Dataset Information

Endoglucanase gene of M42 aminopeptidase/endoglucanase family from thermophilic Bacillus sp. PW1 and PW2 isolated from Tattapani hot spring, Himachal Pradesh, India.

ABSTRACT: BACKGROUND:Thermostable cellulases are in constant demand for several biotechnological applications. Two thermophilic bacterial strains PW1 and PW2 isolated from Tattapani hot spring were found to have cellulolytic activity. Subsequently, PW1 and PW2 were identified and mined for genes encoding cellulase activity. RESULTS:Sequencing of the 16S rDNA of PW1 and PW2 identified them as Bacillus sp. PW1 (Acc no. KU711837) and Bacillus sp. PW2 (Acc no. KU711838), respectively, which clustered in the clades containing thermophilic members of Bacillus sp. and Geobacillus species. Phylogenetic analysis revealed that despite the morphological and sequence identities, Bacillus sp. PW1 and Bacillus sp. PW2 are different at the genetic level. The cellulase genes (~?1.1?kb) of the two bacterial strains were amplified using primers designed against related thermophilic cellulases. Sequencing of the cellulase gene amplicons of PW1 and PW2 revealed that they encode proteins of 280 and 206 amino acid residues, respectively. Sequence and domain analysis of the protein products of PW1 and PW2 revealed that they belong to M42 family of aminopeptidase/endoglucanase. The PW2 endoglucanase coding sequence was submitted to Genbank under accession no. MH049504. The structures of putative endoglucanases of PW1 and PW2 were generated using 1VHE.A as template, which showed the presence of vast proportion of random coils. Molecular docking of the modeled endoglucanase proteins with various substrates and products of cellulases showed that carboxymethyl cellulose and maltose exhibit the highest binding affinity, while xylan and glucose the least. CONCLUSIONS:The two thermophilic bacteria PW1 and PW2 and their endoglucanase gene can be further utilized for recombinant production of thermostable cellulases for their application in industries.

SUBMITTER: Sharma D

PROVIDER: S-EPMC6821146 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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Endoglucanase gene of M42 aminopeptidase/endoglucanase family from thermophilic Bacillus sp. PW1 and PW2 isolated from Tattapani hot spring, Himachal Pradesh, India.

Sharma Divyanshi D Sharma Parul P Dev Kamal K Sourirajan Anuradha A

Journal, genetic engineering & biotechnology 20191002 1

<h4>Background</h4>Thermostable cellulases are in constant demand for several biotechnological applications. Two thermophilic bacterial strains PW1 and PW2 isolated from Tattapani hot spring were found to have cellulolytic activity. Subsequently, PW1 and PW2 were identified and mined for genes encoding cellulase activity.<h4>Results</h4>Sequencing of the 16S rDNA of PW1 and PW2 identified them as Bacillus sp. PW1 (Acc no. KU711837) and Bacillus sp. PW2 (Acc no. KU711838), respectively, which clu ...[more]

PMID: 31659536

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Project description:The harnessing of biocatalysts from extreme environment hot spring niche for biomass conversion is significant and promising owing to the special characteristics of extremozymes attributed by intriguing biogeochemistry and extreme conditions of these environments. Hence, in the present study 38 bacterial isolates obtained from hot springs of Manikaran (~ 95 °C), Kalath (~ 50 °C) and Vasist (~ 65 °C) of Himachal Pradesh were screened for glycosyl hydrolases by in situ enrichment technique using lignocellulosic biomass (LCB). Based on their hydrolytic potential 5 isolates were selected and they were Bacillus tequilensis (VCB1, VCB2 and VSDB4), and B. licheniformis (KBFB2 and KBFB3). Cellulolytic activity assayed by growth under submerged fermentation showed that B. tequilensis VCB1 had maximum FPA activity (3.38 IU ml-1) in 48 h, while B. licheniformis KBFB3 excelled for endoglucanase (EGA of 4.81 IU ml-1 in 24 h) and cellobiase (0.71 IU ml-1 in 48 h) activities. Among all the thermophilic biocatalysts evaluated, highest exoglucanase (0.06 IU ml-1) activity was observed in B. tequilensis VSDB4 while endoglucanase of B. licheniformis KBFB3 showed optimum specific activity at pH 7 and 70 °C. Further, the presence of celS, celB and xlnB genes in the isolates suggest their possible role in biomass conversion. Protein profiling by SDS-PAGE analysis revealed that cellulase isoforms migrated with molecular masses of 75 kDa. The endoglucanase activity of promising strain B. licheniformis KBFB3 was enhanced in the presence of Ca2+, mercaptoethanol and sodium hypochlorite whereas moderately inhibited by Cu2+, Zn2+, urea, SDS and H2O2. The results of this study indicate scope for the possible development of novel biocatalysts with multifunctional thermostable glycosyl hydrolases from hot springs for efficient hydrolysis of the complex lignocellulosic biomass into simple sugars and other derived bioproducts leading to biomass valorization.

Project description:Four closely related facultative anaerobe, moderately thermophilic, Gram positive rods (JS1(T), JS5, JS11, and JS15) were isolated from sediment samples from a hot spring at Suryakund, Jharkhand, India. Colonies were pale yellow, rough surface with uneven edges on TSA after 72 h incubation. Heterotrophic growth was observed at 40-60°C and pH 5.5-11.5; optimum growth occurred at 55°C and pH 7.5. 16S rRNA gene sequence analysis revealed the strains belong to genus Anoxybacillus. DNA-DNA homology values among strains were above 70% and showed distinct ERIC and REP PCR profile. On the basis of morphology and biochemical characteristics, strain JS1(T) was studied further. Strain JS1(T) showed 99.30% sequence similarity with A. flavithermus subsp. yunnanensis, 99.23% with A. mongoliensis, 99.16% with A. eryuanensis, 98.74% with A. flavithermus subsp. flavithermus, 98.54% with A. tengchongensis, 98.51% with A. pushchinoensis, 97.91% with A. thermarum, 97.82% with A. kaynarcensis, 97.77% with A. ayderensis and A. kamchatkensis, 97.63% with A. salavatliensis, 97.55% with A. kestanbolensis, 97.48% with A. contaminans, 97.27% with A. gonensis and 97.17% with A. voinovskiensis. In 16S rRNA secondary structure based phylogenetic comparison, strain JS1(T) was clustered with Anoxybacillus eryuanensis, A. mongoliensis, and A. flavithermus subsp. yunnanensis and showed 15 species specific base substitutions with maximum variability in helix 6. Moreover, DNA-DNA relatedness between JS1(T) and the closely related type strains were well below 70%. The DNA G+C content was 42.1 mol%. The major fatty acids were C(15:0 iso), C(16:0 iso) and C(17:0iso). The polar lipids were a phosphatidylgylycerol, a diphosphatidylglycerol, a phosphatidylethnolamine, a phosphatidylcholine, a phosphatidyl monomethylethanolamine and four unknown lipids. Based on polyphasic approach, strain JS1(T) represent a novel species of the genus Anoxybacillus for which Anoxybacillus suryakundensis sp. nov. is proposed. The type strain is JS1(T) (= DSM 27374(T) = LMG 27616(T) =JCM19211(T)).

Dataset Information

Endoglucanase gene of M42 aminopeptidase/endoglucanase family from thermophilic Bacillus sp. PW1 and PW2 isolated from Tattapani hot spring, Himachal Pradesh, India.

Publications

Endoglucanase gene of M42 aminopeptidase/endoglucanase family from thermophilic Bacillus sp. PW1 and PW2 isolated from Tattapani hot spring, Himachal Pradesh, India.

Similar Datasets

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