| acceptable | DEIRA:Q9RW08 | 2012-04-15 07:06:29 CDT | GO:0016491 oxidoreductase activity (F) | PMID:17464079 | ECO:0000314 direct assay evidence used in manual assertion | Figure 5: Purified DR0861 protein from D. radioduran converted 15,15'-cis-phytoene to lycopene detected by HPLC. This confirmed that the lycopene synthesized from phytoene in D. radioduran was catalyzed by the DR0681 protein.
| challenge |
| acceptable | DEIRA:Y079 | 2012-04-10 21:44:09 CDT | GO:0016787 hydrolase activity (F) | PMID:18512963 | ECO:0000314 direct assay evidence used in manual assertion | Figure 5: NMR spectroscopy was used to follow the
phosphorus resonances for three substrates, CDP, IDP, and CTP, alone and with the addition of DP_0079. This showed the hydrolase activity of DP_0079 by observing the hydrolase products CMP + Pi, IMP + Pi, and CMP + PPi only with the addition of DP_0079.
| challenge |
| acceptable | XANCP:AOTC | 2012-04-13 15:54:26 CDT | GO:0043857 N-acetylornithine carbamoyltransferase activity (F) | PMID:16585758 | ECO:0000314 direct assay evidence used in manual assertion | Table 1: Purified ArgF′ enzyme from Xanthomonas catalyzed the AOTCase reaction which forms N-acetylcitrulline from N-acetylornithine and carbamyl phosphate.
Figure 4: LC-MS analysis of enzymatically synthesized N-acetylcitrulline. The enzymatic product of the X. campestris AOTCase reaction has the same mass ion and retention time as chemically synthesized N-acetylcitrulline.
| challenge |
| requireschanges | CAMJE:ARLY | 2012-04-13 16:33:30 CDT | GO:0004056 argininosuccinate lyase activity (F) | PMID:11683360 | ECO:0000314 direct assay evidence used in manual assertion | E. coli C600 (WT functional argH),E. coli W3678 (no argH gene), C. jejuni TGH9011 (WT functional argH), and transformed E. coli (C. jejuni TGH9011 argH gene) were examined for ASL activity. E. coli C600, which is wild type
with respect to the argH gene, showed a specific activity of 1.1
+ 0.17 ,umol of arginine per h per mg of protein. C. jejuni
TGH9011 showed a specific activity of 1.3 ± 0.32 ,umol of
arginine per h per mg of protein. E. coli W3678 did not possess
any significant ASL activity compared with extract-free controls.
The transformed strain pARGH1-1 had a specific activity
of 5.7 ± 1.2 ,imol of arginine per h per mg of protein. This
result confirms that the protein derived from the recombinant
plasmid possesses argininosuccinate lyase activity activity.
| challenge |
| acceptable | LEUCI:D5FS20 | 2012-04-13 20:27:44 CDT | GO:0047849 dextransucrase activity (F) | PMID:21229247 | ECO:0000314 direct assay evidence used in manual assertion | Figure 2: Highperformance
anion-exchange chromatography with pulsed
amperometric detection was used to compare profiles. L. citreum B/110-1-2 (WT dsrF), DSR-F-DSP-DGBD (recombinant dextransucrase),DSR-S from mesenteroides NRRLB-512F(DSR-S is a dextransucrase). L. citreum B/110-1-2 produces a series of isomaltodextrins
analogs with retention times that were very similar to the ones
produced by the dextransucrase DSR-S from L. mesenteroides NRRLB-512F.
| challenge |
| requireschanges | CITSI:Q84X90 | 2012-04-14 12:02:19 CDT | GO:0046905 phytoene synthase activity (F) | PMID:19813015 | ECO:0000314 direct assay evidence used in manual assertion | Figure 3(a),(b),(c):HPLC analysis revealed that the
GGPP-accumulating E. coli cells transformed with the
CsPSY construct showed a peak (Fig. 3b, peak 1) that was
not detected in the empty vector control, pACCRT-E/pET-28a (Fig. 3a). The retention time and absorption spectrum
of the peak 1 matched well with those of the peak 2
presented in the positive control carrying pACCRT-EB for
phytoene biosynthesis (Fig. 3c, peak 2).
| challenge |
| acceptable | ESCVU:CRTX | 2012-04-15 10:37:53 CDT | GO:0046527 glucosyltransferase activity (F) | PMID:1409639 | ECO:0000314 direct assay evidence used in manual assertion | Figure 3: Extracts from plasmid-containing Es. coli strains were
assayed for in vitro glucosyltransferase activity by incubation
with zeaxanthin and UDP-[14C]glucose. When an extract
from induced Es. coli BL21(DE3) cells containing pAPUX
was used, significant amounts of the yellow-colored radiolabeled
products, zeaxanthin mono- and diglucosides, were
observed. As a negative control, an extract from Es. coli
BL21(DE3) cells containing the pET-3b vector without crtX
was incubated with zeaxanthin and UDP-[14C]glucose in the
in vitro assay, and no product was observed. Thin layer chromatographic analysis of the products of this
assay is shown in the figure.
| challenge |
| requireschanges | 9ASCO:F1T2J9 | 2012-04-15 11:20:48 CDT | GO:0016616 oxidoreductase activity, acting on the CH-OH group of donors, NAD or NADP as acceptor (F) | PMID:21672638 | ECO:0000314 direct assay evidence used in manual assertion | Figure 1: Time dependent changes in PLA levels were monitored using HPLC. The PLA concentration linearly increased (Fig. 1A), indicating that the strain produced PLA at a constant rate. Adding PPA, a potential PLA substrate, to the culture medium increased the amount of PLA produced by strain TK1 (Fig. 1B). After 48-h of culture, 1.5 mM PPA was consumed and 0.8 mM of PLA accumulated in the medium,indicating that 53% of the PPA was converted to PLA. When cultured in MM containing phenylalanine, strain TK1 consumed phenylalanine and produced PLA (Fig. 1C). After the initial 24-h of incubation, 5.0 mMphenylalanine was consumed and 5.7 mMPLA was generated,indicating that most of the added phenylalanine was converted to PLA. We detected phenylalanine aminotransferase (PAT) (1.9±0.2 μmol min−1 mg−1) and PPR (see below) activities in the fungal cell-free extracts.
These results together with the higher levels of PLA
being generated from either PPA or phenylalanine than from glucose when added as a carbon source, indicated that the fungus generates PLA via PPA and/or phenylalanine (Fig. 1D), and that biosynthesized PPA limits PLA production from glucose.
| challenge |
| acceptable | VIBVU:G6PI | 2012-04-15 13:33:58 CDT | GO:0004347 glucose-6-phosphate isomerase activity (F) | PMID:15262231 | ECO:0000314 direct assay evidence used in manual assertion | Figure 3b: Isomerase activity was readily detectable in PGI-LysAP
from V. vulnificus. Assay of commercially available PGI
from rabbit, B. stearothermophilus, and S. cerevisiae also
revealed isomerase activity.
Determination of the pH optima for isomerase activity for PGI from V. vulnificus.
| challenge |
