| US 7,507,574 B2 | ||
| Recombinant Saccharomyces cerevisiae expressing chimeric glucose transporters | ||
| Roslyn Bill, Knowle (United Kingdom); Eckhard Boles, Dreieich (Germany); Lena Gustafsson, Mölndal (Sweden); Stefan Hohmann, Lerum (Sweden); Christer Larsson, Mölndal (Sweden); and Karin Elbing, Göteborg (Sweden) | ||
| Assigned to Gothia Yeast Solutions AB, Kungsbacka (Sweden) | ||
| Appl. No. 10/312,783 PCT Filed Jun. 29, 2001, PCT No. PCT/GB01/03079 § 371(c)(1), (2), (4) Date Jul. 21, 2003, PCT Pub. No. WO02/00880, PCT Pub. Date Jan. 03, 2002. |
||
| Claims priority of application No. 0002460 (SE), filed on Jun. 29, 2000; and application No. 0004241 (SE), filed on Nov. 17, 2000. | ||
| Prior Publication US 2004/0058429 A1, Mar. 25, 2004 | ||
| Int. Cl. C12N 1/19 (2006.01); C12N 15/63 (2006.01); C12N 15/81 (2006.01); C12P 21/02 (2006.01); C12P 1/02 (2006.01); C07H 21/04 (2006.01); C07K 14/395 (2006.01); A23J 1/18 (2006.01); A23J 1/28 (2006.01); C12C 11/00 (2006.01) | ||
| U.S. Cl. 435—254.21 [435/320.1; 435/69.1; 435/69.7; 435/171; 435/483; 536/23.4; 536/23.74; 530/350; 530/371; 426/16; 426/60] | 23 Claims |
| 1. A modified Saccharomyces cerevisiae yeast which produces lower levels of ethanol than wild-type yeast under aerobic conditions and glucose concentrations of 2%,
and which exhibits a growth rate of at least 30% of the wild-type yeast, wherein said yeast contains a chimeric nucleotide
sequence (a chimeric construct) which is stably transformed into the genetic material of the yeast, wherein the construct
comprises a sequence having the form
A-B
wherein
A is a first sequence comprising nucleotide bases w to x;
B is a second sequence comprising nucleotide bases (x+y) to z; wherein
A is from a nucleotide sequence encoding a Saccharomyces cerevisiae glucose transporter selected from HXT-1 (SEQ ID NO:22) or HXT-3 (SEQ ID NO:24) or encoding a sequence which exhibits at least
80% sequence identity to HXT1 (SEQ ID NO:22) or HXT-3 (SEQ ID NO:24), or the sequence complementary to any of the aforesaid
sequences and
B is from a nucleotide sequence encoding a Saccharomyces cerevisiae glucose transporter selected from HXT-2 (SEQ ID NO:23), HXT-4 (SEQ ID NO:25), HXT-6 (SEQ ID NO:26), HXT-7 (SEQ ID NO:27) or
GAL2 (SEQ ID NO:28) or encoding a sequence which exhibits at least 80% sequence identity to HXT-2, HXT-4, HXT-6, HXT-7 or
GAL-2, or the sequence complementary to any of the aforesaid sequences;
w is a first position within the nucleotide sequence from which said first sequence is derived, and is from 1 to 10;
x is a second position within the nucleotide sequence from which said first sequence is derived and is from 400 to 900;
y is an integer less than 3;
x+y is a first position within the nucleotide sequence from which said second sequence is derived;
and z is a second position within the nucleotide sequence from which said second sequence is derived and is greater than x+y
and is from 1600 to 1713;
wherein said values refer to the position within the nucleotide sequence of said glucose transporter,
and wherein said chimeric nucleotide sequence is introduced into a yeast cell which is a null strain which exhibits a glucose
uptake of less than 2 nmol glucose/min/mg biomass to provide said modified Saccharomyces cerevisiae yeast.
|