1. A modified Saccharomyces yeast which produces lower levels of ethanol than wild-type yeast under aerobic conditions and saccharide concentrations of 5 mM or more, preferably 2% glucose, and which exhibits a growth rate of at least 30% of the wild-type yeast.
2. A yeast as claimed in claim 1 wherein said yeast produces less than 0.25 gl, preferably less than 0.12 gl ethanol.
3. A yeast as claimed in claim 1 or 2 wherein said yeast exhibits a growth rate of at least 50%, preferably at least 70% of the wild-type yeast.
4. A yeast as claimed in any one of claims 1 to 3 wherein said yeast is a Saccharomyces cerevisiae.
5. A yeast as claimed in any one of claims 1 to 4 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 at least a first sequence from the nucleotide sequence encoding a first saccharide transporter or a sequence related to or derived therefrom and a second sequence from the nucleotide sequence encoding a different second saccharide transporter or a sequence related to or derived therefrom, wherein said saccharide is preferably glucose.
6. A yeast as claimed in 5 wherein said chimeric construct comprises a sequence having the form
AB
wherein
A is said first sequence comprising nucleotide bases w to x;
B is said second sequence comprising nucleotide bases (xy) to z;
wherein A and B are derived from the nucleotide sequences of said different saccharide transporters or from sequences related to or derived therefrom;
w is a first position within the nucleotide sequence from which said first sequence is derived, which is less than x, preferably 1 to 50;
x is a second position within the nucleotide sequence from which said first sequence is derived and is greater than w and is preferably a number from 500 to 850;
y is an integer, preferably less than 3; xy 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 xy, preferably from 1600 to 1713;
wherein said values refer to the position within the nucleotide sequence of said saccharide transporter, e.g. HXT1 or HXT7 or a comparable sequence thereof.
7. A yeast as claimed in claim 5 or 6 wherein said first saccharide transporter is a high affinity saccharide transporter of the yeast Saccharomyces and said second saccharide transporter is a low affinity saccharide transporter of the yeast Saccharomyces.
8. A yeast as claimed in any one of claims 5 to 7 wherein said transporters are selected from the list consisting of: HXT1 to HXT17, GAL2, SNF3, RGT2, AGT1, YDL247w and YJR160c, preferably from Saccharomyces cerevisiae, or sequences related to or derived from the sequences thereof.
9. A yeast as claimed in any one of claims 5 to 8 wherein the chimeric construct comprises
one or more sequences or portions thereof of the sequences encoding any one of HXT1 to HXT17, GAL2, SNF3, RGT2, AGT1, YDL247w and YJR160c, preferably from Saccharomyces cerevisiae,
or a sequence which hybridizes to said sequence or portion thereof under non-stringent binding conditions of 6SSC50% formamide at room temperature and washing under conditions of high stringency, e.g. 2SSC, 65 C., where SSC0.15 M NaCl, 0.015M sodium citrate, pH 7.2, or a sequence which exhibits at least 95% sequence identity to said sequence or portion thereof,
or a sequence complementary to any of the aforesaid sequences,
which sequence or portion thereof may comprise a first or second sequence as defined in any one of claims 5 to 8.
10. A yeast as claimed in any one of claims 5 to 8 wherein the chimeric construct comprises a sequence which encodes an amino acid sequence which comprises
one or more sequences or portions thereof of the sequences of any one of HXT1 to HXT17, GAL2, SNF3, RGT2, AGT1, YDL247w and YJR160c, preferably from Saccharomyces cerevisiae,
or a sequence which exhibits at least 95% sequence identity to said sequence or portion thereof,
which sequence or portion thereof is encoded by a nucleotide sequence which may comprise a first or second sequence as defined in any one of claims 5 to 8.
11. A yeast as claimed in any one of claims 7 to 10 wherein said low affinity transporter is HXT1 or 3 and said high affinity transporter is HXT2, 4, 6, 7 or GAL2P.
12. A yeast as claimed in any one of claims 5 to 11 wherein said construct comprises sequences from, or related to or derived from HXT 1 and HXT7.
13. A yeast as claimed in any one of claims 6 to 12 wherein x is a position in the nucleotide sequence encompassed by the third to ninth transmembrane stretches of HXT1 or HXT7 or a comparable sequence, preferably x525-575 andor 720-770.
14. A yeast as claimed in any one of claims 6 to 13 wherein x551 or 741, w and y1 and z1713.
15. A yeast as claimed in any one of claims 5 to 14 wherein said chimeric construct encodes a chimeric polypeptide having a total of 10 to 14 transmembrane domains.
16. A yeast as claimed in any one of claims 5 to 15 wherein said chimeric construct contains 2 or more sequences from the nucleotide sequence of at least one transporter molecule or a sequence related to or derived therefrom or contains sequences from the nucleotide sequences of more than 2 transporters or the sequences related to or derived therefrom.
17. A yeast as claimed in any one of claims 5 to 16 wherein the yeast cell into which said chimeric construct is introduced is a null strain without saccharide transporting properties.
18. A yeast as claimed in any one of claims 5 to 17 wherein said chimeric construct comprises the nucleotide sequence encoding Sequence ID Nos. 1 or 2 or comprises the nucleotide sequence of Sequences ID Nos. 3 or 4 or a functional equivalent of said sequences.
19. A yeast as claimed in claim 18 denoted Saccharomyces cerevisiae KOY.TM4P or KOY.TM6P having the deposition number DSM (Deutsche Sammlung Mikroorganismen) 13832 or DSM 13555, respectively or a functional equivalent thereof.
20. A nucleic acid molecule comprising a chimeric nucleotide sequence as defined in any one of claims 5 to 19 or a complementary sequence thereto.
21. A vector or host cell comprising a nucleic acid molecule as defined in claim 20.
22. A polypeptide comprising a sequence encoded by a nucleic acid molecule as defined in claim 20 or a functionally equivalent protein thereof.
23. A method of inserting exogenous genetic material into a yeast cell as defined in any one of claims 1 to 19 to provide a yeast cell which produces a product, said method comprising at least the steps of introducing said material, preferably contained within a vector, into said cell, wherein said exogenous material encodes said product or portion thereof or encodes a polypeptide or part thereof which facilitates the direct or indirect production of said product or portion thereof, or which affects the expression of said polypeptide or product.
24. A method as claimed in claim 23 wherein said product is a high or low molecular metabolite, a bulk or fine chemical, a food stuff, a functional food or a therapeutic agent.
25. A method as claimed in claim 25 wherein said product is an amino acid, a peptide, a polypeptide, a sugar, a small polyol or CO2.
26. A yeast cell obtainable by the method of any one of claims 23 to 25.
27. A method of preparing a product, comprising growing yeast cells as defined in any one of claims 1 to 19 or 26 under aerobic conditions in the presence of high saccharide, preferably glucose, concentrations.
28. A method as claimed in claim 27, additionally comprising the step of isolating the product thus formed.
29. A product obtained by the method of claim 28.
30. A method or product as claimed in any one of claims 27 to 29 wherein said product is a low alcohol or non-alcoholic beverage.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A charging system in a packet switched network for charging packets differently dependent on which service flow the packets belong to, the charging system comprising:
a control system and a serving element associated with a packet forwarding system wherein said control system comprises:
an account function for managing an account of at least one user, and
a charging policy decision point for calculating a different charging policy for each of a plurality of allowed services for the at least one user,
wherein said serving element comprises:
a token bucket associated with the at least one user, said token bucket for storing reservations for a plurality of service flows received from the account function of the at least one user, and
a charging policy enforcement point for performing charging for the plurality of service flows by reducing the stored reservation of the token bucket according to the calculated charging policies.
2. The charging system according to claim 1 wherein the charging policy enforcement point includes means for calculating the charging policies based on historical andor current user specific usage data.
3. The charging system according to claim 1 wherein the serving element comprises means for classifying the services into different service classes based on the tariff plan of the services.
4. The charging system according to claim 3 wherein the allowed subscriber service classes are stored in a Service Class Vector.
5. The charging system according to claim 1 wherein the packet forwarding system is a Gateway GPRS Support Node in a mobile telecommunication network.
6. A charging system in a packet switched network for charging packets differently dependent on which service flow the packets belong to, the charging system comprising:
a control system and a serving element associated with a packet forwarding system, wherein said control system comprises:
an account function adapted to manage an account of at least one user, and
a charging policy decision point arranged to calculate a charging policy for allowed services for the at least one user, wherein the calculated charging policy comprises at least one user rating table and a set of validity conditions,
wherein said serving element comprises:
a token bucket adapted to store reservations received from the account function of a user associated with the token bucket, and
a charging policy enforcement point arranged to perform charging for a plurality of the allowed services by reducing the stored reservation of the token bucket according to the calculated charging policy.
7. The charging system according to claim 6 wherein the set of validity conditions defines the lifetime of the at least one user rating table.
8. The charging system according to claim 6 wherein the charging policy enforcement point includes at least two user rating tables having different time validity conditions.
9. A charging system in a packet switched network for charging packets differently dependent on which service flow the packets belong to, the charging system comprising:
a control system and a serving element associated with a packet forwarding system, wherein said control system comprises:
an account function adapted to manage an account of at least one user, and
a charging policy decision point arranged to calculate a charging policy for allowed services for the at least one user,
wherein said serving element comprises:
a token bucket adapted to store reservations received from the account function of a user associated with the token bucket,
a charging policy enforcement point arranged to perform charging for a plurality of the allowed services by reducing the stored reservation of the token bucket according to the calculated charging policy, and
means for classifying the services into different service classes, said classifying means including a service filter adapted to identify the different service flows.
10. The charging system according to claim 9 wherein the means for classifying the services into different service classes further comprises Protocol Inspection Filters adapted to identify the different service flows, when the service filter is not capable of said identification.
11. A control system of a charging system in a packet switched network, said control system comprising:
an account function for managing an account of at least one user; and
a charging policy decision point for calculating charging policies for a plurality of allowed services for the at least one user, wherein the charging policies are utilized to calculate charges for a plurality of service flows using a single token bucket.
12. The control system according to claim 11 wherein the charging policies are calculated based on historical andor current user specific usage data.
13. The control system according to claim 11 wherein the control system is implemented in a mobile telecommunication network.
14. A control system of a charging system in a packet switched network, said control system comprising:
an account function for managing an account of at least one user; and
a charging policy decision point for calculating a charging policy for the allowed services for the at least one user;
wherein the calculated charging policy includes:
at least one user rating table, and
a set of validity conditions.
15. The control system according to claim 14 wherein the set of validity conditions defines the lifetime of the at least one user rating table.
16. The control system according to claim 14 wherein the calculated charging policy comprises at least two user rating tables having different time validity conditions.
17. A serving element residing in a packet forwarding system of a charging system in a packet switched network, said serving element comprising:
means for receiving reservations for at least one user,
a token bucket for storing the reservations for the user associated with the token bucket,
means for receiving a charging policy for allowed services, and
a charging policy enforcement point for charging for a plurality of the allowed services by reducing the stored reservation of the token bucket according to a received calculated charging policy.
18. The serving element according to claim 17 further comprising a single token bucket per user.
19. The serving element according to claim 17 further comprising means for classifying the services into different service classes based on the tariff plan of the services.
20. The serving element according to claim 19 wherein the allowed subscriber service classes are stored in a Service Class Vector.
21. The serving element according to claim 19 wherein the packet forwarding system is a Gateway GPRS Support Node in a mobile telecommunication network.
22. A serving element residing in a packet forwarding system of a charging system in a packet switched network, said serving element comprising:
means for receiving reservations for at least one user,
a token bucket for storing the reservations for the user associated with the token bucket,
means for receiving a charging policy for allowed services, wherein the charging policy includes at least one user rating table and a set of validity conditions, and
a charging policy enforcement point for charging for a plurality of the allowed services by reducing the stored reservation of the token bucket according to a received calculated charging policy.
23. The serving element according to claim 22 wherein the charging policy is calculated based on historical andor current user specific usage data.
24. The serving element according to claim 22 wherein the set of validity conditions defines the lifetime of the at least one user rating table.
25. The serving element according to claim 22 wherein the charging policy comprises at least two user rating tables having different time validity conditions.
26. A serving element residing in a packet forwarding system of a charging system in a packet switched network, said serving element comprising:
means for receiving reservations for at least one user,
a token bucket for storing the reservations for the user associated with the token bucket,
means for classifying the services into different service classes based on the tariff plan of the services, wherein the means for classifying the services into different service classes includes a service filter adapted to identify different service flows,
means for receiving a charging policy for allowed services, and
a charging policy enforcement point for charging for a plurality of the allowed services by reducing the stored reservation of the token bucket according to a received calculated charging policy.
27. The serving element according to claim 26 wherein the means for classifying the services into different service classes further comprises Protocol Inspection Filters adapted to identify the different service flows, when the service filter is not capable of said identification.