(Below N is a link to NCBI taxonomic web page and E link to ESTHER at designed phylum.) > cellular organisms: NE > Bacteria: NE > Proteobacteria: NE > Gammaproteobacteria: NE > Pseudomonadales: NE > Pseudomonadaceae: NE > Pseudomonas: NE > Pseudomonas aeruginosa group: NE > Pseudomonas mendocina: 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.) Pseudomonas mendocina EGD-AQ5: N, E.
Pseudomonas mendocina ymp: N, E.
Pseudomonas mendocina DLHK: N, E.
Pseudomonas mendocina NK-01: N, E.
Pseudomonas mendocina S5.2: N, E.
Pseudomonas stutzeri A1501: N, E.
Pseudomonadales bacterium RIFCSPHIGHO2_01_FULL_64_12: N, E.
Pseudomonas stutzeri DSM 4166: N, E.
Streptococcus pneumoniae R6: N, E.
Streptococcus pneumoniae: N, E.
Streptococcus pneumoniae SP3-BS71: N, E.
Streptococcus pneumoniae SP19-BS75: N, E.
Streptococcus pneumoniae SP14-BS69: N, E.
Streptococcus pneumoniae Taiwan19F-14: N, E.
Streptococcus pneumoniae MLV-016: N, E.
Streptococcus pneumoniae SP195: N, E.
Streptococcus pneumoniae SP11-BS70: N, E.
Streptococcus pneumoniae SP18-BS74: N, E.
Streptococcus pneumoniae D39: N, E.
Streptococcus pneumoniae CDC0288-04: N, E.
Streptococcus pneumoniae SP9-BS68: N, E.
Streptococcus pneumoniae CDC1873-00: N, E.
Streptococcus pneumoniae G54: N, E.
Streptococcus pneumoniae 70585: N, E.
Streptococcus pneumoniae CGSP14: N, E.
Streptococcus pneumoniae CDC3059-06: N, E.
Streptococcus pneumoniae ATCC 700669: N, E.
Streptococcus pneumoniae CDC1087-00: N, E.
Streptococcus pneumoniae P1031: N, E.
Streptococcus pneumoniae JJA: N, E.
Streptococcus pneumoniae SP6-BS73: N, E.
Streptococcus pneumoniae SP23-BS72: N, E.
Streptococcus pneumoniae Hungary19A-6: N, E.
Streptococcus pneumoniae INV104: N, E.
Streptococcus pneumoniae BS397: N, E.
Streptococcus pneumoniae INV200: N, E.
Streptococcus pneumoniae AP200: N, E.
Streptococcus pneumoniae GA04375: N, E.
Streptococcus pneumoniae SP14-BS292: N, E.
Streptococcus pneumoniae BS458: N, E.
Streptococcus pneumoniae BS455: N, E.
Streptococcus pneumoniae OXC141: N, E.
Streptococcus pneumoniae TCH8431/19A: N, E.
Streptococcus pneumoniae BS457: N, E.
Streptococcus pneumoniae SP-BS293: N, E.
Streptococcus pneumoniae TIGR4: N, E.
Streptococcus pneumoniae 670-6B: N, E.
Streptococcus pneumoniae GA47502: N, E.
Streptococcus pneumoniae GA44500: N, E.
Streptococcus pneumoniae 2070335: N, E.
Streptococcus pneumoniae PCS8203: N, E.
Streptococcus pneumoniae PCS8106: N, E.
Pseudomonas stutzeri B1SMN1: N, E.
Pseudomonas stutzeri: N, E.
Pseudomonas stutzeri ATCC 14405 = CCUG 16156: N, E.
Pseudomonas stutzeri CCUG 29243: N, E.
Pseudomonas stutzeri TS44: N, E.
Pseudomonas stutzeri DSM 10701: N, E.
Pseudomonas stutzeri RCH2: N, E.
Pseudomonas stutzeri NF13: N, E.
Pseudomonas stutzeri MF28: N, E.
Pseudomonas stutzeri KOS6: N, E.
Pseudomonas stutzeri ATCC 17588 = LMG 11199: N, E.
uncultured Pseudomonas sp.: N, E.
Pseudomonas xanthomarina: N, E.
Pseudomonas sp. K35: N, E.
Pseudomonas sp. BICA1-14: 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 MNKNKTLLALCLGSALALSGQAFAATGSGYTATKYPIVLTHGMLGFDSLL GIDYWYGIPSALRRDGAQVYITEVSQLNTSELRGEELLAQVEEIVAISGK PKVNLIGHSHGGPTIRYVAGVRPDLIASVTSVGAPHKGSDVADLIRKVPE GSSGEAIIAGLVNAMGAFINFVSGSSSTAPQNSLGSLESLNSEGAARFNA KFPQGIPTTACGEGAYKVNGVRYYSWSGTSPLTNPLDVSDAMMGAGSLAF SGPNDGLVGRCSSHLGMVIRDDYRMNHLDEVNQFMGLTSLFETDPVSVYR QHANRLKNAGL
References
3 moreTitle: Purification and characterization of an alkaline lipase from a newly isolated Pseudomonas mendocina PK-12CS and chemoselective hydrolysis of fatty acid ester Jinwal UK, Roy U, Chowdhury AR, Bhaduri AP, Roy PK Ref: Bioorganic & Medicinal Chemistry, 11:1041, 2003 : PubMed
Lipase isolated from a soil isolate, Pseudomonas mendocina (PK-12CS) chemoselectively hydrolyzed the fatty ester group in presence of arbamate of compound 5-amino-2,4-dihydro-3H-1,2,4-triazole-3 ones, a class of compounds which are attractive starting materials for the synthesis of triazole annealed heterocycles. The enzymatic method provides an easy access to the synthesis of N-substituted glycine. Under optimized fermentation conditions the culture produced 3510 Lipolytic Units/mL of cell free fermentation broth in 20 h of fermentation. The purified lipase exhibited molecular mass of 80 kDa on SDS polyacrylamide gel electrophoresis. The enzyme was stable at room temperature for more than a month and expressed maximum activity at 37 degrees C and pH 8.
The lipase from Pseudomonas mendocina 3121-1 was found to be homogeneous with a molecular mass of 30 kDa by SDS/PAGE. It is composed of two identical subunits. A molecular mass of 62 kDa was determined by gel chromatography on a Toyopearl HW-55F column. Some physicochemical properties of the lipase were investigated using p-nitrophenyl butyrate (p-NPB), Tween 80 solution and Sigma olive-oil emulsion as substrates. The optimum temperature was determined to be 52 degrees C with p-NPB, in the range 50-60 degrees C with Tween 80 and in the range 50-65 degrees C with olive-oil emulsion. The optimum pH was determined to be in the pH range 7.2-7.5, both with Tween and the emulsion, but was unusually alkaline (pH 9.5) with p-NPB. The enzyme was activated for p-NPB hydrolysis by thermal treatment up to 60 min at 60 degrees C, pH 7.0-8.2, but was rapidly inactivated at 70-80 degrees C and at pH 7.0. The lipase was shown to be more thermolabile at 60 degrees C with respect to other two substrates. Using the emulsified substrate, no activity was obtained after preincubating the enzyme for 30 min at 70 degrees C. The enzyme was found to be pH-tolerant when stored at 20 degrees C, pH 6.3-10.3 (100 mM Briton-Robson buffer) as the half-life (t(1/2)) was more than 240 h when p-NPB was used as the substrate. By contrast, the pH-stability range was more narrow (pH 8.0-10.5) with olive-oil emulsion. The effect of various metal ions and EDTA depended on the nature of the substrate.
Lipase's thermostability and organic solvent tolerance are two crucial properties that enable it to function as a biocatalyst. This study examined the characteristics of two recombinant thermostable lipases (Lk2, Lk3) based on transesterification activity. Conversion of C12-C18 methyl ester with paranitrophenol was investigated in various organic solvent. Both lipases exhibited activity on difference carbon chain length (C12 - C18, C18:1, C18:2) of substrates. The activity of Lk2 was higher in each of substrate compared to that the Lk3. Experimental findings showed that the best substrates for Lk2 and Lk3 are C18:1 and C18:2 respectively, in agreement with the computational analysis. The activity of both enzymes prefers on nonpolar solvent. On nonpolar solvent the enzymes are able to keep its native folding shown by the value of radius gyration, solvent-enzyme interaction and orientation of triad catalytic residues. Lk3 appeared to be more thermostable, with maximum activity at 55 degreesC. The presence of Fe3+ increased the activity of Lk2 and Lk3. However, the activity of both enzymes were dramatically decreased by the present of Ca2+ despite of the enzymes belong to family I.1 lipase known as calcium dependent enzyme. Molecular analysis on His loop of Lk2 and Lk3 on the present of Ca2+ showed that there were shifting on the orientation of catalytic triad residues. All the data suggest that Lk2 and Lk3 are novel lipase on the family I.1 and both lipase available as a biocatalyst candidate.
Mutant of lipase at oxyanion hole (H110 F) was constructed. The gene was highly expressed in Eschericia coli BL21 (DE3) and the recombinant protein was purified using Ni-NTA affinity chromatography. The activity of mutant enzyme was significantly increased compared to that the wild type. Further comparison showed that both of the enzymes exhibited same optimum pH and temperature, and showed highest lipolytic activity on pNP-decanoate (C10). The wild type appeared lost of activity on C14 and C16 substrates meanwhile the mutant still showed activity up to 20 %. In the presence of non polar organic solvent such as n-hexane, the wild type became inactive enzyme meanwhile the mutant still remained 50 % of its activity. The results suggested that mutation at oxyanion hole (H110 F) caused enzyme-substrate interaction change resulting on elevation of activity, better activity toward longer carbon chain substrate and improving the activity in the present of non polar organic solvent.
Title: Purification and characterization of an alkaline lipase from a newly isolated Pseudomonas mendocina PK-12CS and chemoselective hydrolysis of fatty acid ester Jinwal UK, Roy U, Chowdhury AR, Bhaduri AP, Roy PK Ref: Bioorganic & Medicinal Chemistry, 11:1041, 2003 : PubMed
Lipase isolated from a soil isolate, Pseudomonas mendocina (PK-12CS) chemoselectively hydrolyzed the fatty ester group in presence of arbamate of compound 5-amino-2,4-dihydro-3H-1,2,4-triazole-3 ones, a class of compounds which are attractive starting materials for the synthesis of triazole annealed heterocycles. The enzymatic method provides an easy access to the synthesis of N-substituted glycine. Under optimized fermentation conditions the culture produced 3510 Lipolytic Units/mL of cell free fermentation broth in 20 h of fermentation. The purified lipase exhibited molecular mass of 80 kDa on SDS polyacrylamide gel electrophoresis. The enzyme was stable at room temperature for more than a month and expressed maximum activity at 37 degrees C and pH 8.
The lipase from Pseudomonas mendocina 3121-1 was found to be homogeneous with a molecular mass of 30 kDa by SDS/PAGE. It is composed of two identical subunits. A molecular mass of 62 kDa was determined by gel chromatography on a Toyopearl HW-55F column. Some physicochemical properties of the lipase were investigated using p-nitrophenyl butyrate (p-NPB), Tween 80 solution and Sigma olive-oil emulsion as substrates. The optimum temperature was determined to be 52 degrees C with p-NPB, in the range 50-60 degrees C with Tween 80 and in the range 50-65 degrees C with olive-oil emulsion. The optimum pH was determined to be in the pH range 7.2-7.5, both with Tween and the emulsion, but was unusually alkaline (pH 9.5) with p-NPB. The enzyme was activated for p-NPB hydrolysis by thermal treatment up to 60 min at 60 degrees C, pH 7.0-8.2, but was rapidly inactivated at 70-80 degrees C and at pH 7.0. The lipase was shown to be more thermolabile at 60 degrees C with respect to other two substrates. Using the emulsified substrate, no activity was obtained after preincubating the enzyme for 30 min at 70 degrees C. The enzyme was found to be pH-tolerant when stored at 20 degrees C, pH 6.3-10.3 (100 mM Briton-Robson buffer) as the half-life (t(1/2)) was more than 240 h when p-NPB was used as the substrate. By contrast, the pH-stability range was more narrow (pH 8.0-10.5) with olive-oil emulsion. The effect of various metal ions and EDTA depended on the nature of the substrate.
Pseudomonas mendocina lipase PmCut PCLase I