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. 2021 Sep;59(3):325-336.
doi: 10.17113/ftb.59.03.21.7096.

Isolation, Biochemical Characterisation and Identification of Thermotolerant and Cellulolytic Paenibacillus lactis and Bacillus licheniformis

Krzysztof Makowski  1   2 , Martyna Leszczewicz  1 , Natalia Broncel  1 , Lidia Lipińska-Zubrycka  1   3 , Adrian Głębski  1 , Piotr Komorowski  4   5 , Bogdan Walkowiak  4   5
Affiliations

Isolation, Biochemical Characterisation and Identification of Thermotolerant and Cellulolytic Paenibacillus lactis and Bacillus licheniformis

Krzysztof Makowski et al. Food Technol Biotechnol. 2021 Sep.

Abstract

Research background: Cellulose is an ingredient of waste materials that can be converted to other valuable substances. This is possible provided that the polymer molecule is degraded to smaller particles and used as a carbon source by microorganisms. Because of the frequently applied methods of pretreatment of lignocellulosic materials, the cellulases derived from thermophilic microorganisms are particularly desirable.

Experimental approach: We were looking for cellulolytic microorganisms able to grow at 50 °C and we described their morphological features and biochemical characteristics based on carboxymethyl cellulase (CMCase) activity and the API® ZYM system. The growth curves during incubation at 50 °C were examined using the BioLector® microbioreactor.

Results and conclusions: Forty bacterial strains were isolated from fermenting hay, geothermal karst spring, hot spring and geothermal pond at 50 °C. The vast majority of the bacteria were Gram-positive and rod-shaped with the maximum growth temperature of at least 50 °C. We also demonstrated a large diversity of biochemical characteristics among the microorganisms. The CMCase activity was confirmed in 27 strains. Hydrolysis capacities were significant in bacterial strains: BBLN1, BSO6, BSO10, BSO13 and BSO14, and reached 2.74, 1.62, 1.30, 1.38 and 8.02 respectively. Rapid and stable growth was observed, among others, for BBLN1, BSO10, BSO13 and BSO14. The strains fulfilled the selection conditions and were identified based on the 16S rDNA sequences. BBLN1, BSO10, BSO13 were classified as Bacillus licheniformis, whereas BSO14 as Paenibacillus lactis.

Novelty and scientific contribution: We described cellulolytic activity and biochemical characteristics of many bacteria isolated from hot environments. We are also the first to report the cellulolytic activity of thermotolerant P. lactis. Described strains can be a source of new thermostable cellulases, which are extremely desirable in various branches of circular bioeconomy.

Keywords: Bacillus licheniformis; BioLector® microbioreactor; Paenibacillus lactis; carboxymethylcellulose (CMC); cellulolytic activity; thermotolerant bacteria.

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Conflict of interest statement

CONFLICT OF INTEREST The authors declare that they have no conflict of interest.

Figures

Fig. 1
Fig. 1
An exemplary morphology of isolated bacteria at 1000× magnification: a) BWO8, b) BGR9, c) BBLN3, d) BSO7, e) BSO14, and f) BSO8
Fig. 2
Fig. 2
Comparison of growth of bacteria isolated from: a) geothermal karst spring, b) hot spring, c) geothermal pond, and d) and e) fermenting hay (BSO1-8 and BSO9-16 respectively) during 48 h of incubation at 50 °C in the BioLector® bioreactor
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