BSGatlas

lip

BSGatlas-gene-346

BSGatlas

Description	Information
Coordinates	292205..292843
Genomic Size	639 bp
Name	lip
Outside Links	SubtiWiki
	BsubCyc
Strand	+
Type	CDS

SubtiWiki

Description	Information
Alternative Name	lip
	lip
	lipA
Category	SW 2 Metabolism
	SW 2.4 Lipid metabolism
	SW 2.4.1 Utilization of lipids
	SW 2.4.1.4 Utilization of lipids/ other
	SW 6 Groups of genes
	SW 6.12 Secreted proteins
Description	extracellular lipase
Function	lipid degradation
Is essential?	no
Isoelectric point	10.06
Locus Tag	BSU_02700
Molecular weight	22.6449
Name	lip
Product	extracellular lipase

RefSeq

Description	Information
Alternative Locus Tag	BSU02700
Description	Evidence 1a: Function from experimental evidencesin the studied strain; PubMedId: 8396026, 15812018,18053819, 11029590, 24827611, 28963005; Product type e :enzyme
Enzyme Classifications	EC 3.1.1.3: triacylglycerol lipase
Functions	16.11: Scavenge (Catabolism)
	16.8: Protect
Locus Tag	BSU_02700
Name	estA
Title	secreted alkaliphilic lipase
Type	CDS

BsubCyc

Description	Information
Alternative Name	lip
	lipA
Citation	Ahmad S;Kumar V;Ramanand KB;Rao NM Probing protein stability and proteolytic resistance by loop scanning: a comprehensive mutational analysis. Protein Sci 21(3);433-46 (2012) PUBMED: 22246996
	Augustyniak W;Brzezinska AA;Pijning T;Wienk H;Boelens R;Dijkstra BW;Reetz MT Biophysical characterization of mutants of Bacillus subtilis lipase evolved for thermostability: factors contributing to increased activity retention. Protein Sci 21(4);487-97 (2012) PUBMED: 22267088
	Augustyniak W;Wienk H;Boelens R;Reetz MT (1)H, (13)C and (15)N resonance assignments of wild-type Bacillus subtilis Lipase A and its mutant evolved towards thermostability. Biomol NMR Assign 7(2);249-52 (2013) PUBMED: 22996591
	Cao Y;Wu Z;Wang T;Xiao Y;Huo Q;Liu Y Immobilization of Bacillus subtilis lipase on a Cu-BTC based hierarchically porous metal-organic framework material: a biocatalyst for esterification. Dalton Trans 45(16);6998-7003 (2016) PUBMED: 26988724
	Franken B;Eggert T;Jaeger KE;Pohl M Mechanism of acetaldehyde-induced deactivation of microbial lipases. BMC Biochem 12;10 (2011) PUBMED: 21342514
	Frauenkron-Machedjou VJ;Fulton A;Zhu L;Anker C;Bocola M;Jaeger KE;Schwaneberg U Towards understanding directed evolution: more than half of all amino acid positions contribute to ionic liquid resistance of Bacillus subtilis lipase A. Chembiochem 16(6);937-45 (2015) PUBMED: 25786654
	Fulton A;Frauenkron-Machedjou VJ;Skoczinski P;Wilhelm S;Zhu L;Schwaneberg U;Jaeger KE Exploring the protein stability landscape: Bacillus subtilis lipase A as a model for detergent tolerance. Chembiochem 16(6);930-6 (2015) PUBMED: 25773356
	Kamal MZ;Kumar V;Satyamurthi K;Das KK;Rao NM Mutational probing of protein aggregates to design aggregation-resistant proteins. FEBS Open Bio 6(2);126-34 (2016) PUBMED: 27239434
	Kamal MZ;Mohammad TA;Krishnamoorthy G;Rao NM Role of active site rigidity in activity: MD simulation and fluorescence study on a lipase mutant. PLoS One 7(4);e35188 (2012) PUBMED: 22514720
	Kubler D;Bergmann A;Weger L;Ingenbosch KN;Hoffmann-Jacobsen K Kinetics of Detergent-Induced Activation and Inhibition of a Minimal Lipase. J Phys Chem B 121(6);1248-1257 (2017) PUBMED: 28106397
	Kubler D;Ingenbosch KN;Bergmann A;Weidmann M;Hoffmann-Jacobsen K Fluorescence spectroscopic analysis of the structure and dynamics of Bacillus subtilis lipase A governing its activity profile under alkaline conditions. Eur Biophys J 44(8);655-65 (2015) PUBMED: 26224303
	Mugler A;Kittisopikul M;Hayden L;Liu J;Wiggins CH;Suel GM;Walczak AM Noise Expands the Response Range of the Bacillus subtilis Competence Circuit. PLoS Comput Biol 12(3);e1004793 (2016) PUBMED: 27003682
	Ni Z;Jin R;Chen H;Lin X Just an additional hydrogen bond can dramatically reduce the catalytic activity of Bacillus subtilis lipase A I12T mutant: An integration of computational modeling and experimental analysis. Comput Biol Med 43(11);1882-8 (2013) PUBMED: 24209933
	Ni Z;Zhou P;Jin X;Lin XF Integrating In Silico and In vitro approaches to dissect the stereoselectivity of Bacillus subtilis lipase A toward ketoprofen vinyl ester. Chem Biol Drug Des 78(2);301-8 (2011) PUBMED: 21477088
	Nordwald EM;Plaks JG;Snell JR;Sousa MC;Kaar JL Crystallographic Investigation of Imidazolium Ionic Liquid Effects on Enzyme Structure. Chembiochem 16(17);2456-9 (2015) PUBMED: 26388426
	Rathi PC;Jaeger KE;Gohlke H Structural Rigidity and Protein Thermostability in Variants of Lipase A from Bacillus subtilis. PLoS One 10(7);e0130289 (2015) PUBMED: 26147762
	Singh B;Bulusu G;Mitra A Effects of point mutations on the thermostability of B. subtilis lipase: investigating nonadditivity. J Comput Aided Mol Des 30(10);899-916 (2016) PUBMED: 27696241
	Singh B;Bulusu G;Mitra A Understanding the thermostability and activity of Bacillus subtilis lipase mutants: insights from molecular dynamics simulations. J Phys Chem B 119(2);392-409 (2015) PUBMED: 25495458
	Srivastava A;Sinha S Thermostability of In Vitro Evolved Bacillus subtilis Lipase A: A Network and Dynamics Perspective. PLoS One 9(8);e102856 (2014) PUBMED: 25122499
	Tian F;Yang C;Wang C;Guo T;Zhou P Mutatomics analysis of the systematic thermostability profile of Bacillus subtilis lipase A. J Mol Model 20(6);2257 (2014) PUBMED: 24827611
	Yedavalli P;Rao NM Engineering the loops in a lipase for stability in DMSO. Protein Eng Des Sel 26(4);317-24 (2013) PUBMED: 23404771
	Yun HS;Park HJ;Joo JC;Yoo YJ Thermostabilization of Bacillus subtilis lipase A by minimizing the structural deformation caused by packing enhancement. J Ind Microbiol Biotechnol 40(11);1223-9 (2013) PUBMED: 24005991
	Zhao J;Frauenkron-Machedjou VJ;Kardashliev T;Ruff AJ;Zhu L;Bocola M;Schwaneberg U Amino acid substitutions in random mutagenesis libraries: lessons from analyzing 3000 mutations. Appl Microbiol Biotechnol (2017) PUBMED: 28050632
	Zhao J;Jia N;Jaeger KE;Bocola M;Schwaneberg U Ionic liquid activated Bacillus subtilis lipase A variants through cooperative surface substitutions. Biotechnol Bioeng 112(10);1997-2004 (2015) PUBMED: 25899108
Comment	16.8: Protect 16.11: Scavenge (Catabolism)
Description	secreted alkaliphilic lipase
Enzyme Classifications	EC 3.1.1.3: triacylglycerol lipase
Gene Ontology	GO:0004806 triglyceride lipase activity
	GO:0005576 extracellular region
	GO:0006629 lipid metabolic process
	GO:0016042 lipid catabolic process
	GO:0016787 hydrolase activity
Locus Tag	BSU02700
Molecular weight	22.791
Name	estA

Nicolas et al. predictions

Description	Information
Expression neg. correlated with	BSU27785, BSU28490, BSU08620, BSU10490, new_204891_204975, BSU01810, BSU01820, BSU29060, BSU30110, BSU29070
Expression pos. correlated with	new_292874_293464, BSU40170, BSU33710, BSU02750, BSU09890, BSU14350, BSU14750, BSU33280, BSU33730, BSU33720, BSU40140
Highly expressed condition	(aero) Cells were grown in a synthetic medium (E. Härtig, A. Hartmann, M. Schätzle, A. M. Albertini, D. Jahn, Appl Environ Microbiol 72, 5260, 2006) at 37 °C. For aerobic growth, an overnight culture was used to inoculate 100 ml of the synthetic medium to a starting OD578 of 0.05. The culture was then incubated in a 500 ml baffled flask with shaking at 250 rpm [aero]. Anaerobic growth was carried out (i) in the presence of 10 mM potassium nitrate (nitrate respiration) [nit]; or (ii) in the absence of 10 mM postassium nitrate (fermentative growth) [ferm]. The procedure for anaerobic growth was: medium was inoculated to an OD578 nm of 0.1 in flasks completely filled with medium and sealed with rubber stoppers. They were shaken at 100 rpm to minimize cell aggregation. These cultures were inoculated aerobically with an aerobically grown overnight culture. Anaerobic conditions were achieved in the stoppered flasks after a short time through the consumption of residual oxygen. Cells were harvested during the exponential growth phase.
	(GM+120) A culture of LB medium was inocualted from a frozen glycerol stock of B. subtilis. After few hours at 37oC when the culture was growing exponentially, this culture was used to inoculate M9 minimal medium at several different dilutions usually in the range of 500- to 2000-fold. The dilution range was chosen to ensure that at least one of these M9 precultures had reached an OD600 between 0.5 - 1.0 after overnight incubation. These precultures were then used to inoculate 2.5 L of M9 medium in a 3.1 L KLF bioreactor (Bioengineering AG, Wald, Switzerland) to a starting OD600 of 0.03 – 0.05. Condiions in the bioreactor were rigorously controlled as follows: temperature was controlled at 37 °C; the pH was maintained at exactly 7.2 by automatic titration with 2.0 M KOH and 2.0 M H2SO4, and the dissolved oxygen tension was maintained above 50%. In each nutritional shift experiment cells were grown on the single substrate until the OD600 reached 0.50, at which point the second substrate was added instantaneously (4 g/L L-malate or 3 g/L glucose). The nutrient shifts performed were from glucose to glucose+malate [GM] and from malate to malate+glucose [MG] (Buescher et al., accompanying paper). Cell growth during the course was monitored throughout the experiment by measuring OD600.
	(GM+150) A culture of LB medium was inocualted from a frozen glycerol stock of B. subtilis. After few hours at 37oC when the culture was growing exponentially, this culture was used to inoculate M9 minimal medium at several different dilutions usually in the range of 500- to 2000-fold. The dilution range was chosen to ensure that at least one of these M9 precultures had reached an OD600 between 0.5 - 1.0 after overnight incubation. These precultures were then used to inoculate 2.5 L of M9 medium in a 3.1 L KLF bioreactor (Bioengineering AG, Wald, Switzerland) to a starting OD600 of 0.03 – 0.05. Condiions in the bioreactor were rigorously controlled as follows: temperature was controlled at 37 °C; the pH was maintained at exactly 7.2 by automatic titration with 2.0 M KOH and 2.0 M H2SO4, and the dissolved oxygen tension was maintained above 50%. In each nutritional shift experiment cells were grown on the single substrate until the OD600 reached 0.50, at which point the second substrate was added instantaneously (4 g/L L-malate or 3 g/L glucose). The nutrient shifts performed were from glucose to glucose+malate [GM] and from malate to malate+glucose [MG] (Buescher et al., accompanying paper). Cell growth during the course was monitored throughout the experiment by measuring OD600.
	(M9tran) Cells were grown in M9 supplemented with glucose (0.3 %) at 37°C with vigorous shaking. The composition of the M9 minimal medium is (per liter): 8.5 g Na2HPO4.2H20, 3 g KH2PO4, 1 g NH4Cl and 0.5 g NaCl. The following solutions were individually sterilized and added (volumes per liter of medium): 1 ml 0.1 M CaCl2.2H2O, 1 ml 1 M MgSO4.7H2O, 1 ml 50 mM Fe-Citrate. Also added was 10 ml of a trace salts solution containing (per liter): 170 mg ZnCl2, 100 mg MnCl2.4H2O, 60 mg CoCl2.6H2O, 60 mg Na2MoO4.2H2O and 43 mg CuCl2.2H2O. Overnight cultures were diluted 2000-fold in pre-warmed M9 medium and samples were harvested during exponential growth [M9exp], at the transition phase [M9tran] and during stationary phase [M9stat].
	(MG+150) A culture of LB medium was inocualted from a frozen glycerol stock of B. subtilis. After few hours at 37oC when the culture was growing exponentially, this culture was used to inoculate M9 minimal medium at several different dilutions usually in the range of 500- to 2000-fold. The dilution range was chosen to ensure that at least one of these M9 precultures had reached an OD600 between 0.5 - 1.0 after overnight incubation. These precultures were then used to inoculate 2.5 L of M9 medium in a 3.1 L KLF bioreactor (Bioengineering AG, Wald, Switzerland) to a starting OD600 of 0.03 – 0.05. Condiions in the bioreactor were rigorously controlled as follows: temperature was controlled at 37 °C; the pH was maintained at exactly 7.2 by automatic titration with 2.0 M KOH and 2.0 M H2SO4, and the dissolved oxygen tension was maintained above 50%. In each nutritional shift experiment cells were grown on the single substrate until the OD600 reached 0.50, at which point the second substrate was added instantaneously (4 g/L L-malate or 3 g/L glucose). The nutrient shifts performed were from glucose to glucose+malate [GM] and from malate to malate+glucose [MG] (Buescher et al., accompanying paper). Cell growth during the course was monitored throughout the experiment by measuring OD600.
	(SMMPr) Cells were grown in Spizizen’s minimal medium (SMM) (C. Anagnostopoulos, J. Spizizen, J Bacteriol 81, 741, May, 1961) with vigorous agitation. The control culture was grown at 37 °C [SMMPr]. For growth at high or low temperatures, pre-cultures were grown at 37 °C, diluted to an OD578nm of 0.1 and subsequently transferred to 51 °C [HiTm] and 16 °C [LoTm], respectively. For the growth at high salinity, the salinity of the medium was adjusted by adding NaCl (5 M stock solution) to produce a final concentration of 1.2 M [HiOs].
	(T-1.40H) Anon-sporulating B. subtilis strain was grown in a modified M9 medium in batch culture (T. Hardiman, K. Lemuth, M. A. Keller, M. Reuss, M. Siemann-Herzberg, J Biotechnol 132, 359, Dec 1, 2007). Glucose was exhausted when the culture reached an OD600 of approx. 10 and this was designated T0 [T0.0H]. 7 samples were harvested at various times before glucose exhaustion [T-5.40H to T-0.40H] and 10 samples at various times after glucose exhaustion [T0.30H to T5.0H].
	(T-2.40H) Anon-sporulating B. subtilis strain was grown in a modified M9 medium in batch culture (T. Hardiman, K. Lemuth, M. A. Keller, M. Reuss, M. Siemann-Herzberg, J Biotechnol 132, 359, Dec 1, 2007). Glucose was exhausted when the culture reached an OD600 of approx. 10 and this was designated T0 [T0.0H]. 7 samples were harvested at various times before glucose exhaustion [T-5.40H to T-0.40H] and 10 samples at various times after glucose exhaustion [T0.30H to T5.0H].
	(T-3.40H) Anon-sporulating B. subtilis strain was grown in a modified M9 medium in batch culture (T. Hardiman, K. Lemuth, M. A. Keller, M. Reuss, M. Siemann-Herzberg, J Biotechnol 132, 359, Dec 1, 2007). Glucose was exhausted when the culture reached an OD600 of approx. 10 and this was designated T0 [T0.0H]. 7 samples were harvested at various times before glucose exhaustion [T-5.40H to T-0.40H] and 10 samples at various times after glucose exhaustion [T0.30H to T5.0H].
	(T-4.40H) Anon-sporulating B. subtilis strain was grown in a modified M9 medium in batch culture (T. Hardiman, K. Lemuth, M. A. Keller, M. Reuss, M. Siemann-Herzberg, J Biotechnol 132, 359, Dec 1, 2007). Glucose was exhausted when the culture reached an OD600 of approx. 10 and this was designated T0 [T0.0H]. 7 samples were harvested at various times before glucose exhaustion [T-5.40H to T-0.40H] and 10 samples at various times after glucose exhaustion [T0.30H to T5.0H].
Lowely expressed condition	(BC) Cultures were inoculated from frozen glycerol stocks and grown overnight in LB at 37°C. These cultures were thendiluted, plated onto LB plates, and incubated for 16 h at 37°C. Cells were harvested from plates containing individual colonies [BI] andfrom plates with confluen growth [BC].
	(dia15) Diamide was added to an exponentially growing culture (OD600 approx. 0.6) at a sub-lethal concentration(0.5 mM) and growth continued at 37°C with vigorous shaking. Samples were collected 0, 5 and 15 minutes after diamide addition [dia0, dia5 and dia15].
	(LBGstat) Cells were grown in Luria-Bertani medium (Sigma) supplemented with glucose 0.3 % [LBG] at 37°C with vigorous shaking in flasks. Overnight cultures were diluted 2000-fold in fresh pre-warmed medium and samples were collected during the exponential [exp], transition [tran] and stationary [stat] phases of the growth cycle .
	(LBstat) Cells were grown in Luria-Bertani medium (Sigma) [LB] at 37°C with vigorous shaking in flasks. Overnight cultures were diluted 2000-fold in fresh pre-warmed medium and samples were collected during the exponential [exp], transition [tran] and stationary [stat] phases of the growth cycle .
	(M0t90) Cells were grown in LB medium at 37°C with vigorous shaking. An exponentially growing culture (O.D.600 approx. 0.25) was divided: one culture acted as the control [no mitomycin C , M0] while mitomycin was added to the second culture to a final concentration of 40 ng/ml [mitomycin, M40]. Samples were harvested at 0, 45 and 90 minutes after mitomycin addition [t0, t45 and t90].
	(M40t90) Cells were grown in LB medium at 37°C with vigorous shaking. An exponentially growing culture (O.D.600 approx. 0.25) was divided: one culture acted as the control [no mitomycin C , M0] while mitomycin was added to the second culture to a final concentration of 40 ng/ml [mitomycin, M40]. Samples were harvested at 0, 45 and 90 minutes after mitomycin addition [t0, t45 and t90].
	(S6) Cells were grown in CH medium at 37°C and sporulation was induced by resuspension in warm sporulation medium as described by Sterlini and Mandelstam (J. M. Sterlini, J. Mandelstam, Biochem J 113, 29, Jun, 1969). The initiation of sporulation was designated T0, the time of resuspension. Samples were harvested at hourly intervals for 6 hours [S0 to S6] for the first set of experiments and for 8 hours [S0 to S8] for a second set of experiments.
	(S7) Cells were grown in CH medium at 37°C and sporulation was induced by resuspension in warm sporulation medium as described by Sterlini and Mandelstam (J. M. Sterlini, J. Mandelstam, Biochem J 113, 29, Jun, 1969). The initiation of sporulation was designated T0, the time of resuspension. Samples were harvested at hourly intervals for 6 hours [S0 to S6] for the first set of experiments and for 8 hours [S0 to S8] for a second set of experiments.
	(S8) Cells were grown in CH medium at 37°C and sporulation was induced by resuspension in warm sporulation medium as described by Sterlini and Mandelstam (J. M. Sterlini, J. Mandelstam, Biochem J 113, 29, Jun, 1969). The initiation of sporulation was designated T0, the time of resuspension. Samples were harvested at hourly intervals for 6 hours [S0 to S6] for the first set of experiments and for 8 hours [S0 to S8] for a second set of experiments.
	(Sw) Exponentially growing cells were spotted on 1 % agar LB plates and incubated at 37°C. Swarming cells were collected after 16 hours.
Name	estA

KEGG Pathways

Description	Information
Pathway	Glycerolipid metabolism (ko00561)
	Metabolic pathways (ko01100)

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