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The first was reliable nuclear magnetic resonance NMR spectrometers that allowed chemists to detect carbon This isotope of carbon is present naturally in minute quantities — 1. Like hydrogen, carbon has a nuclear spin of one-half, so it can be detected by NMR. This meant that the chemists no longer had to break down the final products in order to determine where in the molecule the labelled atoms were.

With this powerful new tool, Battersby and others made great strides in working out the pathways to the pigments of life. Growing this organism in a medium free of cobalt gave the three intermediates called precorrins 1, 2 and 3, according to how many methyl groups were added at each step.

Researchers were held up at this point for some eight years. Something else was needed. The answer was genetic engineering. InBattersby teamed up with Crouzet and Blanche and their colleagues. The French team had already detected and sequenced the genes that code for the various enzymes that synthesise B12 in P. Used together with spectroscopy, intermediates beyond precorrin-3 were produced.

Inthe French team isolated precorrin-6x, a compound several steps on from precorrin To their surprise they found that ring shrinking, which chemists had believed happened later on in the pathway, had already occurred. Later came a series of steps, all triggered by one enzyme, that led to precorrin 8x.

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The final step — leading to the corrin ring system of vitamin B12 — involved the transfer of the methyl group on carbon atom 11 to its neighbour on carbon atom This left them with another piece to fit into the puzzle: what had happened to the methyl group at carbon in pre-corrin-3? It was not there in precorrin-6x. The answer came earlier this year, when yet another intermediate, precorrin-4, was isolated by the French team; it had been converted to an acetyl CH 3 CO- group attached to carbon 1.

If anaerobic, cobalt appears earlier. Like any good mountaineer, Battersby relishes all of the milestones that he has passed in his 25 years of work on B Plenty of midnight oil has been burned en route, and long weekends spent waiting patiently for microorganisms, enzymes and NMR spectrometers to do their work. Perhaps the milestone that gave him the most satisfaction, and caused the greatest sensation, was the French discovery of precorrin-6x. For example, we used to think that ring contraction to give the corrin system occurred quite late in the pathway.

That had to go. Ring contraction is now seen as an early step. Battersby is at pains to emphasise the importance of the cross-disciplinary collaboration that has brought all these fresh insights since This in turn has enabled chemists to detect the new biosynthetic intermediates, such as precorrin-6x.

He is already looking forward to more detailed work on precorrin-4 and the discovery of the elusive precorrin Like Everest of 40 years ago, the assault on B12 is nearly over. Lionel Milgrom is a lecturer in inorganic chemistry at Brunel University and a freelance science journalist.

The anatomy of all naturally occurring tetrapyrroles is essentially the same. Haem, for example, consists of four pyrrole-type rings joined by carbon bridges to form a larger ring system called a macrocycle which consists of 20 carbon atoms and 4 nitrogen atoms. The easiest way to remember how the carbons are numbered is to remember the bridging carbons.

The topmost is carbon atom 5 C-5the right-hand one is C, the bottom one is C, and the left-hand one is C The rest follow. The way other groups are arranged on the macrocycle shows the family relationship between the pigments of life.

The outermost groups also fine-tune the properties of macrocycles. In chlorophyll, modification of one of the side chains helps to stabilise the reduced chlorin ring in the presence of oxygen chlo-rins without such rings are easy to oxidise.

The crowding of groups around the shrunken cobalt corrin ring of vitamin B12 allows nature to have its chemically reactive cake and eat it. The cobalt-carbon bond of B12 is easily broken to give a radical — a molecule with one unpaired electron.

Such radicals are usually highly reactive, but with high reactivity usually comes indiscrimination. By fencing off the reactive centre of B12 with lots of groups, the molecule keeps its high reactivity and specificity.

This control is the envy of human chemists. To reproduce the range of reactions of the lowliest microbe would need a chemical works the size of Britain. The route to vitamin B12 begins with 5-aminolaevulinic acid, ALA. In animals, ALA is synthesised from the amino acid glycine and succinic acid, that has been made more reactive by joining to coenzyme A or CoA, for short. The next three steps are the same for all the pigments of life. Two molecules of ALA are joined together to give a pyrrole — a five-membered ring consisting of four carbon atoms and one nitrogen atom — called porphobilinogen or PBG.

This reaction is similar to that used by chemists to make pyrroles in the laboratory. Something strange happens during this step. One of the PBG units is turned round so that the order of two large groups is reversed.