Biomaterial Database

Gene cloning, sequence analysis and heterologous expression of a novel cold-active lipase from Pseudomonas sp. PF16. 2015

Xiuling Ji, and Shan Li, and Lianbing Lin, and Qi Zhang, and Yunlin Wei

A pychrotrophic bacterium secreing an extracellular alkaline cold-active lipase was isolated from refrigeratory. Based on morphology and 16S rRNA gene sequence analysis, the isolate which was named as PF16 was identified as a member of Pseudomonas. A novel lip-PF16 gene was obtained from the genomic DNA of strain PF16. Nucleotide sequence analysis showed that open reading frame (ORF) was consisting of 1,431 nucleotides encoding 476 amino acids long protein. Lip-PF16 showed 89.5% and 88% identity with the cold-adapted lipases from Pseudomonas fluorescens and Pseudomonas sp. KB700A, respectively. It was found to be a member of conserved subfamily I.3 bacterial lipase, which contained a lipase consensus sequence GXSXG. The lip-PF16 gene was expressed in Escherichia coli BL21 (DE3) and functional lipase was obtained successfully. The molecular weight was estimated to be 50 KDa by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The cloned lipase was active over broad range of temperature range with optimum activity at 4^°C, and found to be alkaline preferring with optimum activity at pH 9.0. This recombination lipase show high catalytic activity at low temperature.

UI	MeSH Term	Description	Entries
D008049	Lipase	An enzyme of the hydrolase class that catalyzes the reaction of triacylglycerol and water to yield diacylglycerol and a fatty acid anion. It is produced by glands on the tongue and by the pancreas and initiates the digestion of dietary fats. (From Dorland, 27th ed) EC 3.1.1.3.	Triacylglycerol Lipase,Tributyrinase,Triglyceride Lipase,Acid Lipase,Acid Lipase A,Acid Lipase B,Acid Lipase I,Acid Lipase II,Exolipase,Monoester Lipase,Triacylglycerol Hydrolase,Triglyceridase,Triolean Hydrolase,Hydrolase, Triacylglycerol,Hydrolase, Triolean,Lipase A, Acid,Lipase B, Acid,Lipase I, Acid,Lipase II, Acid,Lipase, Acid,Lipase, Monoester,Lipase, Triglyceride
D011549	Pseudomonas	A genus of gram-negative, aerobic, rod-shaped bacteria widely distributed in nature. Some species are pathogenic for humans, animals, and plants.	Chryseomonas,Pseudomona,Flavimonas
D011994	Recombinant Proteins	Proteins prepared by recombinant DNA technology.	Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D003001	Cloning, Molecular	The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.	Molecular Cloning
D003080	Cold Temperature	An absence of warmth or heat or a temperature notably below an accustomed norm.	Cold,Cold Temperatures,Temperature, Cold,Temperatures, Cold
D006863	Hydrogen-Ion Concentration	The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH	pH,Concentration, Hydrogen-Ion,Concentrations, Hydrogen-Ion,Hydrogen Ion Concentration,Hydrogen-Ion Concentrations
D001483	Base Sequence	The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.	DNA Sequence,Nucleotide Sequence,RNA Sequence,DNA Sequences,Base Sequences,Nucleotide Sequences,RNA Sequences,Sequence, Base,Sequence, DNA,Sequence, Nucleotide,Sequence, RNA,Sequences, Base,Sequences, DNA,Sequences, Nucleotide,Sequences, RNA
D012336	RNA, Ribosomal, 16S	Constituent of 30S subunit prokaryotic ribosomes containing 1600 nucleotides and 21 proteins. 16S rRNA is involved in initiation of polypeptide synthesis.	16S Ribosomal RNA,16S rRNA,RNA, 16S Ribosomal,Ribosomal RNA, 16S,rRNA, 16S
D017421	Sequence Analysis	A multistage process that includes the determination of a sequence (protein, carbohydrate, etc.), its fragmentation and analysis, and the interpretation of the resulting sequence information.	Sequence Determination,Analysis, Sequence,Determination, Sequence,Determinations, Sequence,Sequence Determinations,Analyses, Sequence,Sequence Analyses