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Comparison of Fecal Methanogenic Archaeal Community Between Erhualian and Landrace Pigs Using Denaturing Gradient Gel Electrophoresis and Real-Time PCR Analysis

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Journal of Integrative Agriculture 2014年第6期13卷 1340-1348页

作者： SU Yong Hauke Smidt ZHU Wei-Yun laboratory of Gastrointestinal microbiology College of Animal Science and TechnologyNanjing Agricultural University laboratory of microbiology Agrotechnology and Food Sciences GroupWageningen University

Erhualian and Landrace breeds are typical genetically obese and lean pigs, respectively. To compare the fecal methanogenic Archaeal community between these two pig breeds, fecal samples from different growth phase pigs were collected and used for PCR-denaturing gradient gel electrophoresis （DGGE） with two primer pairs （344fGC/519r and 519f/915rGC） and real-time PCR analysis. Results showed that a better separation and higher quality of bands pattern were obtained in DGGE proifles using primers 344fGC/519r as compared with primers 519f/915rGC. Sequencing of DGGE bands showed that the predominant methanogens in the feces of Erhualian and Landrace pigs belonged to Methanobrevibacter spp. and Methanosphaera spp. Real-time PCR analysis revealed that there was no signiifcant difference in the numbers of fecal total methanogens between Erhualian and Landrace pigs;however, pig growth phase affected the numbers of 16S rRNA genes of total methanogens and Methanobrevibacter smithii. Dissociation curves of methyl coenzyme-M reductase subunit A （mcrA） gene fragments ampliifed with real-time PCR showed all samples possessed a single peak at 82＆#176;C, which might be associated with M. smithii. Samples from the same growth phase of each breed showed good replicative dissociation curves. The results suggest that the growth phase （including diet factor） other than genotype of pig may affect the fecal methanogenic Archaeal community of pigs.

关键词： methanogenic Archaea Erhualian pig Landrace pig Methanobrevibacter smithii

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Purification and biochemical characterization of manganesecontaining superoxide dismutase from deep-sea thermophile Geobacillus sp. EPT3

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Acta Oceanologica Sinica 2014年第12期33卷 163-169页

作者： ZHU Yanbing LI Hebin NI Hui LIU Jingwen XIAO Anfeng CAI Huinong College of food and Biological Engineering Jimei University Fujian Provincial Key laboratory of food microbiology and Enzyme Engineering Research Center of food microbiology and Enzyme Engineering Technology of Fujian Province Xiamen Medical College

Thermostable SOD is a promising enzyme in biotechnological applications. In the present study, ***3 was isolated from a deep-sea hydrothermal field in the East Pacific. A thermo-stable superoxide dismutase （SOD） from this strain was purified to homogeneity by steps of fractional am-monium sulfate precipitation, DEAE-Sepharose chromatography, and Phenyl-Sepharose chromatography. SOD was purified 13.4 fold to homogeneity with a specific activity of 3 354 U/mg and 11.1% recovery. SOD fromGeobacillussp. EPT3 was of the Mn-SOD type, judged by the insensitivity of the enzyme to both KCN and H2O2. SOD was determined to be a homodimer with monomeric molecular mass of 26.0 kDa. It had high thermostability at 50＆#176;C and 60＆#176;C. At tested conditions,SOD was relatively stable in the presence of some inhibitors and denaturants, such asβ-mercaptoethanol （β-ME）, dithiothreitol （DTT）, phenylmethylsulfonyl fluoride （PMSF）, urea, and guanidine ***. EPT3 SOD showed striking stability across a wide pH range from 5.0 to 11.0. It could withstand denaturants of extremely acidic and alkaline conditions, which makes it useful in the industrial applications.

关键词： Geobacillus sp. superoxide dismutase thermostability purification

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Rapid biosorption and reduction removal of Cr(Ⅵ) from aqueous solution by dried seaweeds

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Journal of Central South University 2014年第7期21卷 2801-2809页

作者：贾成光张亚平王鹤欧光南刘启明林锦美 College of Biology Engineering Jimei University Fujian Provincial Key laboratory of food microbiology and Enzyme Engineering

Four types of common seaweeds(Laminaria japonica,Undaria pinnatifida,Porphyra haitanensis,and Gracilaria lemaneiformis) were examined to remove Cr(Ⅵ) ions from aqueous *** experimental parameters that affected the biosorption process including pH,biomass dosage,contact time and temperature were investigated via batch *** surface characteristics of seaweeds before and after Cr(Ⅵ) adsorption were studied with scanning electron microscopy and Fourier transform infrared *** results show that an initial solution with the pH of 1.0 is most favorable for Cr(Ⅵ) *** adsorption is observed in the initial stage and adsorption equilibrium state is reached within 1 *** adsorption efficiency by Porphyra haitanensis is the maximum among four types of seaweed powders,followed by Laminaria japonica and Undaria pinnatifida with biosorption efficiency up to 90%.The removal rate of Gracilaria lemaneiformis is less than 60%.The kinetic data obtained using the seaweeds are found to follow pseudo-second order kinetic *** sorption data adequately correlate with the Langmuir *** indicates that amino and carboxyl groups play an important role in the process of Cr(Ⅵ) adsorption and a large percentage of Cr(Ⅵ) ions are reduced by reductive groups on the surface of seaweeds.

关键词： biosorption hexavalent chromium seaweed reduction

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Transformation of coccolithophorid Emiliania huxleyi harboring a marine virus(Coccolithoviruses)serine palmitoyltransferase(SPT)gene by electroporation

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Journal of Oceanology and Limnology 2021年第2期39卷 693-704页

作者： Weicong CAI Xueting WANG Jinjing SU Jian LI Jun ZENG Guiling LI Jingwen LIU College of food and Bioengineering Jimei UniversityXiamen 361021China Fujian Provincial Key laboratory of food microbiology and Enzyme Engineering Xiamen 361021China

Emiliania huxleyi is the most prominent modern coccolithophore,a group of marine unicellular eukaryotes that play a critical role in ocean *** are large double stranded DNA viruses,which is responsible for the demise of large oceanic blooms formed by *** virus(EhVs)acquired a series of enzyme-coding genes predicted to be involved in the sphingolipid biosynthesis by horizontal gene transfer between ***,there is limited experimental validation identifying the functions of these genes in *** transformation of eukaryotic cells is a powerful tool to get an insight into gene functions of the studied *** palmitoyltransferase(SPT)catalyzes the first committed step in de novo sphingolipid biosynthetic ***,a novel vector system for the transformation of *** was *** contained fragments of promoter and terminator sequences of *** endogenic fucoxanthin chlorophyll a/c-binding protein gene“fcp”and harbored EhV-99B1 spt *** resultant recombinant transformation vectors pEhux-I-spt and pEhux-II were co-transferred into *** BOF92 by *** were obtained upon glufosinate-ammonium selection,and confirmed by Southern hybridization,genome PCR,qRT-PCR and Western blot screening of spt gene,which indicated that spt gene was integrated into the nuclear genome and was expressed at the mRNA and protein *** expression of the viral spt gene led to differences in lipid compositions analyzed using thin-layer chromatography(TLC).The results present the genetic transformation system for ***,providing additional genetic resource with potential for exploring basic biological questions such as the virus-host interactions.

关键词： E miliania huxleyi coccolithovirus genetic transformation serine palmitoyltransferase(SPT) total lipid

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Quantum Dot Nanobeads-Labelled Lateral Flow Immunoassay Strip for Rapid and Sensitive Detection of Salmonella Typhimurium Based on Strand Displacement Loop-Mediated Isothermal Amplification

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Engineering 2022年第12期19卷 62-70页

作者： Yuting Shang Shuzhen Cai Qinghua Ye Qingping Wu Yanna Shao Xiaoying Qu Xinran Xiang Baoqing Zhou Yu Ding Moutong Chen Liang Xue Honghui Zhu Jumei Zhang Guangdong Provincial Key laboratory of Microbial Safety and Health&State Key laboratory of Applied microbiology Southern China&Institute of microbiology Guangdong Academy of SciencesGuangzhou 510070China State Key laboratory of food Science and Technology&National Engineering Research Center for Functional foods&Synergetic Innovation Center of food Safety&Joint International Research laboratory on food Safety School of Food Science and TechnologyJiangnan UniversityWuxi 214122China

Rapid,sensitive,point-of-care detection of pathogenic bacteria is important for food *** this study,we developed a novel quantum dot nanobeads-labelled lateral flow immunoassay strip(QBs-labelled LFIAS)combined with strand displacement loop-mediated isothermal amplification(SD-LAMP)for quantitative Salmonella Typhimurium(ST)*** dot nanobeads(QBs)served as fluorescence reporters,providing good detection *** customizable strand displacement(SD)probe was used in LAMP to improve the specificity of the method and prevent by-product *** was based on a sandwich immunoassay.A fluorescence strip reader measured the fluorescence intensity(FI)of the test(T)line and control(C)*** linear detection range of the strip was 10^(2)–10^(8) colony forming units(CFU)·mL^(-1).The visual limit of detection was 10^(3) CFU·mL^(-1),indicating that the system was ten-fold more sensitive than AuNPs-labelled test *** specificity was analyzed in accordance with agarose gel outputs of polymerase chain reaction(PCR)and *** detected ST in foods with an acceptable recovery of 85%–110%.The method is rapid,simple,almost equipment-free,and suitable for bacterial detection in foods and for clinical diagnosis.

关键词： Salmonella Typhimurium Quantum dot nanobeads Lateral flow immunoassay strip Loop-mediated isothermal amplification Strand displacement probe

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Dynamics design of a non-natural transcription factor responding to androst-4-ene-3,17-dione

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Synthetic and Systems Biotechnology 2024年第3期9卷 436-444页

作者： Ming Zhao Mengkai Hu Rumeng Han Chao Ye Xiangfei Li Tianwen Wang Yan Liu Zhenglian Xue Kun Liu Anhui Engineering laboratory for Industrial microbiology Molecular Breeding College of Biology and Food EngineeringAnhui Polytechnic UniversityWuhu 241000China

The production of androst-4-ene-3,17-dione(AD)by the steroidal microbial cell factory requires transcription factors(TFs)to participate in metabolic ***,microbial cell factory lacks effective TFs that can respond to AD in its metabolic ***,finding and obtaining natural TFs that specifically respond to AD is a complex and onerous *** this study,we devised an artificial TF that responds to AD,termed AdT,based on structure-guided molecular dynamics(MD)*** to MD analysis of the conformational changes of AdT after binding to AD,an LBD in which the N-and C-termini exhibited convergence tendencies was used as a microswitch to guide the assembly of a DNA-binding domain lexA,a linker(GGGGS)2,and a transcription activation domain B42 into an artificial *** a proof of design,a AD biosensor was designed and constructed in yeast on the basis of the ligand-binding domain(LBD)of hormone *** addition,the transcription factor activity of AdT was increased by 1.44-fold for its variant ***,we created non-natural TF elements for AD microbial cell factory,and expected that the design TF strategy will be applied to running in parallel to the signaling machinery of the host cell.

关键词： Protein design Artificial transcription factor Molecular dynamics Steriods Site-mutations

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Molecular basis of cross-interactions between Aβ and Tau protofibrils probed by molecular simulations

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Chinese Journal of Chemical Engineering 2023年第3期55卷 173-180页

作者： Fufeng Liu Luying Jiang Jingcheng Sang Fuping Lu Li Li State Key laboratory of food Nutrition and Safety Key Laboratory of Industrial Fermentation Microbiology of Ministry of EducationTianjin Key Laboratory of Industrial MicrobiologyCollege of BiotechnologyTianjin University of Science&TechnologyTianjin 300457China College of Marine and Environmental Science Tianjin University of Science&TechnologyTianjin 300457China

Amyloid β-protein(Aβ) and Tau, two common pathogenic proteins associated with Alzheimer’s disease(AD), cross-interact, and thus co-assemble into hybrid aggregates. However, molecular mechanism of the cross-interactions remains unclear. To explore the issue, docking and molecular dynamics(MD) simulations were coupled to study the cross-interactions between Aβ pentamer and Tau pentamer. Four stable hybrid decamer conformations including double layer, single layer, block, and part-in were obtained by protein-protein docking software HADDOCK 2.2. Then, MD simulations were used to explore the molecular mechanism of cross-interactions between Aβ pentamer and Tau pentamer. The results of MD simulations showed that the part-in structure was the most stable among all the above four representative ones. The binding energy between Aβ and Tau was about-759.77 kJ·mol-1in the part-in structure. Moreover, the part-in conformation would undergo conformational transition, which would improve its hydrophobicity and make the structure more compact. This work offers a structural understanding of cross-interactions between Aβ and Tau linked to AD.

关键词： Alzheimer’s disease Amyloidβ-protein Tau Cross-interactions Protein-protein docking Molecular dynamics simulation

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Cross-linkedα-amylase aggregates on Fe3O4 magnetic nanoparticles modified with polydopamine/polyethyleneimine for efficient hydrolysis of starch

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Particuology 2024年第7期90卷 98-105页

作者： Jiandong Cui Xiuming Tang Qingqing Ma Yuyan Chang Qunli Zhang Shiru Jia State Key laboratory of food Nutrition and Safety Laboratory of Industrial Fermentation MicrobiologyMinistry of EducationTianjin University of Science and TechnologyTianjin 300457China Key laboratory of Biomass Chemical Engineering of Ministry of Education Zhejiang UniversityHangzhou 310027China

In this workα-amylase was immobilized on magnetic Fe3O4 nanoparticles with polyethylenimine(PEI)/polydopamine(PDA)coating or 3-aminopropyl triethoxysilane(APTES)for the first time via adsorption–precipitation–*** with the freeα-amylase,the resultant magnetic cross-linkedα-amylase aggregates(PEI/PDA-M-CLEAs and N-M-CLEAs)exhibited excellent thermal and storage stability as well as pH *** storage at 25°C for 60 days,freeα-amylase only retained 60%of its initial activity,while PEI/PDA-M-CLEAs and N-M-CLEAs retained 80%and 78%of their initial activities,***,N-M-CLEAs and PEI/PDA-M-CLEAs showed good *** 6 repeated uses,PEI/PDA-M-CLEAs and N-M-CLEAs still maintained 65%and 62%of their initial activities,***,PEI/PDA-M-CLEAs and N-M-CLEAs exhibited higher starch hydrolysis efficiency than freeα-*** maximum dextrose equivalent(DE)values of starch hydrolysis by PEI/PDA-M-CLEAs and N-M-CLEAs reached 29.24%and 28.79%within 90 min,***,the maximum DE values of starch hydrolysis by the freeα-amylase was only 27.89%even in 150 *** magnetic cross-linkedα-amylase aggregates could be introduced as effective biocatalyst for industrial applications in production of maltose syrups.

关键词： α-amylase Magnetic nanoparticles Functional modification Magnetic cross-linked enzyme aggregates Hydrolysis of starch

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An Effective Platform for Exploring Rotavirus Receptors by Bacterial Surface Display System

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Virologica Sinica 2020年第1期35卷 103-109页

作者： Danlei Liu Haoran Geng Zilei Zhang Yifan Xing Danlu Yang Zhicheng Liu Dapeng Wang Department of food Science and Technology School of Agriculture and BiologyShanghai Jiao Tong UniversityShanghai 200240China State Key laboratory of Applied microbiology Southern China Guangdong Provincial Key Laboratory of Microbial Culture Collection and ApplicationGuangdong Open Laboratory of Applied MicrobiologyGuangdong Institute of MicrobiologyGuangzhou 510070China Shanghai food Safety and Engineering Technology Research Center School of Agriculture and BiologyShanghai Jiao Tong UniversityShanghai 200240China

Rotavirus(RV) is a major foodborne pathogen. For RV prevention and control, it is a key to uncover the interaction mechanism between virus and its receptors. However, it is hard to specially purify the viral receptors, including histo-blood group antigens(HBGAs). Previously, the protruding domain protein(P protein) of human norovirus(genotype Ⅱ.4) was displayed on the surface of Escherichia coli, and it specifically recognized and captured the viral ligands. In order to further verify the feasibility of the system, P protein was replaced by VP8* of RV(G9 P[8]) in this study. In the system, VP8*could be correctly released by thrombin treatment with antigenicity retaining, which was confirmed by Western blot and Enzyme-Linked Immunosorbent Assays. Type A HBGAs from porcine gastric mucin(PGM) were recognized and captured by this system. From saliva mixture, the captured viral receptor bound with displayed VP8* was confirmed positive with monoclonal antibody against type A HBGAs. It indicated that the target ligands could be easily separated from the complex matrix. These results demonstrate that the bacterial surface display system will be an effective platform to explore viral receptors/ligands from cell lines or food matrix.

关键词： Rotavirus Bacterial surface display system Receptors Histo-blood group antigens(HBGAs) Saliva

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Overexpression and characterization of a thermostable β-agarase producing neoagarotetraose from a marine isolate Microbulbifer ***1

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Acta Oceanologica Sinica 2019年第2期38卷 96-106页

作者： Yanbing Zhu He Gao Hebin Li Hui Ni Zedong Jiang Lijun Li Anfeng Xiao College of food and Biological Engineering Jimei University Fujian Provincial Key laboratory of food microbiology and Enzyme Engineering Research Center of food Biotechnology of Xiamen City Key laboratory of Systemic Utilization and In-depth Processing of Economic Seaweed Xiamen Southern Ocean Technology Center of China Department of Pharmacy Xiamen Medical College

An agarase gene containing 1 302 bp was cloned from Microbulbifer sp. AG1. It encoded a mature protein of 413 amino acids plus a 20-residue signal peptide. The recombinant enzyme without the signal peptide was expressed and purified from Escherichia coli BL21(DE3). When agarose was used as a substrate, the optimal temperature and pH for the enzyme were 60℃ and 7.5, respectively. The recombinant agarase showed excellent thermostability with 67% and 19% of residual activities after incubation at 50℃ and 60℃ for 1 h, *** SDS, the recombinant agarase had a relatively good resistance against the detected inhibitors, detergents and urea denaturant. Thin layer chromatography analysis and enzyme assay using p-nitrophenyl-α/β-Dgalactopyranoside revealed that the recombinant agarase was a β-agarase that degraded agarose into neoagarotetraose as the main end product. The enzymatic hydrolysis products with different degree of polymerization exhibited the antioxidant activities.

关键词： thermostable β-agarase neoagarotetraose Microbulbifer sp.

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