Use of the cell wall protein Pir4 as a fusion partner for the expression of <i>Bacillus</i> sp. BP‐7 xylanase A in <i>Saccharomyces cerevisiae</i>

Andrés, Isabel Mateu; Gallardo, Óscar; Parascandola, Palma; Pastor, F. I. Javier; Zueco, Jesús

doi:10.1002/bit.20375

Cited by 36 publications

(33 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the case of Cwp1, the PIR repeat may be used as an additional wall anchorage point because the protein is attached to the wall by both an alkali-labile and a GPI-dependent linkage (Kapteyn et al 2001). Like GPI-CWP, PIR proteins can be used as carriers to direct surface expression of heterologous proteins fused to them (Andrés et al 2005;Shimma et al 2006).…”

Section: Incorporation Of Pir Proteins Into the Wallmentioning

confidence: 99%

Architecture and Biosynthesis of the Saccharomyces cerevisiae Cell Wall

2012

View full text Add to dashboard Cite

The wall gives a Saccharomyces cerevisiae cell its osmotic integrity; defines cell shape during budding growth, mating, sporulation, and pseudohypha formation; and presents adhesive glycoproteins to other yeast cells. The wall consists of b1,3-and b1,6-glucans, a small amount of chitin, and many different proteins that may bear N-and O-linked glycans and a glycolipid anchor. These components become cross-linked in various ways to form higher-order complexes. Wall composition and degree of cross-linking vary during growth and development and change in response to cell wall stress. This article reviews wall biogenesis in vegetative cells, covering the structure of wall components and how they are cross-linked; the biosynthesis of N-and O-linked glycans, glycosylphosphatidylinositol membrane anchors, b1,3-and b1,6-linked glucans, and chitin; the reactions that cross-link wall components; and the possible functions of enzymatic and nonenzymatic cell wall proteins. TABLE OF CONTENTS Abstract 775Introduction 777

show abstract

Section: Incorporation Of Pir Proteins Into the Wallmentioning

confidence: 99%

Architecture and Biosynthesis of the Saccharomyces cerevisiae Cell Wall

2012

View full text Add to dashboard Cite

show abstract

“…For this, the DNA sequence coding VP8* was fused to the genes coding two cell wall proteins of S. cerevisiae, Icwp (Ssr1p) and Pir4. Icwp had been previously used by our group as a partner for the targeting to the cell wall of the Staphylococcal protein A (Andrés et al, 2003), whilst Pir4 has been recently being proposed as partner to promote the cell wall targeting or the secretion of recombinant enzymes (Andrés et al, 2005).…”

Section: Resultsmentioning

confidence: 99%

“…Constructions V2-V4 involved the fusion of VP8* with Pir4 a cell wall protein that can be extracted from the cell wall by reducing agents (Moukadiri et al, 1999). Also in this case, our group has shown the feasibility of using Pir4 to achieve the targeting to the cell wall or the secretion to the growth medium of recombinant proteins (Andrés et al, 2005). Constructions V2 and V3 consisted in the insertion of the VP8* sequence without affecting the Pir4 domain responsible for the attachment through disulphide bridges, whilst V4 involved replacing the region containing this domain by VP8*.…”

Section: Discussionmentioning

confidence: 98%

Yeast expression of the VP8* fragment of the rotavirus spike protein and its use as immunogen in mice

Andrés

Rodrı́guez-Dı́az

Buesa

et al. 2005

Biotech & Bioengineering

Self Cite

View full text Add to dashboard Cite

The VP8* fragment from the rotavirus spike protein was expressed as a fusion protein with two different cell wall proteins of Saccharomyces cerevisiae, Icwp (Ssr1p) and Pir4, to achieve cell wall targeting or secretion to the growth medium of the fusion proteins. Two different host strains were used for the expression of the fusion proteins, a standard S. cerevisiae strain and a mnn9 glycosylation deficient strain, the later to reduce hyper-glycosylation. The Icwp-VP8* fusion could only be detected in the growth medium, indicating that the presence of the VP8* moiety interferes with the anchorage of Icwp to the cell wall. In the case of the Pir4-VP8* fusion proteins, we achieved cell wall targeting or secretion depending on how the gene fusion had been performed. In all cases, the fusion proteins expressed in the mnn9 strain showed a reduced level of glycosylation. Mice were inoculated intraperitoneally either with Pir4-VP8* or Icwp-VP8* fusion proteins purified from the growth medium of mnn9 strains expressing them or with whole cells of an mnn9 strain expressing a Pir4-VP8 fusion protein on its cell walls. Hundred percent of mice inoculated with the Pir4-VP8* fusion protein and 25% of those inoculated with the Icwp-VP8* fusion protein showed high titers of anti-VP8* antibodies. No specific immune response was detected in those mice inoculated with whole cells. Finally, susceptibility to rotavirus infection of the offspring of immunized dams was determined and protection was found in a percentage of approximately 60% with respect to the control group.

show abstract

“…In the present work, to discriminate the effects given by a heterologous metabolic burden over those related to the cultural conditions, we have compared the growth parameters of CEN.PK113-5D expressing an heterologous product against those of the non-transformed strain. The recombinant strain carries, on the episomal pIA1 derivative [7], the coding sequence for human interleukin-1b (IL-1b) gene fused with the regulatory sequences of PIR4 gene [8] that promotes IL-1b secretion to the growth medium [9].…”

Section: Introductionmentioning

confidence: 99%

High cell density culture with S. cerevisiae CEN.PK113-5D for IL-1β production: optimization, modeling, and physiological aspects

Landi

Paciello

Alteriis

et al. 2014

Bioprocess Biosyst Eng

Self Cite

View full text Add to dashboard Cite

Saccharomyces cerevisiae CEN.PK113-5D, a strain auxotrophic for uracil belonging to the CEN.PK family of the yeast S. cerevisiae, was cultured in aerated fed-batch reactor as such and once transformed to express human interleukin-1β (IL-1β), aiming at obtaining high cell densities and optimizing IL-1β production. Three different exponentially increasing glucose feeding profiles were tested, all of them "in theory" promoting respiratory metabolism to obtain high biomass/product yield. A non-structured non-segregated model was developed to describe the performance of S. cerevisiae CEN.PK113-5D during the fed-batch process and, in particular, its capability to metabolize simultaneously glucose and ethanol which derived from the precedent batch growth. Our study showed that the proliferative capacity of the yeast population declined along the fed-batch run, as shown by the exponentially decreasing specific growth rates on glucose. Further, a shift towards fermentative metabolism occurred. This shift took place earlier the higher was the feed rate and was more pronounced in the case of the recombinant strain. Determination of some physiological markers (acetate production, intracellular ROS accumulation, catalase activity and cell viability) showed that neither poor oxygenation nor oxidative stress was responsible for the decreased specific growth rate, nor for the shift to fermentative metabolism.

show abstract

Use of the cell wall protein Pir4 as a fusion partner for the expression of Bacillus sp. BP‐7 xylanase A in Saccharomyces cerevisiae

Cited by 36 publications

References 40 publications

Architecture and Biosynthesis of the Saccharomyces cerevisiae Cell Wall

Architecture and Biosynthesis of the Saccharomyces cerevisiae Cell Wall

Yeast expression of the VP8* fragment of the rotavirus spike protein and its use as immunogen in mice

High cell density culture with S. cerevisiae CEN.PK113-5D for IL-1β production: optimization, modeling, and physiological aspects

Contact Info

Product

Resources

About