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A quantitative analytical method for valienone and its application in the evaluation of valienone production by a breakthrough microbial process

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Chinese Journal of Natural Medicines 2017年第10期15卷 794-800页

作者： CUI Li Karin Yanagi SHI Ting LIU Zhang-Min BAI Lin-Quan FENG Yan State Key Laboratory of microbial Metabolism School of Life Science & Biotechnology and Joint International Research Laboratory of Metabolic & Developmental Sciences Shanghai Jiao Tong University Shanghai 200240 China

Valienone is a significant natural carbasugar member of the C7-cyclitol family as a valuable precursor for glycosidase inhibitor *** is an intermediate of validamycin A biosynthesis pathway and exhibits minimal accumulation in the fermentation broth of the natural Streptomyces producer.A quantitative analytical method is crucial for the development of a breakthrough microbial process overcoming the consumption of the natural metabolic *** present study was designed to develop a pre-column derivatization high-performance liquid chromatography method for quantification of valienone and to help establish a straightforward fermentation process for valienone production by metabolically engineered Streptomyces hygroscopicus *** was derivatized by 2,4-dinitrophenylhydrazine(DNPH)in 10 mmol×L^(-1) H_3PO_4 at 37℃ for 45 min and the derivatives were separated on Eclipse XDB-C18(5 μm,4.6 mm×150 mm)column at 30 °C eluted with 50% acetonitrile for 18 *** derivatives were detected by diode array detector at 380 nm and the configurations of the derivatives were determined by computational *** method was shown to be effective,sensitive,and *** linearity was found in the range of 5–2 000 μg×mL^(-1).The intra-and inter-day precisions were 1.1%–2.7% and 1.7%–2.2%,*** absolute recovery of the spiked samples was 97.2%–102.6%.To date,this is the first reversed-phase high-performance liquid chromatography detection method for valienone in microbial culture *** method successfully helped evaluate the valienone production capability of the engineered Streptomyces hygroscopicus 5008 and could be promising for C7-cyclitol profiling of different engineered mutants combined with the metabonomics methods.

关键词： Valienone Carbasugar microbial process Derivatization HPLC

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Antibodies Targeting a Conserved Surface Polysaccharide Are Protective Against a Wide Range of microbial Pathogens Producing β-1-6-Linked Poly-N-Acetylglucosamine(PNAG)

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Engineering 2024年第7期38卷 69-76页

作者： Xi Lu Guoqing Li Jing Pang Xinyi Yang Colette Cywes-Bentley Xuefu You Gerald B.Pier Beijing Key Laboratory of Antimicrobial Agents Institute of Medicinal BiotechnologyChinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100050China Division of Infectious Diseases Department of MedicineBrigham and Women’s HospitalHarvard Medical SchoolBostonMA 02115USA

The b-1-6-linked poly-N-acetylglucosamine(PNAG)polymer is a conserved surface polysaccharide produced by many bacteria,fungi,and protozoan(and even filarial)*** wide-ranging expression makes PNAG an attractive target for vaccine development,as it potentially encompasses a broad range of *** progress has been made in discovering important properties of the biology of PNAG expression in recent *** molecular characterization and regulation of operons for the production of PNAG biosynthetic proteins and enzymes have been studied in many *** addition,the physiological function of PNAG has been further ***-based vaccines and PNAG-targeting antibodies have shown great efficacy in preclinical ***,clinical tests for both vaccines and antibodies have been carried out in humans and economically important animals,and the results are *** it is not destined to be a smooth road,we are optimistic about new vaccines and immunotherapeutics targeting PNAG becoming validated and eventually licensed for clinical use against multiple infectious agents.

关键词： Poly-N-acetylglucosamine Conjugate vaccine Monoclonal antibody

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Association between low molecular polypeptide 7 single nucleotide polymorphism and response to therapy in hepatitis C virus infection

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World Journal of Hepatology 2013年第3期5卷 97-103页

作者： Moataza H Omran Basma E Fotouh Samar S Youssef Noha E Ibrahim Wael Nabil EL-Sayed M Mahdy Wafaa G Shosha Mostafa K El-Awady microbial biotechnology Department Genetic Engineering Division National Research Center Chemistry Department Faculty of Science Helwan University

AIM: To investigate the relationship between low molecular polypeptide-7 (LMP-7) gene polymorphism and response to interferon (IFN) therapy in chronic hepatitis C virus (HCV) patients. METHODS: LMP-7 polymorphism at codon 49 with nucleotide substitution from A to C was amplified in 104 chronic HCV patients of genotype 4. The amplicons were digested with restriction endonuclease Bsm I and the produced restriction fragment length polymorphism was analyzed. Patients received IFN + regional blood volume therapy for 48 wk and the frequency of thissingle nucleotide polymorphism (SNP) was statistically correlated with treatment response. The exclusion criteria for these patients were stated by the national health program for treating viral hepatitis. Main exclusion criteria included co-infection with hepatitis B virus or schistosomiasis, thyroid dysfunction, uncontrolled diabetes mellitus, history of long term drug or alcohol intake and autoimmune hepatitis. Multivariate analyses were done to correlate LMP-7 SNP plus several factors such as age, gender, weight, serum alpha-fetoprotein (AFP) and alanine aminotransferase levels, liver activity, fibrosis score and viral load with response to therapy. RESULTS: The data presented in this study clearly demonstrated statistically significant differences between sustained virological response (SVR) (defined as the absence of HCV RNA levels in the patient's sera at least 6 mo after discontinuation of treatment) and non-response (NR) (where HCV RNA levels in the patient's sera never become undetectable for 6 mo during or after treatment). Variables were described as odds ratio with 95%CI. The data were considered significant if P values were ≤ 0.05; highly significant if P < 0.01 and very highly significant if P < 0.001. Current data showed that 91.7% of patients carrying LMP-7 C/C allele were associated with SVR, while the other two genotypes C/A and A/A were associated with NR patients, 83.3% and 64.3% respectively, showing that g

关键词： Hepatitis C virus Interferon therapy Low molecular mass polypeptide Host gene Single nucleotide polymorphism

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The Nitrogen-Cycling Network of Bacterial Symbionts in the Sponge Spheciospongia vesparium

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Journal of Ocean University of China 2021年第4期20卷 999-1012页

作者： HE Liming KARLEP Liisi LI Zhiyong Marine biotechnology Laboratory State Key Laboratory of Microbial MetabolismSchool of Life Sciences and BiotechnologyShanghai Jiao Tong UniversityShanghai 200240China Joint International Research Laboratory of Metabolic&Developmental Sciences Shanghai Jiao Tong UniversityShanghai 200240China Department of Chemistry and biotechnology Division of Gene TechnologyTallinn University of TechnologyEstonia

The microbes associated with sponges play important roles in the nitrogen cycle of the coral reefs ecosystem,e.g.,nitrification,denitrification,and nitrogen ***,the whole nitrogen-cycling network has remained incomplete in any individual sponge *** this study,454 pyrosequencing of the 16S rRNA genes revealed that the sponge Spheciospongia vesparium from the South China Sea has a unique bacterial community(including 12 bacterial phyla),dominated particularly by the genus Shewanella(order Alteromonadales).A total of 10 functional genes,nifH,amoA,narG,napA,nirK,norB,nosZ,ureC,nrfA,and gltB,were detected in the microbiome of the sponge *** by gene-targeted analysis,revealing an almost complete nitrogen-cycling network in this ***,bacterial urea utilization and the whole denitrification pathway were *** analysis suggests that Proteobacteria(e.g.,Shewanella)and Bacteroidetes(e.g.,Bizionia)are probably involved in the nitrogen cycle in the sponge ***.

关键词： Spheciospongia vesparium bacterial symbionts 454 pyrosequencing functional gene analysis nitrogen-cycling net-work

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Radical SAM-Dependent Demetallation of Heme

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Chinese Journal of Chemistry 2022年第9期40卷 1053-1058页

作者： Jinduo Cheng Wei Ding Qi Zhang Department of Chemistry Fudan UniversityShanghai 200433China State Key Laboratory of microbial Metabolism School of Life Sciences&BiotechnologyShanghai Jiao Tong UniversityShanghai 200240China

Demetallation of heme to release iron is a chemical difficult reaction and is highly rare in biochemistry,with apoferritin as the only known enzyme responsible for this *** we show the heme degradation enzyme Chuw catalyzes heme demetallation besides its known methyltransferase activity(which converts heme to a ring-open product anaerobilin).We show the demetallation activity of Chuw is radical SAM-dependent,and likely involves the same set of intermediates involved in the anaerobilin-producing *** ChuW-catalyzed demetallation reaction does not require external reductant,and can occur on several heme analogs with different metal *** findings establish a brand-new chemistry in the radical SAM enzymes,highlighting the remarkable catalytic diversity of this superfamily of enzymes.

关键词： Biosynthesis Biotransformations Cofactors Enzymes Hemin

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Highly efficient tumor-targeted nanomedicine assembled from affibody-drug conjugate for colorectal cancer therapy

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Nano Research 2023年第4期16卷 5256-5264页

作者： Xiaoyuan Yang Xuelin Xia Wei Huang Xiaoxia Xia Deyue Yan School of Chemistry and Chemical Engineering Shanghai Jiao Tong UniversityShanghai 200240China State Key Laboratory of microbial Metabolism School of Life Sciences and BiotechnologyShanghai Jiao Tong UniversityShanghai 200240China

Affibody is a new class of small non-immunoglobulin affinity proteins that possesses high affinity at the picomole level to several tumor overexpressed *** to the simple framework,affibody is flexible for modification with payload,but the relatively low molecular weight of this construction simultaneously results in short half-life time which hinders its application in cancer *** this work,we prepared a nanomedicine self-assembled from the conjugate of affibody(ZPDGFRβ:09591,PDGFRβ:platelet-derived growth factor receptorβ)with monomethyl auristatin E(MMAE)through cathepsin B cleavable dipeptide linker for targeted colorectal cancer *** nanoscale characteristics of ZPDGFRβ:09591-MMAE affibody-drug conjugate nanomedicine(ZPDGFRβ:09591-M ADCN)resulted in enhanced pharmacokinetics,improved drug accumulation,and promoted biosecurity performance than those of free *** a result,ZPDGFRβ:09591-M ADCN exhibited excellent antitumor efficacy with tumor inhibition rates(TIR)over 99.0%in PDGFRβ-positive tumor models with small solid tumors(~150 mm^(3))or large established tumors(~500 mm^(3)),indicating that ZPDGFRβ:09591-MMAE ADCN is promising for the clinic application in colorectal cancer therapy.

关键词： olecular self-assembly ZPDGFRβ:09591 affibody monomethyl auristatin E(MMAE) nanodrug targeted delivery

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Consecutive Methylation Catalyzed by TsrM,an Atypical Class B Radical SAM Methylase

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Chinese Journal of Chemistry 2022年第14期40卷 1693-1698页

作者： Runze Wu Wei Ding Qi Zhang Department of Chemistry Fudan UniversityShanghai 200433China State Key Laboratory of microbial Metabolism School of Life Sciences&BiotechnologyShanghai Jiao Tong UniversityShanghai200240China

TsrM is a cobalamin-dependent radical S-adenosylmethionine(SAM)methyltransferase belonging to the Class B radical SAM methylase(RSM)*** enzyme catalyzes the C-2 methylation of L-tryptophan to produce 2-methyltrytophan(2-MeTrp),an intermediate involved in the biosynthesis of thiostrepton *** this work,we report characterization of an unexpected activity of TsrM,which carries out an additional methylation reaction on the product 2-MeTrp.A series of isotopic labeling studies and assays with different Trp analogs revealed that TsrM is able to transfer a methyl group from SAM to the C4 of 2-MeTrp to produce 2,*** results reveal the intriguing substrate specificity of TsrM,further expanding the reaction promiscuity of the radical SAM superfamily enzymes.

关键词： Biosynthesis Biotransformations Catalytic promiscuity Enzyme Methyltransferase

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Aryl C–H iodination: are there actual flavin-dependent iodinases in nature?

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Science China Chemistry 2021年第10期64卷 1730-1735页

作者： Yuyang Zhang Lin Chen Hongping Chen Tingting Huang Qing Shi Xiaozheng Wang Yan Wang Man-Cheng Tang Ning-Yi Zhou Shuangjun Lin Synthetic Biology Engineering Lab of Henan Province School of Life Science and BiotechnologyXinxiang Medical UniversityXinxiang 453003China State Key Laboratory of microbial Metabolism Joint International Laboratory on Metabolic&Developmental SciencesSchool of Life Sciences&BiotechnologyShanghai Jiao Tong UniversityShanghai 200240China

Flavin-dependent halogenases (FDHs) are well known to introduce carbon halide bonds (mainly C–Cl and C–Br) into natural products with the assistance of a partner protein flavin reductase to generate reduced flavin (FADH_(2)or FMNH_(2)).Compared with the common chloride-and bromide-containing natural products (approximately 5,000 compounds),iodinated natural products(approximately 100 compounds) are very *** iodinases have also rarely been identified in nature to *** study discovered a novel relationship between iodination and flavin reductases for the first *** mechanistic studies,it was identified that peroxide (H_(2)O_(2)) was released from the uncoupling reaction of flavin reductases and then reacted with iodide ions(I^(-)) to produce hypoiodous acid (IOH) for the final ***,this study also unintentionally verified that the recently reported flavin-dependent iodinase Vir X1 from the marine virus and its two homologs (MBG and NCV) did not catalyze iodination in the in vitro biochemical system but likely belonged to a new phylogenetic clade in the tryptophan halogenase *** a consequence,actual flavin-dependent iodinases in nature remain to be discovered by the scientific community in the future.

关键词： flavin-dependent halogenase flavin reductase iodination H_(2)O_(2) iodinase

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Electrobiocorrosion by microbes without outer-surface cytochromes

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mLife 2024年第1期3卷 110-118页

作者： Dawn E.Holmes Trevor L.Woodard Jessica A.Smith Florin Musat Derek R.Lovley Department of Microbiology University of Massachusetts AmherstAmherstMassachusettsUSA Department of Physical and Biological Science Western New England UniversitySpringfieldMassachusettsUSA Department of Biomolecular Sciences Central Connecticut State UniversityNew BritainConnecticutUSA Department of Biology Section for MicrobiologyAarhus UniversityAarhusDenmark Department of Molecular Biology and biotechnology Facuity of Biology and GeologyBabes-Bolyai UniversityCluj-NapocaRomania

Anaerobic microbial corrosion of iron-containing metals causes extensive economic *** microbes are capable of direct metal-to-microbe electron transfer(electrobiocorrosion),but the prevalence of electrobiocorrosion among diverse methanogens and acetogens is poorly understood because of a lack of tools for their genetic *** studies have suggested that respiration with 316L stainless steel as the electron donor is indicative of electrobiocorrosion,because,unlike pure Fe^(0),316L stainless steel does not abiotically generate H_(2) as an intermediary electron ***,we report that all of the methanogens(Methanosarcina vacuolata,Methanothrix soehngenii,and Methanobacterium strain IM1)and acetogens(Sporomusa ovata and Clostridium ljungdahli)evaluated respired with pure Fe^(0)as the electron donor,but only ***,***,and *** were capable of stainless steel *** electrobiocorrosive methanogens re-quired acetate as an additional energy source in order to produce methane from stainless *** of *** and *** demonstrated how acetogens can provide acetate to methanogens during *** only was Meth-anobacterium strain IM1 not capable of electrobiocorrosion,but it also did not accept electrons from Geobacter metal-lireducens,an effective electron-donating partner for direct interspecies electron transfer to all methanogens that can directly accept electrons from Fe^(0).The finding that ***,***,and *** are capable of electrobiocorrosion,despite a lack of the outer-surface c-type cytochromes previously found to be important in other electrobiocorrosive microbes,demonstrates that there are multiple microbial strategies for making electrical contact with Fe^(0).

关键词： acetogen corrosion Fe^(0) methanogen stainless steel

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Emerging biosensors in detection of natural products

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Synthetic and Systems biotechnology 2020年第4期5卷 293-303页

作者： Firoozeh Piroozmand Fatemeh Mohammadipanah Farnoush Faridbod Pharmaceutical biotechnology Lab Department of Microbial BiotechnologySchool of Biology and Center of Excellence in Phylogeny of Living OrganismsCollege of ScienceUniversity of Tehran14155-6455TehranIran Center of Excellence in Electrochemistry School of ChemistryCollege of ScienceUniversity of TehranTehranIran

Natural products(NPs)are a valuable source in the food,pharmaceutical,agricultural,environmental,and many other industrial *** beneficial properties along with their potential toxicities make the detection,determination or quantification of NPs essential for their *** advanced instrumental methods require time-consuming sample preparation and *** contrast,biosensors allow rapid detection of NPs,especially in complex media,and are the preferred choice of detection when speed and high throughput are ***,we review diverse biosensors reported for the detection of *** emerging approaches for improving the efficiency of biosensors,such as microfluidics,nanotechnology,and magnetic beads,are also *** simultaneous use of two detection techniques is suggested as a robust strategy for precise detection of a specific NP with structural complexity in complicated *** parallel detection of a variety of NPs structures or biological activities in a mixture of extract in a single detection phase is among the anticipated future advancements in this field which can be achieved using multisystem biosensors applying multiple flow cells,sensing elements,and detection mechanisms on miniaturized folded chips.

关键词： Natural products Biosensor Rapid detection Molecular biosensors Nanomaterials

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