Phomopsin A

Phomopsis leptostromiformis WA1515 produced 75 to 150 mg of phomopsin A per liter in stationary cultures in a Czapek-Dox medium supplemented with 5 to 10 g of yeast extract per liter. pH and temperature optima were approximately 6.0 and 25 degrees C, respectively. A commercial tryptic digest of casein was a satisfactory alternative to the yeast extract, but poor growth and very little phomopsin were obtained when the yeast was replaced by vitamin-free Casamino Acids or a mixture of 18 amino acids. Approximately 95% of the phomopsin A produced was found in the cutlure liquid. No phomopsin was detected in shaken cultures. No phomopsin B was found in any culture. Methods are described for recovery and estimation of phomopsin A from culture liquids.

Phomopsin A (PHO-A), produced by the fungus Diaporthe toxica, is a mycotoxin known to be responsible for fatal liver disease of lupin-fed sheep. The full spectrum of the toxic secondary metabolites produced by D. toxica is still unknown. PHO-A and the naturally occurring derivatives B-E have been subject to several studies to reveal their structures as well as chemical and toxicological properties. In this work, a methylated derivative (1) of PHO-A isolated from lupin seeds inoculated with D. toxica is described. It was characterized by high-resolution mass and NMR data and shown to be the N-methylated derivative of PHO-A. 1 is cytotoxic against HepG2 cells.

Fungi of Aspergillus and Penicillium genus can infect peas (Pisum sativum), leading to a contamination with the nephrotoxic and carcinogenic ochratoxin A (OTA). Under unfavourable conditions, a fungus primarily found on lupines, Diapothe toxica, may also grow on peas and produce the hepatotoxic phomopsin A (PHOA). To study the effect of processing on OTA and PHOA content, two model products-wheat/rye-mixed bread with pea flour addition and pea pasta-were manufactured at small-business scale from artificially contaminated pea flour. The decrease of OTA and PHOA contents were monitored along the production process as indicators for toxin transformation. Pea bread dough was subjected to proofing for 30-40 min at 32 °C and baked at 250 °C to 230 °C for 40 min. OTA content (LODs < 0.1 μg/kg) showed a reduction in the bread crust (initially 17.0 μg/kg) to 88% and no reduction in the crumb (110%). For PHOA (LODs < 3.6 μg/kg), a decrease to approximately 21% occurred in the bread crust (initially 12.5 μg/kg), whilst for crumb, a less intense decrease to 91% was found. Pea pasta prepared with two toxin levels was extruded at room temperature, dried and cooked for 8 min in boiling water. In pea pasta, OTA was reduced from 29.8 to 13.9 μg/kg by 22% each after cooking, whilst 15% and 10% of the initial toxin amounts were found in the cooking water, respectively. For PHOA, 60% and 78% of initially 14.3 μg/kg and 7.21 μg/kg remained in the cooked pasta. As only the decrease of the initial content was measured and no specific degradation products could be detected, further research is needed to characterise potential transformation products. Heat treatment reduces the initial PHOA content stronger than the OTA content during pasta cooking and bread making. However, significant amounts of both toxins would remain in the final products.