A novel chimaeric flocculation protein enhances flocculation in Saccharomyces cerevisiae

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Westman, Johan,1983Chalmers tekniska högskola,Chalmers University of Technology,Chr. Hansen Holding A/S (author)

A novel chimaeric flocculation protein enhances flocculation in Saccharomyces cerevisiae

Article/chapterEnglish2018

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Elsevier BV,2018
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LIBRIS-ID:oai:research.chalmers.se:85d1b01f-ae25-47d4-9461-b8d15fe6103c
https://doi.org/10.1016/j.meteno.2018.04.001DOI
https://research.chalmers.se/publication/502966URI

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Language:English
Summary in:English

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Subject category:ref swepub-contenttype

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Yeast flocculation is the reversible formation of multicellular complexes mediated by lectin-like cell wall proteins binding to neighbouring cells. Strong flocculation can improve the inhibitor tolerance and fermentation performance of yeast cells in second generation bioethanol production. The strength of flocculation increases with the size of the flocculation protein and is strain dependent. However, the large number of internal repeats in the sequence of FLO1 from Saccharomyces cerevisiae S288c makes it difficult to recombinantly express the gene to its full length. In the search for novel flocculation genes resulting in strong flocculation, we discovered a DNA sequence, FLONF, that gives NewFlo phenotype flocculation in S. cerevisiae CEN.PK 113-7D. The nucleotide sequence of the internal repeats of FLONF differed from those of FLO1. We hypothesized that a chimaeric flocculation gene made up of a FLO1 variant derived from S. cerevisiae S288c and additional repeats from FLONF from S. cerevisiae CCUG 53310 would be more stable and easier to amplify by PCR. The constructed gene, FLOw, had 22 internal repeats compared to 18 in FLO1. Expression of FLOw in otherwise non-flocculating strains led to strong flocculation. Despite the length of the gene, the cassette containing FLOw could be easily amplified and transformed into yeast strains of different genetic background, leading to strong flocculation in all cases tested. The developed gene can be used as a self-immobilization technique or to obtain rapidly sedimenting cells for application in e.g. sequential batches without need for centrifugation.

Subject headings and genre

MEDICIN OCH HÄLSOVETENSKAP Medicinsk bioteknologi Medicinsk bioteknologi hsv//swe
MEDICAL AND HEALTH SCIENCES Medical Biotechnology Medical Biotechnology hsv//eng
NATURVETENSKAP Biologi Genetik hsv//swe
NATURAL SCIENCES Biological Sciences Genetics hsv//eng
TEKNIK OCH TEKNOLOGIER Industriell bioteknik Annan industriell bioteknik hsv//swe
ENGINEERING AND TECHNOLOGY Industrial Biotechnology Other Industrial Biotechnology hsv//eng
Protein engineering
Saccharomyces cerevisiae
Chimaeric
Cell-cell adhesion
Bioethanol

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Nyman, JonasUniversity of Southampton (author)
Manara, Richard M.A.University of Southampton (author)
Mapelli, Valeria,1978Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)mapelli (author)
Franzén, Carl Johan,1966Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)franzen (author)
Chalmers tekniska högskolaChr. Hansen Holding A/S (creator_code:org_t)

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In:Metabolic Engineering Communications: Elsevier BV6, s. 49-552214-0301

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