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. 2020 Jun 4;25(11):2609.
doi: 10.3390/molecules25112609.

Effect of Cultural Conditions on Protease Production by a Thermophilic Geobacillus thermoglucosidasius SKF4 Isolated from Sungai Klah Hot Spring Park, Malaysia

Allison D Suleiman  1   2 , Nor'Aini Abdul Rahman  1 , Hidayat Mohd Yusof  1 , Fairolniza Mohd Shariff  3 , Nur Adeela Yasid  4
Affiliations

Effect of Cultural Conditions on Protease Production by a Thermophilic Geobacillus thermoglucosidasius SKF4 Isolated from Sungai Klah Hot Spring Park, Malaysia

Allison D Suleiman et al. Molecules. .

Abstract

Major progress in the fields of agriculture, industry, and biotechnology over the years has influenced the quest for a potent microorganism with favorable properties to be used in scientific research and industry. This study intended to isolate a new thermophilic-protease-producing bacterium and evaluate its growth and protease production under cultural conditions. Protease producing bacteria were successfully isolated from Sungai Klah Hot Spring Park in Perak, Malaysia, and coded as SKF4; they were promising protease producers. Based on microscopic, morphological, and 16S rRNA gene analysis, isolate SKF4 was identified as Geobacillus thermoglucosidasius SKF4. The process of isolating SKF4 to grow and produce proteases under different cultural conditions, including temperature, pH, NaCl concentration, carbon and nitrogen sources, and incubation time, was explored. The optimum cultural conditions observed for growth and protease production were at 60 to 65 °C of temperature, pH 7 to 8, and under 1% NaCl concentration. Further, the use of casein and yeast extract as the nitrogen sources, and sucrose and fructose as the carbon sources enhanced the growth and protease production of isolate SKF4. Meanwhile, isolate SKF4 reached maximum growth and protease production at 24 h of incubation time. The results of this study revealed a new potent strain of thermophilic bacterium isolated from Sungai Klah Hot Spring Park in Perak, Malaysia for the first time. The high production of thermostable protease enzyme by G. thermoglucosidasius SKF4 highlighted the promising properties of this bacterium for industrial and biotechnological applications.

Keywords: Geobacillus thermoglucosidasius SKF4; Malaysia; hot spring; protease.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Sungai Klah Hot Spring Park located in Perak, Malaysia.
Figure 2
Figure 2
(A) Growth and colony morphology of isolate SKF4 on Luria Bertani agar. (B) Protease activity of isolate SKF4 on skim milk agar.
Figure 3
Figure 3
(A) The agarose gel electrophoresis of 16S rRNA amplified from the genomic DNA of the isolate SKF4 using 27F and 1492R primers. (B) Phylogenetic tree of Geobacillus thermoglucosidasius SKF4. The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The analysis involved 15 nucleotide sequences of some closely related species. Bacillus subtillis was placed as an outer group.
Figure 4
Figure 4
Effects of (A) temperature, (B) pH, (C) carbon source, (D) NaCl concentration, (E) nitrogen source, and (F) incubation time on growth and protease production of G. thermoglucosidasius SKF4. Data are expressed as means (n = 3) ± SDs.
See this image and copyright information in PMC

References

    1. Ibrahim A.S.S., Al-Salamah A.A., Elbadawi Y.B., El-Tayeb M.A., Ibrahim S.S.S. Production of extracellular alkaline protease by new halotolerant alkaliphilic Bacillus sp. NPST-AK15 isolated from hyper saline soda lakes. Electron. J. Biotechnol. 2015;18:236–243. doi: 10.1016/j.ejbt.2015.04.001. - DOI
    1. Yildirim V., Baltaci M.O., Ozgencli I., Sisecioglu M., Adiguzel A., Adiguzel G. Purification and biochemical characterization of a novel thermostable serine alkaline protease from Aeribacillus pallidus C10: A potential additive for detergents. J. Enzym. Inhib. Med. Chem. 2017;32:468–477. doi: 10.1080/14756366.2016.1261131. - DOI - PMC - PubMed
    1. Rao M.B., Tanksale A.M., Ghatge M.S., Deshpande V.V. Molecular and biotechnological aspects of microbial proteases. Microbiol. Mol. Biol. Rev. 1998;62:597–635. doi: 10.1128/MMBR.62.3.597-635.1998. - DOI - PMC - PubMed
    1. Mahajan R.T., Badgujar S.B. Biological aspects of proteolytic enzymes: A Review. J. Pharm. Res. 2010
    1. Subba Rao C., Sathish T., Ravichandra P., Prakasham R.S. Characterization of thermo- and detergent stable serine protease from isolated Bacillus circulans and evaluation of eco-friendly applications. Process Biochem. 2009;44:262–268. doi: 10.1016/j.procbio.2008.10.022. - DOI

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