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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

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 Laboratory 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 CRE15TG was increased to 2817.4±21.7 U m L^(-1),about 1.98-fold compared with that of the wild ***α-arbutin yield of strain CRE15A was increased to 134.7 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

Chemical Synthesis of a Key Precursor Relevant to the Tetrasaccharide Repeating Unit from Treponema medium ATCC 700293

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Chinese Journal of Chemistry 2024年第14期42卷 1615-1622页

作者： Wenbin Sun Guangzong Tian Meiru Ding Chunjun Qin Xiaopeng Zou Jing Hu Jian Yin Key Laboratory of Carbohydrate Chemistry and biotechnology Ministry of EducationSchool of Biotechnology&School of Life Sciences and Health EngineeringJiangnan UniversityLihu Avenue 1800WuxiJiangsu214122 China Wuxi School of Medicine Jiangnan UniversityLihu Avenue 1800WuxiJiangsu214122 China

Treponema is a Gram-negative anaerobic bacterium,among which the pathogenic Treponema can cause various diseases,such as venereal syphilis(Treponema pallidum),yaws(Treponema carateum),and oral diseases(Treponema denticola and Treponema medium).Although different from conventional lipopolysaccharides,the extracellular glycoconjugate of Treponema may still be a potential antigen and provide a candidate for vaccine ***,we completed the first chemical synthesis of Treponema medium ATCC 700293 tetrasaccharide precursor containing L-ornithine(L-Orn)and D-aspartic acid(D-Asp)*** efficiency of non-reducing end disaccharide formation has been improved by optimizing the assembly of the protecting groups in the donors and ***[3+1]glycosylation strategy attempted to reduce the length of the acceptor to increase the nucleophilicity of the hydroxyl group,thereby improving the efficiency of synthesizing the target *** L-Orn derivative was introduced at the final stage due to its influence on the glycosylation stereospecificity and ***,the successful introduction of two amino acid derivatives and the synthesis of a tetrasaccharide precursor with complex functional-group modifications have provided valuable insights for synthesizing other complex bacterial glycans.

关键词： Glycosylation Total synthesis Treponema medium ATCC 700293 Tetrasaccharide precursor Protecting group Oligosaccharides Regioselectivity Neighboring-group effects

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 Laboratory of Carbohydrate Chemistry and biotechnology Ministry of EducationSchool of BiotechnologyJiangnan UniversityWuxi214122China The Key Laboratory 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

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

Carrimycin inhibits coronavirus replication by decreasing the efficiency of programmed–1 ribosomal frameshifting through directly binding to the RNA pseudoknot of viral frameshift-stimulatory element

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Acta Pharmaceutica Sinica B 2024年第6期14卷 2567-2580页

作者： Hongying Li Jianrui Li Jiayu Li Hu Li Xuekai Wang Jing Jiang Lei Lei Han Sun Mei Tang Biao Dong Weiqing He Shuyi Si Bin Hong Yinghong Li Danqing Song Zonggen Peng Yongsheng Che Jian-Dong Jiang CAMS Key Laboratory of Antiviral Drug Research Institute of Medicinal BiotechnologyChinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100050China Beijing Key Laboratory of Antimicrobial Agents Institute of Medicinal BiotechnologyChinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100050China Key Laboratory of biotechnology of Antibiotics the National Health and Family Planning Commission(NHFPC)Institute of Medicinal BiotechnologyChinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100050China

The pandemic of SARS-CoV-2 worldwide with successive emerging variants urgently calls for small-molecule oral drugs with broad-spectrum antiviral ***,we show that carrimycin,a new macrolide antibiotic in the clinic and an antiviral candidate for SARS-CoV-2 in phase III trials,decreases the efficiency of programmed–1 ribosomal frameshifting of coronaviruses and thus impedes viral replication in a broad-spectrum *** binds directly to the coronaviral frameshift-stimulatory element(FSE)RNA pseudoknot,interrupting the viral protein translation switch from ORF1a to ORF1b and thereby reducing the level of the core components of the viral replication and transcription *** carrimycin with known viral replicase inhibitors yielded a synergistic inhibitory effect on *** the FSE mechanism is essential in all coronaviruses,carrimycin could be a new broad-spectrum antiviral drug for human coronaviruses by directly targeting the conserved coronaviral FSE *** finding may open a new direction in antiviral drug discovery for coronavirus variants.

关键词： Carrimycin Coronavirus Broad-spectrum antiviral activity Programmed-1 ribosomal frameshifting RNA pseudoknot Antiviral agent RNA target Synergistic inhibitory effect

Rational engineering of homospermidine synthase for enhanced catalytic efficiency toward spermidine synthesis

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Synthetic and Systems biotechnology 2024年第3期9卷 549-557页

作者： Wenjing Liu Xiaoxiang Hu Yi Yan Yujie Cai The Key Laboratory of Industrial biotechnology Ministry of EducationSchool of BiotechnologyJiangnan University1800 Lihu RoadWuxiJiangsu214122China

Spermidine is a naturally occurring polyamine widely utilized in the prevention and treatment of various *** spermidine biosynthetic methods have problems such as low efficiency and complex multi-enzyme *** on sequence-structure-function relationships,we engineered the widely studied homospermidine synthase from Blastochloris viridis(BvHSS)and obtained mutants that could catalyze the production of spermidine from 1,3-diaminopropane and *** specific activities of BvHSS and the mutants D361E and E232D+D361E(E232D-D)were 8.72,46.04 and 48.30 U/mg,*** optimal pH for both mutants was 9.0,and the optimal temperature was 50◦*** docking and dynamics simulations revealed that mutating aspartic acid at position 361 to glutamic acid narrowed the substrate binding pocket,promoting stable spermidine ***,mutating glutamic acid at position 232 to aspartic acid enlarged the substrate channel entrance,facilitating substrate entry into the active pocket and enhancing spermidine *** whole-cell catalysis lasting 6 h,D361E and E232D-D synthesized 725.3 and 933.5 mg/L of spermidine,*** study offers a practical approach for single-enzyme catalyzed spermidine synthesis and sheds light on the crucial residues influencing homospermidine synthase catalytic activity in spermidine production.

关键词： Homospermidine synthase Polyamine Site-directed mutagenesis Spermidine Structural analysis

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

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