Biochemistry, Biophysics, and Structural Biology Commons^™

Microbial Nad Metabolism: Lessons From Comparative Genomics, Francesca Gazzaniga, Rebecca Stebbins, Sheila Z. Chang, Mark A. Mcpeek, Charles Brenner

Dartmouth Scholarship

NAD is a coenzyme for redox reactions and a substrate of NAD-consuming enzymes, including ADP-ribose transferases, Sir2-related protein lysine deacetylases, and bacterial DNA ligases. Microorganisms that synthesize NAD from as few as one to as many as five of the six identified biosynthetic precursors have been identified. De novo NAD synthesis from aspartate or tryptophan is neither universal nor strictly aerobic. Salvage NAD synthesis from nicotinamide, nicotinic acid, nicotinamide riboside, and nicotinic acid riboside occurs via modules of different genes. Nicotinamide salvage genes nadV and pncA, found in distinct bacteria, appear to have spread throughout the tree of life …

The Primary Structure Of A Fungal Chitin Deacetylase Reveals The Function For Two Bacterial Gene Products., Dimitris Kafetzopoulos, George Thireos, John N. Vournakis, Vassilis Bouriotis

Dartmouth Scholarship

Chitin deacetylase (EC 3.5.1.41) hydrolyzes the N-acetamido groups of N-acetyl-D-glucosamine residues in chitin. A cDNA to the Mucor rouxii mRNA encoding chitin deacetylase was isolated, characterized, and sequenced. Protein sequence comparisons revealed significant similarities of the fungal chitin deacetylase to rhizobial nodB proteins and to an uncharacterized protein encoded by a Bacillus stearothermophilus open reading frame. These data suggest the functional homology of these evolutionarily distant proteins. NodB is a chitooligosaccharide deacetylase essential for the biosynthesis of the bacterial nodulation signals, termed Nod factors. The observed similarity of chitin deacetylase to the B. stearothermophilus gene product suggests that this gene …

Fermentation Of Cellulosic Substrates In Batch And Continuous Culture By Clostridium Thermocellum, Lee R. Lynd, Hans E. Grethlein, Richard H. Wolkin

Dartmouth Scholarship

Fermentation of dilute-acid-pretreated mixed hardwood and Avicel by Clostridium thermocellum was comparedinbatchandcontinuouscultures.Maximumspecificgrowthratesper hourobtainedon cellulosic substrateswere 0.1inbatchcultureand>0.13incontinuousculture.Cellyields(gramsofcellsper gram of substrate)inbatchculturewere 0.17forpretreatedwoodand0.15forAvicel.Ethanolandacetatewere the mainproductsobservedunderalconditions.Ethanol:acetateratios(ingrams)were approximately1.8:1in batchcultureand generallyslightlylessthan 1:1incontinuousculture.Utilizationofcellulosicsubstrateswas essentially complete in batch culture. A prolonged lag phase was initialy observed in batch culture on pretreated wood; the length of the lag phase could be shortened by addition of cell-free spent medium. In continuousculturewith-5g ofglucoseequivalentper literinthefeed,substrateconversionrelativeto theoreticalrangedfrom0.86ata dilutionrate(D)of0.05/hto0.48ata D of0.167/hforAvicelandfrom0.75 ata D of0.05/hto0.43ata D of0.11/hforpretreatedwood.Atfeedconcentrationsof<4.5g ofglucose equivalentperliter,conversionofpretreatedwoodwas80to90%atD= 0.083/h.Lowerconversionwas obtainedathigherfeedsubstrateconcentrations,consistentwitha limitingfactorotherthancellulose.Free Avicelaseactivitiesof12to84mU/mlwere observed,withactivityincreasinginthisorder:batchceliobiose, batchpretreatedwood< batchAvicel,continuouspretreatedwood< continuousAvicel.Freecellulaseactivity was higheratincreasingextentsofsubstrateutilizationforbothpretreatedwoodandAvicelunderal conditions tested. The results indicate that fermentation parameters, with the exception of free cellulase activity,are essentiallythesame forpretreatedmixedhardwoodandAvicelundera varietyofconditions. HydrolysisyieldsobtainedwithC.thermocellumcellulaseactingeitherinvitroor invivowere comparableto thosepreviouslyreportedforTrichodermareeseion thesame substrates.

Fermentation of dilute-acid-pretreated mixed hardwood and Avicel by Clostridium thermocellum was comparedinbatchandcontinuouscultures.Maximumspecificgrowthratesper hourobtainedon cellulosic substrateswere 0.1inbatchcultureand>0.13incontinuousculture.Cellyields(gramsofcellsper gram of substrate)inbatchculturewere 0.17forpretreatedwoodand0.15forAvicel.Ethanolandacetatewere the mainproductsobservedunderalconditions.Ethanol:acetateratios(ingrams)were approximately1.8:1in batchcultureand generallyslightlylessthan 1:1incontinuousculture.Utilizationofcellulosicsubstrateswas …