Identification of key amino acid residues in the catalytic mechanism of diaminopropionate ammonialyase from Salmonella typhimurium

Diaminopropionate ammonialyase (DAPAL), a fold-typeII pyridoxal 5-phosphate-dependent enzyme, catalyzes the ,-elimination of diaminopropionate (DAP) to pyruvate and ammonia. DAPAL was able to utilize both d- and l-DAP as substrates with almost equal efficiency. Mutational analysis of functionally important residues such as Thr385, Asp125 and Asp194 was carried out to understand the mechanism by which the isomers are hydrolyzed. Further, the putative residues involved in the formation of disulfide bond Cys271 and Cys299 were also mutated. T385S, T385D sDAPAL were as active with dl-DAP as substrate as sDAPAL, whereas the later exhibited a threefold increase in catalytic efficiency with d-Ser as substrate. Further analysis of these mutants suggested that DAPAL might follow an anti-E-2 mechanism of catalysis that does not involve the formation of a quinonoid intermediate. Of the two mutants of Asp125, D125E showed complete loss of activity with d-DAP as substrate, whereas the reaction with l-DAP was not affected significantly, demonstrating that Asp125 was essential for abstraction of protons from the d-isomer. By contrast, mutational analysis of Asp194 showed that the residue may not be directly involved in proton abstraction from l-DAP. sDAPAL does not form a disulfide bond in solution, although the position of Cys299 and Cys271 in the modeled structure of sDAPAL favored the formation of a disulfide bond. Further, unlike eDAPAL, sDAPAL could be activated by monovalent cations. Mutation of the cysteine residues showed that Cys271 may be involved in coordinating the monovalent cation, as observed in the case of other fold-typeII enzymes.

Probing the role of highly conserved residues in triosephosphate isomerase-analysis of site specific mutants at positions 64 and 75 in the Plasmodial enzyme

Highly conserved residues in enzymes are often found to be clustered close to active sites, suggesting that functional constraints dictate the nature of amino acid residues accommodated at these sites. Using the Plasmodiumfalciparum triosephosphate isomerase (PfTIM) enzyme () as a template, we have examined the effects of mutations at positions 64 and 75, which are not directly involved in the proton transfer cycle. Thr (T) occurring at position 75 is completely conserved, whereas only Gln (Q) and Glu (E) are accommodated at position 64. Biophysical and kinetic data are reported for four T75 (T75S/V/C/N) and two Q64 (Q64N/E) mutants. The dimeric structure is weakened in the Q64E and Q64N mutants, whereas dimer integrity is unimpaired in all four T75 mutants. Measurement of the concentration dependence of enzyme activity permits an estimate of K-d values for dimer dissociation (Q64N=73.79.2nm and Q64E=44.6 +/- 8.4nm). The T75S/V/C mutants have activities comparable to the wild-type enzyme, whereas a fourfold drop is observed for T75N. All four T75 mutants show a dramatic fall in activity between 35 degrees C and 45 degrees C. Crystal structure determination of the T75S/V/N mutants provides insights into the variations in local interactions, with the T75N mutant showing the largest changes. Hydrogen-bond interactions determine dimer stability restricting the choice of residues at position 64 to Gln (Q) and Glu (E). At position 75, the overwhelming preference for Thr (T) may be dictated by the imperative of maintaining temperature stability of enzyme activity.

The analysis of dieldrin residues in the brain, liver and muscles of the African catfish Clarias gariepinus (Burchell 1822) chronically exposed to dieldrin

Mature adult Clarias gariepinus were obtained at the ABRU hatchery in Sonning (UK), where they had beenbred and reared for several years. These were exposed to two concentrations of dieldrin in water (2.4 mu g super(-1) and 4.0 mu g super(-1). The residue analysis of diedrin in three tissues exposed for on moth at two concentrations was carried out. These were subjected to GLC analytical process. The results indicated significantly (P<0.05) higher residues in liver than in muscle and brain. The results also showed that residue levels were dependant on exposure concentration

Fisheries production in Asia: its role in food security and nutrition

This paper reviews the status and some management issues of fisheries production in Asia, as well as the supply and demand situation. Its food security and nutritional roles and opportunities for value addition are also discussed.

The role of small indigenous fish species in food and nutrition security in Bangladesh

In Bangladesh, only 6% of the daily food intake is animal food of which fish accounts for 50%. Rice is the mainstay, making up 60% of the daily food intake. However, many nutrients such as vitamins A and C, iron, calcium, zinc and iodine are not found in rice and have to be obtained from other sources. Small indigenous fish are a vital contribution to the diet of the rural poor in Bangladesh, where more than 30,000 children go blind every year from vitamin A deficiency and 70% of women and children are iron-deficient. Small fish, which are less than 10 cm in length and usually eaten whole with the organs and bones, contain large amount of calcium and possibly iron and zinc. The largest fish promoted in aquaculture do not contribute significantly to calcium intake. Some species also contain large amount of vitamin A. Much of the small indigenous fish (SIS) of Bangladesh are caught in floodplains and natural waterbodies. Small fish are eaten frequently in small amounts and are more equally distributed among family members than big fish of which men get the larger share. Unfortunately, overfishing and the deterioration of natural habitats have resulted in a decline in SIS. When measures are taken to improve food and nutrition security, there should be a focus on production of small fish so that greater quantities are accessible for consumption by the rural poor.

Fisheries production in Asia: its role in food security and nutrition

This paper reviews the status and some management issues of fisheries production in Asia, as well as the supply and demand situation. Its food security and nutritional roles and opportunities for value addition are also discussed.

Viability of seeds in food products proposed for field trips in Galapagos

Seeds of 14 plant species extracted from food products proposed for field trips in Galapagos were tested for viability. Strawberry Fragaria ananasa and Blackberry Rubus glaucus jams (Snob and Gustadina brands) contained no viable seeds. Schullo brand granola contained inviable Sesame Sesamum indicum seed, but Sesame in granolas prepared in Galapagos was viable. Sesame seed in bread was viable but Flax Linum usitatissimum seed in bread was not. Brown Rice Oryza sativa and Sunflower seeds Helianthus annuus were both viable. Fresh Apple Malus domestica, Naranjilla Solanum quitoense, Cucumber Cucumis sativus, Pineapple Ananas comosus, Pear Pyrus communis, Bell Pepper Capsicum annuum, Tomato Solanum lycopersicum, Grape Vinis vinifera all contained viable seeds. We recommend prohibiting any product with viable seeds from field trips to uninhabited areas. CDF Contribution Number 1009.

Limit of drug residues in aquatic products (NY5071-2002)

Weimin, ZOU, lexian YANG, lan JIANG, Shuqin WU, Qi YI, Jianli WU

Promising practices in food security and nutrition assistance to vulnerable households in the Tonle Sap Region, Cambodia

The CGIAR Research Program on Aquatic Agricultural Systems (AAS) seeks to reduce poverty and improve food security for many small-scale fishers and farmers who are dependent on aquatic agriculture systems by partnering with local, national and international partners to achieve large-scale development impact. This study on promising practices in food security and nutrition assistance to vulnerable households in the Tonle Sap region forms part of the preliminary research that informs AAS work in the highly productive Mekong Delta and Tonle Sap Lake floodplain. The study aims to identify and learn from promising practices that have had a positive impact on the food security and nutrition of vulnerable households in the Tonle Sap region.

Organochlorine insecticide residues in sediments from the coast of Chittagong, Bangladesh

A total of 5 samples of marine sediments were analyzed for residues of organochlorine insecticide from the coast of Chittagong, Bangladesh. The analytical method consisted of 3 phases, extraction, clean-up and, analysis through Gas Chromatography (GC) with Electron Capture Detector (ECD). The concentration ranges were as follows 0.18 - 1.33 ng.g¹ for aldrin, 0.2 - 1.84 ng.g¹ for dielddrin, 0.30 - 1.31 ng.g¹ for endrin, 0.11 - 0.26 ng.g¹ for lindane, 0.56- 3.36 ng.g¹ for heptachlor, 0.2- 1.51 ng.g¹ for P,P' DDE, 0.18- 2.91 ng.g¹ for P,P' DDD, 0.11 - 3.12 ng.g¹ for P,P' DDT. These results reveal that the sediments along coast of Chittagong are slightly contaminated with some of these organochlorine insecticides.

A study on the economic efficiency of hydrogen production from biomass residues in China

As part of Pilot Project of KIP of CAS, a feasibility study of hydrogen production system using biomass residues is conducted. This study is based on a process of oxygen-rich air gasification of biomass in a downdraft gasifier plus CO-shift. The capacity of this system is 6.4 t biomass/d. Applying this system, it is expected that an annual production of 480 billion N m(3) H-2 will be generated for domestic supply in China. The capital cost of the plant used in this study is 1328$/(N m(3)/h) H-2 out, and product supply cost is 0.15$/N m(3) H-2. The cost sensitivity analysis on this system tells that electricity and catalyst cost are the two most important factors to influence hydrogen production cost.