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

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.

Microbial proteases: ubiquitous enzymes with innumerable uses

“…The microorganisms are isolated from a variety of sources/ecosystems followed by selection for the desired character. These sources/ecosystems have specific or adverse feature which reflects the characteristics of the enzyme to be produced; for example, dumping sites, hot springs, soda lake, soil samples from milk processing plants, poultry waste sites, meat waste contaminated soil, tannery waste, detergent industry, leather industry, and wood factory (Anandharaj et al 2016 ; Kshetri et al 2016 ; Suleiman et al 2020 ; Sharma et al 2017 ; Chu 2007 ; Kandasamy et al 2016 ; Ramakodi et al 2020 ). Other sources includes saltern Qingdao at China, Izmir Gulf at Turkey, composting soil, sewage sludge, vermicomposting pit soil, sea mud, marine sediments, southern ocean, and mangrove sediments (Baweja et al 2017 ; Li et al 2017 ; Sonune and Garode 2018 ; Bhatia et al 2021 ).…”

Effects of Bacillus Safensis YY-01 on the Growth of Tomato

“…Sample 1: soil samples for screening B. safensis were taken from the tomato rhizosphere in Zhanjiang city, Guangdong Province, China (109° 35' E, 21° 25' N) and were put into a sterilized glass bottle. Then, it was brought back to the laboratory and stored in a 4℃ refrigerator as required (Suleiman et al 2020).…”

Novel alkaline protease of <em>Staphylococcus cohnii </em>N3 isolated from poultry farm soil used for silver recovery from waste X-ray film

“…Hence the optimum pH for isolate Staphylococcus cohnii N3 strain was found to be pH 9 with maximum proteolytic activity of 107.3±0.5 U/ml in with casein as substrate (Fig 3). Whereas, protease of Geobacillus thermoglucosidasius SKF4 exhibit maximum protease activity at pH 7 to 8 (Suleiman et al, 2020) [18] . Bacillus isronensis strain KD3 produced maximum proteases at 42°C and pH 7-8 after 48h of the incubation period (Patil and Kurhekar, 2020) [19] .…”