Abstract:
:The recombinant acidstable α-amylase (Ba-amy) of acidophilic bacterium Bacillus acidicola TSAS1 has been produced extracellularly using a combination of cloning (E. coli and P. pastoris) and physico-chemical treatment strategies. A total of 150,000 U/L of Ba-amy were attained under constitutive promoter in P. pastoris, which is 15-fold higher than that of the wild strain B. acidicola (10,000 U/L). The recombinant P. pastoris integrated two copies of Ba-amy under GAP promoter. The pure Ba-amy expressed in P. pastoris is a glycoprotein of 66 kDa, which is optimally active at pH 4.0 and 60 °C with a T 1/2 of 25 min at 70 °C. The K m, V max and K cat values of the recombinant Ba-amy are 1.66 mg/mL, 53.6 µmol/mg/min and 106.8/s, respectively. The enzyme generates maltose (30 %), maltotriose (20 %) and other higher maltooligosaccharides from starch, thus, useful in baking as an antistale. This is the first report on the optimization of extracellular production of recombinant acidic α-amylase of an acidophilic bacterium.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Parashar D,Satyanarayana Tdoi
10.1007/s12033-016-9970-xsubject
Has Abstractpub_date
2016-11-01 00:00:00pages
707-717issue
11eissn
1073-6085issn
1559-0305pii
10.1007/s12033-016-9970-xjournal_volume
58pub_type
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