Pichia pastoris is commonly used to express and secrete target proteins, although not all recombinant proteins can be successfully produced. In this study, we used methyl parathion hydrolase (MPH) from Ochrobactrum sp. M231 as a model to study the importance of the N-terminus of the protein for its secretion. While MPH can be efficiently expressed intracellularly in P. pastoris, it is not secre...

BACKGROUND Recombinant antibodies are highly successful in many different pathological conditions and currently enjoy overwhelming recognition of their potential. There are a wide variety of protein expression systems available, but almost all therapeutic antibodies are produced in mammalian cell lines, which mimic human glycosylation. The production of clinical-grade antibodies in mammalian ce...

Gene replacement in yeast is often accomplished by using a counterselectable marker such as URA3. Although ura3 strains of Pichia pastoris have been generated, these strains are inconvenient to work with because they grow slowly, even in the presence of uracil. To overcome this limitation, we have developed an alternative counterselectable marker that can be used in any P. pastoris strain. This...

Recombinant protein production in Pichia pastoris is based on alcohol oxidase promoterswhich are regulated by methanol. However, the use of methanol has several disadvantages,which is why current trends in bioprocess development with Pichia pastoris (P. pastoris) arefocusing on methanol mixed feeding strategies. This work aimed to develop a new experimentalmethod and compare the effect of vario...

Introduction: The development of an effective vaccine against Hepatitis C virus (HCV) is still a target of intensive vaccine research. The HCV envelope proteins E1 and E2 which can induce broadly neutralizing antibodies are the major candidate for this purpose. Different types of expression systems have been used to express these glycoproteins. In this study, an expression system using Pichia p...

Polyploidization plays an important role in plant speciation. The most recent estimates report that up to 15% of angiosperm speciation events and 31% in ferns are accompanied by changes in ploidy level. Polyploids can arise either through autopolyploidy, when the sets of chromosomes originate from a single species, or through allopolyploidy, when they originate from different species. In this s...

Human granulocyte-colony stimulating factor (hG-CSF) cDNA was expressed in the methylotrophic yeast Pichia pastoris under the control of the alcohol oxidase (AOX1) promoter. An expression vector for hG-CSF secretion was constructed using vector pPIC9. Higher levels of hG-CSF was obtained using a P. pastoris Mut+ (methanol utilization fast) phenotype. The effects of environmental factors such as...

Medium-chain volatile flavour esters are important molecules since they have extensive applications in food, fragrance, cosmetic, paint and coating industries, which determine different characteristics of aroma or taste in commercial products. Biosynthesis of these compounds by alcoholysis is catalyzed by acyl-CoA:ethanol O-acyltransferases Eht1 or Eeb1 in Saccharomyces cerevisiae. In this stud...

To express the antimicrobial peptide cecropin D in Pichia pastoris and determine the activity of the expressed product, four oligonucleotide fragments were synthesized in accordance with the available cecropin D sequences and a codon bias suitable for Pichia pastoris. Sequence fragments were phosphorylated, annealed, linked and cloned into the expression vector pGAPZαA and the yeast α-mating fa...

Yeast Pichia pastoris is a widely used system for heterologous protein expression. However, post-translational modifications, especially glycosylation, usually impede pharmaceutical application of recombinant proteins because of unexpected alterations in protein structure and function. The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-...