Acid stable α-amylase from Pseudomonas balearica VITPS19—Production, purification and characterization

• Published in: Biotechnology reports (Amsterdam, Netherlands) Netherlands), 2021-06, Vol.30, p.e00603-e00603, Article e00603
• Main Authors: Kizhakedathil, Moni Philip Jacob, C, Subathra Devi
• Format: Article
• Language: English
• Subjects: Amylase; Characterization; Enzyme kinetics; Optimization; Pseudomonas balearica VITPS19
• ISSN: 2215-017X; 2215-017X
• DOI: 10.1016/j.btre.2021.e00603

•α – Amylase was produced from a rhizobacteria Pseudomonas balearica VITPS19.•One factor at a time method (OFAT) was employed to optimize the α –amylase production.•Three step purification of α – amylase from the fermentation broth.•Determining the optimal conditions for enzyme activity.•Estimation of the enzymatic kinetic parameters of the α-amylase. In the present study, α-amylase from Pseudomonas balearica VITPS19 isolated from Kolathur, Tamil Nadu, India was studied. Initially, one factor at a time (OFAT) approach was used to optimize the medium parameters like pH, temperature, carbon and nitrogen sources and the presence of metal ions to enhance the amylase activity. After the optimization, 6.5-fold increase in the enzyme production was observed. Enzyme purification was carried out in three stages. The molecular weight of purified α-amylase was estimated to be 47 kDa.The optimum activity for the purified enzyme was observed at pH 6 in 0.1 M phosphate buffer at 25 ± 2 °C and the activity is enhanced in the presence of ions like Mn2+, Mo6+, Na+, Mg2+and Zn2+ and was inhibited in the presence of Hg2+ ions. Compounds such as Sodium dodecyl sulfate (SDS), Ethylenediaminetetraacetic acid (EDTA), urea and β- mercaptoethanol reduced the amylase activity. The Km and Vmax of the α-amylase was estimated to be 45.23 mM and 20.83 U/mL, respectively.