BMRB Entry 18575

Chem Shift validation: AVS_full, LACS
BMRB Entry DOI: doi:10.13018/BMR18575

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Title:
1H, 13C and 15N resonance assignments of Bacillus subtilis Lipase A mutant evolved towards thermostability
Deposition date:
2012-07-05
Original release date:
2012-08-29
Authors:
Augustyniak, Wojciech; Wienk, Hans; Boelens, Rolf; Reetz, Manfred
Citation:

Citation: Augustyniak, Wojciech; Brzezinska, Agnieszka; Pijning, Tjaard; Wienk, Hans; Boelens, Rolf; Dijkstra, Bauke; Reetz, Manfred. "Biophysical characterization of mutants of Bacillus subtilis lipase evolved for thermostability: factors contributing to increased activity retention."  Protein Sci. 21, 487-497 (2012).
PubMed: 22267088

Assembly members:

Assembly members:
variant_XI_LipA, polymer, 181 residues, 19192 Da.

Natural source:

Natural source:   Common Name: Firmicutes   Taxonomy ID: 1423   Superkingdom: Bacteria   Kingdom: not available   Genus/species: Bacillus subtilis

Experimental source:

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli   Vector: pET-22b(+)

Entity Sequences (FASTA):

Entity Sequences (FASTA):
variant_XI_LipA: AEHNPVVMVHGIGGASFNFA GIKSYLVSQGWSGDKLYAVD FWDKTGTNYNNGPVLSRFVQ KVLDETGAKKVDIVAHSMGG ANTLYYIKNLDGGNKVANVV TLGGANRLTTGDALPGTDPN QKILYTSIYSSADDIVMNCL SRLDGARNVQIHGVGHMGLL YSSQVNSLIKEGLNGGGQNT N

Data sets:
Data typeCount
13C chemical shifts741
15N chemical shifts195
1H chemical shifts1228

Additional metadata:

  • Assembly
  • Samples and Experiments
  • Software
  • Spectrometers
  • Hide all

Assembly:

Entity Assembly IDEntity NameEntity ID
1variant XI LipA1

Entities:

Entity 1, variant XI LipA 181 residues - 19192 Da.

Full length Lipase A variant

1   ALAGLUHISASNPROVALVALMETVALHIS
2   GLYILEGLYGLYALASERPHEASNPHEALA
3   GLYILELYSSERTYRLEUVALSERGLNGLY
4   TRPSERGLYASPLYSLEUTYRALAVALASP
5   PHETRPASPLYSTHRGLYTHRASNTYRASN
6   ASNGLYPROVALLEUSERARGPHEVALGLN
7   LYSVALLEUASPGLUTHRGLYALALYSLYS
8   VALASPILEVALALAHISSERMETGLYGLY
9   ALAASNTHRLEUTYRTYRILELYSASNLEU
10   ASPGLYGLYASNLYSVALALAASNVALVAL
11   THRLEUGLYGLYALAASNARGLEUTHRTHR
12   GLYASPALALEUPROGLYTHRASPPROASN
13   GLNLYSILELEUTYRTHRSERILETYRSER
14   SERALAASPASPILEVALMETASNCYSLEU
15   SERARGLEUASPGLYALAARGASNVALGLN
16   ILEHISGLYVALGLYHISMETGLYLEULEU
17   TYRSERSERGLNVALASNSERLEUILELYS
18   GLUGLYLEUASNGLYGLYGLYGLNASNTHR
19   ASN

Samples:

sample_1: variant XI LipA, [U-99% 13C; U-99% 15N], 0.5 ± 0.05 mM; DTT, [U-99% 2H], 5.0 ± 0.1 mM; potassium phosphate 10.0 ± 0.1 mM; sodium azide 4.6 ± 0.1 mM; H2O 90 ± 0.1 %; D2O, [U-99% 2H], 10 ± 0.1 %

sample_conditions_1: ionic strength: 40 mM; pH: 7.0; pressure: 1 atm; temperature: 298.0 K

Experiments:

NameSampleSample stateSample conditions
2D 1H-15N HSQCsample_1isotropicsample_conditions_1
2D 1H-13C HSQCsample_1isotropicsample_conditions_1
3D HNCOsample_1isotropicsample_conditions_1
3D CBCA(CO)NHsample_1isotropicsample_conditions_1
3D HNCACBsample_1isotropicsample_conditions_1
3D HBHA(CO)NHsample_1isotropicsample_conditions_1
3D HCCH-TOCSYsample_1isotropicsample_conditions_1
3D 1H-15N NOESYsample_1isotropicsample_conditions_1
3D 1H-13C NOESYsample_1isotropicsample_conditions_1
2D 1H-1H NOESYsample_1isotropicsample_conditions_1

Software:

TOPSPIN v2.1, Bruker Biospin - collection, processing

CARA v1.8.4, Keller and Wuthrich - chemical shift assignment

NMR spectrometers:

  • Bruker Avance 600 MHz
  • Bruker Avance 900 MHz

Related Database Links:

BMRB 18574
PDB
DBJ BAA22231 BAI83725 BAM49200 BAM56470 BAP19127
EMBL CAB12064 CEI55388 CEJ75813 CJR84790 COO28547
GB ABQ08591 ABQ42598 ABQ50891 AFN11641 AFQ56205
REF NP_388152 WP_003246250 WP_014478727 WP_019712347 WP_038428323
SP P37957
AlphaFold P37957

Download HSQC peak lists in one of the following formats:
CSV: Backbone or all simulated peaks
SPARKY: Backbone or all simulated peaks