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Volume 34 Number 4 Volume 35 Number 1 Volume 35 Number 2

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Research articles

ScienceAsia 35 (2009): 17-23 |doi: 10.2306/scienceasia1513-1874.2009.35.017

Over-expression and characterization of the alkalophilic, organic solvent-tolerant, and thermotolerant endo-1,4-β-mannanase from Bacillus licheniformis isolate THCM 3.1

Pornpimon Kanjanavasa, Paisarn Khawsaka, Arda Pakpitcharoena, Supatra Areekita, Thayat Sriyaphaia, Khajeenart Pothivejkulb, Somchai Santiwatanakulc, Kenji Matsuid, Tadahiko Kajiwarad, Kosum Chansiria,*

ABSTRACT:     Gene encoding for endo-1,4-β-mannanase (EC from Bacillus licheniformis THCM 3.1 was cloned and over-expressed in pET 100/D TOPO vector. The molecular weight of the purified enzyme was about 40 kDa. This enzyme had an optimum pH of 9 and an optimum temperature of 45 °C and retained up to 77% of its activity after incubation for 48 h. The activity of the enzyme was inhibited by 10 mM of Pb2+, Ag+, Fe3+, Sn2+, Cu2+, and EDTA. Although partially inhibited, the enzyme retained much of its activity when the reaction solution was mixed with 15% (v/v) of the organic solvents acetone, toluene, benzene, dimethyl sulphoxide, 2-propanol, acetonitrile, or cyclohexane.

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a Department of Biochemistry, Faculty of Medicine, Srinakharinwirot University, Bangkok 10110, Thailand
b Department of Biology, Faculty of Sciences, Srinakharinwirot University, Bangkok 10110, Thailand
c Department of Pathology, Faculty of Medicine, Srinakharinwirot University, Bangkok 10110, Thailand
d Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi, Japan

* Corresponding author, E-mail: kosum@swu.ac.th

Received 29 Apr 2008, Accepted 6 Feb 2009