Evaluation of the antiobesity effects of an amino acid mixture and conjugated linoleic acid on exercising healthy overweight humans: a randomized, double-blind, placebo-controlled trial.

This single-centre, randomized, double-blind, placebo-controlled trial investigated the effects of administering a mixture of four amino acids (lysine, proline, alanine and arginine) with or without conjugated linoleic acid to healthy overweight humans before and after exercising. Forty-one healthy subjects (body mass index >or= 23 to < 30 kg/m(2)) completed the study following randomization to receive either placebo or one of three test supplements: amino acid mixture 0.76 g/day; amino acid mixture 1.52 g/day; or amino acid mixture 1.52 g/day coadministered with conjugated linoleic acid 1.6 g/day. Each of the study treatments was administered 30 min before and immediately after a period of daily exercise, which was delivered by an exercise expert, for a period of 12 weeks. When compared with the placebo group, several indicators, such as waist and hip circumferences, were found to have significantly decreased in the test supplement groups compared with the placebo. These results suggest that ingestion of these supplements might enhance the fat-burning effects of exercise.

Analysis of essential histidine residues of maize branching enzymes by chemical modification and site-directed mutagenesis.

Incubation of maize branching enzyme, mBEI and mBEII, with 100 microM diethylpyrocarbonate (DEPC) rapidly inactivated the enzymes. Treatment of the DEPC-inactivated enzymes with 100500 mM hydroxylamine restored the enzyme activities. Spectroscopic data indicated that the inactivation of BE with DEPC was the result of histidine modification. The addition of the substrate amylose or amylopectin retarded the enzyme inactivation by DEPC, suggesting that the histidine residues are important for substrate binding. In maize BEII, conserved histidine residues are in catalytic regions 1 (His320) and 4 (His508). His320 and His508 were individually replaced by Ala via site-directed mutagenesis to probe their role in catalysis. Expression of these mutants in E. coli showed a significant decrease of the activity and the mutant enzymes had Km values 10 times higher than the wild type. Therefore, residues His320 and His508 do play an important role in substrate binding.