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A homology model predicted that the active-site cavity volume of the F66L/N222G mutant is increased to 748 Å3, from 652 Å3 of the wild-type OKS.
However, the period of the H429A mutant is increased only modestly (28 hours) and the D417A mutant displayed a normal period (Fig. 3); the KaiCTaT mutant is arhythmic as established earlier [21].
Importantly, the amount of the 15 kD P2 fragment of the FLL mutant is increased compared to the P2 fragment generated from wild-type FIT2, indicating a conformational change in FLL relative to wild-type FIT2.
Furthermore, like in M13V, the degree of heme ruffling in the double mutant is increased.
Rather, R347 is important for substrate binding because the KM for the R347A mutant is increased by ∼30-fold.
The most likely explanation is that the copper requirement of this mutant is increased and affects iron acquisition due to impaired siderophore-mediated iron uptake and hampered RIA due to FreB-deficiency.
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The proline content of the mutant was increased by 11.05% while the malondialdehyde content was significantly lower than that of wild type.
In contrast to GA3ox2 gene, the expression levels of both the GA catabolism genes in slr1 mutant were increased compared to wild type.
By engineering the rate-limiting enzyme l-2,4-diaminobutyric acid (DABA) aminotransferase (EctB), ectoine production and the specific activity of the EctB mutant were increased.
The dissociation constant of the double mutant was increased by approximately 10 times (Kd = 4.23 nM±0.27).
Importantly, the amount of P2 fragment generated by digestion of the FLL mutant was increased relative to wild-type FIT2, indicating an altered conformation of this mutant.
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