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

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

Polish Journal of Microbiology

Volume 67 (2018): Issue 2 (June 2018)

By: KATARZYNA W. SKRZYPCZAK, WALDEMAR Z. GUSTAW and ADAM D. WAŚKO

Open Access

|Jun 2018

Figures & Tables

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.

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.

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

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 strain	ΔpH^*
L. helveticus strain	6 h	12 h	18 h	24 h	30 h	36 h
T104	1.31 ± 0.01	1.35 ± 0.01	0.48 ± 0.01	0.03 ± 0.01	0.08 ± 0.01	0
T105	1.79 ± 0.01	1.23 ± 0.02	0.24 ± 0.02	0.05 ± 0.01	0.01 ± 0.01	0.01 ± 0.01
141	0.8 ± 0.01	0.62 ± 0.01	0.76 ± 0.01	0.07 ± 0.02	0.22 ± 0.01	0.13 ± 0.01
B734	0.86 ± 0.02	0.24 ± 0.01	0.75 ± 0.01	0.06 ± 0.01	0.5 ± 0.02	0
80	0.8 ± 0.01	0.21 ± 0.01	0.78 ± 0.01	0.09 ± 0.01	0.49 ± 0.01	0.17 ± 0.01
T159	0.90 ± 0.01	0.15 ± 0.03	0.65 ± 0.02	0.22 ± 0.02	0.53 ± 0.01	0.23 ± 0.01
T15	0.74 ± 0.01	0.16 ± 0.02	1.05 ± 0.01	0.09 ± 0.01	0.51 ± 0.01	0
T199	0.85 ± 0.01	0.23 ± 0.02	0.82 ± 0.01	0.04 ± 0.01	0.49 ± 0.01	0.28 ± 0.01
DSMZ 20075	0.89 ± 0.02	0.66 ± 0.03	0.31 ± 0.01	0.66 ± 0.01	0.22 ± 0.01	0
T103	0.76 ± 0.01	0.33 ± 0.02	1.06 ± 0.02	0.20 ± 0.01	0.25 ± 0.01	0
T80	0.93 ± 0.01	0.89 ± 0.01	0.18 ± 0.01	0.54 ± 0.01	0.07 ± 0.01	0.21 ± 0.01
K1	0.82 ± 0.03	0.24 ± 0.02	0.76 ± 0.01	0.14 ± 0.01	0.56 ± 0.03	0.26 ± 0.01

The proteolytic activity of L_ helveticus strains_

The bacterial strain	Proteolytic activity [mM of released α-aminoacids/l]	Profiles of amplification products of CEPs
L. helveticus T104	87.06^c ± 0.21	I (prtH/prtH2/prtH3)
L. helveticus T105	114.72^a ± 0.64
L. helveticus 141	57.67^d ± 0.54
L. helveticus B734	58.78^d ± 0.52	II (prtH/prtH3)
L. helveticus 80	37.78^h ± 0.68	III (prtH2/prtH3)
L. helveticus T159	42.67^e ± 0.14
L. helveticus T15	40.61^fg ± 0.48
L. helveticus T199	40.61^fg ± 0.34
L. helveticus DSMZ 20075	96.94^b ± 1.1
L. helveticus T103	41.33^ef ± 0.36	IV (prtH3)
L. helveticus T80	39.78^fg ± 0.28
L. helveticus K1	40.11^fg ± 0.42
L. rhamnosus E/N	39.11^gh ± 0.44	–

Sequences of primers used in reaction of amplifications of the fragments of CEPs genes_

Gen	Primer	Sequence (5’→3’)	Tm [°C]	Source
prtH	PrtH-for-1PrtH-rev-1	GGTACTTCAATGGCTTCTCCGATGCGCCATCAATCTTCTT	51.849.7	Genay et al., 2009; Lozo et al., 2011
prtH2	prtH2fprtH2r	AAGCAAAGGATGTTGTTCCAAGTAAGCCACTCTCTTCCTTCTTACCAGTTGATGATTGAACT	58.760.7	Smeianov et al., 2007
prtH3	prtH3fprtH3r	GATGATCAAGCAGATGTAAAACCGGCAGAAGATTTACTGAAGAATTAGTCAAATGACCTGTTGTCGG	61.761.0	Broadbent et al., 2011
prtH4	prtH4fprtH4r	CTGAAGCAGCAACTAATGATCCTGGTGGATTAGGATCCGTTCTGGTTGTCAG	57.759.7	Broadbent et al., 2011

DOI: https://doi.org/10.21307/pjm-2018-026 | Journal eISSN: 2544-4646 | Journal ISSN: 1733-1331

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Language: English

Page range: 203 - 211

Submitted on: Jul 21, 2017

Accepted on: Oct 27, 2017

Published on: Jun 30, 2018

Published by: Polish Society of Microbiologists

In partnership with: Paradigm Publishing Services

Publication frequency: 4 issues per year

Keywords:

cell envelope proteinases (CEPs),

proteolytic activity

Related subjects:

Microbiology and virology

© 2018 KATARZYNA W. SKRZYPCZAK, WALDEMAR Z. GUSTAW, ADAM D. WAŚKO, published by Polish Society of Microbiologists
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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