Supporting Information

A reductase gene mppE controls yellow component production in Monascus azaphilone pigment biosynthesis

Bijinu Balakrishnan ∙Si-Hyung Park∙Hyung-Jin Kwon

B. Biniju ∙H. -J. Kwon ()

Department of Biological Science and Bioinformatics, Myongji University, Yongin-si, Gyunggi-do, 449-728, Republic of Korea

e-mail:

S. -H. Park

Department of Oriental Medicine Resources and Institute for Traditional Korean Medicine Industry, Mokpo National University,Muan-gun, Jeollanam-do, 534-729, Republic of Korea.

Supplementary Table 1—Primers used. The nucleotide sequences introduced for In-Fusion cloning are underlined in italic. The sequence information was retrieved from the M. purpureus genome sequence portal of Department ofEnergy-Joint Genome Institute (

Name / Sequence (5'3')
MppEup-F / CCATGATTACGAATTCCCTTCTGCCCAAGACCTACA
MppEup-R / GCTCGACGTATTTCAGGGGATCAAGTCGAATTTCA
hyg-F / TGAAATACGTCGAGCCTGCT
Hyg-mppE-R / CTGGCTCAAATACCCCCTGTGCATTCTGGGTAAAC
MppEdw-F / GGGTATTTGAGCCAGGTGAA
MppEdw-R / TACCGAGCTCGAATTCGACCAGGCACTGGATGATCT
PtrpC-F / TCGAGCTCGGTACCCTTACCGCCTTTGAGTGAGCT
PtrpC-mppE-R / TGGATGGACACGAATATCGATGCTTGGGTAGAATAGG
MppE-F / ATTCGTGTCCATCCATGATGC
MppE-R / ACGTTAAGTGGATCCGATACAGGAACAAGGCAATGC
TtrpC-F / GGATCCACTTAACGTTACTG
TtrpC-R / TGTCGAAAGATCCCCTGAAGCTTGCATGCCTGCAG

Supplementary Fig 1—LC-MS analysis for identification of MAz compounds in the extract. (a, b)Total ion chromatograms, absorption traces of the extracts from WT (a)and the mppE mutant (b). For absorption chromatograms, the wavelengths characteristic of MAz compounds are shown; 390, 470, and 520 nm for yellow, orange, and red components, respectively. The absorption traces are drawn to the same scale. MAz compounds are marked on relevant traces with the numbers used in the main text. (c)Absorption spectra of MAz peaks in LC separation. Each spectrum from WT and mppEare marked as aand b, respectively, to indicate the correlation to the LC-MS chromatograms data of (a)and (b). The marked numbers following a or b are the compound numbers used in the LC-MS chromatograms data of (a)and (b). (d - g) Mass spectra and their collision-induced fragmented data are provided for 5 (d) and 6 (e) from WT, and R-1 (f) and R-2 (g) from mppE.

Supplementary Fig. 2—Pigment content and profile of theorganic extracts from PDB cultures.(a)Absorptionspectra were collected with proper dilutions in methanol and the values were converted to represent the values in the original culture volume. WT, mppE, and OV-mppE are represented by a bold line, unfilled circles, and filled circles, respectively. The wavelengths characteristic of MAzPs are indicated. (b) A410 and A500are obtained from biological triplets. A410/A500 ratios are shown in the top of the panel.

Supplementary Fig. 3—Structures of the compounds other than 16. The acetate units and S-adenosyl-L-methionine-derived carbons are denoted with bars and black dots, respectively.