Gene Cloning and Characterization of Pcal_0222, α-Amylase from Pyrobaculum calidifontis
Gene Cloning and Characterization of Pcal_0222, α-Amylase from Pyrobaculum calidifontis
Sadaf Ashraf1, Masood Ahmed Siddiqui1*, Kanwal Nisa1, Samar Ali1 and Naeem Rashid2
ABSTRACT
The gene encoding Pcal_0222 from hyperthermophilic archaeon Pyrobaculum calidifontis was cloned and expressed in Escherichia coli. Pcal_0222 was composed of 529 amino acids with a theoretical molecular mass of 58 kDa. The amino acid sequence contained the four conserved regions that are a characteristic of GH13 family members. Recombinant Pcal_0222 was purified to apparent homogeneity using cation exchange and gel filtration column chromatographies. Purified Pcal_0222 exhibited optimal α-amylase activity at 85°C and pH 5.5. The activity was not dependent on any metal ion. The hyperthermophilic nature and metal ion independence make it a suitable candidate for both basic as well as applied research.
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April 2022
Vol. 54, Iss. 2, Pages 503-1000
