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Category:Team Grizzly Ninjas

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StatusPageUserDate/TimeGO Term (Aspect)ReferenceEvidenceNotesLinks
requireschangesPSESD:ATZCRolfecat, Team Grizzly Ninjas2012-04-15 13:21:23 CDTGO:0018764 N-isopropylammelide isopropylaminohydrolase activity (F)PMID:9422605ECO:0000314 direct assay evidence used in manual assertion

AtzC codes for the enzyme which transforms N-isopropylammelide to cyanuric acid and isopropylamine by hydrolytic deamination.

Table 1 shows the control strain of Pseduomonas had this activity. The E.coli strains which had a plasmid insterted containing the AtzC gene also transformed N-isopropylammelide to cyanuric acid and isopropylamine.

requireschangesECOLI:FABHRolfecat, Team Grizzly Ninjas2012-04-15 13:04:52 CDTGO:0004315 3-oxoacyl-(acyl-carrier-protein) synthase activity (F)PMID:1551888ECO:0000314 direct assay evidence used in manual assertion

FabH catalyzes the condensation of acetyl-CoA and initiation of fatty acid synthesis.

Table 2 shows the level of FabH correlates with the membrane fatty acid composition. Chromatography was used to analyze the fatty acid methyl esters.

requireschangesPSEMV:MVAARolfecat, Team Grizzly Ninjas2012-04-15 12:58:38 CDTGO:0042282 hydroxymethylglutaryl-CoA reductase activity (F)PMID:2656635ECO:0000314 direct assay evidence used in manual assertion

mvaA codes for the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase.

Figure 5 shows E.coli cells transformed with the plasmid containing mvaA and induced with IPTG. Assayed for HMG-CoA reductase activity. Cells induced with IPTG resulted in HMG-CoA reductase activity shown in lane 1.

acceptableYEAST:ALG5Millstea, Team Grizzly Ninjas2012-04-15 00:04:56 CDTGO:0004581 dolichyl-phosphate beta-glucosyltransferase activity (F)PMID:8076653ECO:0000315 mutant phenotype evidence used in manual assertion

Mutations of ALG5 result in loss of enzyme function and underglycosylation of proteins. Fig. 2 shows ALG5 where complementations restored the ability to glycosylate carboxypeptidase Y. Also, Table 2 further illustrates that mutated ALG5 genes resulted in loss of function to synthesize DolP-Glc.

requireschangesASPTE:B3IUN8Millstea, Team Grizzly Ninjas2012-04-14 20:35:33 CDTGO:0047613 aconitate decarboxylase activity (F)PMID:18584171ECO:0000315 mutant phenotype evidence used in manual assertion

The CAD1 gene was cloned from and then expressed in S. cerevisiae where activity was verified. Fig. 3 shows how the recombinant activity (measured by direct detection of itaconic acid product) was detected by HPLC.

requireschangesCUPPJ:TFDARolfecat, Team Grizzly Ninjas2012-04-14 17:54:42 CDTGO:0018602 2,4-dichlorophenoxyacetate alpha-ketoglutarate dioxygenase activity (F)PMID:3036764ECO:0000314 direct assay evidence used in manual assertion

TfdA codes for the 2,4-D monooxygenase.

Figure 4 shows omega mutagenesis of the tfdA gene. An omega fragment containing stop signals was inserted into the coding region of a tfdA plasmid. The results were a truncated tfdA product for the plasmids which the omega fragment was inserted, in turn disrupting the enzymatic activity for the 2,4-D monooxygenase.

acceptableHELTU:O81986Millstea, Team Grizzly Ninjas2012-04-14 17:02:48 CDTGO:0050306 sucrose 1F-fructosyltransferase activity (F)PMID:12226250ECO:0000314 direct assay evidence used in manual assertion

Fig.2 shows the sucrose (substrate) dependent rate of GF2 by 1-SST from the plant tubers. Fig. 3 is a representation of the efficiency of this same conversion with an added step to eliminate the possibility of any 1-FFT impurities.

requireschangesPSEOL:RURERolfecat, Team Grizzly Ninjas2012-04-14 16:56:12 CDTGO:0015044 rubredoxin-NAD+ reductase activity (F)PMID:10727934ECO:0000314 direct assay evidence used in manual assertion

alkT encodes the 3rd component of the alkane hydroxylase complex, the rubredoxin reductase. Transfers electrons from NADH to rubredoxin.

Figure 7 shows the highest specific activity for alkT in P.oleovorans, lower in E.coli HB101 and lowest in E.coli W3110. This concluded that the rubredoxin reductase is present

in excess in the E. coli alk+ recombinants

requireschangesHELTU:O81985Millstea, Team Grizzly Ninjas2012-04-14 16:40:54 CDTGO:0047207 1,2-beta-fructan 1F-fructosyltransferase activity (F)PMID:9753774ECO:0000315 mutant phenotype evidence used in manual assertion

Fig. 6 illustrates transferred fructosyltransferase activity after incorporating the fft-1 gene product into transgenic Petunia that does not naturally have fructosyltransferase function.

requireschanges9PSED:Q7WSJ4Rolfecat, Team Grizzly Ninjas2012-04-14 16:17:20 CDTGO:0034013 aliphatic aldoxime dehydratase activity (F)PMID:12773527ECO:0000314 direct assay evidence used in manual assertion

OxdA encodes for aldoxime dehydratase activity.

Figure 3As shows the activity between the purified enzyme, the enzyme with Na2S2O4-reduced, and the enzyme with Na2S2O4-reduced plus CO. As a result, oxdA was shown to have much higher activity in the presence of Na2S2O4. This shows the Na2S2O4 reagent assists in the already present reducing activity.

requireschangesIDEDE:CLRARolfecat, Team Grizzly Ninjas2012-04-14 15:53:52 CDTGO:0047143 chlorate reductase activity (F)PMID:12957948ECO:0000314 direct assay evidence used in manual assertion

ClrA is one of three subunits which contribute to the chlorate reductase activity.

Figure 1 uses SDS-PAGE to purify the chlorate reductase. The results were 3 major bands which shows it is a complex of 3 different subunits in a 1:1:1 ratio.

requireschangesZOBGA:Q9RGX9Rolfecat, Team Grizzly Ninjas2012-04-14 15:38:39 CDTGO:0033916 beta-agarase activity (F)PMID:15456406ECO:0000314 direct assay evidence used in manual assertion

Assists in cleaving the β-(1→4) linkages of agarose.

Figure 3 shows the hydrolysis of agarose by AgaA. After 24 hours,neoagarotetraose and neoagarohexaose were produced.

acceptableSYNE7:ALDECMillstea, Team Grizzly Ninjas2012-04-14 09:06:38 CDTGO:0071771 aldehyde decarbonylase activity (F)PMID:20671186ECO:0000315 mutant phenotype evidence used in manual assertion

Aldehyde decarbonylase, purified from Synechococcus elongatus PCC 7942 was able to decarbonylate fatty aldehydes when expressed in E.coli. According to Fig. 1, when introduced along with acyl-[acp] reductase, the host organism was conferred the ability to produce alkane.

acceptableARATH:CESA8Millstea, Team Grizzly Ninjas2012-04-13 23:09:04 CDTGO:0016760 cellulose synthase (UDP-forming) activity (F)PMID:9165747ECO:0000315 mutant phenotype evidence used in manual assertion

CesA8 mutants have a reduced cellulose content and resulting irregular xylem shape. This suggests that this cellulose synthase isoform is involved in secondary wall synthesis of Arabidopsis thaliana.Fig. 6 demonstrates the results through electron microscopy of thinned cellular walls in mutants.Fig. 7 also illustrates how mutants (irx is synonym for CesA8) are weaker/more susceptible to bending.


MSU Internal Competition

StatusPageUserDate/TimeGO Term (Aspect)ReferenceEvidenceNotesLinks
requireschangesPENCH:AAAARolfecat, Team Grizzly Ninjas2012-04-01 16:05:09 CDTGO:0050640 isopenicillin-N N-acyltransferase activity (F)PMID:8344300ECO:0000314 direct assay evidence used in manual assertion

Responsible for isopenicillin-N amidohydrolase, 6-aminopenicillanic acid acyltransferase and penicillin amidase activities in P. chrysogenum.

Mutants npe6, npe8 and npe10 with no acyltransferase activity were transformed with a plamsid, pULB101, containing penDE gene. Mutants were then prepared and assayed. Npe6, npe8 and npe10 showed AAT, IAT, IPN amidohydrolase, penicillin transacylase and penicllin amidase activities. This indicated the penDE gene recovered all acyltransferase activity. Table 6 shows the results of an assay for the comparison between P. chrysogenum and C. acremonium. C. acremonium does not contain the penDE gene and recorded zero activity for Acyl-CoA:CAPA acyltransferase activity, extracellular accumulation of 6-APA, and production of penicillin. Whereas, P. chrysogenum does contain the penDE gene and showed positive results for these activities.

requireschangesBACGL:UGLRolfecat, Team Grizzly Ninjas2012-03-31 17:37:50 CDTGO:0052788 d-4,5 unsaturated beta-glucuronyl hydrolase activity (F)PMID:10441389ECO:0000314 direct assay evidence used in manual assertion

Release of unsaturated glucuronic acids from oligosaccharides produced from polysaccharide lyase reactions.

Fig. 2 shows the transformation from a tetrasaccharide to a trisaccharide without unsaturated glucuronic acid using enzyme fraction. This shows an enzyme, unsaturated

glucuronyl hydrolase, is present in Bacillus sp. GL1. Also, the absorbance level at 235nm was shown to decrease due to the double bond diminishing over a 15min period.

requireschangesPELCA:Q60154Millstea, Team Grizzly Ninjas2012-03-31 13:59:18 CDTGO:0004148 dihydrolipoyl dehydrogenase activity (F)PMID:1898934ECO:0000021 physical interaction evidence

Codes for a subunit(dihydrolipoamide dehydrogenase)that dimerizes to become the E3 subunit dihydrolipoamide dehydrogenase (DHLDH) in the acetoin dehydrogenase complex that catalyzes the conversion of acetoin to acetyl-CoA and acetaldehyde. Table 6 indicates DHLDH is only present in high activity in acetoin utilizing cultures. Fig. 4 illustrates growth of P. carbinolicus in double immunodiffusion tests that gave strong cross-reactions only between organisms capable of utilizing and degrading acetoin


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