Benanomicin A

Cell wall mannan of the pathogenic yeast Candida krusei was prepared using the antibiotic Benanomicin A, which has a lectin-like function. The chemical structure of this molecule was found to be similar to that of mannan prepared from the same yeast by the conventional method using Fehling reagent. Only a few degradation products were detected when the mannan prepared using Fehling reagent was subjected to alkali treatment (β-elimination), but multiple α-1,2-linked oligosaccharides were detected when the mannan purified with Benanomicin A was treated with alkali. These results indicate that most of the O-linked sugar chains in mannan were lost under conventional conditions when exposed to the strongly alkaline Fehling reagent. In contrast, the O-glycosidic bond in mannan was not cleaved and the O-linked sugar chains were maintained and almost intact following treatment with the mild novel preparation method using Benanomicin A. Therefore, we argue that the new mannan preparation method using Benanomicin A is superior to conventional methods. In addition, our study suggests that some yeast mannans, whose overall structure has already been reported, may contain more O-linked sugar chains than previously recognized.

The effects of benanomicin A, a mannose-binding antifungal antibiotic, on yeast cells of Saccharomyces cerevisiae were studied by electron microscopy. Cytological studies using vital stain with methylene blue demonstrated that benanomicin A at 20 and 80 microg/ml killed buds in preference to parent cells. In confirmation, examination by TEM revealed that benanomicin A at 80 microg/ml damaged buds more severely than parent cells. The major effect on the ultrastructure was characterized by severe damage to the cell membrane. In addition, it caused expansion and vacuolation of the endoplasmic reticulum (ER), and partial fragmentation and disappearance of nuclear membranes. The membrane-disruptive activity of benanomicin A may be closely associated with its membrane affinity.

The key intermediate tri-substituted alpha-tetralone (8) has been synthesized, either via tandem Michael addition-Dieckmann condensation reaction between dienolate and methyl crotonate in a low yield or via Barton's radical decarboxylation of diester (9) without 4-dimethylaminopyridine in 75% yield, and applied to the synthesis of the substituted 5,6-dihydrobenzo[a]naphthacenequinone.