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Robust production of N-acetyl-glucosamine in engineered Escherichia coli from glycerol-glucose mixture

Synthetic and Systems Biotechnology 2025年第3期10卷 1014-1026页

作者： Sun, Wenchao Tang, Yulin Tian, Yahao Liu, Zhengkai Xiong, Wenwen Zhang, Hengshuai Chen, Liang Wu, Heyun Ma, Qian Xie, Xixian Key Laboratory of Industrial Fermentation Microbiology Ministry of Education Tianjin University of Science and Technology Tianjin 300457 China College of Biotechnology Tianjin University of Science and Technology Tianjin 300457 China National and Local United Engineering Lab of Metabolic Control Fermentation Technology Tianjin University of Science and Technology Tianjin 300457 China

Achieving a balanced cell growth and biosynthesis of target products is a pivotal challenge confronted in the realm of green biomanufacturing of chemicals, and it holds the promise of significantly enhancing the titer/yield/productivity of the target products. The co-utilization of carbon sources has emerged as a proven approach for the precise regulation of cell growth and biosynthesis processes, thereby serving as a potent strategy to expedite the production of chemicals. In our previous study, we successfully demonstrated the efficient production of N-acetyl-glucosamine in engineered Escherichia coli by leveraging an appropriate catabolic division of labor, utilizing a mixture of glycerol and glucose as the carbon source. In this study, we further refined the division of labor between these two carbon sources by meticulously regulating the expression of the zwf gene, which encodes glucose-6-phosphate dehydrogenase and is crucial in diverting carbon source to the pentose phosphate pathway (PPP). After comparing three strategies for balancing cell growth and production, the engineered strain NAG-1 with zwf knocked out aided by the optimization of the glycerol-to-glucose ratio and the feeding mode of carbon sources resulted in robust production of N-acetyl-glucosamine. Remarkable production in a 5 L bioreactor was achieved, obtaining 249 g/L of N-acetyl-glucosamine with a yield of 0.684 g/g of total carbon sources (specifically, 0.791 g/g of glucose) and a productivity of 3.46 g/L/h. These results establish NAG-1 as the most efficient microbial cell factory reported thus far for the bioproduction of N-acetyl-glucosamine. The robust production of N-acetyl-glucosamine in E. coli using a mixture of glycerol and glucose suggests the immense potential of mixed carbon sources in the industrial green biomanufacturing of chemicals. © 2025 The Authors

关键词： Escherichia coli Glycerol-glucose mixture N-Acetyl-glucosamine Robust production

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Gut liver brain axis in diseases:the implications for therapeutic interventions

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Signal Transduction and Targeted Therapy 2024年第1期9卷 79-104页

作者： Mengyao Yan Shuli Man Benyue Sun Long Ma Lanping Guo Luqi Huang Wenyuan Gao 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&Technology300457 TianjinChina National Resource Center for Chinese Materia Medica China Academy of Chinese Medical Sciences100700 BeijingChina Tianjin Key Laboratory for Modern Drug Delivery&High-Efficiency School of Pharmaceutical Science and TechnologyTianjin UniversityWeijin Road300072 TianjinChina

Gut-liver-brain axis is a three-way highway of information interaction system among the gastrointestinal tract,liver,and nervous *** the past few decades,breakthrough progress has been made in the gut liver brain axis,mainly through understanding its formation mechanism and increasing treatment *** this review,we discuss various complex networks including barrier permeability,gut hormones,gut microbial metabolites,vagus nerve,neurotransmitters,immunity,brain toxic metabolites,β-amyloid(Aβ)metabolism,and epigenetic regulation in the gut-liver-brain *** therapies containing antibiotics,probiotics,prebiotics,synbiotics,fecal microbiota transplantation(FMT),polyphenols,low FODMAP diet and nanotechnology application regulate the gut liver brain ***,some special treatments targeting gut-liver axis include farnesoid X receptor(FXR)agonists,takeda G protein-coupled receptor 5(TGR5)agonists,glucagon-like peptide-1(GLP-1)receptor antagonists and fibroblast growth factor 19(FGF19)*** gut-brain axis embraces cognitive behavioral therapy(CBT),antidepressants and tryptophan metabolism-related *** liver-brain axis contains epigenetic regulation and Aβmetabolism-related *** the future,a better understanding of gut-liver-brain axis interactions will promote the development of novel preventative strategies and the discovery of precise therapeutic targets in multiple diseases.

关键词： metabolism therapeutic immunity

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Rhizoma paridis saponins protected against liver injury in diethylnitrosamine-induced mice

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Traditional Medicine Research 2023年第2期8卷 7-12页

作者： Chen Luo Jing-Wen Yao Shu-Li Man Wen-Yuan Gao 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/College 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

Background:Diethylnitrosamine,one of food additives,possessed a strong carcinogenic effect in *** paridis saponins,as the main active components of Paris polyphylla,have a good anti-cancer effect in our previous *** verify their inhibitory effect on diethylnitrosamine-induced liver cancer,we carried out this ***:We established diethylnitrosamine-induced mouse hepatocarcinoma models to evaluate antitumor of Rhizoma paridis ***,gas chromatography-mass spectrometry was applied to analyze the metabolites in the urine and serum ***:Rhizoma paridis saponins alleviated diethylnitrosamine-induced *** the one hand,Rhizoma paridis saponins down-regulated the levels of liver function markers,such as alanine aminotransferase,aspartate transaminase and alpha *** the other hand,Rhizoma paridis saponins reduced metabolic disorders by increasing fructose and mannose metabolism,and decreasing pentose and glucuronate interconversion,inositol phosphate metabolism,and the process of saturated fatty acids transforming to unsaturated fatty acids,which based on the regulating mRNA expression of glucose transporter type 4,lactate dehydrogenase A,fatty acid synthetas,acetyl-CoA carboxylase and apolipoprotein ***:Rhizoma paridis saponins has the potential application to inhibit chemical-induced hepatocarcinogenesis in the future.

关键词： Rhizoma paridis saponins liver injury diethylnitrosamine metabolites

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Platycladus orientalis shells promote melanogenesis via p38,AKT and Wnt/β-catenin signaling pathways

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Traditional Medicine Research 2022年第6期7卷 56-64页

作者： Fu-Rui Liu Hao Xiang Qian-Bei Guo Shu-Li Man Long Ma 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

Background:Platycladus orientalis,which has been employed in traditional Chinese medicine for cool blood,antibacterial,promotion of hair growth and therapy of poliosis for ***,there have been few reports focusing on Platycladus orientalis shells treating pigmentary ***:In present study,gas chromatography–mass spectrometry was applied to analyzing the volatile composition of Platycladus orientalis shells and cellular *** in vivo were carried out to evaluate the effect of Platycladus orientalis shells treating pigmentation disorders in C57BL/6J ***:Our results indicated that cedrol occupied the largest percentage in Platycladus orientalis shells(17.1%).Meanwhile,Platycladus orientalis shells up-regulated the content of palmitic acid and increased melanin content and tyrosinase *** mechanism possibly involved in the inhibiting phosphorylation of AKT and β-catenin,increasing phosphorylation of p38 to promote microphthalmia-associated transcription factor *** animal experiment also proved that Platycladus orientalis shells promoted melanogensis and hair blacken in C57BL/6J ***:All in all,Platycladus orientalis shells potentially promoted melanogenesis in vitro and in *** was regarded as the main active substance in Platycladus orientalis ***,it could be used for treatment of pigmentary disorders under safe concentration in the prospective application.

关键词： GC-MS Platycladus orientalis shells melanogenesis tyrosinase pigmentary disorders

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Multi-component quantitative and feed-forward neural network for pattern classification of raw and wine-processed Corni Fructus

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Traditional Medicine Research 2023年第1期8卷 12-19页

作者： Yu Liu Ying-Fang Cui Dan-Dan Shi Shu-Li Man Xia Li Wen-Yuan Gao 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 NutritionSafety and Medicinal Chemistry/College of BiotechnologyTianjin University of Science&TechnologyTianjin 300457China Tianjin Key Laboratory for Modern Drug Delivery&High-Efficiency School of Pharmaceutical Science and TechnologyTianjin UniversityTianjin 300072China.

Background:To promote the quality evaluation,clarify the processing mechanism and distinguish origins of Corni Fructus(cornus)from different ***:This study developed a high performance liquid chromatography method for simultaneous determination of 5-hydroxymethylfurfural,2 phenolic acids and 4 iridoid glycosides and the reference fingerprint of cornus from different *** addition,the feedforward neural network model provided a pattern classification of sample ***:The content of morroniside and loganin were the highest in all raw cornus samples ranging from 9.45μg/mg to 16.3μg/mg and 6.64μg/mg to 13.7μg/mg,*** level of sweroside in raw cornus from Henan(0.83μg/mg^(-1).39μg/mg)and Zhejiang(0.64μg/mg^(-1).17μg/mg)were greater than other *** wine-processing,the glucose or fructose were dehydrated to increase the levels of *** C-4 position of-COOCH3 of hot-sensitive iridoid glycosides was hydrolyzed to generate-COOH as stable *** derivatives may be degraded to increase the content of phenolic ***,an excellent feedforward neural network model for identification of raw cornus and wine-prepared cornus was established which could distinguish the sample ***:This work provided a trustworthy method to evaluate the quality and distinguish the sources of ***,the clear processing mechanism provided a scientific foundation for controlling the cornus quality during wine-processing.

关键词： Cornus officinalis Sieb.et Zucc. quality evaluation fingerprints processing mechanism feedforward neural network

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Lycorine derivative effectively inhibits the replication of coronaviruses both in vitro and in vivo

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hLife 2024年第2期2卷 75-87页

作者： Liang Shen Jianzhong Zhao Ying Xia Junjie Lu Jiali Sun Jian Tang Hui Xing Lijuan Yin Yang Yang Chunhua Wang Department of Clinical Laboratory Xiangyang Central HospitalAffiliated Hospital of Hubei University of Arts and ScienceHubeiChina Shenzhen Key Laboratory of Pathogen and Immunity National Clinical Research Center for Infectious DiseaseState Key Discipline of Infectious DiseaseShenzhen Third People’s HospitalSecond Hospital Affiliated to Southern University of Science and TechnologyGuangdongChina Department of Clinical Laboratory Xiangyang No.1 People’s HospitalHubei University of MedicineHubeiChina 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&TechnologyTianjinChina

The established and ongoing prevalence of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)and seasonal human coronaviruses(HCoV)like HCoV-OC43,HCoV-NL63,and HCoV-229E,pose a continuous threat to public ***,it is urgently needed to explore antiviral drugs with broad-spectrum anti-coronavirus *** previous studies have revealed that lycorine is a potent broad-spectrum anti-coronavirus drug,a natural alkaloid extracted from Amaryllidaceae with various pharmacological and microbiological ***,it is unsafe to directly use lycorine as a clinical antiviral drug due to the cytotoxicity and induction of cell *** this study,a series of lycorine derivatives were designed and *** of them,named Ly-8,was found to effectively inhibit the replication of different coronavirus strains in vitro,including ***,Ly-8 was also shown to effectively inhibit HCoVOC43 replication in the central nervous system,and provide effective protection against HCoV-OC43 infection in mice with low drug ***,Ly-8-resistant mutants were not observed during the 30 times sequential passages in cell ***,these findings suggest that Ly-8 may be a potential candidate drug for the future development of broad-spectrum anti-coronavirus drugs.

关键词： broad-spectrum antiviral drug lycorine derivative Ly-8

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Diosgenin alleviates neuropathic pain based on the enhancement of autophagy and promotion of M2 polarization

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International Immunopharmacology 2025年 159卷 114936页

作者： Man, Shuli Zhou, Yaxue Zhang, Xinghao Tang, Xiaoqin Xie, Lu Ma, Long Gao, Wenyuan 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 Tianjin Key Laboratory for Modern Drug Delivery and High Efficiency School of Pharmaceutical Science and Technology Faculty of Medicine Tianjin University Tianjin 300072 China

Neuropathic pain has serious impacts on the quality of people's life. Diosgenin extracted from yams has shown significant potential in the treatment of nervous system diseases. To investigate the potential mechanism of diosgenin fighting against neuropathic pain, we established a chronic compression injury (CCI) model. The pain-related behaviors were determined by Von Frey, hot plate, and sciatic nerve index test. As a result, diosgenin significantly lowered the pain threshold, improved the sciatic nerve index, and decreased the damage of sciatic nerve structure in CCI model mice. It increased the protein levels of LC3-II and Beclin-1, the number of the lysosomes, decreased the protein level of P62 and the number of the autophagosomes, and therefore activated autophagy. Diosgenin also inhibited the proliferation of microglia cells and the translocation of NF-κB p65, while promoting promoted M2 polarization of microglia cells. Autophagy inhibitor like chloroquine inhibited the M2 polarization of microglia cells, and reversed the therapeutic effect of diosgenin. All in all, diosgenin alleviated neuropathic pain in CCI mice based on the enhancement of autophagy and the promotion of M2 polarization. © 2024

关键词： Autophagy Diosgenin Microglia Neuropathic pain

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A magnetic CRISPR/Cas12a-SERS nanobiosensor for amplification-free and ultrasensitive detection of norovirus in water and food samples

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Analytica Chimica Acta 2025年 1363卷 344133页

作者： Zhou, Caihong Zhang, Yongzhao Yang, Xuejing Zhao, Ziqian Xia, Menglei Wei, Chunhui Wu, Qiankun Chang, Zisong Ma, Long Yin, Lijuan 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 Tianjin300457 China Sichuan University of Science & Engineering Liquor Making Biological Technology and Application of Key Laboratory of Sichuan Province Yibin644000 China Institute of Quantum Materials and Physics Henan Academy of Sciences Zhengzhou450046 China Academy of National Food and Strategic Reserves Administration Beijing100037 China Tianjin Odrei Biomedical Technology Co. LTD Tianjin300457 China

Background: Norovirus (NoV) is the leading cause of foodborne disease outbreaks worldwide, typically spreading via contaminated food and water. Rapid, sensitive, and portable detection of NoV is crucial. Results: Here, we presented a magnetic CRISPR/Cas12a-SERS nanobiosensor capable of detecting NoV with high sensitivity, accuracy, speed, and portability. In this nanobiosensor, SERS nanoprobes linked to magnetic nanoprobes via linker single-stranded DNAs (ssDNAs). The presence of NoV nucleic acid triggered Cas12a′s trans-cleavage activity, degrading the linker ssDNA. After magnetic separation, the dissociated SERS nanoprobes were efficiently separated from the magnetic nanoprobes. This enhanced the SERS signal in the supernatant, detectable using a portable Raman spectrometer. The detection limit for NoV is 100 copies/mL within 60 min. The nanobiosensor was further assessed in real-world settings, demonstrating excellent sensitivity and selectivity for detecting trace NoV in complex food samples. Significance: This approach not only broadens CRISPR-based pathogen detection but also provides a reliable tool for monitoring foodborne viruses. Its potential extends beyond NoV, promising enhanced surveillance of various pathogens in food safety, environmental monitoring, and public health sectors. © 2025 Elsevier B.V.

关键词： Enzyme sensors

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A Ratiometric Fluorescent Biosensing Platform for Ultrasensitive Detection of Salmonella Typhimurium Via Crispr/Cas12a and Silver Nanoclusters

SSRN

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SSRN 2022年

作者： Ma, Long Wang, Jiajing Li, Yaru Liao, Dan Han, Xiao 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

Rapid, sensitive and specific detection of bacteria is of great importance. Herein, we developed a versatile biosensing platform for ultrasensitive detection of pathogenic bacteria, termed as SCENT-Cas (Silver nanoCluster Empowered Nucleic acids Test using CRISPR/Cas12a). Simply, the species-specific invA gene of Salmonella typhi was isothermally amplified using LAMP, which subsequently triggered the trans-cleavage of CRISPR/Cas12a. The trans-cleavage degraded any single-stranded DNA (ssDNA) non-specifically. A DNA-regulated AgNCs probe was then employed, in which green fluorescence emissive AgNCs eﬀectively converted to red fluorescence-emissive AgNCs when placed in close vicinity to a pre-designed converter ssDNA. As such, the trans-cleavage was utilized for shredding converter ssDNA, enabling the green-to-red fluorescent change to form a ratiometric biosensing platform. With this strategy, target nucleic acid was dexterously converted into ratiometric fluorescence that was recorded to detect as low as 1 CFU/mL Salmonella typhimurium with a dynamic range from 1 to 10 8 CFU/mL. To our knowledge, this is the first report regarding the use of ratiometric fluorescence in CRISPR/Cas-based detection, which minimizes interference and improves reliability. Lastly, this proposed strategy was challenged by detecting S. typhi contamination in real food samples. Our work enriches CRISPR/Cas toolbox in biosensing by providing a desirable method for bacterial detection. © 2022, The Authors. All rights reserved.

关键词： DNA

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Lycorine derivative effectively inhibits the replication of coronaviruses both in vitro and in vivo

微生物与宿主健康（英文）

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微生物与宿主健康（英文） 2024年第2期2卷 75-87页

作者： Liang Shen Jianzhong Zhao Ying Xia Junjie Lu Jiali Sun Jian Tang Hui Xing Lijuan Yin Yang Yang Chunhua Wang Department of Clinical Laboratory Xiangyang Central HospitalAffiliated Hospital of Hubei University of Arts and ScienceHubeiChina Department of Clinical Laboratory Xiangyang No.1 People's HospitalHubei University of MedicineHubeiChina 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 &TechnologyTianjinChina Shenzhen Key Laboratory of Pathogen and Immunity National Clinical Research Center for Infectious DiseaseState Key Discipline of Infectious DiseaseShenzhen Third People's HospitalSecond Hospital Affiliated to Southern University of Science and TechnologyGuangdongChina

The established and ongoing prevalence of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)and sea-sonal human coronaviruses(HCoV)like HCoV-OC43,HCoV-NL63,and HCoV-229E,pose a continuous threat to public ***,it is urgently needed to explore antiviral drugs with broad-spectrum anti-coronavirus *** pre-vious studies have revealed that lycorine is a potent broad-spectrum anti-coronavirus drug,a natural alkaloid extracted from Amaryllidaceae with various pharmacological and microbiological ***,it is unsafe to directly use ly-corine as a clinical antiviral drug due to the cytotoxicity and induction of cell *** this study,a series of lycorine derivatives were designed and *** of them,named Ly-8,was found to effectively inhibit the replication of different coronavirus strains in vitro,including ***,Ly-8 was also shown to effectively inhibit HCoV-OC43 replication in the central nervous system,and provide effective protection against HCoV-OC43 infection in mice with low drug ***,Ly-8-resistant mutants were not observed during the 30 times sequential passages in cell ***,these findings suggest that Ly-8 may be a potential candidate drug for the future develop-ment of broad-spectrum anti-coronavirus drugs.

关键词： broad-spectrum antiviral drug lycorine derivative Ly-8

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