Maximum energy is liberated by the hydrolysis of

Step 8 of Glycolysis - i. 2-phosphoglycerate is conveed to phosphoenol pyruvate by the enzyme enolase. One water molecule is removed. ii. A high energy phosphate bond is produced,so when it is hydrolysed max.energy is liberated. The reaction is reversible. iii. Enolase requires Mg++, and by removing magnesium ions, fluoride will irreversibly inhibit this enzyme. Thus, fluoride will stop the whole glycolysis. So when taking blood for sugar estimation, fluoride is added to blood. If not, glucose is metabolized by the blood cells, so that lower blood glucose values are obtained. Ref:DM VASUDEVAN TEXTBOOK OF BIOCHEMISTRY,EIGHTH EDITION,PG.NO.,131.