Biomolecular engineering最新文献

Molecular evolution of Fome lignosus laccase by ethyl methane sulfonate-based random mutagenesis in vitro 甲烷磺酸乙酯随机诱变木质素漆酶的分子进化

Biomolecular engineering Pub Date : 2007-12-01 DOI: 10.1016/j.bioeng.2007.08.020

Mei-Rong Hu , Ya-Peng Chao , Guo-Qing Zhang , Xiu-Qing Yang , Zhi-Quan Xue , Shi-Jun Qian

{"title":"Molecular evolution of Fome lignosus laccase by ethyl methane sulfonate-based random mutagenesis in vitro","authors":"Mei-Rong Hu , Ya-Peng Chao , Guo-Qing Zhang , Xiu-Qing Yang , Zhi-Quan Xue , Shi-Jun Qian","doi":"10.1016/j.bioeng.2007.08.020","DOIUrl":"10.1016/j.bioeng.2007.08.020","url":null,"abstract":"<div><p>In order to improve the laccase activity, mutant libraries are constructed through ethyl methane sulfonate-based (EMS) random mutagenesis. Mutagenesis improved expression 3.7-fold to 144mgl<sup>−1</sup> laccase in yeast, together with a 1.4-fold increase in <em>K</em><sub>cat</sub>. Thus, the total activity is enhanced 5-fold for 2,2′-azino-bis 3-ethylbenzothiaoline-6-sulfonic acid (ABTS). In the presence of 0.6mM copper, the highest activity value reached 30Uml<sup>−1</sup> after a 3-day cultivation at a temperature of 30°C<sub>.</sub> In comparison with the wild type, the best mutant enzymatic properties (<em>K</em><sub>m</sub> for ABTS and guaiacol, thermo- and pH stability, optimal pH) are not changed. Moreover, amino acid sequence analysis indicates that there are four substitutions in the best mutant laccase (Gly160Asp, Ala167Thr, Gly174Asp, and Glu234Gly). The best mutant laccase model showed that the Gly160 and Ala167 are to be found near the water channel; especially the distance of Ala167 to the Cu3a is 14.46Å. This implies that it is likely involved in the formation of water channel and that it helps facilitate the easy incoming and outgoing of water.</p></div>","PeriodicalId":80259,"journal":{"name":"Biomolecular engineering","volume":"24 6","pages":"Pages 619-624"},"PeriodicalIF":0.0,"publicationDate":"2007-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.bioeng.2007.08.020","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27036617","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 33

Development of a heat shock inducible and inheritable RNAi system in silkworm 家蚕热激诱导遗传RNAi系统的建立

Biomolecular engineering Pub Date : 2007-12-01 DOI: 10.1016/j.bioeng.2007.10.004

Hongjiu Dai , Rongjing Jiang , Jue Wang , Guojiang Xu , Meixun Cao , Zhugang Wang , Jian Fei

{"title":"Development of a heat shock inducible and inheritable RNAi system in silkworm","authors":"Hongjiu Dai , Rongjing Jiang , Jue Wang , Guojiang Xu , Meixun Cao , Zhugang Wang , Jian Fei","doi":"10.1016/j.bioeng.2007.10.004","DOIUrl":"10.1016/j.bioeng.2007.10.004","url":null,"abstract":"<div><p>A heat shock inducible and inheritable RNA interference (RNAi) system was developed in the silkworm (<em>Bombyx mori</em>). RNAi transgenic silkworms were generated by injecting silkworm eggs with a piggyBac transposon plasmid carrying RNAi sequence against target gene driven by the <em>Drosophila</em> heat shock protein 70 (HSP70) promoter and the helper plasmid expressing piggyBac transposase. The transgenic EGFP gene and the endogenous eclosion hormone (EH) gene were chosen respectively as the target genes. In the RNAi transgenic silkworms, heat shock at 42°C significantly and specifically reduced the expression of EGFP or EH gene in silkworms according to the corresponding RNAi targeting sequence but not in silkworms with the irrelevant RNAi sequence demonstrating the efficiency and specificity of the RNAi effect. Heat shock in the pupal stage hampered pupal–adult eclosion and reduced egg fertility in EH RNAi transgenic silkworms but not in the wild type or EGFP RNAi transgenic silkworms. The establishment of this heat inducible and inheritable conditional RNA interference system in silkworms provided an approach for the first time to dissect the functions of target genes in silkworms at different stages.</p></div>","PeriodicalId":80259,"journal":{"name":"Biomolecular engineering","volume":"24 6","pages":"Pages 625-630"},"PeriodicalIF":0.0,"publicationDate":"2007-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.bioeng.2007.10.004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27095681","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 21

CIDB: Chlamydia Interactive Database for cross-querying genomics, transcriptomics and proteomics data 衣原体交互数据库，用于交叉查询基因组学、转录组学和蛋白质组学数据

Biomolecular engineering Pub Date : 2007-12-01 DOI: 10.1016/j.bioeng.2007.08.017

Yan Chen , Peter Timms , Yi-Ping Phoebe Chen

引用次数: 7

Effect of different growth factors on human osteoblasts activities: A possible application in bone regeneration for tissue engineering 不同生长因子对人成骨细胞活性的影响:在组织工程骨再生中的可能应用

Biomolecular engineering Pub Date : 2007-12-01 DOI: 10.1016/j.bioeng.2007.08.019

Michela Bosetti , Francesca Boccafoschi , Massimiliano Leigheb , Mario F. Cannas

{"title":"Effect of different growth factors on human osteoblasts activities: A possible application in bone regeneration for tissue engineering","authors":"Michela Bosetti , Francesca Boccafoschi , Massimiliano Leigheb , Mario F. Cannas","doi":"10.1016/j.bioeng.2007.08.019","DOIUrl":"10.1016/j.bioeng.2007.08.019","url":null,"abstract":"<div><p>Cultured human primary osteoblasts reproduce the phenotypic differentiation and maturation of cells <em>in vivo</em>. We have investigated the influence of three isoforms of transforming growth factor beta (TGF-β1, TGF-β2 and TGF-β3), three fibroblast growth factors (FGF-2, FGF-4 and FGF-6) and the active metabolite of Vitamin D [1,25-(OH)<sub>2</sub>D3] on proliferation, alkaline phosphatase activity and mineralization of human osteoblasts during a period of 24 days of culture. TGF-β isoforms and three FGFs examined have been proved to be inducers of osteoblasts proliferation (higher extent for TGF-β and FGF-2) and inhibitors of alkaline phosphatase activity and osteoblasts mineralization. Combination of these growth factors with the active form of Vitamin D induced osteodifferentiation. In fact Vitamin D showed an additive effect on alkaline phosphatase activity and calcium content, induced by FGF-2 and TGF-β in human osteoblast. These results highlight the potential of proliferating cytokines’ combination with mineralizing agents for <em>in vitro</em> bone growth induction in bone tissue engineering.</p></div>","PeriodicalId":80259,"journal":{"name":"Biomolecular engineering","volume":"24 6","pages":"Pages 613-618"},"PeriodicalIF":0.0,"publicationDate":"2007-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.bioeng.2007.08.019","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27047189","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 66

Effect of site-directed PEGylation of trichosanthin on its biological activity, immunogenicity, and pharmacokinetics 位点定向聚乙二醇化天花粉蛋白对其生物活性、免疫原性和药代动力学的影响

Biomolecular engineering Pub Date : 2007-12-01 DOI: 10.1016/j.bioeng.2007.10.002

Qunxing An , Yingfeng Lei , Ning Jia , Xianqing Zhang , Yinlan Bai , Jing Yi , Rui Chen , Aijun Xia , Jing Yang , Sanhua Wei , Xiaodong Cheng , Ailing Fan , Shijie Mu , Zhikai Xu

{"title":"Effect of site-directed PEGylation of trichosanthin on its biological activity, immunogenicity, and pharmacokinetics","authors":"Qunxing An , Yingfeng Lei , Ning Jia , Xianqing Zhang , Yinlan Bai , Jing Yi , Rui Chen , Aijun Xia , Jing Yang , Sanhua Wei , Xiaodong Cheng , Ailing Fan , Shijie Mu , Zhikai Xu","doi":"10.1016/j.bioeng.2007.10.002","DOIUrl":"10.1016/j.bioeng.2007.10.002","url":null,"abstract":"<div><p>Trichosanthin (TCS) is a type I ribosome-inactivating protein (RIP) with multiple biological and pharmacological activities. It has been approved effective in the clinical treatment of AIDS and tumor, but its strong immunogenicity and short plasma half-life have limited the clinical administration. To reduce the immunogenicity and prolong the plasma half-life of this compound, three TCS muteins (M<sub>1</sub>, M<sub>2</sub> and M<sub>3</sub>) and two PEGylated TCS muteins (PM<sub>1</sub> and PM<sub>2</sub>) were constructed by site-directed mutagenesis and PEGylation, respectively. Compared with the unmodified TCS, both PEGylated TCS showed a 3- to 4-fold decrease in immunogenicity, a 0.5- to 0.8-fold decrease in non-specific toxicity, and a 4.5- to 6-fold increase in plasma half-life. But there is a problem of activity reduction. The increased circulating half-life <em>in vivo</em> may compensate for the reduced activity. Together with the other benefits of PEGylation such as reduced immunogenicity and toxicity, it is worthwhile to further explore the potential application of the PEGylated TCS as a better therapeutic agent for AIDS and tumor.</p></div>","PeriodicalId":80259,"journal":{"name":"Biomolecular engineering","volume":"24 6","pages":"Pages 643-649"},"PeriodicalIF":0.0,"publicationDate":"2007-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.bioeng.2007.10.002","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27095680","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 34

Direct haplotyping of bi-allelic SNPs using ARMS and RFLP analysis techniques 使用ARMS和RFLP分析技术对双等位基因SNPs进行直接单倍型分析

Biomolecular engineering Pub Date : 2007-12-01 DOI: 10.1016/j.bioeng.2007.08.018

M. Najafi , M. Firoozrai , H.L. Gohari , A. Zavarehie , G. Basiri

引用次数: 8

Overexpression of GLT1 in fps1ΔgpdΔ mutant for optimum ethanol formation by Saccharomyces cerevisiae 在fps1ΔgpdΔ突变体中过表达GLT1使酿酒酵母产生最佳乙醇

Biomolecular engineering Pub Date : 2007-12-01 DOI: 10.1016/j.bioeng.2007.10.003

Limin Cao, Aili Zhang, Qingxue Kong, Xiaojing Xu, Tchouopou Lontchi Josine, Xun Chen

{"title":"Overexpression of GLT1 in fps1ΔgpdΔ mutant for optimum ethanol formation by Saccharomyces cerevisiae","authors":"Limin Cao, Aili Zhang, Qingxue Kong, Xiaojing Xu, Tchouopou Lontchi Josine, Xun Chen","doi":"10.1016/j.bioeng.2007.10.003","DOIUrl":"10.1016/j.bioeng.2007.10.003","url":null,"abstract":"<div><p>Glycerol is the main byproduct produced under anaerobic ethanol fermentations by <em>Saccharomyces cerevisiae</em> and consumes a considerable amount of substrate. To verify the metabolic phenotype predications for increasing ethanol formation, two engineered <em>S</em>. <em>cerevisiae</em> KAM-14, KAM-15 strains were constructed for possible redirection of glycerol carbon flux into ethanol by overexpression of <em>GLT1</em> in the <em>fps1</em>Δ<em>gpd</em>Δ mutant. The engineered strains KAM-14 and KAM-15 compared to the control strain KAM-2, produced 12.24% and 10.42% higher ethanol, 39.72% and 31.03% lower glycerol yield during anaerobic batch fermentations, respectively. The maximum specific growth rates of KAM-14 and KAM-15 were found to be relatively lower than that of KAM-2 during the exponential growth phase. In the meantime, the biomass concentrations of both KAM-14 and KAM-15 were similar to KAM-2. Acetate and pyruvate concentrations of KAM-14 and KAM-15 were greatly decreased comparing to those of KAM-2, respectively. These experimental results approved the metabolic pathway strategies to improve ethanol formation.</p></div>","PeriodicalId":80259,"journal":{"name":"Biomolecular engineering","volume":"24 6","pages":"Pages 638-642"},"PeriodicalIF":0.0,"publicationDate":"2007-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.bioeng.2007.10.003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27105346","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 21

Real-time study of the effect of different stress factors on lactic acid bacteria by electrochemical optical waveguide lightmode spectroscopy 电化学光波导光模光谱法实时研究不同应激因素对乳酸菌的影响

Biomolecular engineering Pub Date : 2007-12-01 DOI: 10.1016/j.bioeng.2007.10.001

Edina Németh , Nóra Adányi , Anna Halász , Mária Váradi , István Szendrő

{"title":"Real-time study of the effect of different stress factors on lactic acid bacteria by electrochemical optical waveguide lightmode spectroscopy","authors":"Edina Németh , Nóra Adányi , Anna Halász , Mária Váradi , István Szendrő","doi":"10.1016/j.bioeng.2007.10.001","DOIUrl":"10.1016/j.bioeng.2007.10.001","url":null,"abstract":"<div><p>Lactic acid bacteria play an important role in the fermentation of different food products. During the fermentation processes, lactobacilli are confronted with many inhibitor factors. These factors by themselves or in combination can influence the growth of lactic acid bacteria and their acidification capacity. The subject of our study was to monitor with a newly developed biosensing technique how the different chemical stress factors influence the survival of lactic acid bacteria. Electrochemical optical waveguide lightmode spectroscopy combines evanescent-field optical sensing with electrochemical control of surface adsorption processes. For optical sensing, a layer of indium tin oxide served as a high refractive index waveguide and as a conductive electrode, as well.</p><p><em>Lactobacillus plantarum</em> 2142 suspended in Jerusalem artichoke syrup was used in the experiments. Electrochemical optical waveguide lightmode spectroscopy measurements were undertaken by using OW 2400c indium tin oxide coated waveguide sensors (MicroVacuum, Budapest, Hungary) and were performed in a flow-injection analyzer system. The bacterial cells were adsorbed in native form without any chemical binding on the surface of the sensor by ensuring polarizing potential (1V) and were exposed to different concentration of acetic acid/Jerusalem artichoke syrup, lactic acid/Jerusalem artichoke syrup and hydrogen peroxide/Jerusalem artichoke syrup solution for 1h, respectively, and the effect on bacteria cells was monitored. Results were compared to the traditional micro-assay method, and it can be assumed that after further investigations this new technique could be used in real-time application.</p></div>","PeriodicalId":80259,"journal":{"name":"Biomolecular engineering","volume":"24 6","pages":"Pages 631-637"},"PeriodicalIF":0.0,"publicationDate":"2007-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.bioeng.2007.10.001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27095201","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 16

Advances in SELEX and application of aptamers in the central nervous system SELEX及其适配体在中枢神经系统中的应用研究进展

Biomolecular engineering Pub Date : 2007-12-01 DOI: 10.1016/j.bioeng.2007.06.003

Yan Yang , Dongliang Yang , Hermann J. Schluesener , Zhiren Zhang

{"title":"Advances in SELEX and application of aptamers in the central nervous system","authors":"Yan Yang , Dongliang Yang , Hermann J. Schluesener , Zhiren Zhang","doi":"10.1016/j.bioeng.2007.06.003","DOIUrl":"10.1016/j.bioeng.2007.06.003","url":null,"abstract":"<div><p>SELEX (Systematic Evolution of Ligands by Exponential Enrichment) is a screening technique that involves the progressive selection of highly specific ligands by repeated rounds of partition and amplification from a large combinatorial nucleic acid library. The products of the selection are called aptamers, which are short single stranded DNA or RNA molecules, binding with high affinity, attributed to their specific three-dimensional shapes, to a large variety of targets, ranging from small molecules to complex mixtures. Various improvement of the original SELEX method described in 1990 have been obtained recently, such as capillary electrophoresis SELEX, Toggle-SELEX, Tailored-SELEX, Photo-SELEX, and others. These new variants greatly shorten time of selection and improve aptamer affinity and specificity. Such aptamers have great potential as detecting and/or diagnostic reagents. Furthermore, some aptamers specifically inhibit biological functions of targeted proteins, and are considered as potent therapeutic lead structures evaluated in preclinical disease models. Recently, one aptamer has been approved by Food and Drug Administration of US for treating age-related macular degeneration. This review presents recent advances in the field of SELEX with special emphasis on applications of aptamers as analytical, diagnostic and therapeutic tools in the central nervous system.</p></div>","PeriodicalId":80259,"journal":{"name":"Biomolecular engineering","volume":"24 6","pages":"Pages 583-592"},"PeriodicalIF":0.0,"publicationDate":"2007-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.bioeng.2007.06.003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26874127","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 78

Growth factors upregulate deposition and remodeling of ECM by endothelial cells cultured for tissue-engineering applications 生长因子上调内皮细胞在组织工程中的沉积和重塑

Biomolecular engineering Pub Date : 2007-12-01 DOI: 10.1016/j.bioeng.2007.07.010

P. Divya, P.R. Sreerekha, Lissy K. Krishnan

{"title":"Growth factors upregulate deposition and remodeling of ECM by endothelial cells cultured for tissue-engineering applications","authors":"P. Divya, P.R. Sreerekha, Lissy K. Krishnan","doi":"10.1016/j.bioeng.2007.07.010","DOIUrl":"10.1016/j.bioeng.2007.07.010","url":null,"abstract":"<div><p>Appropriate matrix formation, turnover and remodeling in tissue-engineered small diameter vascular conduits are crucial for their long-term function. The interaction between cells and extra-cellular components is indispensable in determining cellular behavior in tissues and on biomaterials. The fibrin that contains fibronectin shows promise in most aspects as a tissue engineering scaffold, whereas, deposition of elastin and collagen by endothelial cells grown in the lumen of the construct is desirable to improve post implant retention, mechanical stability and vaso-responsiveness. So far there is no report on production of extra-cellular matrix (ECM) proteins, elastin and collagen by endothelial cells (EC) in <em>in vitro</em> culture conditions. In this study, we have used a biomimetic approach of providing multiple growth factors (GF) in the fibronectin (FN)-containing fibrin matrix to induce production of elastin and collagen by the endothelial cells for application in vascular tissue engineering. Deposition of elastin and collagens with matrix remodeling is demonstrated through qualitative analysis of the matrices that were recovered after growing cells on the initial fibrin–FN–GF matrix. Expressions of mRNA for both proteins were assessed by real time polymerase chain reaction (RT-PCR) to estimate the effects of multiple growth factor compositions. Marked deposition of elastin and collagen was evidenced by staining the recovered matrix after different culture intervals. Obviously, the biomimetic environment created by adding angiogenic and platelet growth factors in the fibrin–fibronectin–gelatin matrix can induce deposition of collagens and elastin by EC.</p></div>","PeriodicalId":80259,"journal":{"name":"Biomolecular engineering","volume":"24 6","pages":"Pages 593-602"},"PeriodicalIF":0.0,"publicationDate":"2007-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.bioeng.2007.07.010","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40974583","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 25