Supplementary Material
Production of ginsenoside aglycons and Rb1 deglycosylation
pathway profiling by HPLC and ESI-MS/MS using
Sphingobacterium multivorum GIN723
Eun-Mi Kim1, Joo-Hyun Seo1, Juhan Kim1, 3, Jun-Seong Park2, Duck-Hee Kim2, Byung-Gee
Kim1,*
1
School of Chemical and Biological Engineering, Seoul National University
2
R & D Center, Amore-Pacific Corporation, Kyounggi-do, Korea
3
Present address: UCB 216, Cooperative Institute for Research in Environmental Sciences,
CIRES 318, University of Colorado, Boulder, CO 80302
*
To whom correspondence should be addressed.
Telephone: +82-2-880-6774, Fax: +82-2-872-7528, E-mail: byungkim@snu.ac.kr
Fig. S1. 16S rRNA sequence
Figure S2. Purification result from S. multivorum GIN 723
a. protein 1
b. protein 2.
(continued)
(continued)
Figure S3. Coding gene and protein sequences. a. protein 2, b. protein 1
Figure S4. Optimum pH. a. whole cell. b. cell extract
Figure S5. The MS/MS fragmentation spectrum of peak at m/z 807 using ESI-MS/MS.The
peak at m/z 365 indicates Na+ adduct of serial two glucose moieties
.
Figure S6. Analysis of time-course reaction profile for F2 deglcosylation. a. HPLC analysis
of reaction mixtures according to reaction time. b. proposed reaction pathway from F2 to
PPD(S), “a” pathway is dominant and “b” pathway doesn’t work.
Table S1. Peptided from De Novo sequencing and degenerate primers for finding of coding
gene sequence. Underline is indicated primer region
Peptide Sequence
(De Novo sequencing)
Forward degenerate primer
Backward degenerate primer
GYTMNFTDDNLLC
GGI TAY ACI ATG AAY TTY ACI GAY GA
TC RTC IGT RAA RTT CAT IGT RTA ICC
FANEDFTNEVYLDK
TTY GCI AAY GAR GAY TTY ACI AAY GA
TC RTT IGT RAA RTC YTC RTT IGC RAA
VGEYYGLTLR
GTI GGI GAR TAY TAY GGI MTI AC
GT IAK ICC RTA RTA YTC ICC IAC
CDGFNGFEANSR
TGY GAY GGI TTY AAY GGI TTY GA
TC RAA ICC RTT RAA ICC RTC RCA
LNNTFSLSASR
AAY AAY ACI TTY WSI HT
AD ISW RAA IGT RTT RTT
SPVDFNLADYK
GAY TTY AAY HTI GCI GAY TAY AA
TT RTA RTC IGC IAD RTT RAA RTC
NLSFSFLWDFR
TTY WSI TTY HTI TGG GAY TT
AA RTC CCA IAD RAA ISW RAA
LGFDFTYYKKK
TTY GAY TTY ACI TAY TAY AA
TT RTA RTA IGT RAA RTC RAA
Glu3
Glu4
Table S2. Components of media
Components (g/L)
Name
C-source
N-source
YE
5 g Yeast Extract
BE
5 g Beef Extract
ST
5 g Soytone
TT
5 g Tryptone
PT
5 g Peptone
AC
5 g NH2Cl
Additive
10 g sodium chloride
1 mM MgCl2
1% Glucose
YEG
5 g Yeast Extract + 1g Ginseng powder
BEG
5 g Beef Extract + 1g Ginseng powder
STG
5 g Soytone + 1g Ginseng powder
TTG
5 g Tryptone + 1g Ginseng powder
PTG
5 g Peptone + 1g Ginseng powder
ACG
5 g NH2Cl + 1g Ginseng powder
0.1 mM CaCl2
Table S3. Substrates and products list