Chemistry: molecular biology and microbiology – Treatment of micro-organisms or enzymes with electrical or... – Modification of viruses
Patent
1993-02-01
1996-04-23
Moody, Patricia R.
Chemistry: molecular biology and microbiology
Treatment of micro-organisms or enzymes with electrical or...
Modification of viruses
4351721, 435 691, 435 701, 536 236, 536 245, 800205, 800DIG69, C12N 1529, C12N 1582, C12N 1510, C12N 1552
Patent
active
055102557
DESCRIPTION:
BRIEF SUMMARY
FIELD OF INVENTION
The present invention is directed to synthase enzymes relevant to fatty acid synthesis in plants, protein preparations, amino acid and nucleic acid sciences related thereto, and methods of use for such compositions.
INTRODUCTION
BACKGROUND
Plant oils are used in a variety of industrial and edible uses. Novel vegetable oils compositions and/or improved means to obtain oils compositions, from biosynthetic or natural plant sources, are needed. Depending upon the intended oil use, various different fatty acid compositions are desired.
For example, in some instances having an oilseed with a higher ratio of oil to seed meal would be useful to obtain a desired oil at lower cost. This would be typical of a high value oil product. In some instances, having an oilseed with a lower ratio of oil to seed meal would be useful to lower caloric content. In other uses, edible plant oils with a higher percentage of unsaturated fatty acids are desired for cardio-vascular health reasons. And alternatively, temperate substitutes for high saturate tropical oils such as palm and coconut, would also find uses in a variety of industrial and food applications.
One means postulated to obtain such oils and/or modified fatty acid compositions is through the genetic engineering of plants. However, in order to genetically engineer plants one must have in place the means to transfer genetic material to the plant in a stable and heritable manner. Additionally, one must have nucleic acid sequences capable of producing the desired phenotypic result, regulatory regions capable of directing the correct application of such sequences, and the like. Moreover, it should be appreciated that in order to produce a desired phenotype requires that the Fatty Acid Synthetase (FAS) pathway of the plant is modified to the extent that the ratios of reactants are modulated or changed.
Higher plants appear to synthesize fatty acids via a common metabolic pathway. In developing seeds, where fatty acids attached to triglycerides are stored as a source of energy for further germination, the FAS pathway is located in the proplastids. The first step is the formation of acetyl-ACP (acyl carrier protein) from acety-CoA and ACP catalyzed by the enzyme, acetyl-CoA:ACP transacylase (ATA). Elongation of acetyl-ACP to 16- and 18-carbon fatty acids involves the cyclical action of the following sequence of reactions: condensation with a two-carbon unit from malonyl-ACP to form a .beta.-ketoacyl-ACP (.beta.-ketoayl-ACP synthase), reduction of the keto-function to an alcohol (.beta.-ketoacyl-ACP reductase), dehydration to form an enoyl-ACP (.beta.-hydroxyacyl-ACP dehydrase), and finally reduction of the enoyl-ACP to form the elongated saturated acyl-ACP (enoyl-ACP reductase). .beta.-ketoacyl-ACP synthase I, catalyzes elongation up to palmitoyl-ACP (C16:0), whereas .beta.-ketoacyl-ACP synthase II catalyzes the final elongation to stearoyl-ACP (C18:0). Common plant unsaturated fatty acids, such as oleic, linoleic and .alpha.-linolenic acids found in storage triglycerides, originate from the desaturation of stearoyl-ACP to form oleoyl-ACP (C18:1) in a reaction catalyzed by a soluble plastid .DELTA.-9 desaturase (also often referred to as "stearoyl-ACP desaturase"). Molecular oxygen is required for desaturation in which reduced ferredoxin serves as an electron co-donor. Additional desaturation is effected sequentially by the actions of membrane bound .DELTA.-12 desaturase and .DELTA.-15 desaturase. These "desaturases" thus create mono- or polyunsaturated fatty acids respectively.
A third .beta.-ketoacyl-ACP synthase has been reported in S. oleracea leaves having activity specific toward very short acyl-ACPs. This acetoacyl-ACP synthase or ".beta.-ketoacyl-ACP" synthase III has a preference to acetyl-CoA over acetyl-ACP, Jaworski, J. G., et al., Plant Phys. (1989) 90:41-44. It has been postulated that this enzyme may be an alternate pathway to begin FAS, instead of ATA.
Obtaining nucleic acid sequences capable of producing a phenotypic result in FAS, de
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