Page 11 - Read Online

Kikukawa et al. Microbiome Res Rep 2023;2:4 https://dx.doi.org/10.20517/mrr.2022.24 Page 5 of 12

st
a
Table 1. The C16:1 content of Bifidobacterium strains (1 screening)
b
TFA C16:1 C16:1 TFA b C16:1 C16:1
Strain Strain
(mg/L) (%) (mg/L) (mg/L) (%) (mg/L)
B. bifidum JCM 1255 24.7 0 0 B. breve JCM 1192 67.8 0.5 0.3
B. adolescentis JCM 1275 43.4 0 0 B. catenulatum subsp. catenulatum 36.6 0 0
JCM 1194
B. adolescentis 12451 55.8 1.8 1.0 B. dentium JCM 1195 44.4 0.6 0.3
B. adolescentis 3-117 54.4 0.5 0.3 B. gallunarum subsp. gallunarum 20.2 0.7 0.1
JCM 6291
B. adolescentis 12-111 106.0 2.3 2.4 B. indicum JCM 1302 23.4 0 0
B. adolescentis 12-114 39.4 0.9 0.4 B. longum subsp. longum JCM 65.4 0 0
31944
B. adolescentis 4-2 57.4 0 0 B. longum subsp. infantis JCM 1222 56.0 0 0
B. adolescentis 4-16 19.6 0 0 B. longum NCC 2705 93.5 0 0
B. adolescentis 4-58 39.0 0.6 0.2 B. pseudocatenulatum JCM 1200 40.9 0 0
B. adolescentis 9-124 33.4 0.3 0.1 B. ruminantium JCM 8222 74.9 0.7 0.5
B. animalis subsp. animalis 44.7 0.0 0 B. thermophilum JCM 1207 68.1 0.8 0.5
JCM 1190
B. animalis subsp. lactis JCM 35.2 0.6 0.2 Bifidobacterium sp. JCM 7042 106.6 2.3 2.4
10602
B. boum JCM 1211 77.5 2.7 2.1

a b
C16:1: unknown C16:1, not including 9-cis-16:1. C16:1: C16 monounsaturated fatty acid; TFA: total fatty acids. The values obtained from a single
technical replicate are shown.

nd
Figure 1. The C16:1 content of the selected Bifidobacterium strains (2 screening). The gray bar indicates C16:1 content (mg/L of broth),
while the black bar indicates the C16:1 percentage in total fatty acids (TFA). Data are mean ± SD of three independent assays,
represented by the bars and whiskers. C16:1: unknown C16:1, not including 9-cis-16:1. C16:1: C16 monounsaturated fatty acid.

Chemical identity of the C16:1 extracted from Bifidobacterium sp. JCM 7042 cells
To examine the chemical identity of unknown C16:1, the DMOX derivatives of TFA extracted from the
JCM 7042 strain were analyzed by GC-MS [Figure 2A]. The analysis revealed that the molecular mass of the
C16:1 was the same as that of C16:1 authentic standards (m/z 307). The elution time of the C16:1
corresponded to that of the 7-cis-C16:1 authentic standard but not to that of 6-cis-C16:1 or 9-cis-C16:1.

To determine the position of the double bond in the C16:1 obtained from the JCM 7042 strain, FFAs
prepared from TFA were analyzed by LC-MS. For that analysis, FFAs were not derivatized. Via the plasma-