(Below N is a link to NCBI taxonomic web page and E link to ESTHER at designed phylum.) > cellular organisms: NE > Bacteria: NE > Terrabacteria group: NE > Firmicutes: NE > Bacilli: NE > Bacillales: NE > Bacillaceae: NE > Lysinibacillus: NE > Lysinibacillus sphaericus: NE
Warning: This entry is a compilation of different species or line or strain with more than 90% amino acide identity. You can retrieve all strain data
(Below N is a link to NCBI taxonomic web page and E link to ESTHER at designed phylum.) Lysinibacillus sphaericus: N, E.
Lysinibacillus sphaericus CBAM5: N, E.
Lysinibacillus sphaericus OT4b.31: N, E.
Lysinibacillus sphaericus C3-41: N, E.
LegendThis sequence has been compared to family alignement (MSA) red => minority aminoacid blue => majority aminoacid color intensity => conservation rate title => sequence position(MSA position)aminoacid rate Catalytic site Catalytic site in the MSA MNQITNWRPRGFLQWLLTILSVIVCFIIIIVAYYIFNPSNMDKIMSLLAW GTSIFPIMLVVTAFIIIVLLALSFWKKTFIALTVLFPILLLLMFLTVQPI STMKSYAKSENVSVALSSHFFYNQNISTKPSVDVVYGKTTDGIELKLDVW PAKKKSEDVLTPVIVQVHGGGWVSGDKGQVQDWNQWMNDQGYTVFDVQYR MPPVAGWKDEVGDVKSAIGWIVQHADTYKIDPNRIILMGESAGGNLAMLA AYSLGDKHLPPSTDVPDVPIKAVINMYGPSDMTAFYKNNPSKRYVQDVLD QYIGGSPSDYPARYKKLSPISYIQEHTPPTIMFLGTGDRIVPVEQANVLD DKLTTSGVAHELYLLPKVDHGFDANPGSLSTQFAKEKVKAFLQKYNK
References
Title: Impact of signal peptide and transmembrane segments on expression and biochemical properties of a lipase from Bacillus sphaericus 205y Masomian M, Jasni AS, Rahman RNZRA, Salleh AB, Basri M Ref: J Biotechnol, 264:51, 2017 : PubMed
A total of 97 amino acids, considered as the signal peptide and transmembrane segments were removed from 205y lipase gene using polymerase chain reaction technique that abolished the low activity of this enzyme. The mature enzyme was expressed in Escherichia coli using pBAD expression vector, which gave up to a 13-fold increase in lipase activity. The mature 205y lipase (without signal peptide and transmembrane; -SP/TM) was purified to homogeneity using the isoelectric focusing technique with 53% recovery. Removing of the signal peptide and transmembrane segments had resulted in the shift of optimal pH, an increase in optimal temperature and tolerance towards more water-miscible organic solvents as compared to the characteristics of open reading frame (ORF) of 205y lipase. Also, in the presence of 1mM inhibitors, less decrease in the activity of mature 205y lipase was observed compared to the ORF of the enzyme. Protein structure modeling showed that 205y lipase consisted of an alpha/beta hydrolase fold without lid domain. However, the transmembrane segment could effect on the enzyme activity by covering the active site or aggregation the protein.
Title: A novel organic solvent tolerant lipase from Bacillus sphaericus 205y: extracellular expression of a novel OST-lipase gene Sulong MR, Rahman RNZRA, Salleh AB, Basri M Ref: Protein Expr Purif, 49:190, 2006 : PubMed
UNLABELLED: An organic solvent tolerant (OST) lipase gene from Bacillus sphaericus 205y was successfully expressed extracellularly. The expressed lipase was purified using two steps purification; ultrafiltration and hydrophobic interaction chromatography (HIC) to 8-fold purity and 32% recovery. The purified 205y lipase revealed homogeneity on denaturing gel electrophoresis and the molecular mass was at approximately 30 kDa. The optimum pH for the purified 205y lipase was 7.0-8.0 and its stability showed a broad range of pH value between pH 5.0 to 13.0 at 37 degrees C. The purified 205y lipase exhibited an optimum temperature of 55 degrees C. The activity of the purified lipase was stimulated in the presence of Ca2+ and Mg2+. Ethylenediaminetetraacetic acid (EDTA) has no effect on its activity; however inhibition was observed with phenylmethane sulfonoyl fluoride (PMSF) a serine hydrolase inhibitor. Organic solvents such as dimethylsulfoxide (DMSO), methanol, p-xylene and n-decane enhanced the activity. Studies on the effect of oil showed that the lipase was most active in the presence of tricaprin (C10). The lipase exhibited 1,3 positional specificity. KEYWORDS: Bacter
Title: Cloning and expression of a novel lipase gene from Bacillus sphaericus 205y Rahman RNZRA, Chin JH, Salleh AB, Basri M Ref: Mol Genet Genomics, 269:252, 2003 : PubMed
A Bacillus sphaericus strain (205y) that produces an organic solvent-tolerant lipase was isolated in Port Dickson, Malaysia. The gene for the lipase was recovered from a genomic library and sequenced. Phylogenetic analysis was performed based on an alignment of thirteen microbial lipase sequences obtained from the NCBI database. The analysis suggested that the B. sphaericus lipase gene is a novel gene, as it is distinct from other lipase genes in Families I.4 and I.5 reported so far. Expression in Escherichia coli under the control of the lacZ promoter resulted in an eight-fold increase in enzyme activity after a 3-h induction with 1 mM IPTG. The crude enzyme thus obtained showed a slight (10%) enhancement in activity after a 30-min incubation in 25% (v/v) n-hexane at 37 degrees C, and retained 90% of its activity after a similar period in 25% (v/v) p-xylene.
