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Enhanced production of poly-3-hydroxybutyrate by recombinant Escherichia coli containing NAD kinase and phb CAB operon

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Science China Chemistry 2016年第11期59卷 1390-1396页

作者： Shaoqin Zhang Lei Fang Zhengjun Li Yingying guo guo-qiang chen Center for Synthetic and Systems Biology School of Life Science Tsinghua-Peking Center for Life SciencesTsinghua University Beijing Key Laboratory of Bioprocess College of Life Science and Technology Beijing University of Chemical Technology Center for Nano and Micro Mechanics Tsinghua University

With the help of Tn5 transposon technique, gene yfj B encoding NAD kinase in Escherichia coli(E. coli) was inserted into chromosome of recombinant E. coli polyhydroxybutyrate(PHB) containing PHB synthesis operon integrated in the host genome. After successful transposition of an extra yfj B gene copy into genome, the selected recombinant named E. coli PHBTY4 showed stronger NAD kinase activity than that of E. coli PHB. Shake flask studies suggested that both cell dry weight and PHB accumulation were significantly increased in E. coli PHBTY4 compared with that of the control. E. coli PHBTY4 produced approximately 23 g/L PHB compared with its control which synthesized only 10 g/L PHB when grown under the same conditions in a 6 L fermentor after 32 h of cultivation. In addition, E. coli PHBTY4 maintained high genetic stability during the cultivation processes. These results revealed a practical method to construct genetically stable strains harboring extra NAD kinase gene to enhance NADP(H)-dependent bio-reactions.

关键词： PHB polyhydroxyalkanoates Tn5 transposon NAD kinase Escherichia coli

Engineering bacteria for enhanced polyhydroxyalkanoates(PHA)biosynthesis

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Synthetic and Systems Biotechnology 2017年第3期2卷 192-197页

作者： guo-qiang chen Xiao-Ran Jiang School of Life Sciences Tsinghua UniversityBeijing 100084China Center for Synthetic and Systems Biology Tsinghua UniversityBeijing 100084China Tsinghua-Peking Center for Life Sciences Tsinghua UniversityBeijing 100084China Center for Nano and Micro Mechanics Tsinghua UniversityBeijing 100084China MOE Key Lab of Industrial Biocatalysis Dept Chemical EngineeringTsinghua UniversityBeijing 100084China

Polyhydroxyalkanoates(PHA)have been produced by some bacteria as bioplastics for many *** their commercialization is still on the way.A few issues are related to the difficulty of PHA commercialization:namely,high cost and instabilities on molecular weights(Mw)and structures,thus instability on thermo-mechanical *** high cost is the result of complicated bioprocessing associated with sterilization,low conversion of carbon substrates to PHA products,and slow growth of microorganisms as well as difficulty of downstream *** engineering on PHA producing microorganisms should be focused on contamination resistant bacteria especially extremophiles,developments of engineering approaches for the extremophiles,increase on carbon substrates to PHA conversion and controlling Mw of *** concept proof studies could still be conducted on *** or Pseudomonas *** are easily used for molecular *** this review,we will use *** and halophiles as examples to show how to engineer bacteria for enhanced PHA biosynthesis and for increasing PHA competitiveness.

关键词： PHB Polyhydroxyalkanoates Extremophiles Halophiles Next generation industrial biotechnology NGIB Metabolic engineering Pathway engineering Morphology engineering Contents

Synthetic biology of microbes synthesizing polyhydroxyalkanoates (PHA)

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Synthetic and Systems Biotechnology 2016年第4期1卷 236-242页

作者： guo-qiang chen Xiao-Ran Jiang Yingying guo School of Life Sciences Tsinghua UniversityBeijing 100084China Center for Synthetic and Systems Biology Tsinghua UniversityBeijing 100084China Tsinghua-Peking Center for Life Sciences Tsinghua UniversityBeijing 100084China Center for Nano and Micro Mechanics Tsinghua UniversityBeijing 100084China MOE Key Lab of Industrial Biocatalysis Dept Chemical EngineeringTsinghua UniversityBeijing 100084China

Microbial polyhydroxyalkanoates(PHA)have been produced as bioplastics for various *** the support of China National Basic Research 973 Project,we developed synthetic biology methods to diversify the PHA structures into homo-,random,block polymers with improved properties to better meet various application *** the same time,various pathways were assembled to produce various PHA from glucose as a simple carbon *** the end,Halomonas bacteria were reconstructed to produce PHA in changing morphology for low cost production under unsterile and continuous *** synthetic biology will advance the PHA into a bio-and material industry.

关键词： Polyhydroxyalkanoates 973 Bioprocesses Pathways

Phenotype and target-based chemical biology investigations in cancers

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National Science Review 2019年第6期6卷 1111-1127页

作者： guo-qiang chen Ying Xu Shao-Ming Shen Jian Zhang Department of Pathophysiology Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of EducationShanghai Jiao Tong University School of Medicine (SJTU-SM)

Chemical biology has been attracting a lot of attention because of the key roles of chemical methods and techniques in helping to decipher and manipulate biological systems. Although chemical biology encompasses a broad field, this review will focus on chemical biology aimed at using exogenous chemical probes to interrogate, modify and manipulate biological processes, at the cellular and organismal levels,in a highly controlled and dynamic manner. In this area, many advances have been achieved for cancer biology and therapeutics, from target identification and validation based on active anticancer compounds(forward approaches) to discoveries of anticancer molecules based on some important targets including protein-protein interaction(reverse approaches). Herein we attempt to summarize some recent progresses mainly from China through applying chemical biology approaches to explore molecular mechanisms of carcinogenesis. Additionally, we also outline several new strategies for chemistry to probe cellular activities such as proximity-dependent labeling methods for identifying protein-protein interactions, genetically encoded sensors, and light activating or repressing gene expression system.

关键词： chemical biology chemical compounds target cancer leukemia

Applications of synthetic biology in medical and pharmaceutical fields

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Signal Transduction and Targeted Therapy 2023年第6期8卷 2638-2670页

作者： Xu Yan Xu Liu Cuihuan Zhao guo-qiang chen School of Life Sciences Tsinghua University100084BeijingChina PhaBuilder Biotech Co.Ltd. Shunyi DistrictZhaoquan Ying101309BeijingChina Center for Synthetic and Systems Biology Tsinghua University100084BeijingChina MOE Key Lab for Industrial Biocatalysis Dept Chemical EngineeringTsinghua University100084BeijingChina

Synthetic biology aims to design or assemble existing bioparts or bio-components for useful *** the past decades,progresses have been made to build delicate biocircuits,standardized biological building blocks and to develop various genomic/metabolic engineering tools and *** and pharmaceutical demands have also pushed the development of synthetic biology,including integration of heterologous pathways into designer cells to efficiently produce medical agents,enhanced yields of natural products in cell growth media to equal or higher than that of the extracts from plants or fungi,constructions of novel genetic circuits for tumor targeting,controllable releases of therapeutic agents in response to specific biomarkers to fight diseases such as diabetes and ***,new strategies are developed to treat complex immune diseases,infectious diseases and metabolic disorders that are hard to cure via traditional *** general,synthetic biology brings new capabilities to medical and pharmaceutical *** review summarizes the timeline of synthetic biology developments,the past and present of synthetic biology for microbial productions of pharmaceutics,engineered cells equipped with synthetic DNA circuits for diagnosis and therapies,live and auto-assemblied biomaterials for medical treatments,cell-free synthetic biology in medical and pharmaceutical fields,and DNA engineering approaches with potentials for biomedical applications.

关键词： diagnosis synthetic equipped

Poly-β-hydroxybutyrate alleviated diarrhea and colitis via Lactobacillus johnsonii biofilm-mediated maturation of sulfomucin

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Science China(Life Sciences) 2023年第7期66卷 1569-1588页

作者： Ning Ma Pingting guo Jingyu chen Zengkai Qi Chunchen Liu Jiakun Shen Yiwei Sun Xiyue chen guo-qiang chen Xi Ma State Key Laboratory of Animal Nutrition College of Animal Science and TechnologyChina Agricultural UniversityBeijing100193China Beijing Laboratory for Food Quality and Safety College of Food Science and Nutritional EngineeringChina Agricultural UniversityBeijing100083China Center for Synthetic and Systems Biology MOE Key Lab for Industrial BiocatalysisSchool of Life Sciences and Dept Chemical EngineeringTsinghua UniversityBeijing100084China

Maintainance of sulfomucin is a key end point in the treatment of diarrhea and inflammatory bowel disease(IBD).However,the mechanisms underlying the microbial sense to sulfomucin are poorly understood,and to date,there are no therapies targeting the secretion and maturation of sulfomucin in ***,we biosynthesized poly-β-hydroxybutyrate(PHB)and found that PHB could alleviate inflammation caused by diarrhea and colitis by enhancing the differentiation of *** transplantation and clearance together demonstrate that PHB promoting sulfomucin is mediated by Lactobacillus johnsonii(***).Further studies revealed that PHB provides a favorable niche for *** biofilm formation to resist disturbance and support its ***-biofilm alleviates colitis by regulating fucose residues to promote goblet cell differentiation and subsequent sulfomucin ***,PHB alleviates colitis by enhancing sulfomucin secretion and maturation in a ***-dependent *** represents a class of guardians,acting as a safe probiotic-biofilm delivery system that significantly promotes probiotic ***,this study adds weight to the possible role of probiotics and functional materials in the treatment of intestinal *** application of PHB and biofilm self-coating *** carries high translational potential and may be of clinical relevance.

关键词： poly-β-hydroxybutyrate diarrhea colitis Lactobacillus johnsonii biofilm goblet cell sulfomucin

Molecular mechanisms of leukemia-associated proteindegradation

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Frontiers of Medicine 2010年第4期4卷 363-370页

作者： Ying-Li WU Hu-chen ZHOU guo-qiang chen Department of Pathophysiology and Chemical Biology Division of Shanghai Universities E-Institutes Key laboratory of Cell Differentiation and Apoptosis of the Ministry of Education of ChinaShanghai JiaoTong University School of MedicineShanghai 200025China The School of Pharmacy and E-Institute of Chemical Biology Shanghai JiaoTong UniversityShanghai 200240China

Chemical biology,using small molecules as probes to study the cellular signaling network,has developed rapidly in recent *** interaction between chemistry and biology not only provides new insight into the understanding of cellular activities,but also generates new lead compounds for the treatment of *** factors and kinases such as retinoic acid receptor-alpha(RARα),acute myeloid leukemia 1(AML1),CAAT/enhancer-binding proteinα(C/EBPα),c-myc,and c-abl play important roles in the differentiation of hematopoietic stem/progenitor *** in these proteins may cause the dysregulation of hematopoi-esis and even the occurrence of ***-mediated protein degradation represents a critical mechan-ism in regulating the cellular levels and functions of these ***,targeting protein degradation has been emerging as an important strategy to conquer malignant *** this review,we will summarize the recent advances in the understanding of the roles of protein degradation in leukemia,with an emphasis on the mechanisms revealed by small molecules.

关键词： protein degradation leukemia chemical biol-ogy transcription factors oncoprotein

Construction of sustainable, colored and multifunctional protein silk fabric using biomass riboflavin sodium phosphate

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Frontiers of Chemical Science and Engineering 2023年第8期17卷 1131-1139页

作者： Wen-Jie Jin Yu Xin Xian-Wei cheng Jin-Ping Guan guo-qiang chen Key Laboratory of Flame Retardancy Finishing of Textile Materials(CNTAC) College of Textile and Clothing EngineeringSoochow UniversitySuzhou 215123China

Riboflavin sodium phosphate has been confirmed as a promising biomass product derived from natural *** this paper,a novel method of dyeing and multifunctional modification of silk fabric by impregnation with riboflavin sodium phosphate was proposed,such that protein silk fabric can be endowed with bright yellow color and *** results of this paper confirmed that the pH and concentration of riboflavin sodium phosphate solution are critical factors for dyeing and multifunctional ***,the photochromic performance was one of the most distinctive features of the modified silk fabric,and the dyed silk fabric turned into fluorescent green from original yellow under 365 nm ultraviolet ***,the modified silk fabric exhibited good antibacterial properties with a high inhibition rate of 92%for Escherichia ***,the flame retardancy of silk fabric was significantly improved after *** damaged length of modified silk fabric with 40%owf riboflavin sodium phosphate was lower than 10.4 cm and passed the B1 *** revealed by the result of this paper,riboflavin sodium phosphate is sufficiently effective in serving as an eco-friendly multifunctional agent for strengthening the add-value of silk textiles.

关键词： biomass riboflavin sodium phosphate silk protein multifunctional modification flame retardant

3-Hydroxybutyrate ameliorates insulin resistance by inhibiting PPARγSer273 phosphorylation in type 2 diabetic mice

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Signal Transduction and Targeted Therapy 2023年第6期8卷 3158-3167页

作者： Yudian Zhang Zihua Li Xinyi Liu Xinyu chen Shujie Zhang Yuemeng chen Jiangnan chen Jin chen Fuqing Wu guo-qiang chen School of Life Sciences Tsinghua UniversityBeijing100084P.R.China Department of Medical Genetics and Cell Biology School of Basic Medical Science of Ningxia Medical UniversityYinchuanNingxia750004P.R.China Center for Synthetic and Systems Biology Tsinghua UniversityBeijing100084China MOE Key Lab of Industrial Biocatalysis Dept of Chemical EngineeringTsinghua UniversityBeijing100084P.R.China

3-Hydroxybutyrate(3HB)is a small ketone body molecule produced endogenously by the body in the *** studies have shown that 3HB can reduce blood glucose level in type 2 diabetic(T2D)***,there is no systematic study and clear mechanism to evaluate and explain the hypoglycemic effect of *** we demonstrate that 3HB reduces fasting blood glucose level,improves glucose tolerance,and ameliorates insulin resistance in type 2 diabetic mice through hydroxycarboxylic acid receptor 2(HCAR2).Mechanistically,3HB increases intracellular calcium ion(Ca^(2+))levels by activating HCAR2,thereby stimulating adenylate cyclase(AC)to increase cyclic adenosine monophosphate(cAMP)concentration,and then activating protein kinase A(PKA).Activated PKA inhibits Raf1 proto-oncogene serine/threonine-protein kinase(Raf1)activity,resulting in a decrease in extracellular signal-regulated kinases 1/2(ERK1/2)activity and ultimately inhibiting peroxisome proliferator-activated receptorγ(PPARγ)Ser273 phosphorylation in *** of PPARγSer273 phosphorylation by 3HB altered the expression of PPARγregulated genes and reduced insulin ***,3HB ameliorates insulin resistance in type 2 diabetic mice through a pathway of HCAR2/Ca^(2+)/cAMP/PKA/Raf1/ERK1/2/PPARγ.

关键词： PPARγ thereby ketone