Characterization and sequence analyses of hyper-cellulase producing Bacillus paramycoides

• Authors: Ameer Khusro , Ramakrishnan Venkatesh Prabu
• Languages of publication: EN

Abstracts

EN

The present study was investigated to isolate and identify a hyper-cellulase producing bacterium from soil. Initially, a total of 11 cellulase producing bacteria were isolated from the collected soil sample using serial dilution technique, followed by cellulose hydrolysis plate assay method. The hyper-cellulase producing bacterium was identified as Bacillus paramycoides strain KASHA using different biochemical and molecular characterization tools. Furthermore, 16S rRNA sequences of strain KASHA were used to determine the secondary structure of RNA using RNAfold web server. As per the results obtained, the minimum free energy value for strain KASHA was calculated as -199.69 kcal/mol. Mountain plot and entropy depicted the hierarchical representation of RNA secondary structure. The GC content of sequence for strain KASHA was calculated as 53%. The evolutionary relationship of the strain was determined using Neighbor Joining algorithm which showed close resemblance with other bacilli strains.

Keywords

EN

16S rRNA sequencing; Bacillus paramycoides; Cellulase; Characterization; Phylogenetic tree

Discipline

• MICROBIOLOGY: MICROBIOLOGY

• Publisher: Scientific Publishing House „DARWIN”
• Journal: World News of Natural Sciences
• Year: 2025
• Volume: 61
• Issue: 2
• Pages: 65-74

Contributors

author

Ameer Khusro

• Department of Research Analytics, Saveetha Dental College and Hospitals, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai – 600077, India

author

Ramakrishnan Venkatesh Prabu

• Department of Microbiology, Sona College of Arts and Science, Salem - 636005, Tamil Nadu, India

References

• [1] Imadi SR, Babar MM, Hasan H, Gul A (2016). Soil microflora – An extensive research. In: Hakeem K, Akhtar J, Sabir M (eds) Soil Science: Agricultural and Environmental Prospectives. Springer. 303-316. ISBN: 978-3-319-34449-2
• [2] Archana J, Thilo E, Ruth F, Philippe B, Wilfred O (2015). Quantification of spatial distribution and spread of bacteria in soil at microscale. EGU General Assembly. Vienna, Austria. Id:15420
• [3] Khusro A, Aarti C, Almutairi MH, Almutairi BO (2024). Production and statistical optimization of invertase-free exoinulinase from Glutamicibacter arilaitensis using goat dung as ideal feedstock. BioResources 19, 3249-3270
• [4] Aarti C, Khusro A, Agastian P, Darwish NM, Al Farraj DA (2020). Molecular diversity and hydrolytic enzymes production abilities of soil bacteria. Saudi J. Biol. Sci. 27, 3235-3248
• [5] Lugani Y, Rai R, Prabhu AA, Maan P, Hans M, Kumar V, et al. (2020). Recent advances in bioethanol production from lignocelluloses: A comprehensive review with a focus on enzyme engineering and designer biocatalysts. Biofuel Res. J. 28, 1267-1295
• [6] Binod P, Palkhiwala P, Gaikaiwari R, Nampoothiri KM, Duggal A, Dey K, et al. (2013). Industrial enzymes – present status and future perspectives for India. J. Sci. Ind. Res. 72, 271-286
• [7] Zhou J, Wang YH, Chu J, Zhuang YP, Zhang SL, Yin P (2008). Identification and purification of the main components of cellulases from a mutant strain of Trichoderma viride T 100-14. Bioresour. Technol. 99, 6826-6833
• [8] Aarti C, Khusro A, Agastian P, Kuppusamy P, Al Farraj DA (2022). Synthesis of gold nanoparticles using bacterial cellulase and its role in saccharification and bioethanol production from aquatic weeds. J. King Saud Univ. Sci. 34, 101974, doi.org/10.1016/j.jksus.2022.101974
• [9] Sadhu S, Ghosh PK, Aditya G, Maiti TK (2014). Optimization and strain improvement by mutation for enhanced cellulase production by Bacillus sp. (MTCC10046) isolated from cow dung. J. King Saud Univ. Sci. 26, 323-332
• [10] Aarti C, Khusro A, Agastian P (2018). Carboxymethyl cellulase production optimization from Glutamicibacter arilaitensis strain ALA4 and its application in lignocellulosic waste biomass saccharification. Prep. Biochem. Biotechnol. 48, 853-866
• [11] Sneath PHA (1994). Gram positive rods. In Bergeys Manual of Systematic Bacteriology, 9th ed.; Hensyl, W.M., Ed.; Williams and Wilkins: Philadelphia, PA, USA. 2106-2111.
• [12] Gruber AR, Lorenz R, Bernhart SH, Neuböck R, Hofacker IL (2008). The Vienna RNA website. Nucleic Acids Res. 36 (Web Server issue): W70-4. doi: 10.1093/nar/gkn188
• [13] Rosano GL and Ceccarelli EA (2009). Rare codon content affects the solubility of recombinant proteins in a codon bias-adjusted Escherichia coli strain. Microb. Cell Fact. 8, 41. doi.org/10.1186/1475-2859-8-41
• [14] Tamura K, Dudley J, Nei M, Kumar S (2007). MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol. Biol. Evol. 24, 1596-1599
• [15] Jacoby R, Peukert M, Succurro A, Koprivova A, Kopriva S (2017). The role of soil microorganisms in plant mineral nutrition-current knowledge and future directions. Front. Plant Sci. 8, 1-19
• [16] Sankari D and Khusro A (2014). Biochemical, molecular characterization and sequence analysis of keratinase producing novel strain of Bacillus licheniformis isolated from poultry farm. Int. J. Pharm. Pharm. Sci. 6, 457-461
• [17] Khusro A, Preetamraj JP, Panicker SG (2014). Multiple heavy metals response and antibiotic sensitivity pattern of Bacillus subtilis strain KPA. J. Chem. Pharm. Res. 6, 532-538
• [18] Saraswathi K, Mahalakshmi S, Khusro A, Arumugam P, Al Farraj DA, Alkufeidy RM (2020). In vitro biological properties of Streptomyces cangkringensis isolated from the floral rhizosphere regions. Saudi J. Biol. Sci. 27, 3249-3257
• [19] Tiwari S, Yadav J, Gaur R, Singh R, Verma T, Yadav JS, et al. (2022). Multistep structural and chemical evaluation of sugarcane baggase, pretreated with alkali for enhancing the enzymatic saccharification by cellulase and xylanase of the Pseudomonas sp. CVB-10 (MK443365) and Bacillus paramycoides T4 (MN370035) mix-culture system. Front. Energy Res. 9, 726010.
• [20] Khusro A, Aarti C, Barbabosa-Pilego A, Hernández SR (2019). Anti-pathogenic, antibiofilm, and technological properties of fermented food associated Staphylococcus succinus strain AAS2. Prep. Biochem. Biotechnol. 49, 176-183
• [21] Aarti C, Khusro A (2019). Functional and technological properties of exopolysaccharide producing autochthonous Lactobacillus plantarum strain AAS3 from dry fish based fermented food. LWT Food Sci. Technol. 114, 108387, doi.org/10.1016/j.lwt.2019.108387
• [22] Khusro A, Aarti C (2015). Molecular identification of newly isolated Bacillus strains from poultry farm and optimization of process parameters for enhanced production of extracellular amylase using OFAT method. Res. J. Microbiol. 10, 393-420
• [23] Ntushelo K, Harrison NA, Elliott ML (2012). Comparison of the ribosomal RNA operon from Texas Phoenix decline and lethal yellowing phytoplasmas. Eur. J. Plant Pathol. 133, 779-782
• [24] Aarti C and Khusro A (2015). Discovery of polygalacturonase producing Bacillus tequilensis strain ARMATI using 16S rRNA gene sequencing. Asian J. Pharm. Clin. Res. 8, 58-62
• [25] Zuker M and Stiegler P (1981). Optimal computer folding of large RNA sequences using thermodynamics and auxiliary information. Nucleic Acids Res. 9, 133-148

• Document Type: article