Fig. 5: Elucidation of the biosynthetic pathway to the shunt product 8.
From: A severe leakage of intermediates to shunt products in acarbose biosynthesis
a HPLC-TOF/MS analysis of the reduction products of 12 by the crude cell-free extract of Actinoplanes sp. SE50/110 (abbreviated as SE50/110), SE50/110ΔacbN and QQ-3, purified ACPL_7966, ACPL_7511 and ACPL_5736 proteins, and the reaction with boiled crude cell-free extract (BCF) was set as a negative control. All the chromatograms show the extraction of calculated ion m/z = 255.0275 [M-H]−. b GC-QMS confirmation of the configuration of the reaction products after dephosphorylation by AcbJ. AcbJ was added to the reaction products of crude cell-free extract of Actinoplanes sp. SE50/110, SE50/110ΔacbN, and QQ-3, purified ACPL_7966, ACPL_7511, ACPL_5736 proteins, and control reaction. The standards (std) 8 and 9 were also analyzed by GC-QMS after derivatization by BSTFA. For more details, see also Supplementary Fig. 11. All the chromatograms show the extraction of unique product ion m/z = 332.2. TMS is the abbreviation of trimethyl silicyl. Source data are provided as a Source Data file.
