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Dive into the research topics where Bianca Elisabeth Maria Gielesen is active.

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Featured researches published by Bianca Elisabeth Maria Gielesen.


Archive | 2013

Transformation of Filamentous Fungi in Microtiter Plate

Bianca Elisabeth Maria Gielesen; Marco van den Berg

The polyethylene glycol (PEG) protoplast method for transformation of filamentous fungi is developed for use in microtiter plates, enabling one to perform a multitude of transformations in parallel.


Metabolic Engineering | 2018

Iterative algorithm-guided design of massive strain libraries, applied to itaconic acid production in yeast

Eric M. Young; Zheng Zhao; Bianca Elisabeth Maria Gielesen; Liang Wu; D. Benjamin Gordon; Johannes Andries Roubos; Christopher A. Voigt

Metabolic engineering requires multiple rounds of strain construction to evaluate alternative pathways and enzyme concentrations. Optimizing multigene pathways stepwise or by randomly selecting enzymes and expression levels is inefficient. Here, we apply methods from design of experiments (DOE) to guide the construction of strain libraries from which the maximum information can be extracted without sampling every possible combination. We use Saccharomyces cerevisiae as a host for a novel six-gene pathway to itaconic acid, selected by comparing alternative shunt pathways that bypass the mitochondrial TCA cycle. The pathway is distinctive for the use of acetylating acetaldehyde dehydrogenase to increase cytosolic acetyl-CoA pools, a bacterial enzyme to synthesize citrate in the cytosol, and an itaconic acid exporter. Precise control over the expression of each gene is enabled by a set of promoter-terminator pairs that span a 174-fold range. Two large combinatorial libraries (160 variants, 2.4 Mb and 32 variants, 0.6 Mb) are designed where the expression levels are selected by statistical methods (I-optimal response surface methodology, full factorial, or Plackett-Burman) with the intent of extracting different types of guiding information after the screen. This is applied to the design of a third library (24 variants, 0.5 Mb) intended to alleviate a bottleneck in cis-aconitate decarboxylase (CAD) expression. The top strain produces 815 mg/l itaconic acid, a 4-fold improvement over the initial strain achieved by iteratively balancing pathway expression. Including a methylated product in the total, the strain produces 1.3 g/l combined itaconic acids. Further, a regression analysis of the libraries reveals the optimal expression level of CAD as well as pairwise interdependencies between genes that result in increased titer and purity of itaconic acid. This work demonstrates adapting algorithmic design strategies to guide automated yeast strain construction and learn information after each iteration.


Archive | 2009

A pentose sugar fermenting cell

Paul Klaassen; Van Der Jan Metske Laan; Bianca Elisabeth Maria Gielesen; Van Gijsberdina Pieternella Suylekom


Archive | 2010

FERMENTATIVE PRODUCTION OF ETHANOL FROM GLUCOSE, GALACTOSE AND ARABINOSE EMPLOYING A RECOMBINANT YEAST STRAIN

Paul Klaassen; Van Gijsberdina Pieternella Suylekom; Bianca Elisabeth Maria Gielesen; Nicolette Jasmijn Broers; Beate Wiedemann; De Wilhelmus Theodorus Antonius Maria Laat


Archive | 2011

CELL SUITABLE FOR FERMENTATION OF A MIXED SUGAR COMPOSITION

Paul Klaassen; Bianca Elisabeth Maria Gielesen; Van Gijsberdina Pieternella Suylekom; Wilbert Herman Marie Heijne


Archive | 2011

Process for the production of cells which are capable of converting arabinose

Paul Klaassen; Bianca Elisabeth Maria Gielesen; Wilbert Herman Marie Heijne; Van Gijsberdina Pieternella Suylekom


Archive | 2012

Yeast cell capable of converting sugars including arabinose and xylose

Paul Klaassen; Bianca Elisabeth Maria Gielesen; Gijsberdina Pieternella Van Suylekom; Panagiotis Sarantinopoulos; Wilbert Herman Marie Heijne; Aldo Greeve


Archive | 2007

High throughput transfection of filamentous fungi

Marco van den Berg; Bianca Elisabeth Maria Gielesen; Adriana Marina Riemens


Archive | 2016

A CRISPR-CAS SYSTEM FOR A LIPOLYTIC YEAST HOST CELL

Bernard Meijrink; René Verwaal; Bianca Elisabeth Maria Gielesen; Johannes Andries Roubos


Archive | 2016

A crispr-cas system for a filamentous fungal host cell

Bernard Meijrink; René Verwaal; Bianca Elisabeth Maria Gielesen; Johannes Andries Roubos; Yvonne Nygård; Roelof Arij Lans Bovenberg; Arnold J. M. Driessen; Carsten Pohl

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