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the authors compared different strategies to optimize the production of bovine β…


Biology Articles » Microbiology » Microbial Physiology » Efficient production and secretion of bovine beta-lactoglobulin by Lactobacillus casei » Tables

Tables
- Efficient production and secretion of bovine beta-lactoglobulin by Lactobacillus casei

Table 1
Bacterial strains and plasmids
Strain or plasmid
Characteristics
Reference
L. casei


      BL23
L. casei ATCC 393 (pLZ15-)
[26]
      BL23(int:nisRK)
BL23 containing the nisRK genes integrated to the tRNASer locus; obtained by transformation with pMEC10
This work
Plasmids


      pVE3655
Cmr, ori(pWV01), carries the nisin-inducible promoter PnisA

      pCYT:BLG
Cmr, ori(pWV01), with a DNA fragment encoding the BLG mature moiety expressed under PnisA transcriptional control
[6]
      pSEC:BLG
Cmr, ori(pWV01), with a DNA fragment encoding the precursor SPUsp45-BLG expressed under PnisA transcriptional control
[6]
      pSEC:LEISS-BLG
Cmr, ori(pWV01), with a DNA fragment encoding the precursor SPUsp45-LEISS-BLG expressed under PnisA transcriptional control
[8]
      pSEC:LEISS-Nuc-BLG
Cmr, ori(pWV01), with a DNA fragment encoding the precursor SPUsp45-LEISS-Nuc-BLG expressed under PnisA transcriptional control
[8]
      pSEC:Nuc
Cmr, ori(pWV01), with a DNA fragment encoding the precursor SPUsp45-Nuc expressed under under PnisA transcriptional control
[18]
      pNZ9520
Emr, nisRK cloned in pIL253
[17]
      pMEC10
Emr, integration plasmid containing the nisRK genes and the int-attP cassette for integration to the tRNASer locus.
[21]
Table 2
Quantitative assays of BLG in different fractions of recombinant L. casei strains
Fractionb
Concentration of BLG (μg/L)a


BL23 + pNZ9520c
BL23(int:nisRK)d



pSEC-Nuc
pCYT:BLG
pSEC:BLG
pSEC:LEISS-BLG
pSEC:LEISS-Nuc-BLG
pSEC:LEISS-Nuc-BLG
S (% BLGn)e
ndf
1.4 ± 2.2 (43%)
1.4 ± 1.8 (0%)
8.7 ± 2.3 (40%)
27 ± 3.7 (46%)
29.4 ± 6.4 (46%)
Cs (% BLGn)
nd
57.6 ± 10.3 (27%)
57.6 ± 27.3 (1%)
38.4 ± 12 (1%)
44.1 ± 12 (8%)
57.7 ± 6.3 (8%)
Ci
nd
19.2 ± 2.6
647.4 ± 301.1
975 ± 238.1
342.9 ± 128.7
526.9 ± 131.4
Total
nd
78.2 ± 6.5
706.4 ± 303.9
1022.1 ± 327.5
413.9 ± 116
613.9 ± 144.1
a Values represent the means ± SD of at least three independent experiments; b S, supernatant; Cs, soluble cytoplasmic fraction; Ci, insoluble cytoplasmic fraction; c L. casei BL23 cells transformed with pNZ9520 plasmid; d L. casei BL23 cells with nisRK genes integrated in the chromosome; e Percentage of BLG in its native form; f nd, not detectable.
Table 3
Quantitative assays of BLG in different fractions of BL23(int:nisRK):LEISS-Nuc-BLG
Fractionb
Concentration (ng BLG/mg total protein)a


Protocol Ac
Protocol Bc

Cs (% BLGn)d
725,8 ± 33,7 (8%)
2481,6 ± 407,2 (1%)
Ci
6573,6 ± 631,6
884,7 ± 283,4
Total
7299.4 ± 597.9
3366.8 ± 356.4
a Values represent the means ± SD of at least two independent experiments; b Cs, soluble cytoplasmic fraction; Ci, insoluble cytoplasmic fraction; c see nisin induction in Materials and Methods; d Percentage of BLG in its native form.

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