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Supplementary material for chapter 3

2% 6%0%4%

3%1%

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7%

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9% 4%

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4%

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11%

8%

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D M N O T U V A J K L C E F G H I P Q R S

TMC 3115 BF3 PRI 1 S6

NCTC13001

MGYG-HGUT-02396 JCM 1255

BGN4 PRL2010 S17 Inner ring

Outer ring

a

D, 1%

M, 4%

N, 0%

O, 3%

T, 3% U, 1%

V, 2%

W, 0%

Y, 0%

Z, 0%A, 0%

B, 0%

J, 7%

K, 5%

L, 7%

C, 3%

E, 7%

F, 3%

G, 7%

H, 3%

I, 2%

P, 3%

Q, 1%

R, 8%

S, 6%

_ , 23%

[D] Cell cycle control, cell division, chromosome partitioning [M] Cell wall/membrane/envelope biogenesis

[N] Cell motility

[O] Post-translational modification, protein turnover, and chaperones [T] Signal transduction mechanisms

[U] Intracellular trafficking, secretion, and vesicular transport [V] Defense mechanisms

[W] Extracellular structures [Y] Nuclear structure [Z] Cytoskeleton

[A] RNA processing and modification [B] Chromatin structure and dynamics

[J] Translation, ribosomal structure and biogenesis [K] Transcription

[L] Replication, recombination and repair [C] Energy production and conversion [E] Amino acid transport and metabolism [F] Nucleotide transport and metabolism [G] Carbohydrate transport and metabolism [H] Coenzyme transport and metabolism [I] Lipid transport and metabolism [P] Inorganic ion transport and metabolism

[Q] Secondary metabolites biosynthesis, transport, and catabolism [R] General function prediction only

[S] Function unknown [-] Not assigned

0 100 200 300 400 500 600

1

METABOLISM NOT ASSIGNED

INFORMATION STORAGE AND PROCESSING

POORLY CHARACTERIZED CELLULAR PROCESSING AND SIGNALING

Supplementary Figure 3.1. Distribution of Cluster of Orthologues (COG) functional categories in TMC3115 genome. (a) The COG subcategories distribution. (b) Top four COG categories distribution.

a

b

98

BF3, 40

PRI 1, 147

S6, 0

TMC3115, 105 NCTC13001, 10

MGYG-HGUT-02396, 0 JCM1255, 4

BGN4, 49 PRL2010, 110

S17, 93

b

Supplementary Figure 3.2. Comparative genomics of B. bifidum TMC3115. (a) Distribution of COG categories among the strains. The numbers highlighted in black shows the average percentage of genes for each category while the number in red shows the percentage for the TMC3115 strain. COG classification: [D] Cell cycle control, cell division, chromosome partitioning; [M] Cell wall/membrane/envelope biogenesis;[N] Cell motility;[O] Post-translational modification, protein turnover, and chaperones;[T] Signal transduction mechanisms;[U] Intracellular trafficking, secretion, and vesicular transport;[V] Defense mechanisms;[A] RNA processing and modification; ;[J] Translation, ribosomal structure and biogenesis;[K] Transcription;[L] Replication, recombination and repair;[C] Energy production and conversion;[E] Amino acid transport and metabolism;[F] Nucleotide transport and metabolism;[G] Carbohydrate transport and metabolism;[H] Coenzyme transport and metabolism;[I] Lipid transport and metabolism;[P] Inorganic ion transport and metabolism;[Q] Secondary metabolites biosynthesis, transport, and catabolism;[R] General function prediction only;[S] Function unknown. (b) The number of unique genes present in each strain.

2% 6%0%4%

3%1%

3%0%

10%

7%

10%

9% 4%

4%

8%

3%

2%

4%

1%

11%

8%

2% 5%0%3%

3%1%

4%0%

10%

7%

9%

10% 4%

4%

8%

3%

2%

4%

1%

11%

8%

D M N O T U V A J K L C E F G H I P Q R S

TMC 3115 BF3 PRI 1 S6

NCTC13001

MGYG-HGUT-02396 JCM 1255

BGN4 PRL2010 S17 Inner ring

Outer ring

a

99

Supplementary Table 3.1. Sortase dependent pili clusters in B. bifidum strains. The strains are grouped in four groups based on number of pili and their pilin motifs.

CWSS Pilin Motif E box CWSS Pilin Motif E box CWSS Pilin Motif E box

PRL2010 3 LPGTGGNATLTVSTK YTLTETEAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

KGALPTVVKK DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

NCIMB 41171 3 LPGTGKGALPTVVKK YTLTETEAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

GNATLTVSTK DTLKVTVDNK GGAAATVYAK

GKTLLTVTMK VGKNVTVEYK

IGAGVTVGVK

BGN4 3 LPGTGKGALPTVVKK YTLTETEAPAGYLPLTG KGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

NNNTLTVAMK GTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

MJR8628B 3 LPGTGKGALPTVVKK YTLTETEAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVGTAATVTFK YTVTETAVADGY

DNTLLTVAMK DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

A8 3 LPGTGKGNLPTVDKK YTLTETEAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

NNNTLTVAMK DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

324B 3 LPGTGKGNLPTVDKK YTLTETEAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

NNNTLTVAMK DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

BF3 3 LPGTGKGNLPTVDKK YTLTETEAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

NNNTLTVAMK DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

Bbif1887B 3 LPGTGKGNLPTVDKK YTLTETEAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

NNNTLTVAMK DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

LMG 11582 3 LPGTGKGDLPTVDKK YTLTETEAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

NNNTLTVAMK DTLKVTVDNK GGAAATVYAK

IGAGVTVGVK VGKTVTVEYK

LMG 13195 3 LPGTGKGDLPTVDKK YTLTETEAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

NNNTLTVAMK DTLKVTVDNK GGAAATVYAK

IGAGVTVGVK VGKTVTVEYK

Calf96 3 LPGTGKGDLPTVDKK YTLTETEAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

DNTLLTVAMK DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

S6 3 LPGTGKGNLPTVDKK YTLTETKAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

NNNTLTVAMK DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

HGUT02396 3 LPGTGKGNLPTVDKK YTLTETKAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

NNNTLTVAMK DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK STRAIN

No of Pili

MAJOR PILINS

fimA fimA fimP Groups

G1

100

CWSS Pilin Motif E box CWSS Pilin Motif E box CWSS Pilin Motif E box IGAGVTVGVK

S17 3 LPGTGKGDLPTVDKK YTLTETEAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

ATCC 29521 3 LPGTGKGDLPTVDKKYTLTETEAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

LMG 11041 3 LPGTGKGDLPTVDKKYTLTETEAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

DTLKVTVDNK GGAAATVYAK

IGAGVTVGVK VGKTVTVEYK

DSM 20456 3 LPGTGKGDLPTVDKKYTLTETEAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

JCM 1255 3 LPGTGKGDLPTVDKKYTLTETEAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

DTLKVTVDNK GGAAATVYAK

IGAGVTVGVK VGKTVTVEYK

NCTC13001 3 LPGTGKGDLPTVDKK YTLTETKAPAGYLPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

TMC3115 3 LPGTGKGALPTVVKK YTLTETEAPAGYLPLTG IGAGVTVGVK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

NNNTLTVAMK GGAAATVYAK

JCM 1254 3 LPGTGNNNTLTVAMKYTLTETEAPAGYLKYTGNGYQFTVSDK YTIEEIAAPNGY LPKTGGGAAATVYAKYTVTETAVADGY DTLKVTVDNK

3 LPGTGKGDLPTVDKKYTLTETEAPAGYLKYTGNGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

NNNTLTVAMK DTLKVTVDNK GGAAATVYAK

156B 2 LPGTGKGALPTVVKK YTLTETEAPAGY LPKTGVDTAATVTFK YTVTETAVADGY

NNNTLTVAMK GGAAATVYAK

ICIS-310 2 LPGTGKGDLPTVDKKYTLTETEAPAGY LPKTGVDTAATVTFK YTVTETAVADGY

NNNTLTVAMK GGAAATVYAK

2789STDY56088772 LPGTGKGNLPTVDKKYTLTETEAPAGY LPKTGVDTAATVTFK YTVTETAVADGY

NNNTLTVAMK GGAAATVYAK

791 2 LPGTGKGNLPTVDKKYTLTETEAPAGY LPKTGVDTAATVTFK YTVTETAVADGY

NNNTLTVAMK GGAAATVYAK

BI-14 2 LPGTGKGNLPTVDKKYTLTETEAPAGY LPKTGVDTAATVTFK YTVTETAVADGY

NNNTLTVAMK GGAAATVYAK

IPLA 20015 2 LPGTGKGDLPTVDKKYTLTETEAPAGY LPKTGVDTAATVTFK YTVTETAVADGY

GNATLTVSTK GGAAATVYAK

85B 2 LPGTGGNATLTVSTK YTLTETEAPAGY LPKTGVDTAATVTFK YTVTETAVADGY

KGDLPTVDKK GGAAATVYAK

IPLA 20017 2 LPGTGKGDLPTVDKKYTLTETEAPAGY LPKTGVDTAATVTFK YTVTETAVADGY

GKTLLTVAMK GGAAATVYAK

LMG 11583 2 LPGTGDNATLTVSTK YTLTETEAPAGY LPKTGVDTAATVTFK YTVTETAVADGY

KGALPTVVKK GGAAATVYAK

G1971 2 LPGTGKGDLPTVDKKYTLTETEAPAGY LPKTGVDTAATVTFK YTVTETAVADGY

GGAAATVYAK

62-13 2 LPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

ASM157686v1 2 LPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

CAG234 2 LPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

PRI1 2 LPLTG NGYQFTVSDK YTIEEIAAPNGY LPKTGVDTAATVTFK YTVTETAVADGY

DTLKVTVDNK GGAAATVYAK

VGKNVTVEYK IGAGVTVGVK

ASM157689v1 1 LPKTGVDTAATVTFK YTVTETAVADGY

GGAAATVYAK STRAIN

No of Pili

MAJOR PILINS

Groups

fimA fimA fimP

G2

G3

G4

G1

101