Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or... – The polynucleotide alters fat – fatty oil – ester-type wax – or...
Patent
1995-06-09
1998-10-20
Fox, David T.
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
The polynucleotide alters fat, fatty oil, ester-type wax, or...
800DIG42, 435 691, 435101, 4351723, 435194, 4352523, 4352542, 4353201, 435325, 435366, 435419, 536 232, 536 236, 426438, A01H 500, C12N 1529, C12N 1554, C12N 1582
Patent
active
058248625
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
This invention relates to a DNA encoding ATP-dependent fructose 6-phosphate 1-phosphotransferase (EC 2.7.1.11) (hereinafter referred to as "PFK"), a recombinant vector containing the same and a method for changing sugar content in plant cells under low temperature using the recombinant vector.
BACKGROUND ART
PFK is an enzyme which catalyzes the phosphorylation of fructose 6-phosphate (F6P) to fructose 1,6-bisphosphate, a key regulatory step in the glycolytic pathway.
It is well-known that when plant tissues are exposed to a low temperature, the content of sugars such as sucrose, glucose, fructose and the like is generally increased. For example, this phenomenon is observed in potato tubers. Accumulation of reducing sugars such as glucose and fructose in potato tubers during storage at low temperature is undesirable in industry because they cause excess browning of potato chips during processing. As for the cause of the accumulation of the reducing sugars, there are various hypotheses. However, it is thought that the activity of the glycolytic pathway is greatly reduced at low temperatures, so that the flow of the breakdown products of starch to sucrose is increased, thereby increasing the accumulation of the reducing sugars. PFK is thought to be one of the key enzymes of the glycolytic pathway, and it is well-known that PFK is cold labile. Therefore, it is thought that the drastic reduction of the activity of the glycolytic pathway at low temperature is caused by the drastic reduction of the activity of PFK.
PFK genes have been isolated from prokaryotes such as Escherichia coli, thermophilic bacteria, Bacillus subtilis and mycoplasma, and from eukaryotic tissues such as human muscle, human liver, rabbit muscle and mouse liver. However, the isolation of a plant PFK gene has not been reported, and the isolation of a gene encoding cold stable PFK has also not been reported. Therefore, it has been difficult hitherto to develop a potato variety resistant to Cold Induced Sweetening by introducing PFK genes. Nor has it been possible to reduce the activity of the glycolytic pathway in plant tissues by expressing an anti-sense PFK gene, thereby developing a plant having a high sugar content, which has a new taste.
European Patent Publication 0 438 904 (Japanese Laid-open Patent Application (Kokai) No. 4-341126) discloses an example of introducing a PFK gene into a plant. In the invention described in this publication, the E. coli PFK gene is expressed in potato and rice and it is shown that the amount of several intermediates of the pathways of the carbohydrate metabolism are changed. In particular, it is shown that the sucrose content in the tubers immediately after harvest is significantly decreased. However, in the invention of the publication, the amount of glucose and fructose in the tubers during storage at low temperature, which is an industrial problem, is not mentioned. In view of the fact that E. coli PFK is an enzyme which is cold labile (Kruger, N. J. (1989) Biochemical Society Transaction 629th Meeting, London Vol. 17, 760-761), it is expected that a potato variety resistant to Cold Induced Sweetening cannot be obtained by introducing the E. coli PFK gene into potato and expressing it in the tubers. To attain this objective, a gene encoding PFK which is cold stable is necessary. However, a gene for a cold stable PFK has not yet been isolated.
In order to store potato tubers for a long time, storage at low temperature after harvest is very important for suppressing diseases, sprouting and aging.
However, when tubers stored at low temperature are directly used to make potato chips or French fries, browning of the products occurs during processing and the product values are greatly decreased (especially in potato chips). It is known that this browning is caused by the Maillard reaction between amino acids and reducing sugars contained in potato tubers, which occurs during processing in a hot cooking oil (Schallenberger, R. S. et al., (1959) J. Agric. Fd Chem., 7, 274). It is known that
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Hiyoshi Toru
Kasaoka Keisuke
Mine Toshiki
Page Anthony Miles John
Tyson Robert Huw
Fox David T.
Japan Tobacco Inc.
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