Abstract
Bio-based industries require stable enzymes in a broad range of environmental conditions. Extremophiles have attracted more interests as the source of such enzymes, one of which is α-amylase. This study aimed to purify and characterize α-amylase produced by a thermo-halophilic bacterium PLS 75 isolated from underwater fumaroles. Ammonium sulfate precipitation results showed that the highest specific α-amylase activity (21.7 U/mg) obtained at 40-60% saturation level, with a purity of 7.7-fold of the crude extract with 16.2% yield. Further purification using DEAE Sepharose column chromatography increased the enzyme purity 11.1-fold of the crude extract with 7.1% yield. Specific activity after column chromatography purification was 31.3 U/mg. The pure enzyme had a low molecular weight of 14 kDa. The enzyme showed the highest activity at 80 °C and pH 5. The activity increased to 126% when in methanol, while decreased when in ethyl acetate and chloroform. The characteristics of α-amylase with low molecular weight, which was active in acidic condition, stable in polar and non-polar solvents, may be used for for specific industrial needs.
Highlights
Enzymatic bioprocess in industries has continued to develop in the last two decades because enzyme-catalyzed reactions are specific, selective and controlled
One of important enzymes used in industries is α-amylase, which contributes to 30% of the global enzyme market (Sivaramakrishnan et al, 2006). α-amylases catalyze the hydrolysis of α-1,4 glycosidic bonds in starch molecules and produce dextrins, oligosaccharides and glucose with various chain lengths (Kuriki and Imanaka, 1999). α-amylase has a very broad spectrum of applications, including starch, bioethanol, food, detergent, textile and paper processing industries (Rana et al, 2013)
The enzymes can be produced from various sources such as plants, animals and microorganisms, but bacteria remain the main source of the enzymes (Sivaramakrishnan et al, 2006)
Summary
Enzymatic bioprocess in industries has continued to develop in the last two decades because enzyme-catalyzed reactions are specific, selective and controlled. One of important enzymes used in industries is α-amylase, which contributes to 30% of the global enzyme market (Sivaramakrishnan et al, 2006). Industrial processes sometimes require extreme temperature and pH, the use of extremozymes that are active in extreme conditions is preferred (Elleuche et al, 2014). While the exploration of new microorganisms from extreme environments to produce more stable enzymes has been done for many years, studies to find new α-amylases continue being carried out (Krüger et al, 2018). Owing to the importance of finding new variants of the enzyme, here we purified and characterized α-amylase produced by an extremophilic bacterial isolate from underwater fumaroles in Pria Laot Sabang area, Weh Island, Indonesia. The results of this study may be used as the basis of further research to make the enzyme applicab-
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