No abstract

Metabolic engineering to enhance production of isoprenoid metabolites for industrial and medical purposes is an important goal. The substrate for isoprenoid synthesis in plants is produced by the mevalonate pathway (MEV) in the cytosol and by the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway in plastids. A multi-gene approach was employed to insert the entire cytosolic MEV pathway into the tobacco chloroplast genome. Molecular analysis confirmed the site-specific insertion of seven transgenes and homoplasmy. Functionality was demonstrated by unimpeded growth on fosmidomycin, which specifically inhibits the MEP pathway. Transplastomic plants containing the MEV pathway genes accumulated higher levels of mevalonate, carotenoids, squalene, sterols, and triacyglycerols than control plants. This is the first time an entire eukaryotic pathway with six enzymes has been transplastomically expressed in plants. Thus, we have developed an important tool to redirect metabolic fluxes in the isoprenoid biosynthesis pathway and a viable multigene strategy for engineering metabolism in plants.

Cereal Chem. 77(4):518-521

Cereal Chem. 80(3):260-263The in vitro bile acid binding by rice bran, oat bran, dehulled barley, and β-glucan enriched barley was determined using a mixture of bile acids at a duodenal physiological pH of 6.3. Six treatments and two blank incubations were conducted testing substrates on an equal protein basis. The relative in vitro bile acid binding of the cereal brans on an equal total dietary fiber (TDF) and insoluble dietary fiber (IDF) basis considering cholestyramine as 100% bound was rice bran 45 and 49%; oat bran 23 and 30%; dehulled barley 33 and 57%; and β-glucan enriched barley 20 and 40%, respectively. Bile acid bindings on equal protein basis for the respective cereals were 68, 26, 41, and 49%. Bile acid binding by rice bran may account to a great extent for its cholesterol-lowering properties, while bile acid binding by oat bran suggests that the primary mechanism of cholesterol lowering by oat bran is not due to the bile acid binding by its soluble fiber. Bile acid binding was not proportional to the soluble fiber content of the cereal brans tested. Except for dehulled barley, bile acid binding for rice bran, oat bran, and β-glucan enriched barley appear to be related to their IDF content. Highest relative bile acid binding values for rice bran and β-glucan enriched barley were observed on an equal protein basis, whereas highest values for dehulled barley were based on IDF. Data suggest that of all four cereals tested, bile acid binding may be related to IDF or protein anionic, cationic, physical and chemical structure, composition, metabolites, or their interaction with active binding sites.