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Distribution of Cell Envelope Proteinases Genes among Polish Strains of Lactobacillus helveticus

KATARZYNA W. SKRZYPCZAK
KATARZYNA W. SKRZYPCZAK
, 
WALDEMAR Z. GUSTAW
WALDEMAR Z. GUSTAW
 y 
ADAM D. WAŚKO
ADAM D. WAŚKO
   | 30 jun 2018
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Article Category: original-paper
Publicado en línea: 30 jun 2018
Páginas: 203 - 211
Recibido: 21 jul 2017
Aceptado: 27 oct 2017
DOI: https://doi.org/10.21307/pjm-2018-026
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© 2018 Katarzyna W. Skrzypczak et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Agarose gel electrophoresis of Multiplex PCR products obtained for Lactobacillus helvetisus strains: 1 – 80; 2 – T104; 3 – T105; 4 – T159; 5 – 14; 6 – B734; 7 – T103; 8 – T15; 9 – T199; 10 – T80; 11 – K1; 12 – DSMZ 20075; M – DNA molecular marker 100 bp.
Agarose gel electrophoresis of Multiplex PCR products obtained for Lactobacillus helvetisus strains: 1 – 80; 2 – T104; 3 – T105; 4 – T159; 5 – 14; 6 – B734; 7 – T103; 8 – T15; 9 – T199; 10 – T80; 11 – K1; 12 – DSMZ 20075; M – DNA molecular marker 100 bp.

Fig. 2.

Results of amplification genes encoding CEPs: prtH (A); prtH2 (B); prtH3 (C) in Lactobacillus helvetisus strains: Line: 1 – 80; 2 – T104; 3 – T105; 4 – T159; 5 – 141; 6 – B734; 7 – T103; 8 – T15; 9 – T199; 10 – T80; 11 – K1; 12 – DSMZ 20075; line 13: Lactobacillus rhamnosus E/N; M – DNA molecular marker.
Results of amplification genes encoding CEPs: prtH (A); prtH2 (B); prtH3 (C) in Lactobacillus helvetisus strains: Line: 1 – 80; 2 – T104; 3 – T105; 4 – T159; 5 – 141; 6 – B734; 7 – T103; 8 – T15; 9 – T199; 10 – T80; 11 – K1; 12 – DSMZ 20075; line 13: Lactobacillus rhamnosus E/N; M – DNA molecular marker.

Fig. 3.

Sequence alignment for prtH of chosen strains and Lactobac illus helveticus CRZN32 (no. AF133727). Stars indicate residues that are similar in all sequences.
Sequence alignment for prtH of chosen strains and Lactobac illus helveticus CRZN32 (no. AF133727). Stars indicate residues that are similar in all sequences.

Fig. 4.

Phylogenetic tree of prtH3 gene sequences of analyzed Polish L. helveticus strains and L. helveticus CNRZ32 (no. HQ602769.1).
Phylogenetic tree of prtH3 gene sequences of analyzed Polish L. helveticus strains and L. helveticus CNRZ32 (no. HQ602769.1).

Dynamics of decrease of skim milk pH value during fermentation conducted by L. helveticus strains.

L. helveticus strainΔpH*
6 h12 h18 h24 h30 h36 h
T1041.31 ± 0.011.35 ± 0.010.48 ± 0.010.03 ± 0.010.08 ± 0.010
T1051.79 ± 0.011.23 ± 0.020.24 ± 0.020.05 ± 0.010.01 ± 0.010.01 ± 0.01
1410.8 ± 0.010.62 ± 0.010.76 ± 0.010.07 ± 0.020.22 ± 0.010.13 ± 0.01
B7340.86 ± 0.020.24 ± 0.010.75 ± 0.010.06 ± 0.010.5 ± 0.020
800.8 ± 0.010.21 ± 0.010.78 ± 0.010.09 ± 0.010.49 ± 0.010.17 ± 0.01
T1590.90 ± 0.010.15 ± 0.030.65 ± 0.020.22 ± 0.020.53 ± 0.010.23 ± 0.01
T150.74 ± 0.010.16 ± 0.021.05 ± 0.010.09 ± 0.010.51 ± 0.010
T1990.85 ± 0.010.23 ± 0.020.82 ± 0.010.04 ± 0.010.49 ± 0.010.28 ± 0.01
DSMZ 200750.89 ± 0.020.66 ± 0.030.31 ± 0.010.66 ± 0.010.22 ± 0.010
T1030.76 ± 0.010.33 ± 0.021.06 ± 0.020.20 ± 0.010.25 ± 0.010
T800.93 ± 0.010.89 ± 0.010.18 ± 0.010.54 ± 0.010.07 ± 0.010.21 ± 0.01
K10.82 ± 0.030.24 ± 0.020.76 ± 0.010.14 ± 0.010.56 ± 0.030.26 ± 0.01

The proteolytic activity of L. helveticus strains.

The bacterial strainProteolytic activity [mM of released α-aminoacids/l]Profiles of amplification products of CEPs
L. helveticus T10487.06c ± 0.21I (prtH/prtH2/prtH3)
L. helveticus T105114.72a ± 0.64
L. helveticus 14157.67d ± 0.54
L. helveticus B73458.78d ± 0.52II (prtH/prtH3)
L. helveticus 8037.78h ± 0.68III (prtH2/prtH3)
L. helveticus T15942.67e ± 0.14
L. helveticus T1540.61fg ± 0.48
L. helveticus T19940.61fg ± 0.34
L. helveticus DSMZ 2007596.94b ± 1.1
L. helveticus T10341.33ef ± 0.36IV (prtH3)
L. helveticus T8039.78fg ± 0.28
L. helveticus K140.11fg ± 0.42
L. rhamnosus E/N39.11gh ± 0.44–

Sequences of primers used in reaction of amplifications of the fragments of CEPs genes.

GenPrimerSequence (5’→3’)Tm [°C]Source
prtHPrtH-for-1PrtH-rev-1GGTACTTCAATGGCTTCTCCGATGCGCCATCAATCTTCTT51.849.7Genay et al., 2009; Lozo et al., 2011
prtH2prtH2fprtH2rAAGCAAAGGATGTTGTTCCAAGTAAGCCACTCTCTTCCTTCTTACCAGTTGATGATTGAACT58.760.7Smeianov et al., 2007
prtH3prtH3fprtH3rGATGATCAAGCAGATGTAAAACCGGCAGAAGATTTACTGAAGAATTAGTCAAATGACCTGTTGTCGG61.761.0Broadbent et al., 2011
prtH4prtH4fprtH4rCTGAAGCAGCAACTAATGATCCTGGTGGATTAGGATCCGTTCTGGTTGTCAG57.759.7Broadbent et al., 2011
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