Supplemental Materials

[Title]
Poly--glutamic Acid Synthesis Using a Novel Catalytic Activity
of RimK from Escherichia coli K-12

[Author]
Kuniki Kino,* Toshinobu Arai, and Yasuhiro Arimura

*Corresponding author. Mailing address: Department of Applied

Chemistry, Faculty of Science and Engineering, Waseda
University, 3-4-1 Ohkubo, Shinjuku-ku, Tokyo 169-8555, Japan.
Phone: +81-3-5286-3211. Fax: +81-3-3232-3889. E-mail:
kkino@waseda.jp
 Supplememtal TABLE 1. LC-ESI-MS analysis of the tripeptide synthesis catalyzed by RimK
+
Substrate Phosphate
a [M+H]
Supplemental 1 2
Deduced product
(mM) Calcd
b
Found
c

TABLE 1. Glu-Glu Val Glu-Glu-Val 0.5 376.2 376.0

Leu Glu-Glu-Leu 0.4 390.2 390.1
Ile Glu-Glu-Ile 0.3 390.2 390.0
Ala Glu-Glu-Ala 0.9 348.1 348.0
Gly Glu-Glu-Gly 1.1 334.1 334.0
Ser Glu-Glu-Ser 1.3 364.1 364.1
Thr Glu-Glu-Thr 0.9 378.2 378.1
Met Glu-Glu-Met 0.6 408.1 408.1
Cys Glu-Glu-Cys 1.0 380.1 n.d.
d
Glu-Glu-Cys-S-S-Cys-Glu-Glu 757.2 757.1
Arg Glu-Glu-Arg 0.1 433.2 n.d.
Lys Glu-Glu-Lys 0 405.2 n.d.
His Glu-Glu-His 0.2 414.2 n.d.
Trp Glu-Glu-trp 0.2 463.2 463.1
Phe Glu-Glu-Phe 0.2 424.2 424.0
Tyr Glu-Glu-Tyr 0.2 440.2 439.9
Asp Glu-Glu-Asp 1.0 392.1 392.1
Asn Glu-Glu-Asn 0.5 391.2 391.1
Gln Glu-Glu-Gln 0.5 405.2 405.1
Pro Glu-Glu-Pro 0 374.2 n.d.
a
Average of 2 measurements. All data were adjusted using the blank value (2.3 mM). When Glu-Glu and Glu was
used, 8.5 mM Pi was detected.
b +
[M+H] value was calculated on the basis of chemical formula.
c +
[M+H] value are shown when m/z peak corresponding to deduced peptide was detected. n.d., not detected.
d
Glu-Glu-Cys-S-S-Cys-Glu-Glu indicates that Glu-Glu-Cys was connected to one another with disulfide bond.
 Supplemental FIG. 1.

100
[RE10+H]+
90 m/z 1465.6 [RE11+H]+
m/z 1594.6
80
Relative abundance

70 [RE12+H]+
[RE9+H]+ m/z 1723.6
60 m/z 1336.5

50

40 [RE13+H]+
[RE8+H]+ m/z 1852.7
30 [RE7+H]+ m/z 1207.5
m/z 1078.4 [RE14+H]+
20 m/z 1981.7

10

0
200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700 1800 1900 2000

m/z

Supplemental FIG. 1. LC-ESI-MS analysis of the hetero-peptide synthesis catalyzed by RimK. Arg-Glu and Glu were used as substrates.
Peptides are shown in brackets as REX, where R is Arg; E is Glu; and X is peptide length of Glu. For example, RE7 indicates Arg-Glu-Glu-Glu-
Glu-Glu-Glu-Glu. The peaks were assigned as follows: m/z 1078.4, [RE7+H]+; m/z 1207.5, [RE8+H]+; m/z 1336.5, [RE9+H]+; m/z 1465.6,
[RE10+H]+; m/z 1594.6, [RE11+H]+; m/z 1723.6, [RE12+H]+; m/z 1852.7, [RE13+H]+; and m/z 1981.7, [RE14+H]+.