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| Registros recuperados: 14 | |
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| Ping,Ruo; Sin Teoh,Leong; Kwai Chan,Mooi; Miswan,Noorizan; Yin Khoo,Boon. |
| Background: Human is an essential cellular enzyme that is found in all human cells. As this enzyme is upregulated in cancer cells exceedingly, it is used as a target for cancer chemotherapeutic drug development. As such, producing the in-house enzyme for the purpose to speed up the search for more cost-effective and target specific hTopoI inhibitors is warranted. This study aims to compare the optimised conditions for the expression of hTopoI in KM71H (MutS) and X33 (Mut+) strains of Pichia pastoris. P. pastoris transfected with an hTopoI recombinant vector was used for the optimization of a higher level of hTopoI expression. Results: In the process, fed-batch cultivation parameters that influence the expression of hTopoI, such as culture temperature,... |
| Tipo: Journal article |
Palavras-chave: Cell density; Pichia pastoris; Total protein concentration; Recombinant protein expression. |
| Ano: 2016 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000300001 |
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| Zhou,Min; Huang,Zhiqing; Zhou,Bo; Luo,Yanliu; Jia,Gang; Liu,Guangmang; Zhao,Hua; Chen,Xiaoling. |
| Background Aspergillus niger has the ability to secrete feruloyl esterase. However, for economically viable industrial applications, it is necessary to increase their catalytic activities and/or protein yields to satisfy the increasing needs for feruloyl esterases. Results The gene AnFaeA that encodes a type A feruloyl esterase was successfully expressed in Pichia pastoris by a two-copy engineered yeast. After a screen in shaker flask, a one-copy strain GSKFA3 having the highest feruloyl esterase activity of 2.4 U/mL was obtained. Then, the pPICZaA-AnFaeA plasmid was transformed into GSKFA3 and the transformants were grown on YPDS plates with antibiotic Zeocin. After cultivation, a two-copy strain GSKZaFA20 with the highest feruloyl esterase activity of... |
| Tipo: Journal article |
Palavras-chave: Aspergillus niger; Feruloyl esterase; Pichia pastoris. |
| Ano: 2015 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000500002 |
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| Arias,Cesar Andres Diaz; Marques,Daniela de Araujo Viana; Malpiedi,Luciana Pellegrini; Maranhão,Andrea Queiroz; Parra,Dulcineia Abdalla Saes; Converti,Attilio; Pessoa Junior,Adalberto. |
| Abstract Antibodies and antibody fragments are nowadays among the most important biotechnological products, and Pichia pastoris is one of the most important vectors to produce them as well as other recombinant proteins. The conditions to effectively cultivate a P. pastoris strain previously genetically modified to produce the single-chain variable fragment anti low density lipoprotein (-) under the control of the alcohol oxidase promoter have been investigated in this study. In particular, it was evaluated if, and eventually how, the carbon source (glucose or glycerol) used in the preculture preceding cryopreservation in 20% glycerol influences both cell and antibody fragment productions either in flasks or in bioreactor. Although in flasks the volumetric... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Pichia pastoris; ScFv antibody fragment; Cryopreservation; Expression; Carbon source. |
| Ano: 2017 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822017000300419 |
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| Teh,Ser Huy; Fong,Mun Yik; Mohamed,Zulqarnain. |
| The Pichia pastoris expression system was used to produce recombinant human erythropoietin, a protein synthesized by the adult kidney and responsible for the regulation of red blood cell production. The entire recombinant human erythropoietin (rhEPO) gene was constructed using the Splicing by Overlap Extension by PCR (SOE-PCR) technique, cloned and expressed through the secretory pathway of the Pichia expression system. Recombinant erythropoietin was successfully expressed in P. pastoris. The estimated molecular mass of the expressed protein ranged from 32 kDa to 75 kDa, with the variation in size being attributed to the presence of rhEPO glycosylation analogs. A crude functional analysis of the soluble proteins showed that all of the forms were active in... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Erythropoietin; Glycosylation; Pichia pastoris; SOE-PCR. |
| Ano: 2011 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000300017 |
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| Craveiro,R.B.; Ramalho,J.D.; Chagas,J.R.; Wang,P.H.M.; Casarini,D.E.; Pesquero,J.L.; Araújo,R.C.; Pesquero,J.B.. |
| Carboxypeptidase M (CPM) is an extracellular glycosylphosphatidyl-inositol-anchored membrane glycoprotein, which removes the C-terminal basic residues, lysine and arginine, from peptides and proteins at neutral pH. CPM plays an important role in the control of peptide hormones and growth factor activity on the cell surface. The present study was carried out to clone and express human CPM in the yeast Pichia pastoris in order to evaluate the importance of this enzyme in physiological and pathological processes. The cDNA for the enzyme was amplified from total placental RNA by RT-PCR and cloned in the vector pPIC9, which uses the methanol oxidase promoter and drives the expression of high levels of heterologous proteins in P. pastoris. The cpm gene, after... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Kinins; Human carboxypeptidase M; Recombinant protein; Pichia pastoris. |
| Ano: 2006 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2006000200007 |
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| An,Hongde; Xiao,Tingting; Fan,Huan; Wei,Dongsheng. |
| Background Laccase has been considered important for the degradation of lignocellulose by wood rot fungi. The properties and functions of laccase in white rot fungi have been investigated extensively, but those from brown rot fungi remain largely unknown. In this paper, a laccase isoform Pplcc2 from the brown rot fungus Postia placenta MAD-698-R was expressed heterologously in Pichia pastoris GS115, purified and the properties of the enzyme were determined. Results The molecular weight of the protein was determined to be 67 kDa using SDS-PAGE. It cannot oxidize syringaldazine (SGZ), but it can oxidize 2,2'-azino-di-(3-ethylbenzothialozin-6-Sulfonic acid) (ABTS) and 2,6-dimethoxyphenol (DMP). Specific activity for ABTS was 1960 ± 19 Unit/mg. The catalytic... |
| Tipo: Journal article |
Palavras-chave: Brown rot fungi; Pichia pastoris; Postia placenta; Thermostable laccase. |
| Ano: 2015 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000600011 |
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| Sánchez-Castrejón,Edna; Ponce-Rivas,Elizabeth; Aguilar,Manuel B; Díaz,Fernando. |
| Crustacean hyperglycemic hormone (CHH) is the most abundant and best studied member of the CHH/MIH/GIH neuropeptide hormone family. CHH plays a major role in controlling glucose levels in the hemolymph, and it also has significance in regulating molting, reproduction, and osmoregulation. In contrast, molt-inhibiting hormone (MIH) is responsible for maintaining animals in an intermolt stage. In this study, Liv-MIH-1 cDNA, which encodes a mature neuropeptide from the eyestalk of white shrimp, Litopenaeus vannamei, was expressed in methylotrophic yeast (Pichia pastoris KM71) under the control of an alcohol oxidase promoter. Recombinant Liv-MIH-1 was secreted into the culture medium using the α-factor prepro-sequence without Glu-Ala repeats. The... |
| Tipo: Journal article |
Palavras-chave: CHH; Litopenaeus vannamei; Pichia pastoris; Recombinant protein. |
| Ano: 2008 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000400009 |
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| Pinto, Vilson Borba; Costenaro-Ferreira, Cristiano; Oliveira, Paulo Leonardo Silva; Oliveira, Rodrigo Ribeiro Bezerra de; Piedras, Sérgio Renato Noguez; Pouey, Juvêncio Luis Osório Fernandes. |
| Since probiotics have proved to be a viable alternative to antibiotics as enhancers of animal growth, the performance, uniformity and mortality rates of the jundiá (Rhamdia quelen) larvae fed on diets with different probiotics were evaluated. Jundiá larvae, aged four days post hatching, were fed during 21 days with the following diets, in four replicates, namely, CO: control feed, without probiotics; PP: feed with Pichia pastoris; SB: feed with Saccharomyces boulardii; BT: feed with Bacillus cereus var. toyoi. Among the tested probiotic, Bacillus cereus var. toyoi had the best results due to the fact that the larvae were 25% heavier than CO at the end of the first week; the difference increased to 28% by the end of the trial. Further, BT also improved... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: 5.06.03.04-3 fish; Uniformity; Bacillus cereus var. toyoi; Saccharomyces boulardii; Pichia pastoris. |
| Ano: 2015 |
URL: http://periodicos.uem.br/ojs/index.php/ActaSciAnimSci/article/view/27106 |
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| Coimbra,E.C.; Gomes,F.B.; Campos,J.F.; D’arc,M.; Carvalho,J.C.; Mariz,F.C.; Jesus,A.L.S.; Stocco,R.C.; Beçak,W.; Freitas,A.C.. |
| Human papillomavirus (HPV) infection is the most common sexually transmitted disease in the world and is related to the etiology of cervical cancer. The most common high-risk HPV types are 16 and 18; however, the second most prevalent type in the Midwestern region of Brazil is HPV-33. New vaccine strategies against HPV have shown that virus-like particles (VLP) of the major capsid protein (L1) induce efficient production of antibodies, which confer protection against the same viral type. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system for the production of high levels of heterologous proteins stably using a wild-type gene in combination with an integrative vector. It was recently demonstrated that P. pastoris can... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Human papillomavirus; Cervical cancer; Pichia pastoris; L1. |
| Ano: 2011 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2011001200003 |
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| Monte,Leonardo Garcia; Leal,Fernanda Munhoz Dos Anjos; Hartwig,Daiane Drawanz; Vasconcellos,Sílvio Arruda; Brihuega,Bibiana; Dellagostin,Odir Antonio; Hartleben,Cláudia Pinho. |
| The production of recombinant LipL32 protein using Escherichia coli has been used extensively for the development of vaccines and diagnostic tests for leptospirosis. However, E. coli has demonstrated limitations, including low yield and lack of post-translational modifications. In this study, rLipL32 was produced in eukaryotic expression system (Pichia pastoris) and evaluated the antigen by enzyme-linked immunosorbent assay (ELISA). The yield obtained from the culture supernatant reached 270 mg/L and ELISA showed an accuracy of 95.34%. In summary, the production of rLipL32 using P. pastoris did not impair the antigenic characteristics of this antigen and ensured its use for detecting the leptospiral antibodies in swine sera. |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Pichia pastoris; LipL32; ELISA; Leptospirosis. |
| Ano: 2014 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132014000300007 |
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| Yang,Gang; Chang,Juan; Yin,Qingqiang; Wang,Erzhu; Zhu,Qun; Wang,Ping; Dang,Xiaowei; Lu,Fushan. |
| Four kinds of neutral and alkaline protease genes from Aspergillus oryzae and Bacillus subtilis were isolated and shuffled. The shuffled genes were selected, inserted into pGAPZαA plasmid and transformed into Escherichia coli. The gene which could express high-activity protease was selected by screening the sizes of transparent zones around the colonies on casein plates. After an ideal protease gene was selected, it was sequenced and then transformed into Pichia pastoris X33. The result showed that the base in 1022th position of shuffled protease gene was changed from thymine to cytosine, inferring that cysteine was changed to arginine in the mutant protease. After 48 h incubation for the transformed P. pastoris with the mutant or native protease genes,... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Protease gene; DNA shuffling; Pichia pastoris; Gene expression; Protease characterization. |
| Ano: 2015 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132015000300337 |
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| Registros recuperados: 14 | |
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