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Anti-asthmatic mechanism of the Huashanshen dripping pill via suppressing contraction of the airway smooth muscle

Traditional Medicine Research 2022年第3期7卷 19-27页

作者： Yu Liu Ni-Na Cui Xuan-Shuo Liu Peng-Cheng Lin Lorenzo Pecoraro Giuseppe Venturella Shu-Li Man Wen-Yuan Gao The Innovation Platform for the Development and Construction of Special Project of Key Laboratory for Tibet Plateau Phytochemistry of Qinghai Province(2017-ZJ-Y19) College of pharmacyQinghai Nationalities UniversityXining 810007China State Key Laboratory of Food Nutrition and Safety Key Laboratory of Industrial MicrobiologyMinistry of EducationTianjin Key Laboratory of Industry MicrobiologyNational and Local United Engineering Lab of Metabolic Control Fermentation TechnologyChina International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal ChemistryCollege of BiotechnologyTianjin University of Science&TechnologyTianjin 300457China Tianjin Key Laboratory for Modern Drug Delivery and High Efficiency School of Pharmaceutical Science and TechnologyTianjin UniversityTianjin 300072China Department of Agricultural Food and Forest SciencesUniversity of PalermoPalermoItaly

Background:The Huashanshen(HSS)dripping pill has been widely used in asthma for a long time in ***,the relaxant mechanism of HSS is not well ***:In this report,high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was used to identify the constituents in rat plasma after oral administration of ***-sensitized allergic asthma and isolated trachea were studied for the anti-asthmatic mechanism of ***:D-anisodamine,L-anisodamine,scopolamine and atropine were detected in the rat plasma containing *** was clear that the HSS inhibited the release of inflammatory mediators,regulated the balance of T-helper 1 and T-helper 2 to reduce the airway inflammation,and relaxed the tracheal smooth muscle by controlling the KCa channel,Ca^(2+)influx and release to reduce the airway ***:Atropine,anisodamine and scopolamine might be active compounds of HSS which inhibited the release of inflammatory mediators,regulated the balance of Th1/Th2,and relaxed the tracheal smooth muscle to reduce airway hyperresponsiveness.

关键词： Huashanshen dripping pill HPLC-QTOF-MS allergic asthma airway remodeling inflammation airway smooth muscle

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Genomic characteristics of a Streptococcus suis of ST353 resulting in severe endophthalmitis with bilateral deafness

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Infectious Diseases & Immunity 2025年第1期5卷 36-43页

作者： Liang Shen Yanfei Tong Shichao Li Yuda Chen Ruhan A Zhen Li Wenling Wang Lijuan Yin Jiuming Zou Ziyong Sun Changcheng Wu Wenjie Tan Xiangyang Central Hospital Affiliated Hospital of Hubei University of Arts and ScienceXiangyang 441021Hubei ProvinceChina National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases National Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China School of Public Health Baotou Medical CollegeBaotou 014030Inner MongoliaChina Collaborative Innovation Centre of Regenerative Medicine and Medical BioResourse Development and Application Co-constructed by the Province and Ministry Guangxi Medical UniversityNanning 530021Guangxi ProvinceChina State Key Laboratory of Food Nutrition and Safety Key Laboratory of Industrial MicrobiologyMinistry of EducationTianjin Key Laboratory of Industry MicrobiologyNational and Local United Engineering Lab of Metabolic Control Fermentation TechnologyChina International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal ChemistryCollege of BiotechnologyTianjin University of Science&TechnologyTianjin 300457China Department of Laboratory Medicine Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430030Hubei ProvinceChina

Background:Streptococcus suis(***)can be transmitted to humans through exposure to pigs or consumption of raw pork and causes serious *** infection through skin abrasions is considered an important route of transmission,few studies have reported skin injuries in patients during exposure or before *** study explored a rare instance of intraocular infection and bilateral deafness caused by *** and emphasized the importance of timely diagnosis and treatment to prevent disease ***:The diagnosis was made on the basis of clinical symptoms,imaging,molecular detection,and isolated culture *** susceptibility testing was conducted to determine the effectiveness of ***-genome sequencing was used to identify the strain’s sequence type and serotype as well as the presence of pathogenicity islands(PAIs).Results:A patient with intraocular infection and bilateral deafness was diagnosed with an *** *** infection was hypothesized to result from interspecies transmission due to the patient’s occupation and recent skin *** isolated strain was sensitive to ampicillin,ceftriaxone,vancomycin,linezolid,levofloxacin,and ***-genome sequencing revealed that the strain belonged to sequence type 353 and serotype 2,showing close similarity to strains isolated from wild boars in Chinese *** strain lacked the 89 complete kb PAI but had a 33 kb PAI with high similarity,potentially indicating variable ***:This study highlights the importance of employing multiple diagnostic strategies for the timely identification and treatment of *** *** presence of a potentially less virulent strain emphasizes the need for continuous surveillance and monitoring of emerging *** infections,particularly in Asian countries with high-risk populations associated with the pig farming industry.

关键词： Streptococcus suis Endophthalmitis Meningitis Genome sequencing Comparative genomics

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Portable 3d-Printed Apta-Biosensor for Sensitive Atp Detection Based on Hybridization Chain Reaction Amplification and Crispr/Cas12a

SSRN

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SSRN 2022年

作者： Ma, Long Liao, Dan Zhao, Zhiying Kou, Jun Guo, Haoyu Xiong, Xin Man, Shuli State Key Laboratory of Food Nutrition and Safety Key Laboratory of Industrial Microbiology Ministry of Education Tianjin Key Laboratory of Industry Microbiology National and Local United Engineering Lab of Metabolic Control Fermentation Technology China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry College of Biotechnology Tianjin University of Science & Technology Tianjin300457 China College of Artificial Intelligence Tianjin University of Science and Technology 3000457 China

The next generation biosensing tools based on CRISPR/Cas have revolutionized molecular detection. A number of CRISPR/Cas based biosensors have been reported for the detection of nucleic acid targets. The establishment of efficient methods for non-nucleic acid target detection would further broaden the scope of this technique, but up to now the concerning research is limited. In current study, we reported a versatile biosensing platform for non-nucleic acid called FIND-Cas (Functional and Integrated Nucleic acid mediated Detection using HCR amplification and CRISPR/Cas12a). Simply, HCR cascade signal amplification was firstly trigged by functional nucleic acid (aptamer) through target binding. Then, CRISPR/Cas system was integrated to recognize the amplified products followed by the activation of the trans-cleavage. As such, the target can be ingeniously converted to nucleic acid signals and then fluorescent signals that can be readily visualized and analyzed by a customized 3D-printed biosensor with the help of a home-made App-enabled smartphone. ATP was selected as a model target, and under the optimized conditions, we achieved fine analytical performance with a linear range from 0.1 to 750 μM and a detection limit (LOD) of 1.0 nM. The satisfactory selectivity and recoveries that we have obtained further demonstrated this method suitable for a complex sample environment. The sample-to-answer time was less than 100 min. Our work not only expanded the reach of the CRISPR-Cas system in biosensing, but also provided a prototype method that can be generalized for detecting a wider range of analytes with desirable adaptability, sensitivity, specificity and on-site capability. © 2022, The Authors. All rights reserved.

关键词： Biosensors

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Preparation of curcumin loaded carboxymethyl chitosan nanoparticles and their anti-hepatoma effects through liver-gut circulation

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International Journal of Biological Macromolecules 2025年 318卷

作者： Ning Zhang Wenwen Xie Wei Liu Long Ma Shuli Man State Key Laboratory of Food Nutrition and Safety Key Laboratory of Industrial Microbiology Ministry of Education Tianjin Key Laboratory of Industry Microbiology National and Local United Engineering Lab of Metabolic Control Fermentation Technology China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry College of Biotechnology Tianjin University of Science & Technology Tianjin 300457 China

Curcumin, a natural compound extracted from the rhizomes of Curcuma longa , has gained much attention due to its high food safety profile and broad biological activities, especial for anti-cancer activities. However, its clinical application is limited by poor bioavailability. In this study, we develop liver-targeted carboxymethyl chitosan (CMCS) nanoparticles named curcumin (CUR)-loaded dual ligand nanoparticles (DL NPs). CMCS is modified with deoxycholic acid (DCA), lactobionic acid (LA) and glycyrrhetinic acid (GA), which are regarded as safe common food. Transmission electron microscopy, 1 H NMR and Fourier-transform infrared spectroscopy are used for the characterization of DL NPs. As a result, CMCS possesses hydrophilic groups, and is pH-sensitive. DCA acts as the hydrophobic group and promotes the formation of self-assembled nanoparticles. LA and GA are used as the liver specific delivery of CUR. The drug-loaded DL NPs exhibit stronger cell internalization, and anticancer efficacies than the free CUR. Additionally, DL NPs improved the gut dysbiosis, and increased the abundance of Lachnospiraceae_ NK4A136 _group and the content of butyric acid, which further increased the expression of caspase 3 and induced apoptosis. Overall, it's proved that DL NPs would become promising pharmaceutical formulations for selectively overcoming hepatocellular carcinoma, with great potential for clinical applications.

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Identification of Predictive Biomarkers for the Progression to Severe COVID-19 Caused by Omicron Variants Using Metabolomics and Machine Learning Models

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SSRN 2024年

作者： Zhang, Shuaijie Wang, Chunhua Zhang, Lu Ye, Fan Mao, Xiaogang Xia, Ying Lu, Junjie Yu, Tingyu Zhao, Jianzhong Jia, Yanlong Tang, Mengjun Yin, Lijuan Yang, Yang Xing, Hui Tan, Wenjie Shen, Liang Cao, Fengsheng Department of Central Laboratory Xiangyang Central Hospital Affiliated Hospital of Hubei University of Arts and Science Hubei Province Xiangyang441021 China Department of Clinical Laboratory Xiangyang No. 1 People's Hospital Hubei University of Medicine Hubei Province Xiangyang441000 China Department of Critical Care Medicine Xiangyang Central Hospital Affiliated Hospital of Hubei University of Arts and Science Hubei Province Xiangyang441021 China State Key Laboratory of Food Nutrition and Safety Key Laboratory of Industrial Microbiology Ministry of Education Tianjin Key Laboratory of Industry Microbiology National and Local United Engineering Lab of Metabolic Control Fermentation Technology China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry College of Biotechnology Tianjin University of Science & Technology Tianjin300457 China Shenzhen Key Laboratory of Pathogen and Immunity National Clinical Research Center for infectious disease State Key Discipline of Infectious Disease Shenzhen Third People's Hospital Second Hospital Affiliated to Southern University of Science and Technology Shenzhen China NHC Key Laboratory of Biosafety National Institute for Viral Disease Control and Prevention Chinese Center for Disease Control and Prevention Beijing102206 China Xiangyang Central Hospital China

Background: The ongoing pandemic of COVID-19 has entered a new phase. Despite the reduced pathogenicity of the currently prevalent Omicron variant of SARS-CoV-2, there is still a risk of severe illness and death. Currently, there are no specific biomarkers available to accurately predict the progression and outcomes of Omicron induced COVID-19. Previous studies have mainly focused on untargeted or lipid metabolism analysis of individuals infected with the original strain of SARS-CoV-2 or Omicron with mild to moderate symptoms. Methods: We conducted a comprehensive targeted serum metabolomics analysis to analyze the metabolic profiles of COVID-19 patients infected with Omicron. We also correlated differentially expressed metabolites with laboratory test parameters. Finally, we developed a machine learning model that 'predict ' the progression and prognosis of severe COVID-19. Findings: We identified 450 DEMs between the non-severe and severe groups of COVID-19 patients. These differentially regulated metabolites are closely related to laboratory indicators such as inflammatory response, immune function. The results of the KEGG enrichment analysis on DEMs imply the association between the severity of COVID-19 in patients and host carbon homeostasis and resistance to oxidative stress, such as butyric acid metabolism, fructose and mannose metabolism. Subsequently, this study was able to accurately predict the progression of COVID-19 infection caused by the Omicron variant using three metabolites, Moreover, we first reported using five specific metabolites for predicting the outcomes of severe COVID-19 patients caused by the Omicron variant. Interpretation: 450 metabolites differ significantly between severe and non-severe Omicron-induced COVID-19 groups. We accurately predict the progression of COVID-19 infection caused by the Omicron variant using three metabolites. Funding: This research was supported by national Natural Science Foundation of China (82002192, 32302218

关键词： COVID-19

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Multiplexed CRISPR/Cas and Argonaute detection technology: strategies, challenges, and perspectives

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TrAC Trends in Analytical Chemistry 2025年 191卷

作者： Yaru Li Lu Zhao Wen Kang Long Ma Yunfeng Bai Feng Feng School of Agriculture and Life Science Shanxi Datong University Datong 037009 China School of Chemistry and Chemical Engineering Shanxi Provincial Key Laboratory of Chemical Biosensing Shanxi Datong University Datong 037009 China State Key Laboratory of Food Nutrition and Safety Key Laboratory of Industrial Microbiology Ministry of Education Tianjin Key Laboratory of Industry Microbiology National and Local United Engineering Lab of Metabolic Control Fermentation Technology China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry College of Biotechnology Tianjin University of Science and Technology Tianjin 300457 China

An increasing number of applications demand the simultaneous detection of multiple nucleic acid targets within a single reaction, which results in a reduction in both the time required for the assay and the overall cost. The programmable nucleases Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated (Cas) system and Argonaute have evolved into state-of-the-art molecular tools for the development of bioanalytical assays. In the review, the classification of the CRISPR/Cas and Argonaute were introduced followed by barriers of current multiplexed detection. Then the review systematically summarized the application of multiplexed CRISPR/Cas and Argonaute detection according to spatial/local separation, color-labelling differentiation, programmable nucleases sequence preference, sequence specific recognition. Finally, the review put forward the existing challenges and prospects of emerging field. The advancement of multiplexed CRISPR/Cas and Argonaute detection is ushering in a new era in bioanalysis with high sensitivity as well as specificity, offering great promise and desirability.

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CRISPR/Cas-based SERS detection: A win-win integration towards real-world applications

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TrAC Trends in Analytical Chemistry 2025年 191卷

作者： Ruibo Zhao Yaru Li Xinyue Li Myunghee Kim Xingjie Li Shichun Pei Shuli Man Weipan Peng Long Ma State Key Laboratory of Food Nutrition and Safety Key Laboratory of Industrial Microbiology Ministry of Education Tianjin Key Laboratory of Industry Microbiology National and Local United Engineering Lab of Metabolic Control Fermentation Technology China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry College of Biotechnology Tianjin University of Science & Technology Tianjin 300457 China School of Agriculture and Life Science Shanxi Datong University Datong 037009 China Department of Food Science & Technology Yeungnam University Gyeongsan Gyeongsangbuk-do 38541 South Korea Jilin Shuangzheng Diagnostic Monoclonal Antibody Scientist Studio Surge Medical Inc. Tonghua Jilin China Antibody Development Jilin Province University Enterprise Joint Technology Innovation Labs Tonghua Normal University Tonghua Jilin China

In recent years, the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated nuclease (Cas) system has risen as a powerful tool for genome editing. Beyond its editing capabilities, CRISPR/Cas system has garnered significant interest in molecular diagnostics, especially for nucleic acid detection due to its high sensitivity and specificity. Surface-enhanced Raman spectroscopy (SERS), reliant on plasmonic nanoparticles or nanostructures, was extensively employed in biosensing owing to its exceptional sensitivity and distinctive spectral bands. The integration of SERS with CRISPR/Cas technology led to the development of numerous biosensing approaches aimed at achieving ultra-sensitive detection. This review provided an overview of both the CRISPR/Cas and SERS technology, followed by a comprehensive summary of their combined application for nucleic acid detection. Furthermore, it delved into the current landscape of CRISPR/Cas-based SERS detection, addressing existing challenges and prospects for future advancements.

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Systems metabolic engineering strategies for the production of amino acids

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Synthetic and Systems Biotechnology 2017年第2期2卷 87-96页

作者： Qian Ma Quanwei Zhang Qingyang Xu Chenglin Zhang Yanjun Li Xiaoguang Fan Xixian Xie Ning Chen National and Local United Engineering Lab of Metabolic Control Fermentation Technology Tianjin University of Science and TechnologyTianjin300457China Key Laboratory of Industrial Fermentation Microbiology Ministry of EducationTianjin University of Science and TechnologyTianjin300457China College of Biotechnology Tianjin University of Science and TechnologyTianjin300457China

Systems metabolic engineering is a multidisciplinary area that integrates systems biology,synthetic biology and evolutionary *** is an efficient approach for strain improvement and process optimization,and has been successfully applied in the microbial production of various chemicals including amino *** this review,systems metabolic engineering strategies including pathwayfocused approaches,systems biology-based approaches,evolutionary approaches and their applications in two major amino acid producing microorganisms:Corynebacterium glutamicum and Escherichia coli,are summarized.

关键词： Systems metabolic engineering Amino acid Corynebacterium glutamicum Escherichia coli

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Global, regional, and national burden of diabetes from 1990 to 2021, with projections of prevalence to 2050: a systematic analysis for the Global Burden of Disease Study 2021

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The Lancet 2023年第10397期402卷 203-234页

作者： Ong, Kanyin Liane Stafford, Lauryn K. McLaughlin, Susan A. Boyko, Edward J. Vollset, Stein Emil Smith, Amanda E. Dalton, Bronte E. Duprey, Joe Cruz, Jessica A. Hagins, Hailey Lindstedt, Paulina A. Aali, Amirali Abate, Yohannes Habtegiorgis Abate, Melsew Dagne Abbasian, Mohammadreza Abbasi-Kangevari, Zeinab Abbasi-Kangevari, Mohsen ElHafeez, Samar Abd Abd-Rabu, Rami Abdulah, Deldar Morad Abdullah, Abu Yousuf Md Abedi, Vida Abidi, Hassan Aboagye, Richard Gyan Abolhassani, Hassan Abu-Gharbieh, Eman Abu-Zaid, Ahmed Adane, Tigist Demssew Adane, Denberu Eshetie Addo, Isaac Yeboah Adegboye, Oyelola A. Adekanmbi, Victor Adepoju, Abiola Victor Adnani, Qorinah Estiningtyas Sakilah Afolabi, Rotimi Felix Agarwal, Gina Aghdam, Zahra Babaei Agudelo-Botero, Marcela Arriagada, Constanza Elizabeth Aguilera Agyemang-Duah, Williams Ahinkorah, Bright Opoku Ahmad, Danish Ahmad, Rizwan Ahmad, Sajjad Ahmad, Aqeel Ahmadi, Ali Ahmadi, Keivan Ahmed, Ali Ahmed, Luai A. Ahmed, Syed Anees Ajami, Marjan Akinyemi, Rufus Olusola Al Hamad, Hanadi Al Hasan, Syed Mahfuz AL-Ahdal, Tareq Mohammed Ali Alalwan, Tariq A. Al-Aly, Ziyad AlBataineh, Mohammad T. Alcalde-Rabanal, Jacqueline Elizabeth Alemi, Sharifullah Ali, Hassam Alinia, Tahereh Aljunid, Syed Mohamed Almustanyir, Sami Al-Raddadi, Rajaa M. Alvis-Guzman, Nelson Amare, Firehiwot Ameyaw, Edward Kwabena Amiri, Sohrab Amusa, Ganiyu Adeniyi Andrei, Catalina Liliana Anjana, Ranjit Mohan Ansar, Adnan Ansari, Golnoosh Ansari-Moghaddam, Alireza Anyasodor, Anayochukwu Edward Arabloo, Jalal Aravkin, Aleksandr Y. Areda, Demelash Arifin, Hidayat Arkew, Mesay Armocida, Benedetta Arnlov, Johan Artamonov, Anton A. Arulappan, Judie Aruleba, Raphael Taiwo Arumugam, Ashokan Aryan, Zahra Asemu, Mulu Tiruneh Asghari-Jafarabadi, Mohammad Askari, Elaheh Asmelash, Daniel Astell-Burt, Thomas Athar, Mohammad Athari, Seyyed Shamsadin Atout, Maha Moh'd Wahbi Avila-Burgos, Leticia Awaisu, Ahmed Azadnajafabad, Sina Darshan, B.B. Babamohamadi, Hassan Badar, Muhammad Badawi, Alaa Badiye, Ashish D. Baghcheghi, Nayereh Baghe Institute for Health Metrics and Evaluation School of Medicine General Medicine Service Department of Veterans Affairs Seattle WA United States Department of Health Metrics Sciences School of Medicine Faculty of Medicine France Department of Clinical Governance and Quality Improvement Aleta Wondo Hospital Aleta Wondo Ethiopia Department of Nursing Woldia University Woldia Ethiopia Department of Nursing Department of Orthopedic Surgery Orthopaedic Surgery Social Determinants of Health Research Center Non-communicable Diseases Research Center Epidemiology Department Evidence-Based Practice Center Community and Maternity Nursing Unit University of Duhok Duhok Iraq School of Planning Faculty of Environment Department of Public Health Sciences Penn State College of Medicine Pennsylvania State University Hershey PA United States Biocomplexity Institute Virginia Tech Blacksburg VA United States College of Medicine Department of Public Health Laboratory Technology Sciences Department Department of Family and Community Health University of Health and Allied Sciences Ho Ghana Research Center for Immunodeficiencies Department of Biosciences and Nutrition Karolinska University Hospital Huddinge Sweden Clinical Sciences Department Department of Surgery Department of Surgery Department of Obstetrics & Gynecology College of Graduate Health Sciences University of Tennessee Memphis TN United States Department of Obstetrics & Gynecology Department of Clinical and Psychosocial Epidemiology Department of Anesthesia and Critical Care Centre for Social Research in Health Quality and Systems Performance Unit Cancer Institute NSW Sydney NSW Australia Department of Public Health and Tropical Medicine HIV and Infectious Diseases Department Jhpiego Abuja Nigeria Adolescent Research and Care Department Adolescent Friendly Research Initiative and Care Ado Ekiti Nigeria Department of Epidemiology Department of Epidemiology and Medical Statistics Department of Family Medicine McMaster University

Background: Diabetes is one of the leading causes of death and disability worldwide, and affects people regardless of country, age group, or sex. Using the most recent evidentiary and analytical framework from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD), we produced location-specific, age-specific, and sex-specific estimates of diabetes prevalence and burden from 1990 to 2021, the proportion of type 1 and type 2 diabetes in 2021, the proportion of the type 2 diabetes burden attributable to selected risk factors, and projections of diabetes prevalence through 2050. Methods: Estimates of diabetes prevalence and burden were computed in 204 countries and territories, across 25 age groups, for males and females separately and combined;these estimates comprised lost years of healthy life, measured in disability-adjusted life-years (DALYs;defined as the sum of years of life lost [YLLs] and years lived with disability [YLDs]). We used the Cause of Death Ensemble model (CODEm) approach to estimate deaths due to diabetes, incorporating 25 666 location-years of data from vital registration and verbal autopsy reports in separate total (including both type 1 and type 2 diabetes) and type-specific models. Other forms of diabetes, including gestational and monogenic diabetes, were not explicitly modelled. Total and type 1 diabetes prevalence was estimated by use of a Bayesian meta-regression modelling tool, DisMod-MR 2.1, to analyse 1527 location-years of data from the scientific literature, survey microdata, and insurance claims;type 2 diabetes estimates were computed by subtracting type 1 diabetes from total estimates. Mortality and prevalence estimates, along with standard life expectancy and disability weights, were used to calculate YLLs, YLDs, and DALYs. When appropriate, we extrapolated estimates to a hypothetical population with a standardised age structure to allow comparison in populations with different age structures. We used the comparative r

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Strategy for enhancing adenosine production under the guidance of transcriptional and metabolite pool analysis

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Biotechnology Letters 2015年第7期37卷 1361-1369页

作者： Zhang, Chenglin Du, Shanshan Liu, Yue Xie, Xixian Xu, Qingyang Chen, Ning College of Biotechnology Tianjin University of Science and Technology Tianjin300457 China National and Local United Engineering Lab of Metabolic Control Fermentation Technology Tianjin300457 China Tianjin Engineering Lab of Efficient and Green Amino Acid Manufacture Tianjin300457 China

Objectives: To rationally identify targets for enhancing adenosine production, transcription level of genes involved in adenosine synthesis of Bacillus subtilis XGL was detected during the fermentation process, complemented with metabolite pool analysis. Results: PurR-regulated genes (pur operon and purA) and prs were down-regulated and 5-phosphoribosyl 1-pyrophosphate (PRPP) decreased considerably after 24 h when adenosine significantly accumulated. Since PRPP could strongly antagonize the binding of PurR to its targets, it was inferred that down-regulation of pur operon and purA might be due to a low PRPP pool, which was confirmed by metabolite analysis. So desensitized prs responsible for PRPP synthesis was overexpressed, resulting in increased PRPP concentration and pur operon transcription. To further enhance the adenosine production, desensitized purF and prs were co-overexpressed with integrating additional copy of purA to B. subtilis XGL genome, resulting in 24.3 % (1.29 g/g DCW) higher adenosine production than that by B. subtilis XG. Conclusions: Overexpression of prs, purF and purA under the guidance of transcriptional and metabolite pool analysis significantly increased adenosine production. Strategies used in this study have potential applications for rational modification of industrial microorganisms. © 2015, Springer Science+Business Media Dordrecht.

关键词： Transcription

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