Production of Ca2+-Independent and Acidstable Recombinant α-Amylase of Bacillus acidicola Extracellularly and its Applicability in Generating Maltooligosaccharides.

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 Biotechnol

journal_title

Molecular biotechnology

authors

Parashar D,Satyanarayana T

doi

10.1007/s12033-016-9970-x

subject

Has Abstract

pub_date

2016-11-01 00:00:00

pages

707-717

issue

11

eissn

1073-6085

issn

1559-0305

pii

10.1007/s12033-016-9970-x

journal_volume

58

pub_type

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