Open AccessHydrolysis and Isomerization of trans,trans ‐Farnesyl Diphosphate by Andrographis Tissue‐Culture Enzymes
Author(s) -
MACKIE Hugh,
OVERTON Karl H.
Publication year - 1977
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1977.tb11646.x
Subject(s) - farnesol , chemistry , phosphatase , hydrolysis , stereochemistry , enzyme , biochemistry
1. Incubation of (3 R ,5 S )‐[5‐ 3 H 1 ]mevalonate + (3 RS )‐[2‐ 14 C]mevalonate with Andrographis cell‐free extract leads to trans,trans ‐farnesol and cis,trans ‐farnesol which both totally retain tritium. 2. This conflicts with our previous results which predict one third tritium loss in the cis,trans ‐farnesol. Inversion at C‐1 during hydrolysis of trans,trans ‐farnesyl diphosphate to trans,trans ‐farnesol could explain this anomaly. 3. (1 S )‐ trans,trans ‐[1‐ 3 H 1 ]Farnesyl diphosphate and phosphate and (1 R )‐ trans,trans ‐[1‐ 3 H 1 ]‐farnesyl diphosphate and phosphate, all prepared chemically, were hydrolysed with Andrographis phosphatase, and alkaline phosphatase and hydrogenolysed with lithium aluminium hydride and the product alcohols exchanged with liver alcohol hydrogenase. 4. Both Andrographis phosphatase and alkaline phosphatase hydrolyse trans,trans ‐farnesyl diphosphate and trans,trans ‐farnesyl phosphate with retention. 5. Hydrolysis of trans,trans ‐[1‐ 18 O]farnesyl diphosphate in H 2 18 O with both phosphatases supports P‐O fission. 6. The C‐1 configuration in (1 S )‐ trans,trans ‐[1‐ 3 H 1 ]farnesyl diphosphate and phosphate and (1 R )‐ trans,trans ‐[1‐ 3 H 1 ]farnesyl diphosphate and phosphate is progressively racemised in 0.01 M NH 4 OH/MeOH (1/9) at –20°C.