Lipase from Geobacillus sp. strain T1

Details Lipase from Geobacillus sp. strain T1 Lipase from Geobacillus sp. strain T1

2008-01-15

2008-01-15

Woitach, Joseph (Department: 1633)

Chemistry: molecular biology and microbiology

Micro-organism, per se ; compositions thereof; proces of...

Bacteria or actinomycetales; media therefor

C435S019000, C435S020000, C435S198000

Reexamination Certificate

active

11193282

ABSTRACT:
A biologically pure culture ofGeobacillusStrain T1 bacteria isolated from Palm Oil Mill Effluent capable of producing thermostable lipase T1 gene. The production of thermostable lipase T1 gene from a novelGeobacillusbacterium 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 genusGeobacillus, 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.

REFERENCES:
Gupta et al., Appl Microbiol Biotechnol. Jun. 2004;64(6):763-81. Epub Feb. 14, 2004.Bacterial lipases: an overview of production, purification and biochemical properties.
Leow al., High Level of Thermostable Lipase fromGeobacillussp. Strain T1, Biosci Biotechnol Biochem. Jan. 2004;68(1):96-103).
Amershambiosciences. Map of the glutathione S-transferase fusion vectors. Life Science News, Knaust et al., 2000 pp. 1-2.
Frangioni, John V. et al., “Solubilization and Purification of Enzymatically Active Glutathione S-Transferase (pGEX) Fusion Proteins”,Analytical Biochemistry, 1993, pp. 179-187, Molecular Medicine Unit, Beth Israel Hospital, Boston, Massachusetts.

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