Rabbit Anti-Human C4D Polyclonal antibody

3-Hydroxybenzoate 6-hydroxylase (3HB6H) from Rhodococcus jostii RHA1 is an NADH-specific flavoprotein monooxygenase that contains FAD as a redox-active cofactor. The enzyme catalyzes para-hydroxylation of 3-hydroxybenzoate (3HB) to form 2,5-dihydroxybenzoate (2,5-DHB). Based on the enzyme crystal structure, residue His213 is located close to the hydroxyl moiety, whereas Tyr217 is close to the carboxylate group of 3HB. Y217A and Y217S did not show any perturbation of flavin absorption upon addition of 3HB, whereas Y217F has a K-d value for 3HB binding of 7.5 mm, which is similar to 50-fold larger than that found for wild-type enzyme. The results clearly indicate that Tyr217 is necessary for substrate binding. All His213 variants can bind to 3HB with similar affinity as the wild-type enzyme and form C4a-hydroperoxy intermediate. H213S, H213D and H213E produce 2,5-DHB with yields of 28 +/- 5%, 52 +/- 7% and 92 +/- 6%, respectively, whereas H213A cannot catalyze hydroxylation. The results indicate that the interaction between the hydroxyl group of 3HB and residue 213 is important for substrate hydroxylation. Interestingly, the hydroxylation rate constant of H213E (35 s(-1)) is similar to that of wild-type enzyme (36 s(-1)) and this variant has an efficiency of hydroxylation (92 +/- 6%) similar to the wild-type enzyme (86 +/- 2%). Difference spectra of enzyme-bound substrate suggest that 3HB binds to H213E in the phenolic form. The results indicate that His213 and Glu213 in H213E may act as a catalytic base to initiate the substrate deprotonation and facilitate the electrophilic aromatic substitution of 3HB.

White grubs are largely unsolved problems in vegetable and tuber production in East Africa. Novel Rwandan as well as international entomopathogenic nematodes (EPNs) were screened in nine laboratory bioassays and two small-scale field trials in 2014 and 2015. Soil-based laboratory bioassays revealed that all EPNs were able to infect Anomala graueri larvae (Coleoptera: Scarabeidae), although a relatively high number of infective juveniles were needed. Rwandan EPNs were as infectious as their corresponding international strains. At 100 infective juveniles per larva, the Heterorhabditis bacteriophora strains (Rwanda14-N-C4a and international H06) as well as the international Steinernema carpocapsae All caused 18 to 22% grub mortality within 7 days compared to the control. At 1000 infective juveniles, both H. bacteriophora strains as well as the Rwandan S. carpocapsae RW14-G-R3a-2 killed 34 up to 58%. The Rwandan Steinernema RW14-M-C2a-3 least performed in the bioassays (2 to 6%). In two Irish potato fields, the into-soil-applied 1 x 10(9) infective juvenile Steinernema RW14-M-C2b-1 per hectare reduced 29 +/- 33% and 96 +/- 3% of grubs within 30 and 60 days, respectively. About 1 up to 2.5 x 10(9) infective juvenile Steinernema longicaudum X7 per hectare reduced 77 up to 85% of grubs until day 30, respectively, and 82 up to 95% until day 60. Avermectin + Chlorpyrifos tuber coatings reduced 39% of grubs, and Fipronil + Chlorpyrifos reduced 27%, whereas handpicking did not help much. In conclusion, at least Steinernema RW14-M-C2b-1 and S. longicaudum X7 are promising for managing white grubs in tuber production, this is at a rate of at least 1.5 x 10(9) EPNs per hectare, but H. bacteriophora RW14-N-C4a needs further field research. Findings will support the biocontrol product development in Rwanda, including registration if any would be needed for native macrobial biocontrol agents.