1. Volume 22, Issue 11, Pages 1425-1430 (November 2015)
Substrate Flexibility of a Mutated Acyltransferase Domain and Implications for Polyketide Biosynthesis 
Kenny Bravo-Rodriguez, Stephan Klopries, Kyra R.M. Koopmans, Uschi Sundermann, Samir Yahiaoui, Julia Arens, Susanna Kushnir, Frank Schulz, Elsa Sanchez-Garcia 
Chemistry & Biology 
Volume 22, Issue 11, Pages (November 2015)
DOI: /j.chembiol
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2. Chemistry & Biology 2015 22, 1425-1430DOI: (10. 1016/j. chembiol. 2015
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3. Figure 1
Snapshots of Molecular Dynamics Simulations of the Wild-Type AT6 Domain and of AT6 Carrying the V295A Mutation with Native and Non-native Substrates
(A) Alanine allows for greater flexibility of the binding pocket since this residue adopts positions not accessible to valine (WT-AT6, rose; V295A AT6, ochre).
(B) Allyl-MSNAC (1) is placed inside the active site of V295A AT6 with the side group of the malonyl moiety accommodated below Ala295 instead of side by side.
(C) The triple bond of propargyl-MSNAC (2) does not readily fit in the active site of WT-AT6. Notice the positioning of Arg222 outside the active site; Met235 and Val295 do not flank the substituent in the malonyl region.
(D) Improved positioning of 2 in the active site of the V295A variant.
(E) V295A AT6 with iProp-MSNAC (4) and (F) with Hex-MSNAC (6).
The substrate is colored according to the atom type, WT-AT6 is colored in rose and V295A AT6 in ochre.
Chemistry & Biology  , DOI: ( /j.chembiol )
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4. Figure 2
Results of Feeding Experiments with S. erythraea DEBS V295A AT6
(A and B) General scheme for the experimental outline. The malonic acid derivatives 1–3 were supplied to S. erythraea to give rise to the corresponding erythromycin derivatives (see table in B).
(C) LC-ESI-MS total ion chromatogram of a fermentation extract from a culture supplied with 1. It shows the corresponding 2-allylerythromycin A (1e) next to the WT product erythromycin A (9).
Chemistry & Biology  , DOI: ( /j.chembiol )
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5. Figure 3 Comparison of V295A AT6 with the New Triple Variant
WT DEBS does not incorporate the synthetic building block 2 in detectable quantities, and V295A AT6 yields low amounts. The triple variant shows a 3.79-fold increase in productivity. For this figure, the abundance of all signals was normalized to 9, which was set to 1; the chromatogram is shown from 6.55 to 6.95 min. Productivity was quantified based on LC-ESI-MS signals. a, V295A-9; b, triple variant 9; c, V295A 2e; d, triple variant 2e.
Chemistry & Biology  , DOI: ( /j.chembiol )
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