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Food Science and Human Wellness 2024年

作者： Xinwei Gao Hongkang Zhu Litao Hu Jieyu Sun Wenjie Zuo Ping Liu Yang Wang Guocheng Du Zhen Kang The Science Center for Future Foods， Jiangnan University The key {1. of Carbohydrate Chemistry and biotechnology， ministry of education， Jiangnan University The key {1. of industrial biotechnology， ministry of education， School of biotechnology， Jiangnan University School of Food Science and Technology， Jiangnan University

The prebiotic effects of hyaluronan (HA) are widely recognized， contributing to improved gut health and immune modulation. Despite its extensive use as dietary supplement， the specific interactions between HA oligosaccharides (o-HAs) and the gut microbiome remain largely unexplored. To investigate its role and metabolic fate in gut homeostasis， 200 mg/day of o-HAs (average molecule weight 1. Da) were added to an automated computer-controlled SIMulator of the Gastrointestinal tract (SIMGI). The results revealed a significant reshaping of the intestinal flora composition by o-HAs， notably reducing the Firmicutes/Bacteroides ratio. Fermentation of o-HAs by gut microbiota significantly increased the abundance of Bifidobacteria， Prevotellaceae＿Prevotella， Dialister， Eubacterium， and Sutterella， but decreased that of Catenibacterium， Oscillospira， Klebsiella， and Citrobacter (P < 0.05). This corresponded with significant enhancements in the content of short-chain fatty acids (SCFAs) such as acetic acid， propionic acid and n-butyric acid， highlighting the significant impact of o-HAs at the genus level. Furthermore， analysis of microbial function predicted the downregulation of pathological events in nine human diseases， particularly infectious ones (parasitic and bacterial). Potential inhibitions were observed in metabolic pathways associated with pentose and glucuronate interconversions as well as cationic antimicrobial peptide resistance. These findings underscore the in vitro prebiotic effects of o-HAs and their potential relevance in managing diverticular diseases or preventing metabolic disorders through the regulation of gut microbiota.

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