Novel Key Metabolites Reveal Further Branching of the Roquefortine/Meleagrin Biosynthetic Pathway

Metabolic profiling and structural elucidation of novel secondary metabolites obtained from derived deletion strains of the filamentous fungus Penicillium chrysogenum were used to reassign various previously ascribed synthetase genes of the roquefortine/meleagrin pathway to their corresponding products. Next to the structural characterization of roquefortine F and neoxaline, which are for the first time reported for P. chrysogenum, we identified the novel metabolite roquefortine L, including its degradation products, harboring remarkable chemical structures. Their biosynthesis is discussed, questioning the exclusive role of glandicoline A as key intermediate in the pathway. The results reveal that further enzymes of this pathway are rather unspecific and catalyze more than one reaction, leading to excessive branching in the pathway with meleagrin and neoxaline as end products of two branches.     

A Branched Biosynthetic Pathway Is Involved in Production of Roquefortine and Related Compounds in Penicillium chrysogenum

Profiling and structural elucidation of secondary metabolites produced by the filamentous fungus Penicillium chrysogenum and derived deletion strains were used to identify the various metabolites and enzymatic steps belonging to the roquefortine/meleagrin pathway. Major abundant metabolites of this pathway were identified as histidyltryptophanyldiketopiperazine (HTD), dehydrohistidyltryptophanyldi-ketopiperazine (DHTD), roquefortine D, roquefortine C, glandicoline A, glandicoline B and meleagrin. Specific genes could be assigned to each enzymatic reaction step. The nonribosomal peptide synthetase RoqA accepts L-histidine and L-tryptophan as substrates leading to the production of the diketopiperazine HTD. DHTD, previously suggested to be a degradation product of roquefortine C, was found to be derived from HTD involving the cytochrome P450 oxidoreductase RoqR. The dimethylallyltryptophan synthetase RoqD prenylates both HTD and DHTD yielding directly the products roquefortine D and roquefortine C without the synthesis of a previously suggested intermediate and the involvement of RoqM. This leads to a branch in the otherwise linear pathway. Roquefortine C is subsequently converted into glandicoline B with glandicoline A as intermediates, involving two monooxygenases (RoqM and RoqO) which were mixed up in an earlier attempt to elucidate the biosynthetic pathway. Eventually, meleagrin is produced from glandicoline B involving a methyltransferase (RoqN). It is concluded that roquefortine C and meleagrin are derived from a branched biosynthetic pathway.     

Sustainable Utilization and Economic Development of Some Plant Resources in Northern Pakistan

Overshadowed by the vast impact that habitat loss has on targeted plant species survival, it is easy to overlook the fact that utilization and harvesting of species can often result in their genetic depletion and in an often rapid downward trend of their population status. In this regard an ethnobotanical study on the medicinal plants of Shawar valley, District Swat was conducted with the aims to document medicinal plants and to examine the current status of the medicinal plants trade as well as the market chain starting from collectors to consumers. The survey reported 50 species of plants belonging to 33 families as ethnomedicinally important. These species were used as crud drugs by the local people and folk-lore for treating diseases in traditional system of medicine. The detailed local uses, recipe preparation along with their local names and diseases treated were recorded for each species.Market survey revealed that the structure of medicinal plant trade is complex and heterogeneous, involving many players. The collectors are often not aware of the high market prices and medicinal values, and most of the collected material is sold to local middlemen at a very low price. There was an increase of 3 to 5 folds in prices from collectors to the national market.Recommendations were given in the spheres of training sustainable harvesting and post harvesting of wild medicinal plant resources, trade monitoring, equitable sharing of benefits of wild resources, improved control on harvesting and trade for the conservation of resources, enhancement of cultivation efforts, future research into trade in wild harvested plants, community participation in natural resource management and value addition in the herbal products to maximize the benefits.     

Allelopathic effect of aqueous extracts of three weed species on the growth and leaf chlorophyll content of bread wheat

The purpose of this study was to evaluate the allelopathic effect of weeds (Avena fatua, Melilotus officinalis and Polypogon hissaricus) on germination, growth, dry biomass and chlorophyll concentration of three cultivars of wheat (Ata Habib, Pirsabaq and Serin). In germination test, different concentrations of aqueous extracts (5, 10 and 15?g/l) of the three weeds significantly reduced percent germination; however, 15?g/l extract of M. officinalis resulted in complete failure of germination of cultivar Pirsabaq. In pot culture, root and shoot length, chlorophyll concentration and seedling dry biomass of the three wheat varieties showed differential responses to different weeds. Aqueous extract at 15?g/l of A. fatua increased root and shoot length and dry biomass of cultivar Pirsabaq; however, these parameters were significantly retarded in other two wheat cultivars by extract of weeds. Moisture content of the cultivars did not show any response to allelopathic stress of the weeds. In contrast, chlorophyll concentration in Pirsabaq and Serin was significantly increased by aqueous extract of all the weeds but reduced it in cultivar Ata Habib by 50%. In general, Ata Habib was found to be the most sensitive cultivar to the imposed allelopathic stress. The phytotoxic potential of three weeds was found in the order of A. fatua?>?M. officinalis?>?P. hissaricus.     

Synthesis of new bergenin derivatives as potent inhibitors of inflammatory mediators NO and TNF-α

Bergenin is an isocoumarin natural product which aides in fat loss, healthy weight maintenance, enhancing the lipolytic effects of norepinephrine, inhibiting the formation of interleukin 1α and cyclooxygenases-2. Here we describe the anti-inflammatory activity of new bergenin derivatives 1-15 in the respiratory burst assay. Bergenin was isolated from the crude extract of Mallotus philippenensis after repeated column chromatography and was then subjected to chemical derivatization. The structures of all compounds were elucidated by NMR and mass spectroscopic techniques. Compound 2 was also studied using single crystal X-ray diffraction. Compounds 4, (54.5±2.2%) 5 (47.5±0.5%) 5, and 15 (86.8±1.9%) showed significant (P≤0.005) NO inhibitory activities whereas 6, 7, 11, 12 and 13 displayed moderate inhibitory activities that ranges between 16% and 31%. Furthermore compounds 4 and 15, were discovered as significant (P≤0.005) TNF-α inhibitors with 98% and 96% inhibition, respectively, while compounds 3, 5, 7, 8, 11, and 12 showed low level of TNF-α inhibition (0.4-28%). Compounds 8, 13 and 15 exhibited moderate anti-inflammatory IC(50) activities with 212, 222, and 253 μM, respectively, compared to the standard anti-inflammatory drug indomethacin as well as the parent bergenin compound. No cytotoxic effects could be detected when the compounds were tested on 3T3 cells up to concentrations of 100 μM.     

Assessment of potentially toxic heavy metals and health risk in water,sediments, and different fish species of River Kabul,Pakistan

Contamination of freshwater bodies and consequently freshwater fish with toxic heavy metals is a serious environmental issue. The trophic transfer of potentially toxic heavy metals in the human food chains, especially in fish has important implications for human health. The present research study was designed to assess the concentrations of the heavy metals Cr, Ni, Cd, and Pb in the water, sediments, and different freshwater fish species of River Kabul, Pakistan. The heavy metals were quantified in the samples with Atomic Absorption Spectrophotometer. Heavy metal contamination in fish muscles was characterized in terms of metal pollution index and biota-sediments accumulation factor, while human health risk was assessed through calculation of estimated weekly intake. The average concentrations of Cr, Ni, Cd, and Pb in muscle samples of the analyzed fish species at different sampling sites of the river ranged from 12.3 to 33.0, 33.2 to 109.2, 0.98 to 1.5, and 13.9 to 29.6 mg kg^?1 wet weight, respectively. Based on the current study data, consumption of the analyzed freshwater fish species from River Kabul was generally safe in terms of potential risk from Cd and Pb but the observed Ni accumulation may pose a potential health risk to regular/excessive fish consumers.     

Effect of environmental factors on the yield of selected mushroom species growing in two different agro ecological zones of Pakistan

Mushrooms are a rich source of protein and vitamins as human food. In view of the growing importance of mushroom in Pakistan, a research study was initiated with the objective to examine the suitability of Oyster mushroom cultivation and to compare the growth and yield of Oyster mushroom in two different areas (Peshawar and Swat, North-West region of Pakistan) with different ecological conditions. Spawn running time, number of crops, stalk height, stalk diameter, cap size, fresh weight, number of production days, and the interval in days between the time of bag opening and the time of starting fruiting bodies formation were among the important parameters investigated in the current study. Stalk height, stalk diameter, cap size and fresh weight of mushrooms were found higher in Peshawar region as compared to those growing in Swat region. On the other side, the spawn running time, formation of fruiting bodies and the number of productions were higher in Swat region as compared to the mushroom under study in Peshawar region. Mild winter temperatures of Peshawar region, and low summer temperatures in Swat, were found most suitable for growth and yield of Pleurotus ostreatus.