The University of Southampton
University of Southampton Institutional Repository

A novel chimaeric flocculation protein enhances flocculation in Saccharomyces cerevisiae

A novel chimaeric flocculation protein enhances flocculation in Saccharomyces cerevisiae
A novel chimaeric flocculation protein enhances flocculation in Saccharomyces cerevisiae

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.

Bioethanol, Cell-cell adhesion, Chimaeric, Protein engineering, Saccharomyces cerevisiae
49-55
Westman, Johan O.
5cb658f5-0f0f-4fa0-8432-08f30488cce6
Nyman, Jonas
0feff679-4e4f-4205-9ef9-6a4458ff2fc9
Manara, Richard M.A.
33709a59-e423-40dc-a1b8-0880df386cb4
Mapelli, Valeria
a32941b3-2859-45e3-ad3c-a1f1daa546ec
Franzén, Carl Johan
86f93705-288d-471d-afd1-42b58cf295ef
Westman, Johan O.
5cb658f5-0f0f-4fa0-8432-08f30488cce6
Nyman, Jonas
0feff679-4e4f-4205-9ef9-6a4458ff2fc9
Manara, Richard M.A.
33709a59-e423-40dc-a1b8-0880df386cb4
Mapelli, Valeria
a32941b3-2859-45e3-ad3c-a1f1daa546ec
Franzén, Carl Johan
86f93705-288d-471d-afd1-42b58cf295ef

Westman, Johan O., Nyman, Jonas, Manara, Richard M.A., Mapelli, Valeria and Franzén, Carl Johan (2018) A novel chimaeric flocculation protein enhances flocculation in Saccharomyces cerevisiae. Metabolic Engineering Communications, 6, 49-55. (doi:10.1016/j.meteno.2018.04.001).

Record type: Article

Abstract

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.

Text
1-s2.0-S2214030117300172-main - Version of Record
Download (512kB)

More information

Accepted/In Press date: 7 April 2018
e-pub ahead of print date: 9 April 2018
Published date: 1 June 2018
Keywords: Bioethanol, Cell-cell adhesion, Chimaeric, Protein engineering, Saccharomyces cerevisiae

Identifiers

Local EPrints ID: 420423
URI: http://eprints.soton.ac.uk/id/eprint/420423
PURE UUID: 5c03f976-61c5-4597-a8a7-c8f2eed7dc19

Catalogue record

Date deposited: 08 May 2018 16:30
Last modified: 07 Oct 2020 00:12

Export record

Altmetrics

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of http://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×