Supplementary Material
Table S1 Plasmids
Name
Properties
Reference
pJOE4786.1
pBR322
pVWEx1
pJeM1
pUC8:16
pUC57hasA
pJOE7706.1
pJH122.1
pJH174.1
pJH181.3
pJH182.1
pJH183.2
pJH195.2
pJH196.2
pJH197.1
ApR,
lacPOZ‘
ApR, TcR, rop, rep (pMB1)
KmR, rep. Ptac, lacIq
KmR, rhaPBAD, rhaRS, mob, His-tag-eGFP
ApR, lacPOZ, uvrA‘, vgb
ApR, rep, lacZ::hasA
KmR, rop, rep, Ptac lacIq, rrnB, eGFP
pJeM1; ΔeGFP, hasA
pJOE7706.1; ΔeGFP, hasA
pJH174.1; hasC
pJH174.1; vgb
pJH181.3; hasB
pJH174.1; glmU
pJH181.3; glmU
pJH195.2; hasC
(Jeske and Altenbuchner 2010)
(Bolivar et al. 1977)
(Peters-Wendisch et al. 2001)
(Jeske and Altenbuchner 2010)
(Liu et al. 1994)
this work
this work
this work
this work
this work
this work
this work
this work
this work
this work
Table S2 Oligonucleotides used for the construction of hyaluronic acid production plasmids
Name
S8137
S8450
S8140
S8141
S8142
S8143
S8506
S8507
S8614
S8615
Sequence (5'→3')
AAAAAAACTAGTGAACAATTCTTAAGAAGGAGA
AAAAAATGTACAGCTAGCTTACAGCAGTTTTTTACGGG
AAAAAAACTAGTCACAGGCCTAGCCTTTTTG
AAAAAAAAGCTTTCTAGATCAGTAAGCCTTGCCAGTC
AAAAAAACTAGTTCAGCTTTCCTATCAAGGAG
AAAAAATCTAGATCAAGCTGGCGCAATCTTG
AAAAAAACTAGTATGTTAGACCAGCAAACCAT
AAAAAAGCTAGCTTATTCAACCGCTTGAGCG
AAAAAAACTAGTTCACTGACCGCCCAGGAT
AAAAAAGCTAGCTTAGCTCTTCTTGATCTTCTCC
restriction
site(s)
amplified
gene(s)
SpeI
BsrGI, NheI
SpeI
HindIII, XbaI
SpeI
XbaI
SpeI
NheI
SpeI
NheI
hasA
hasA
hasC
hasC
hasB
hasB
vgb
vgb
glmU
glmU
(a)
(b)
(c)
(d)
(e)
(f)
(g)
(h)
(i)
(j)
Figure S1 Microscopy of negative stained C. glutamicum cultures or culture supernatants (48 h of cultivation). (a)
wild type, MEK700 minimal medium. (b) wild type, CGXII minimal medium. (c) pJH174.1, CGXII medium,
uninduced culture. (d) pJH174.1, CGXII medium, induced culture. (e) pJH174.1, CGXII medium, uninduced
culture supernatant. (f) pJH174.1, CGXII medium, induced culture supernatant. (g) pJH182.1, CGXII medium,
uninduced culture. (h) pJH182.1, CGXII medium, induced culture. (i) pJH182.1, CGXII medium, uninduced culture
supernatant. (j) pJH182.1, CGXII medium, induced culture supernatant.
CGXII culture supernatants and the MEK700 wild type culture (OD600 = 8) were not diluted before staining (5 µl
supernatant or culture + 5 µl nigrosin dye). CGXII cultures were diluted with H2O to an OD600 of 1.5 prior to
staining. CGXII wilt type culture was diluted to an OD600 of 8 prior to staining.
References
Bolivar, F., Rodriguez, R.L., Greene, P.J., Betlach, M.C., Heyneker, H.L., Boyer, H.W.,
Crosa, J.H. and Falkow, S. (1977) Construction and characterization of new cloning vehicles.
II. A multipurpose cloning system. Gene 2, 95-113.
Jeske, M. and Altenbuchner, J. (2010) The Escherichia coli rhamnose promoter rhaP(BAD) is
in Pseudomonas putida KT2440 independent of Crp-cAMP activation. Appl Microbiol
Biotechnol 85, 1923-1933.
Liu, S.C., Liu, Y.X., Webster, D.A. and Stark, B.C. (1994) Sequence of the region
downstream of the Vitreoscilla hemoglobin gene: vgb is not part of a multigene operon. Appl
Microbiol Biotechnol 42, 304-308.
Peters-Wendisch, P.G., Schiel, B., Wendisch, V.F., Katsoulidis, E., Möckel, B., Sahm, H. and
Eikmanns, B.J. (2001) Pyruvate carboxylase is a major bottleneck for glutamate and lysine
production by Corynebacterium glutamicum. J Mol Microbiol Biotechnol 3, 295-300.