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Kluyveromyces lactis ^ a retrospective

Hiroshi Fukuhara
Institut Curie, Section de Recherche, UMR2027, Centre Universitaire Paris XI, Orsay, France.
Tel.: 133 1 69 86 3063; fax: 133 1 69 86 9429; e-mail:
Received 27 April 2005; accepted 31 May 2005.
First published online 28 November 2005.
Editor: Lex Scheffers
Kluyveromyces lactis; model yeast; nonconventional yeast.

The use of Kluyveromyces lactis for research started in

early 1960s. In contrast to most cases of yeast research, the
study of this particular species was initially motivated by a
purely academic question, that is, possible adaptive regulation of sugar metabolism in a lower eukaryote. Biotechnological interest in K. lactis came much later. Until about
1980, K. lactis research was barely visible in the shadow of
the formidable development of the Saccharomyces cerevisiae
The early 1960s were the era of lactose regulation in
Escherichia coli, which led to the birth of molecular biology.
Following the achievements of the investigators of the E. coli
system, a number of laboratories were trying to find an
inducible enzyme system in a eukaryotic organism in order
to evaluate the general significance of the operon concept.
Harlyn O. Halvorson at Madison, Wisconsin, was one of
those people. He contacted the great taxonomist L. J.
Wickerham (USDA, Peoria) who knew how different yeast
species assimilated various sugars. Apparently, it was he who
suggested the use of K. lactis, a species that assimilated bglucosides in an adaptive mode. Halvorson and his colleagues have thus started to work on this yeast (then called
Saccharomyces lactis), using two isolates obtained from
Peoria, NRRL Y-1140 (CBS 2359 [Mat a]) and Y-1118 (CBS
6315 [Mat a]). A mating system for genetic analysis was
elaborated. The b-glucosidase system of K. lactis turned out
to be complicated by the fact that it was paraconstitutive
(half-inducible, half-constitutive). After this pioneer work
of the Madison group, only a few laboratories continued to
use K. lactis, essentially in the field of mitochondrial genetics
and biogenesis, in comparison with the S. cerevisiae system.
After all, yeast species available for formal genetic analysis
were rare and are still few even now: apart from the fission
yeast Schizosaccharomyces pombe and the nonfermenting
Yarrowia lipolytica, we have practically only K. lactis.
In the early 1980s, two lines of study, specific to K. lactis,
revived interest in this yeast. One was the series of works on
FEMS Yeast Res 6 (2006) 323324

the regulation of lactose metabolism, and the other was the

discovery of the new killer system involving DNA plasmids.
The studies on the lactose regulon in K. lactis illustrated how
the regulatory system of this unicellular eukaryote differed
from the bacterial lactose operon. The lactose regulon, a
close variation of the galactose regulon of S. cerevisiae, was
shown to involve many genetically unlinked positive and
negative regulatory genes. In parallel, the works of the killer
system involving the linear DNA plasmids pGKL1 and 2 had
also played an important role in the development of K. lactis
biology, because at that time the only plasmids known in
yeast were the 2 mm circular DNA and the double-stranded
killer RNA of S. cerevisiae. In addition to their new killing
mechanism and their linear mode of replication, pGKL
plasmids were interesting in many aspects. The fact that K.
lactis secreted a high-molecular weight killer toxin retained
the attention of a few people who were looking for an
efficient system to produce recombinant proteins in a
secreted form. Indeed the 1980s were years that were full of
enthusiasm for gene engineering for biotechnology. Thus,
the milk-coagulating enzyme chymosin was produced industrially from K. lactis. Such achievements by a few
industrial companies encouraged research on this particular
yeast system. The successive biotechnology programs
funded by the European Commission were a timely support,
and it was through these programs that the first network of
K. lactis research was set up in 1988. Since then the workshop, Biology of Kluyveromyces, continues to be an important instrument of communication and collaboration of
the research community.
Collaboration between K. lactis workers has been facilitated by two circumstances. First, the researchers have used,
from the beginning, only a very small number of K. lactis
isolates, including those used by the Madison group, thus a
relatively homogeneous genetic system has rapidly emerged
and has been shared by most laboratories, allowing the
efficient exchange of strains. Second, after a deliberate
2005 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved


search, a 2 mm type circular plasmid pKD1 was discovered in

the species Kluyveromyces drosophilarum (hence the name of
the plasmid), which was capable of replicating in K. lactis,
offering a replicating vector system equivalent to the 2 mm
vectors. Kluyveromyces drosophilarum was later incorporated
into K. lactis.
The yeast species that assimilate lactose aerobically are
widespread, but those that ferment lactose are rather rare.
Beside K. lactis, Kluyveromyces fragilis is one of such lactosefermenting yeasts, and is well known in industry. This
species is now incorporated into K. marxianus (E. C.
Hansen) van der Walt (1971), distinct from K. lactis. At
present the taxonomists recognize two varieties in K. lactis.
One is K. lactis (Dombrowski) van der Walt var. lactis
(1986), the other is K. lactis (Dombrowski) van der Walt
var. drosophilarum (1986). Whereas the former is heterothallic and ferments lactose, the latter is homothallic and
does not assimilate lactose (Lachance, 1998). Nearly all the
published works on K. lactis concern the variety lactis.
If S. cerevisiae stands in a very exceptional position among
yeasts because of its fermentation-oriented Crabtree-positive physiology, K. lactis appears to be a good model of the
large number of more aerobic species that are used in todays
yeast biotechnology. At the opposite extreme, the fermentation-less Yarrrowia lipolytica may be a model for highly
aerobic species, with its well-established genetic system.

2005 Federation of European Microbiological Societies

Published by Blackwell Publishing Ltd. All rights reserved

H. Fukuhara

While continuously stimulated by industrial interests, the

present research in K. lactis mostly focuses on fundamental
aspects of physiology and gene regulation. The themes cover
a wide range of topics, and the articles gathered in this issue
do not represent the diversity of the ongoing research in this
The total DNA sequence of the K. lactis genome has been
established in 2004 through the Ge nolevures project. Thus,
K. lactis has become, after S. cerevisiae, a most useful
instrument of yeast study, combining both genetic and
genomic information. In view of the distinctive physiologies
of the two species, and their relatively recent common origin
suggested by genomic sequence analyses, the comparison of
these two species will be particularly useful to unveil details
of the process of evolution of gene regulation and genome
organisation. Such a comparative approach is a basic
practice of K. lactis workers, as most of them are active in S.
cerevisiae research and also exploring the biology of other
nonconventional species.

Lachance MA (1998) The Yeasts, A Taxonomic Study. 4th edn
(Kurtzman CP & Fell JW, eds), pp. 227247. Elsevier Science B.
V., Amsterdam.

FEMS Yeast Res 6 (2006) 323324