0000000000162161
AUTHOR
Palma Parascandola
A multidomain xylanase from a Bacillus sp. with a region homologous to thermostabilizing domains of thermophilic enzymes
The gene xynC encoding xylanase C from Bacillus sp. BP-23 was cloned and expressed in Escherichia coli. The nucleotide sequence of a 3538 bp DNA fragment containing xynC gene was determined, revealing an open reading frame of 3258 bp that encodes a protein of 120,567 Da. A comparison of the deduced amino acid sequence of xylanase C with known beta-glycanase sequences showed that the encoded enzyme is a modular protein containing three different domains. The central region of the enzyme is the catalytic domain, which shows high homology to family 10 xylanases. A domain homologous to family IX cellulose-binding domains is located in the C-terminal region of xylanase C, whilst the N-terminal r…
A novel process-based model of microbial growth: self-inhibition in Saccharomyces cerevisiae aerobic fed-batch cultures
Microbial population dynamics in bioreactors depend on both nutrients availability and changes in the growth environment. Research is still ongoing on the optimization of bioreactor yields focusing on the increase of the maximum achievable cell density. A new process-based model is proposed to describe the aerobic growth of Saccharomyces cerevisiae cultured on glucose as carbon and energy source. The model considers the main metabolic routes of glucose assimilation (fermentation to ethanol and respiration) and the occurrence of inhibition due to the accumulation of both ethanol and other self-produced toxic compounds in the medium. Model simulations reproduced data from classic and new expe…
Bread making with Saccharomyces cerevisiae CEN.PK113-5D expressing lipase A from Bacillus subtilis: Leavening characterisation and aroma enhancement
Summary Lipase A from Bacillus subtilis was expressed in the yeast Saccharomyces cerevisiae CEN.PK113-5D strain as a cell wall-immobilised enzyme. The recombinant yeast was used in bread making to test the effect of lipase A on the bread properties such as rheological and aromatic properties. The results were compared to the not transformed strain and the commercial baker's yeast. The recombinant strain resulted a good leavening agent comparable to the commercial baker's yeasts provided 1% sucrose was added to the dough. Whereas, during the leavening, the trend of the rheological properties (cohesivness, gumminess and rigidity) differed from the commercial and the nontransformed (NT) strain…
Immobilization of <i>Saccharomyces cerevisiae</i> Cells to Protein G-Sepharose by Cell Wall Engineering
In this work, we explored the possibility of using the targeting of a heterologous protein to the cell wall of <i>Saccharomyces cerevisiae</i>, by fusing it to a cell wall protein, to construct yeast strains whose cells display on their surface proteins that bind to a matrix, so as to achieve the immobilization of the whole cells. With this aim, we created a gene fusion that comprises the region responsible for attachment of a cell wall protein to the cell wall, and the IgG binding region of staphylococcal protein A, and expressed it in the <i>mnn1mnn9</i> strain of <i>S. cerevisiae</i>. The surface display of the protein A-Icwp fusion protein was positiv…
Performance of the auxotrophic Saccharomyces cerevisiae BY4741 as host for the production of IL-1β in aerated fed-batch reactor: role of ACA supplementation, strain viability, and maintenance energy
Abstract Background Saccharomyces cerevisiae BY4741 is an auxotrophic commonly used strain. In this work it has been used as host for the expression and secretion of human interleukin-1β (IL1β), using the cell wall protein Pir4 as fusion partner. To achieve high cell density and, consequently, high product yield, BY4741 [PIR4-IL1β] was cultured in an aerated fed-batch reactor, using a defined mineral medium supplemented with casamino acids as ACA (auxotrophy-complementing amino acid) source. Also the S. cerevisiae mutant BY4741 Δyca1 [PIR4-IL1β], carrying the deletion of the YCA1 gene coding for a caspase-like protein involved in the apoptotic response, was cultured in aerated fed-batch rea…