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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
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Bacterial strains and plasmids
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Strain or plasmid
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Characteristics
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Reference
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L. casei
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L. casei ATCC 393 (pLZ15-)
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[26]
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BL23 containing the nisRK genes integrated to the tRNASer locus; obtained by transformation with pMEC10
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This work
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Plasmids
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Cmr, ori(pWV01), carries the nisin-inducible promoter PnisA
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Cmr, ori(pWV01), with a DNA fragment encoding the BLG mature moiety expressed under PnisA transcriptional control
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[6]
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Cmr, ori(pWV01), with a DNA fragment encoding the precursor SPUsp45-BLG expressed under PnisA transcriptional control
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[6]
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Cmr, ori(pWV01), with a DNA fragment encoding the precursor SPUsp45-LEISS-BLG expressed under PnisA transcriptional control
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[8]
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Cmr, ori(pWV01), with a DNA fragment encoding the precursor SPUsp45-LEISS-Nuc-BLG expressed under PnisA transcriptional control
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[8]
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Cmr, ori(pWV01), with a DNA fragment encoding the precursor SPUsp45-Nuc expressed under under PnisA transcriptional control
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[18]
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Emr, nisRK cloned in pIL253
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[17]
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Emr, integration plasmid containing the nisRK genes and the int-attP cassette for integration to the tRNASer locus.
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[21]
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Table 2
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Quantitative assays of BLG in different fractions of recombinant L. casei strains
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Fractionb
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Concentration of BLG (μg/L)a
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BL23 + pNZ9520c
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BL23(int:nisRK)d
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pSEC-Nuc
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pCYT:BLG
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pSEC:BLG
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pSEC:LEISS-BLG
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pSEC:LEISS-Nuc-BLG
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pSEC:LEISS-Nuc-BLG
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S (% BLGn)e
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ndf
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1.4 ± 2.2 (43%)
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1.4 ± 1.8 (0%)
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8.7 ± 2.3 (40%)
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27 ± 3.7 (46%)
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29.4 ± 6.4 (46%)
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Cs (% BLGn)
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nd
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57.6 ± 10.3 (27%)
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57.6 ± 27.3 (1%)
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38.4 ± 12 (1%)
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44.1 ± 12 (8%)
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57.7 ± 6.3 (8%)
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Ci
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nd
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19.2 ± 2.6
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647.4 ± 301.1
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975 ± 238.1
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342.9 ± 128.7
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526.9 ± 131.4
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Total
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nd
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78.2 ± 6.5
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706.4 ± 303.9
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1022.1 ± 327.5
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413.9 ± 116
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613.9 ± 144.1
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| 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
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Quantitative assays of BLG in different fractions of BL23(int:nisRK):LEISS-Nuc-BLG
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Fractionb
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Concentration (ng BLG/mg total protein)a
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Protocol Ac
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Protocol Bc
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Cs (% BLGn)d
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725,8 ± 33,7 (8%)
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2481,6 ± 407,2 (1%)
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Ci
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6573,6 ± 631,6
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884,7 ± 283,4
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Total
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7299.4 ± 597.9
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3366.8 ± 356.4
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| 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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