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A Visualized Fusion Protein Based on Self-assembly hydrophobin HGFI

Chemical Research in Chinese Universities 2015年第5期31卷 781-786页

作者： ZHAO Liqiang LIU Jinyuan SONG Dongmin WANG Xiangxiang TAI Feifei XU Haijin QIAO Mingqiang Key Laboratory of Bioaetive Materials Ministry of Education College of Life Sciences Nankai University Tianjin 300071 P. R. China

hydrophobins are a type be used to modify material surfaces of small amphipathic proteins with a and adsorb enzymes, antibodies and unique self-assembly property, which can even cells. In this study, a fusion protein consisting of hydrophobin HGFI and green fluorescent protein（GFP） was successfully obtained from Pichia patoris （P. pastoris）. Water contact angle（WCA） measurement proves that the wettability of the surfaces of different materials was changed. We further demonstrated the self-assembly ability of HGFI-GFP, which can be used to disperse the multi-walled carbon nanotubes（MWCNTs）. Finally, the adsorption of HGFI-GFP onto the surface of the tissue engineering material poly（ε-caprolactone）（PCL） was evaluated by detecting the fluorescence of the fusion protein itself. The resalt demonstrates that both the basic self-assembly activity of the HGFI domain and the functional activity of the GFP domain were still remained.

关键词： Self-assembly hydrophobin Fusion protein Dispersion Modification

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Engineered hydrophobin for Biomimetic Mineralization of Functional Calcium Carbonate Microparticles

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Journal of Biomaterials and Nanobiotechnology 2014年第1期5卷 1-7页

作者： Hanna Heinonen Paivi Laaksonen Markus B.Linder Hans-Peter Hentze VTT—Technical Research Centre of Finland EspooFinland

In this study, the modified hydrophobin, engineered for biomimetic mineralization, has been employed as a structure-directing agent for mineralization of calcium carbonate. For the first time amphiphilic calcium carbonate particles have been obtained, using engineered proteins. The mineral microparticles have been characterized by optical microscopy, scanning electron microscopy (SEM) and X-ray diffraction (XRD). While mineralization in the presence of non-modified hydrophobin results in polymorph mineral structures, uniform microspheres with an average particle diameter of one micron are obtained by employing hydrophobin which has been modified with an additional ceramophilic protein sequence. Owing to the tri-functionality of the modified hydrophobin (hydrophilic, hydrophobic and ceramophilic), the obtained mineral microparticles exhibit amphiphilic properties. Potential applications are in the areas of functional fillers and pigments, like biomedical and composite materials. Pickering emulsions have been prepared as a demonstration of the emulsion-stabilizing properties of the obtained amphiphilic mineral microspheres. The structure-directing effects of the studied engineered hydrophobins are compared with those of synthetic polymers (i.e. polycarboxylates) used as crystallization and scaling inhibitors in industrial applications.

关键词： Biomimetic Mineralization Engineered Proteins hydrophobin Calcium Carbonate Functional Microparticles

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hydrophobin HFBI promoting active expression of pediocin PA-1 in Pichia pastoris and extending its application in emulsion

Hydrophobin HFBI promoting active expression of pediocin PA-...

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中国生物化学与分子生物学会第十二届全国会员代表大会暨2...

作者： Bo Wang Xiangxiang Wang Lu Ge Xiaotian Yang Haijin Xu Mingqiang Qiao The Key Laboratory of Molecular Microbiology and Technology Ministry of EducationCollege of Life SciencesNankai University

Pediocin PA-1,originating from Pediococcus acidilactici,is regarded as a broad-spectrum bacteriocin in lactic acid bacteria.It was reported that PA-1 could highly expressed in Pichia pastoris but without bioactivity,resulting from covalent binding by "collagen-like" materials presented in the supernatant.In this study,P.pastoris was used as the host for expression of fusion protein PA-1-linker-HFBI(PB),and Ultra-filtration and aqueous two-phase system(ATPs) extraction were employed to purify the recombinant protein PB.Antibacterial activity assays showed the fusion protein PB retained the biological properties of native PA-1,indicated HFBI can overcome the unfunctional expression of PA-1 in P.pastoris.In addition,Water contact angle,oil emulsion antibacterial activity assay indicated PB also retained the biological properties of HFBI.The method of active expression of PA-1 in P.pastoris and the applications of PA-1 in food emulsion and antiseptic explored a novel technology to be used for other bacteriocins with the same feature of pediocin PA-1 molecule.

关键词： PA-1 hydrophobin Fusion protein Antibacterial activity Emulsion

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hydrophobin Protein from Trichoderma harzianum Induced Maize Resistance to Maize Leaf Spot Pathogen Curvularia lunata

Hydrophobin Protein from Trichoderma harzianum Induced Maize...

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中国植物病理学会2016年学术年会

作者： YU Chuan-jin LU Zhi-xiang XIA Hai GAO Jin-xin DOU Kai WANG Meng LI Ya-qian CHEN Jie Key Laboratory of Urban Agriculture(South) Ministry of AgricultureSchool of Agriculture and BiologyShanghai Jiao Tong University State Key Laboratory of Microbial Metabolism Shanghai Jiao Tong University

Corn Curvularia leaf spot disease was a frequently occurring disease to maize leaf,which caused by Curvularia lunata leaf spot pathogen,this disease was very common in corn planting area in China.Therefore,the prevention and treatment of the disease was of great significance.hydrophobins,low molecular mass(about 10 kDa) secreted proteins of fungi,were characterized by moderate to high levels of hydrophobic material(hydrophobic substances) to cover on the surface of the microbial cells and the presence of eight conserved cysteine(Cys) residues and formed 4 disulfide bond.A class Ⅱ hydrophobin gene hyd1 was cloned.A 2kb fragment of the hyd1 promoter sequence upstream hyd1,hyd1 and DsRed were ligated together.The expression cassete P:DsRed and P:hyd1:DsRed were introduced into the KO 16(hyd1 disruption transformant) and showed that hyd1 was localized in the plasma membrane by the lazer confocal fluorescense microscope.B73 seeds that treated with the conidia of different hyd1 transformants and grew in greenhouse,the 4-leaf stage leaves were inoculated C.lunata in vitro and in vivo.The results demonstrated that the maize leaf lesion size that treated by KO 16 was larger than WT(wild type),the disease index of the overexpression treatment(OE3 and OE5) maize was lowest.The callose accumulation was increased in the maize leaves infiltrated with in vitro expressed Hyd1 protein comparied with control.It also indicated that hyd1 influenced Trichoderma colonization in maize roots by the asssy of SEM and qPCR.Meanwhile,Hyd1 protein was detected on the plasma membrane of maize root by the IEM and it was demonstrated that hyd1 acted as MAMPs(Microbial-associated molecular patterns) to induce the maize resistance.The yeast bait plasmid pGBKT7-hyd1 was constructed and screened the maize root cDNA Library and found that hyd1 interacted with ubiquilin1.We verified the two interaction proteins again by BiFC and pulll down method and also identified ubiquilinl and hyd1 interaction motif in yeast cells.At the same time,it was proved t

关键词： Trichoderma harzianum hydrophobin Ubiquilinl MAMPs Protein interaction Induce systemic resistance

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Mutations of Class I hydrophobin HGFI have different adsorption properties as their isoelectric point changed

Mutations of Class I hydrophobin HGFI have different adsorpt...

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中国生物化学与分子生物学会第十二届全国会员代表大会暨2...

作者： Jiuxia Yang Lu Ge Bo Wang Xiaotian Yang Haijin Xu Mingqiang Qiao The Key Laboratory of Molecular Microbiology and Technology Ministry of EducationCollege of Life SciencesNankai University

HGFI is a small surface-active hydrophobin protein found in edible mushroom Grifola frondosa.It can form amphipathic protein films on hydrophobic-hydrophilic material interface.It has a low isoelectric point(pI) because there were no basic amino acid among its protein sequence.Here we constructed two HGFI mutants using the site-point mutation method which were named E24 K and D66 K,respectively.E24 K was generated by replacing three acidic amino acids Glu,Asp and Asp instead of basic amino acid Lys.While D66 K was characterized by altering all the six negatively charged amino acids with six positively charged amino acids Lys.Zeta-potential analysis reveals that the pI of E24 K was between 5 and 6,while D66 K was between 4 and 5.Water contact angle,atomic force microscope and quartz-crystal microbalance analysis were performed in this study.Circular dichroism measurement and Thioflavin T assay showed that both E24 K and D66 K have lost the ability to form rodlets.In vitro drug releases were performed using the indometacin(IND) as the drug model.Results implied that both E24 K and D66 K can modify IND into water-soluble nano-particles(NPs).The hydrophobin-coated NPs were more stable and exhibited lower released speed in pH1.2 and pH7 solution at 37℃ especially the D66 K-IND NPs.These results suggest new applications of the mutated hydrphobins in sustained delivery of oral drug in gastrointestinal tract.

关键词： hydrophobin Mutation Nano-particles Adsorption capability Drug release

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Functional modification of poly(ε-caprolactone) with fusion protein containing HGFI and PA-1 improved antibacterial activity

Functional modification of poly(ε-caprolactone) with fusion...

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中国生物化学与分子生物学会2016年全国学术会议

作者： Xiangxiang Wang Jiwei Mao Dongmin Song Jiuxia Yang Bo Wang Xiuming Zhang Yanling Bai Haijin Xu Mingqiang Qiao The Key Laboratory of Molecular Microbiology and Technology Ministry of Education College of Life SciencesNankai University

Pediocin PA-1 is the most extensively studied class IIa bacteriocin,which shows a broad-spectrum activity and has been well characterized to be applied as a food biopreservative.Herein,an easy-to-do approach is designed to extend the application of pediocin PA-1 in improving antibacterial activity of electrospun PCL grafts through combining PA-1 with HGFI,a kind

关键词： hydrophobin Antibacterial PA-1 Poly（ε-caprolactone） Medical device

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