Journal of the American Chemical Society
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arising from the malleable nature of PLP chemistry, would
allow access to unpredictable new biological processes.
ASSOCIATED CONTENT
Supporting Information
Supplementary Methods, Results, Figures and Tables. This
material is available free of charge via the Internet at
http://pubs.acs.org.
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AUTHOR INFORMATION
Corresponding Author
Notes
The authors declare no competing financial interests.
Figure 5. In vitro combinatorial biosynthesis of lincosamide antibiotics.
The ESI m/z [M + H] modes of target compounds are indicated in the
biosynthetic routes (top) and colored in the HPLC-MS traces (bottom).
+
ACKNOWLEDGMENT
(A) Generation of Bu2545 and related products bearing a lincomycin-like
We thank Dr. Wei Deng at NCPSS for technical support. This
work was supported in part by grants from NSFC (81302674
for Q. Z. and 21520102004, 31430005 and 21472231), STCSM
sulfur appendage. The transformations were conducted by incubating the
celesticetin intermediate 1 with LmbF (i), LmbF and LmbG (ii), LmbF and
Ccb4 (iii), or LmbF, Ccb4 and Ccb5 (iv). (B) Generation of the products
with a celesticetin-like sulfur appendage. The transformations were con-
ducted by incubating the lincomycin intermediate 5 with CcbF (i) or
CcbF and Ccb5 (ii).
(
14JC1407700 and 15JC1400400), and MST (2012AA02A706) of
China (all for W.L.).
We therefore extended the idea of in vitro combinatorial
biosynthesis to process the cysteinyl group of the lincomycin
intermediate 5 to generate a two-carbon S-linker characteris-
tic of celesticetin (Figure 5B). Incubation with CcbF for 2-h
resulted in the partial conversion of 5 into aldehyde 10; how-
ever, the further transformation of 10 to alcohol 11 by the
NADPH-dependent oxidoreductase Ccb5 was complete (Fig-
ure 5B and Figure S28-29). Compared to its counterpart
LmbF, CcbF appeared to be more stringent in substrate tol-
erance and its associated decarboxylation-coupled oxidative
deamination reaction could be a limiting step in processing
cysteine S-conjugated surrogates.
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1e
H230-5 . The tool box of synthetic biology is thus enriched,
as the ability to design variable sulfur appendage for the pro-
duction of sulfur-containing active molecules has been estab-
lished. CcbF and LmbF are functionally distinct but highly
homologous to each other, implying that phylogenetically
they evolved from a common PLP-dependent prototype pro-
tein. A genome sequence-based survey revealed a number of
homologs of CcbF and LmbF that remain functionally unas-
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