A biologically pure culture of Geobacillus Strain T1 bacteria isolated
from Palm Oil Mill Effluent capable of producing thermostable lipase T1
gene. The production of thermostable lipase T1 gene from a novel
Geobacillus bacterium having a designated as strain T1, was aerobic, gram
positive and endospore-forming, rod-shaped. The G+C content of the
genomic DNA was 52.6%. On the basic of physiological data, phenotypic
traits and molecular analysis, strain T1 represents a novel species
within the genus Geobacillus, The thermostable T1 lipase gene was
subcloned into the pGEX-4T1 vector with and without signal peptide in
prokaryotic system. The 28 amino acid residues signal peptide alter the
conformation of GST moiety and preventing it from bind to affinity
glutathione Sepharose column. By expression and simplification of the
purification through single step affinity chromatography shows a specific
activity of 292.929 U/mg and 72.55% of fusion lipase was recovered from
crude cell lysate. T1 mature lipase was successfully purified with a
final recovery of 51.49% and specificity activity of 959.033 U/mg.