The Influence of Temperature and Nitrogen Source on Cellulolytic Potential of Microbiota Isolated from Natural Environment

AGNIESZKA WITA; WOJCIECH BIAŁAS; RADOSŁAW WILK; KATARZYNA SZYCHOWSKA; KATARZYNA CZACZYK

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The Influence of Temperature and Nitrogen Source on Cellulolytic Potential of Microbiota Isolated from Natural Environment

| 28 mar 2019

Polish Journal of Microbiology

Volume 68 (2019): Numero 1 (March 2019)

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Article Category: original-paper

Pubblicato online: 28 mar 2019

Pagine: 105 - 114

DOI: https://doi.org/10.21307/pjm-2019-012

Parole chiave
spp., cellulose-degrading microorganisms, nitrogen source

© 2019 Agnieszka Wita et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

CMC agar plate after staining with Congo red and NaCl.(a) determination of the supernatant activity; (b) determination of the biomass activity.

The influence of temperature on cellulolytic activity of the strains examined.(a) 4/3; (b) 4/7; (c) 29.b; (d) 4/11; (e) 4/18; (f) 26.b. (■) CMC-Na; (•) Tissue paper; (▲) Straw.

Comparison of the activity of cellulose-degrading enzymes from B. subtilis (2a) and B. licheniformis (2b). An initial casein peptone, (NH4)2SO4, yeast extract and NH4Cl concentrations were 1.25 g/l.

Contour plot depicting the relation between casein peptone, (NH4)2SO4 and yeast extract and the cellulolytic activity of both strains, Bacillus subtilis (a) and Bacillus licheniformis (b).

Regression coefficients and the selected fit statistics for two strains investigated.

Component in model	4/7 – Bacillus subtilis			4/18 – Bacillus licheniformis
Component in model	CMC-Na	Tissue paper	Straw	CMC-Na	Tissue paper	Straw
A	14.010	13.758	15.113	16.453	9.146	11.111
B	9.885	8.696	11.233	9.128	9.797	3.789
C	15.096	15.718	13.633	16.565	8.661	6.956
D	8.724	7.120	7.468	9.490	3.248	4.099
A×B	3.277	3.000	0.597	−3.483	0.494	−2.712
A×C	−0.808	−5.549	−3.786	−6.638	3.085	−3.472
A×D	1.294	−0.602	−5.714	−2.414	1.225	−2.800
B×C	4.399	−2.491	−1.287	−6.412	−1.193	−0.983
B×D	1.619	4.134	3.687	−4.134	−0.166	3.289
C×D	1.161	2.324	−1.994	−4.717	1.665	−0.530
A×B×C	−5.876	−4.340	−31.119	18.744	−8.735	−6.175
B×C×D	5.785	9.842	24.012	8.648	8.648	8.648
Adj-R²	0.886			0.941
Model p-value	< 0.0001			< 0.0001

Composition of the nitrogen sources blends according to the mixture design.

Run	A: Casein peptone (g/l)	B: (NH₄)₂SO₄(g/l)	C: Yeast extract (g/l)	D: NH₄Cl (g/l)
1	5.00	0.000	0.000	0.000
2	0.00	5.000	0.000	0.000
3	0.00	0.000	5.000	0.000
4	0.00	0.000	0.000	5.000
5	1.25	1.250	1.250	1.250
6	3.13	0.625	0.625	0.625
7	0.63	3.125	0.625	0.625
8	0.63	0.625	3.125	0.625
9	0.63	0.625	0.625	3.125
10	2.50	0.000	0.000	2.500
11	2.50	0.000	2.500	0.000
12	2.50	2.500	0.000	0.000
13	0.00	2.500	0.000	2.500
14	0.00	2.500	2.500	0.000
15	0.00	0.000	2.500	2.500
16	2.50	0.000	2.500	0.000
17	5.00	0.000	0.000	0.000
18	0.00	5.000	0.000	0.000
19	0.00	0.000	5.000	0.000
20	0.00	0.000	0.000	5.000

The response data for the cellulose-degrading enzymes activity [U] of each experimental point after the cultivation of the selected strains on the media containing various substrates for 120 h.

Run	4/7 – Bacillus subtilis			4/18 – Bacillus licheniformis
Run	CMC-Na	Tissue paper	Straw	CMC-Na	Tissue paper	Straw
1	73.341	69.931	76.155	84.545	45.651	54.975
2	51.870	46.036	57.331	47.838	48.956	18.901
3	78.858	81.562	70.540	86.859	43.448	33.492
4	44.842	37.608	38.008	49.592	16.792	20.351
5	86.250	69.578	90.502	67.032	47.400	25.125
6	67.130	59.652	22.006	70.089	42.723	51.931
7	63.610	61.600	55.361	59.664	45.89	21.751
8	75.801	61.600	31.052	72.537	46.864	31.312
9	65.790	70.978	64.362	52.856	35.44	30.849
10	64.633	45.841	18.583	47.315	40.568	21.191
11	67.991	46.718	48.692	40.523	69.297	25.819
12	81.805	74.096	67.183	41.298	50.323	21.398
13	58.227	64.584	67.386	17.805	33.017	41.822
14	91.841	46.815	55.102	22.799	38.85	20.497
15	66.825	71.173	41.523	32.724	42.395	23.676
16	67.451	30.849	48.943	40.674	58.946	20.022
17	66.886	66.850	74.287	80.405	45.374	56.023
18	48.983	41.919	54.057	43.417	48.097	19.754
19	72.537	76.958	68.496	78.736	43.021	34.637
20	42.529	32.785	35.983	43.71	16.822	20.972

The results of identification of cellulose-degrading bacteria, based on the 16 rRNA gene sequencing.

Number of strain	Identity (%)	Strain of the closest match
4/3	86.09	Bacillus pseudomycoides
4/3	85.84	Bacillus cereus
4/7	99.86	Bacillus subtilis
4/11	99.93	Bacillus cereus
4/11	99.86	Bacillus thuringiensis
4/18	99.86	Bacillus licheniformis
6/5	100.00	Achromobacter piechaudi
	100.00	Alcaligenes faecalis
	99.93	Achromobacter xylosoxidans
6/6	99.97	Lactococcus lactis

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The Influence of Temperature and Nitrogen Source on Cellulolytic Potential of Microbiota Isolated from Natural Environment

Article Category: original-paper

Pubblicato online: 28 mar 2019

Pagine: 105 - 114

DOI: https://doi.org/10.21307/pjm-2019-012

Parole chiave
spp., cellulose-degrading microorganisms, nitrogen source

© 2019 Agnieszka Wita et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Regression coefficients and the selected fit statistics for two strains investigated.

Composition of the nitrogen sources blends according to the mixture design.

The response data for the cellulose-degrading enzymes activity [U] of each experimental point after the cultivation of the selected strains on the media containing various substrates for 120 h.

The results of identification of cellulose-degrading bacteria, based on the 16 rRNA gene sequencing.

The Influence of Temperature and Nitrogen Source on Cellulolytic Potential of Microbiota Isolated from Natural Environment

Article Category: original-paper

Pubblicato online: 28 mar 2019

Pagine: 105 - 114

DOI: https://doi.org/10.21307/pjm-2019-012

Parole chiavespp., cellulose-degrading microorganisms, nitrogen source

© 2019 Agnieszka Wita et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Regression coefficients and the selected fit statistics for two strains investigated.

Composition of the nitrogen sources blends according to the mixture design.

The response data for the cellulose-degrading enzymes activity [U] of each experimental point after the cultivation of the selected strains on the media containing various substrates for 120 h.

The results of identification of cellulose-degrading bacteria, based on the 16 rRNA gene sequencing.

Parole chiave
spp., cellulose-degrading microorganisms, nitrogen source