Open Access. Powered by Scholars. Published by Universities.®
- Keyword
-
- Chlamydomonas reinhardtii (2)
- Anaplerosis. (1)
- C/N-assimilation (1)
- CO2-concentrating mechanism (1)
- Green microalgae (1)
-
- Low-CO2 responsive gene (1)
- PEP carboxylase (PEPC [Ppc]) (1)
- Pgp1-1 mutation (1)
- Pichia pastoris; Ion-exchange chromatography; Hydrophobic charge induction chromatography; Homogenized; Filtration; (1)
- Pichia pastoris; protein secretion; protein disulfide isomerase; Necator americanus secretory protein; Na-ASP1; protein overexpression; secretion bottleneck (1)
Articles 1 - 5 of 5
Full-Text Articles in Engineering
Enhancement Of Protein Secretion In Pichia Pastoris By Overexpression Of Protein Disulfide Isomerase, Mehmet Inan, Dinesh Aryasomayajula, Jayanta Sinha, Michael M. Meagher
Enhancement Of Protein Secretion In Pichia Pastoris By Overexpression Of Protein Disulfide Isomerase, Mehmet Inan, Dinesh Aryasomayajula, Jayanta Sinha, Michael M. Meagher
Papers in Biochemical Engineering
A potential vaccine candidate, Necator americanus secretory protein (Na-ASP1), against hookworm infections, has been expressed in Pichia pastoris. Na- ASP1, a 45 kDa protein containing 20 cysteines, was directed outside the cell by fusing the protein to the preprosequence of the a-mating factor of Saccharomyces cerevisiae. Most of the protein produced by single copy clones was secreted outside the cell. However, increasing gene copy number of Na-ASP1 protein in P. pastoris saturated secretory capacity and therefore, decreased the amount of secreted protein in clones harboring multiple copies of Na-ASP1 gene. Overexpression of the endoplasmic reticulum (ER) resident, homologous chaperone protein, …
Purification And Scale-Up Of A Recombinant Heavy Chain Fragment C Of Botulinum Neurotoxin Serotype E In Pichia Pastoris Gs115, Michael P. Dux, Rick Barent, Jayanta Sinha, Mark Gouthro, Todd Swanson, Ardis Barthuli, Mehmet Inan, John T. Ross, Leonard A. Smith, Theresa J. Smith, Robert Webb, Bonnie Loveless, Ian Henderson, Michael M. Meagher
Purification And Scale-Up Of A Recombinant Heavy Chain Fragment C Of Botulinum Neurotoxin Serotype E In Pichia Pastoris Gs115, Michael P. Dux, Rick Barent, Jayanta Sinha, Mark Gouthro, Todd Swanson, Ardis Barthuli, Mehmet Inan, John T. Ross, Leonard A. Smith, Theresa J. Smith, Robert Webb, Bonnie Loveless, Ian Henderson, Michael M. Meagher
Papers in Biochemical Engineering
A recombinant C-terminus heavy chain fragment from botulinum neurotoxin serotype E (BoNT/E) is proposed as a vaccine against the serotype E neurotoxin. This fragment, rBoNTE(Hc), was produced intracellular in Pichia pastoris GS115 by a three-step fermentation process, i.e., glycerol batch phase and a glycerol fed-batch phase to achieve high cell densities, followed by a methanol fed-batch induction phase. The rBoNTE(Hc) protein was purified from the soluble fraction of cell lysates using three ion-exchange chromatography steps (SP Sepharose Fast Flow, Q Sepharose Fast Flow, Sp Sepharose High Performance) and polished with a hydrophobic charge induction chromatography step (MEP HyperCel). Method development …
The Phosphoglycolate Phosphatase Gene And The Mutation In The Phosphoglycolate Phosphatase-Deficient Mutant (Pgp1-1) Of Chlamydomonas Reinhardtii, Kensaku Suzuki, Hidenobu Uchida, Tarlan G. Mamedov
The Phosphoglycolate Phosphatase Gene And The Mutation In The Phosphoglycolate Phosphatase-Deficient Mutant (Pgp1-1) Of Chlamydomonas Reinhardtii, Kensaku Suzuki, Hidenobu Uchida, Tarlan G. Mamedov
Papers in Biochemical Engineering
The sequences of the phosphoglycolate phosphatase (PGPase) gene Pgp1 and the 5′-upstream region from Chlamydomonas reinhardtii wildtype 2137 and the pgp1-1 mutant N142 that lacks the activity of PGPase (PGP1) were determined. The comparison revealed the alteration of a G to A at position 98 relative to the start codon. This destroyed the “GT” splice donor site at the beginning of the first intron of this gene, resulting in an extension of the first exon to 49 translatable codons followed by a stop codon, containing the codons corresponding to whole transit peptide for the chloroplast stroma and the first four …
Identification And Expression Analysis Of Two Inorganic C-And N-Responsive Genes Encoding Novel And Distinct Molecular Forms Of Eukaryotic Phosphoenolpyruvate Carboxylase In The Green Microalga Chlamydomonas Reinhardtii, Tarlan G. Mamedov, Eric R. Moellering, Raymond Chollet
Identification And Expression Analysis Of Two Inorganic C-And N-Responsive Genes Encoding Novel And Distinct Molecular Forms Of Eukaryotic Phosphoenolpyruvate Carboxylase In The Green Microalga Chlamydomonas Reinhardtii, Tarlan G. Mamedov, Eric R. Moellering, Raymond Chollet
Papers in Biochemical Engineering
Phosphoenolpyruvate carboxylase (PEPC [Ppc]) has been previously purified and characterized in biochemical and immunological terms from two green microalgae, Chlamydomonas reinhardtii and Selenastrum minutum. The findings indicate that these algae possess at least two distinct PEPC enzyme-forms, homotetrameric Class1 and heteromeric Class-2 that differ significantly from each other and their plant and prokaryotic counterparts. Surprisingly, however, green-algal PEPC has been unexplored to date in molecular terms. This study reports the molecular cloning of the two Ppc genes in C. reinhardtii (CrPpc1, CrPpc2), each of which is transcribed in vivo and encodes a fully active, recombinant PEPC that lacks the regulatory, …
Crystallization And Preliminary X-Ray Analysis Of Na-Asp-1, A Multi-Domain Pathogenesis-Related-1 Protein From The Human Hookworm Parasite Necator Americanus, Dr.Mehmet Inan, Oluwatoyin A. Asojo, Alex Lucas, Rick Barent, Jicai Huang, Brad Plantz, Amber Swanson, Mark Gouthro, Michael M. Meagher, Peter J. Hotez
Crystallization And Preliminary X-Ray Analysis Of Na-Asp-1, A Multi-Domain Pathogenesis-Related-1 Protein From The Human Hookworm Parasite Necator Americanus, Dr.Mehmet Inan, Oluwatoyin A. Asojo, Alex Lucas, Rick Barent, Jicai Huang, Brad Plantz, Amber Swanson, Mark Gouthro, Michael M. Meagher, Peter J. Hotez
Papers in Biochemical Engineering
Human hookworm infection is a major cause of anemia and malnutrition in the developing world. In an effort to control hookworm infection, the Human Hookworm Vaccine Initiative has identified candidate vaccine antigens from the infective larval stage (L3) of the parasite, including a family of pathogenesis related- 1 (PR-1) proteins known as the ancylostoma-secreted proteins (ASPs). The functions of the ASPs are unknown. In addition, it is unclear why some ASPs have one while others have multiple PR-1 domains. There are no known structures of a multi-domain ASP and in an effort to remedy this situation, recombinant Na-ASP-1 has been …