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Characterization of an organic solvent-tolerant α-amylase from a halophilic isolate, Thalassobacillus sp. LY18

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Abstract

A halophilic isolate Thalassobacillus sp. LY18 producing extracellular amylase was isolated from the saline soil of Yuncheng Salt Lake, China. Production of the enzyme was synchronized with bacterial growth and reached a maximum level during the early stationary phase. The amylase was purified to homogeneity with a molecular mass of 31 kDa. Major products of soluble starch hydrolysis were maltose and maltotriose, indicating an α-amylase activity. Optimal enzyme activity was found to be at 70°C, pH 9.0, and 10 % NaCl. The α-amylase was highly stable over broad temperature (30–90°C), pH (6.0–12.0), and NaCl concentration (0–20 %) ranges, showing excellent thermostable, alkalistable, and halotolerant nature. The enzyme was stimulated by Ca2+, but greatly inhibited by EDTA, indicating it was a metalloenzyme. Complete inhibition by diethyl pyrocarbonate and β-mercaptoethanol revealed that histidine residue and disulfide bond were essential for enzyme catalysis. The surfactants tested had no significant effects on the amylase activity. Furthermore, it showed high activity and stability in the presence of water-insoluble organic solvents with log P ow ≥ 2.13.

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Abbreviations

EDTA:

Ethylenediamine tetraacetic acid

PAO:

Phenylarsine oxide

DEPC:

Diethyl pyrocarbonate

PMSF:

Phenylmethylsulfonyl fluoride

SDS:

Sodium dodecyl sulfate

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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Acknowledgments

This work was financially supported by Shanxi Provincial Science and Technology Foundation (grants no. 20110021) and Natural Science Fund of Shanxi Province (grants no. 2011021031-4).

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  1. Life Science College, Yuncheng University, Yuncheng, 044000, China

    Xin Li & Hui-Ying Yu

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  1. Xin Li
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  2. Hui-Ying Yu
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Correspondence to Xin Li.

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Li, X., Yu, HY. Characterization of an organic solvent-tolerant α-amylase from a halophilic isolate, Thalassobacillus sp. LY18. Folia Microbiol 57, 447–453 (2012). https://doi.org/10.1007/s12223-012-0160-3

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  • DOI: https://doi.org/10.1007/s12223-012-0160-3

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