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Construction of lactose-fermenting a...

Farahnak, Fereydoon.

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Construction of lactose-fermenting and high-ethanol-producing hybrid yeast by protoplast fusion of Saccharomyces cerevisiae and Kluyveromyces fragilis and the analysis of beta-galactosidase gene in the hybrid yeast.

Record Type:	Language materials, printed : Monograph/item
Title/Author:	Construction of lactose-fermenting and high-ethanol-producing hybrid yeast by protoplast fusion of Saccharomyces cerevisiae and Kluyveromyces fragilis and the analysis of beta-galactosidase gene in the hybrid yeast./
Author:	Farahnak, Fereydoon.
Description:	105 p.
Notes:	Source: Dissertation Abstracts International, Volume: 49-09, Section: B, page: 3583.
Contained By:	Dissertation Abstracts International49-09B.
Subject:	Biology, Microbiology. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=8821699

Construction of lactose-fermenting and high-ethanol-producing hybrid yeast by protoplast fusion of Saccharomyces cerevisiae and Kluyveromyces fragilis and the analysis of beta-galactosidase gene in the hybrid yeast.
Farahnak, Fereydoon.

Construction of lactose-fermenting and high-ethanol-producing hybrid yeast by protoplast fusion of Saccharomyces cerevisiae and Kluyveromyces fragilis and the analysis of beta-galactosidase gene in the hybrid yeast. - 105 p.

Source: Dissertation Abstracts International, Volume: 49-09, Section: B, page: 3583.

Thesis (Ph.D.)--University of California, Davis, 1988.

The availability of a yeast strain which is capable of fermenting lactose and at the same time is tolerant to high concentrations of ethanol would be useful for the production of ethanol from lactose. Kluyveromyces fragilis is capable of fermenting lactose, but it is not as tolerant as Saccharomyces cerevisiae to high concentrations of ethanol. In this study, we have used the protoplast fusion technique to construct hybrids between auxotrophic strains of Saccharomyces cerevisiae having high ethanol tolerance and an auxotrophic strain of lactose fermenting Kluyveromyces fragilis isolated by ethyl methanesulfonate mutagenesis. The fusants obtained were prototrophic and capable of assimilating lactose and producing ethanol in excess of 13% (v/v). The complementation frequency of fusion was about 0.7%. Formation of fusants was confirmed by the increased amount of chromosomal DNA per cell. Fusants contained 8-16 Subjects--Topical Terms:

1017734
Biology, Microbiology.

Construction of lactose-fermenting and high-ethanol-producing hybrid yeast by protoplast fusion of Saccharomyces cerevisiae and Kluyveromyces fragilis and the analysis of beta-galactosidase gene in the hybrid yeast.
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008 130515s1988 ||||||||||||||||| ||eng d
035 $a (UMI)AAI8821699
035 $a AAI8821699
040 $a UMI $c UMI
100 1 $a Farahnak, Fereydoon. $3 1683701
245 1 0 $a Construction of lactose-fermenting and high-ethanol-producing hybrid yeast by protoplast fusion of Saccharomyces cerevisiae and Kluyveromyces fragilis and the analysis of beta-galactosidase gene in the hybrid yeast.
300 $a 105 p.
500 $a Source: Dissertation Abstracts International, Volume: 49-09, Section: B, page: 3583.
502 $a Thesis (Ph.D.)--University of California, Davis, 1988.
520 $a The availability of a yeast strain which is capable of fermenting lactose and at the same time is tolerant to high concentrations of ethanol would be useful for the production of ethanol from lactose. Kluyveromyces fragilis is capable of fermenting lactose, but it is not as tolerant as Saccharomyces cerevisiae to high concentrations of ethanol. In this study, we have used the protoplast fusion technique to construct hybrids between auxotrophic strains of Saccharomyces cerevisiae having high ethanol tolerance and an auxotrophic strain of lactose fermenting Kluyveromyces fragilis isolated by ethyl methanesulfonate mutagenesis. The fusants obtained were prototrophic and capable of assimilating lactose and producing ethanol in excess of 13% (v/v). The complementation frequency of fusion was about 0.7%. Formation of fusants was confirmed by the increased amount of chromosomal DNA per cell. Fusants contained 8-16 $\ times $ 10 $\ sp8 $ $\ mu $g DNA/cell as compared to about 4 $\ times $ 10 $\ sp8 $ $\ mu $g DNA/cell for the parental strains suggesting that multiple fusions had taken place.
520 $a The fusant obtained by protoplast fusion of Saccharomyces cerevisiae and Kluyveromyces fragilis was further studied. The gene coding for the enzyme $\ beta $- alactosidase was isolated by cloning in E. coli YMC9 using recombinant DNA techniques. The K. fragilis chromosomal DNA was partially digested with the restriction endonuclease Sau3A and the fragments were ligated into BamHI-digested pBR322 plasmid. The plasmid library pFR-1 was transformed into $l ac\sp{-} $ E. coli strain YMC9 and $l ac\sp{+} $ transformants were selected. Restriction mapping of the inserted DNA fragment revealed that the size of the cloned fragment was 7.6 Kbp. The recombinants were able to synthesize active $\ beta $- galactosidase enzyme, which showed that the cloned gene was expressed in the prokaryotic host, albeit not very efficiently. Furthermore, the DNA-DNA hybridization study revealed that the probe (the cloned gene from the fusant yeast) hybridized to five fragments in the yeast genomic DNA. This further confirmed that the $\ beta $- galactosidase gene which was cloned came from the parent strain of yeast and not from E. coli.
590 $a School code: 0029.
650 4 $a Biology, Microbiology. $3 1017734
690 $a 0410
710 2 $a University of California, Davis. $3 1018682
773 0 $t Dissertation Abstracts International $g 49-09B.
790 $a 0029
791 $a Ph.D.
792 $a 1988
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=8821699