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Methylotrophic Yeast Pichia Pastoris - Crossref

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Last Updated: 15 May 2022

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The methylotrophic yeast Pichia pastoris synthesizes a functionally active chromophore precursor of the plant photoreceptor phytochrome.

A bilin chromophore is covalently linked to the algal phytochrome apoprotein, according to zincc blot results. P. pastoris is also known for its discovery of phytochromobilin synthase, the enzyme that converts biliverdin IX alpha to phytochromobilin, was discussed in this study. When assembly with oat apophytochrome A, HPLC results show that biliverdin was converted to both 3E- and 3Z-isomers of phytochromobilin.

Source link: https://doi.org/10.1073/pnas.93.17.8989


Expression of Functional Influenza Virus RNA Polymerase in the Methylotrophic Yeast Pichia pastoris

Influenza virus RNA polymerase (PB1-PB2-PA), a subunit component of the same chemical synthases and cleavage of RNA, is involved in both transcription and replication of the viral genome. The addition of methanol resulted in a Simultaneous expression of all three P proteins in the yeast P. pastoris. To purify the P protein complexes, a sequence coding for a histidine tag was added to the PB2 protein gene at its N terminus. In addition, we find that functional influenza virus RNA polymerase with the catalytic activity of a transcriptase is present in P. pastoris' methylotrophic yeast.

Source link: https://doi.org/10.1128/jvi.74.9.4074-4084.2000


Catabolite Inactivation in the Methylotrophic Yeast Pichia pastoris

The inactivation of the alcohol oxidase enzyme system of Pichia pastoris During the whole-cell bioconversion of ethanol to acetaldehyde was due to catabolite inactivation. Acetaldehyde in the presence of O 2 was the agent of catabolite activation. A rise in the level of cyclic AMP was initiated by the onset of free acetaldehyde and was characterized by an increase in the amount of cyclic AMP, which coincided with a 55% decline in alcohol oxidase activity. The rate of catabolite activation was dependent on acetaldehyde concentrations up to 0. 14 mM. Alcohol oxidase production was psychrotolerant, with just 17% decrease in initial specific activity as a result of a temperature decrease from 37 to 3°C. 123 g of acetaldehyde per liter was produced at 3°C, relative to 58 g/liter at 30°C. 140. 6 g of acetaldehyde was produced by using 0. 5 M Tris in a cyclic-batch process.

Source link: https://doi.org/10.1128/aem.56.8.2378-2383.1990


Comparative biochemical and pharmacological characterization of the mouse 5HT5A 5-hydroxytryptamine receptor and the human β2-adrenergic receptor produced in the methylotrophic yeast Pichia pastoris

Pichia pastoris has evolved into a useful expression system for a variety of international genes over the past few years. Here, we show that the P. pastoris expression system in high-level manufacture of two G-protein-coupled receptors, the mouse 5HT5A 5-hydroxytryptamine receptor, and the human 2-adrenergic receptor have similar functionality to the baculovirus expression system in the bacterial expression system. The two receptors' cDNAs were cloned under transcriptional control of the highly expressed promoter of the P. pastoris alcohol oxidase 1 gene in a manner that differing expression plasmids were produced. Recombinant clones were tested and tested for the manufacture of the 5HT5A receptor and the 2-adrenergic receptor by radioligand binding using [N-methyl-3H] respectively, following the conversion into P. pastoris strains KM71 and SMD 1163. paraphrasergic receptor-producing clones that produce the 2-adrenergic receptor. The addition of the adrenergic antagonist alprenololololololololololololololololololololololololololololololololololololololo had clones in clones producing the clono clonodrenergic receptor alprenodrenergic receptor adrenergic receptor's adrenergic receptor alprenolox adrenergic receptor alprenolololololololololololololololololololololololololololololololololo In crude membrane preparations, 40 pmol of 5HT5A receptor per mg of membrane protein was produced per mg of membrane protein. The 5HT5A receptor's apparent molecular mass was about 20 kDa higher than expected from the amino acid sequence, according to an immunoblot analysis. In comparison, the fusion of the 2-adrenergic receptor fusion to the -factor leader was correctly handled by the internal Kex2 endopeptidase. In conclusion, the two receptors in P. pastoris' high-volume manufacturing will allow their purification in quantities sufficient for subsequent biophysical and structural studies.

Source link: https://doi.org/10.1042/bj3301137


Genome-scale metabolic reconstruction and in silico analysis of methylotrophic yeast Pichia pastoris for strain improvement

For improved insight into the methylotrophic yeast's metabolism, it is extremely important to establish a systems biotechnological framework, in which a comprehensive in silico model of P. pastoris can be employed alongside high-throughput experimental data analysis for better understanding of the methylotrophic yeast's metabolism. P. pastoris' cellular phenotype was found during chemostat experiments. The constraints-based flux analysis was then used to forecast a realistic growth rate that is consistent with the cellular phenotype of P. pastoris found during chemostat experiments. The impact of various carbon sources on cell growth was detailed in silico research, with sorbitol emerging as a promising candidate for culturing recombinant P. pastoris strains that produce heterologous proteins. Hence, we investigated the applicability of P. pastoris' device to whole-cell biotransformation as well as identified key metabolic engineering goals that have been experimentally verified.

Source link: https://doi.org/10.1186/1475-2859-9-50


High yield expression of leptospirosis vaccine candidates LigA and LipL32 in the methylotrophic yeast Pichia pastoris

Leptospirosis, a zooonosis caused by Leptospira spp. , has been classified as an emergent infectious disease by Leptospira spp. Proteins produced in Escherichia coli's Recombinant protein have shown promising results, but with variable efficacy. Pichia pastoris is an alternative host with many advantages over the production of recombinant proteins. As Results The vaccine candidates LigANI and LipL32 were cloned and expressed in P. pastoris as unidentified proteins. Conclusions and Conclusions The expression of LigANI and LipL32 in P. pastoris resulted in a significant rise in yield relative to expression in E. coli.

Source link: https://doi.org/10.1186/1475-2859-9-98


Recombinant production of bacterial toxins and their derivatives in the methylotrophic yeast Pichia pastoris

Purpose The methylotrophic yeast Pichia pastoris is a common heterologous expression host for the recombinant production of a variety of prokaryotic and eukaryotic proteins. The rapid emergence of P. pastoris as a robust heterologous expression host was aided by the ease with which it can be controlled and propagated, which is similar to that of Escherichia coli and Saccharomyces cerevisiae. Here, we review the recombinant production of bacterial toxins and their derivatives in P. pastoris, with particular attention on their potential clinical use. We also provide general advice to this end considering that de novo design and manufacture of synthetic toxin genes have been often needed to produce optimal heterologous expression in P. pastoris.

Source link: https://doi.org/10.1186/1475-2859-4-33


Operational strategies, monitoring and control of heterologous protein production in the methylotrophic yeast Pichia pastoris under different promoters: A review

Abstract The methylotrophic yeast Pichia pastoris has been widely used as a good expression system for heterologous protein production. According to produce a robust process for the final industrial application, the use of various phenotypes under PAOX promoter, other alternative promoters, culture medium, and organizational plans with the aim of increasing either yield or productivity of the heterologous protein has been reported, but also the production of a reproducible product batch to batch to ensure a consistent process for the final industrial application has been established.

Source link: https://doi.org/10.1186/1475-2859-5-17


Metabolic Engineering of the Methylotrophic Yeast Pichia Pastoris (Komagataella phaffii) for the Production of Β-alanine From Methanol

Fermentative production of -alanine from renewable feedstocks such as glucose has sparked a lot of attention in recent years. In this study, we established the possibility of methanol production from Pichia pastoris as a methylotrophic cell factory. Different sources were tested and expressed in P. pastoris, followed by the optimization of aspartate decarboxylation by raising the ADC copy number and C4 precursor supply by the overexpression of aspartate dehydrogenase. This is the highest chemical production titer ever achieved in P. pastoris using methanol as the substrate, according to our best information.

Source link: https://doi.org/10.21203/rs.3.rs-651971/v1


Analysis of Wild-Type and Mutant Plant Nitrate Reductase Expressed in the Methylotrophic Yeast Pichia pastoris

Recombinant Arabidopsis thaliana NADH: neutritrate reductase was produced in the methylotrophic yeast Pichia pastoris and purified to near-electrophoretic homogeneity, according to the author. Purified enzymes had the spectral and kinetic characteristics of highly purified NR from natural plant sources, as shown by the kinetic and kinetic characteristics. When the invariant cysteine residue, cysteine-191, was replaced by serine or alanine in the A. thaliana NIA2 protein's molybdo-pterin region, the NR protein was still produced but was inactive, showing that this residue is essential for enzyme activity. P. pastoris is a cost-effective and reliable method for obtaining recombinant holo-NR from plants, according to these findings.

Source link: https://doi.org/10.1104/pp.115.3.1135

* Please keep in mind that all text is summarized by machine, we do not bear any responsibility, and you should always check original source before taking any actions

* Please keep in mind that all text is summarized by machine, we do not bear any responsibility, and you should always check original source before taking any actions