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3.4.21.62: Subtilisin

This is an abbreviated version!
For detailed information about Subtilisin, go to the full flat file.

Word Map on EC 3.4.21.62

Reaction

Hydrolysis of proteins with broad specificity for peptide bonds, and a preference for a large uncharged residue in P1. Hydrolyses peptide amides =

Synonyms

AcpII, Ak.1 protease, Alcalase, Alcalase 0.6L, Alcalase 2.5L, ALE1 subtilase, ALK-enzyme, Alkaline mesentericopeptidase, Alkaline protease, alkaline serine protease, ALP1, Alzwiprase, aprE, AprE51, aqualysin, Arp, AsES, Asp v 13, AtSBT1.9, Bacillopeptidase A, Bacillopeptidase B, Bacillus gibsonii alkaline protease, Bacillus subtilis alkaline proteinase Bioprase, BgAP, Bioprase AL 15, Bioprase APL 30, BLS, BPN', BprB, BprV, C1 subtilase, cold active subtilisin-like serine proteinase, Colistinase, EC 3.4.21.14, EC 3.4.4.16, Esperase, Fe protease, Fe prtS8A, Genenase I, intracellular subtilisin protease, ISP, IvaP, Kazusase, Maxatase, mesenteroicopeptidase, More, Nagarse, Opticlean, ORF2, Orientase 10B, P69 subtilase, PBANKA_1106900, PbSOPT, Peptidase, subtilo-, A, PF3D7_0507300, phytophase, PIMMS2, Protease S, Protease VIII, Protease XXVII, Proteinase K, Proteinase, Bacillus subtilis alkaline, Protin A 3L, PSP-3, psychrophilic subtilisin-like protease, S1P subtilase, SAP, SAS-1, SASP, saspase, Savinase, Savinase 16.0L, Savinase 32.0 L EX, Savinase 4.0T, Savinase 8.0L, savinaseTM, SBc, SBL, SDD1 subtilase, senescence-associated subtilisin protease, SES7, SISBT3 subtilase, SOPT, SP 266, Sspa, SSU0757, SUB1, SUB2, subC, subtilase, subtilase subfamily 1 member 9, subtilase-like protease, subtilisin, subtilisin 72, subtilisin A, Subtilisin amylosacchariticus, Subtilisin BL, subtilisin BPN’, subtilisin C., subtilisin Carlsberg, subtilisin DJ-4, Subtilisin DY, Subtilisin E, subtilisin E-S7, Subtilisin GX, subtilisin JB1, subtilisin Karlsberg, Subtilisin Novo, subtilisin Pr1-like protease, subtilisin protease, subtilisin QK, Subtilisin S41, subtilisin S4I, subtilisin S88, Subtilisin Sendai, subtilisin Sph, subtilisin-like ookinete protein, subtilisin-like ookinete protein important for transmission, subtilisin-like protease, subtilisin-like protease AprV2, subtilisin-like serine protease, Subtilisn J, Subtilopeptidase, Superase, thermitase, thermo-active subtilisin-like serine protease, Thermoase, Thermoase PC 10, thermophilic thermitase, thermostable subtilisin, ThSS45, Tk-subtilisin, trans-cinnamoyl-subtilisin, V. cholerae-secreted serine protease, VC_0157, vPR

ECTree

     3 Hydrolases
         3.4 Acting on peptide bonds (peptidases)
             3.4.21 Serine endopeptidases
                3.4.21.62 Subtilisin

Cloned

Cloned on EC 3.4.21.62 - Subtilisin

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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
AkP gene cloned and expressed in Escherichia coli PB5517
almost every property of subtilisin has been altered by protein engineering, e.g. substrate specificity, pH-profile and stability to oxidative, thermal and alkaline inactivation
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aprE gene rraprE
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Bacillus amyloliquefaciens gene expressed in Bacillus subtilis I-168 (Marburg strain)
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Bacillus stearothermophilus NCIMB10278 gene expressed in Bacillus subtilis DB104/pZS101
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cloned and amino acid sequence deduced from its cDNA
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cloned and expressed in Escherichia coli
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cloning in Escherichia coli strain DH5alpha and Saccharomyces cerevisiae strain YPH499, recombinant expression of wild-type and mutant enzymes in Bacillus megaterium strain WH320 from a shuttle expression vector carrying a ribosome-binding site and a signal sequence of extracellular metalloprotease from Bacillus megaterium downstream of xylose isomerase promoter
DNA and amino acid sequence determination and analysis
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DNA and amino acid sequence determination and analysis, expression in Escherichia coli strain DH5alpha
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Escherichia coli HB101 transformed with plasmid pBR1560 containing the wild-type prepro-subtilisin. G56S-propeptide and His-tagged G56S-propeptide overexpressed in Escherichia coli BL21-CodonPlus(DE3)-RIL transformed with vector pET25b and in Escherichia coli BL21(DE3) with vector pET28a, respectively
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expressed in Escherichia coli
expressed in the protease deficient host Bacillus subtilis ISW1214
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expression of full-length enzyme and mature form, expression of mature form does not give enzymatic activity
expression of wild-type and mutant enzymes in Bacillus subtilis strain DB104, subcloning in Escherichia coli strain DH5alpha
GN111900.1
full-length SUB2 gene from strain GS115 transformed into Escherichia coli. Overexpressed in Pichia pastoris KM71
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gene aprE, construction of error-prone PCR libraries, recombinant expression of wild-type and mutant enzymes in Bacillus subtilis strain WB600 and secretion to the culture medium
gene aprE, genetic structure, recombinant expression of wild-type and mutant enzymes in Bacillus subtilis strain 168
gene aprE, recombinant expression of wild-type and mutant enzymes in Bacillus subtilis strain WB600 and secretion to the culture medium
gene At3g14067, DNA and amino acid sequence determination and analysis, phylogenetic analysis, recombinant expression in an enzyme-knockout line sasp-1 or SALK_147962 and functional complementation
Q0WWH7
gene Fe prtS8A, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of the enzyme under control of the Trichoderma reesei cbh1 (cel7A) promoter in Escherichia coli strain XL10-Gold, and functional recombinant expression in Trichoderma reesei protoplasts, the enzyme is secreted
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gene ORF2, from a gene library of the psychrophilic bacterium Shewanella arctica, DNA and amino acid sequence determination and analysis, phylogenetic analysis, recombinant expression in Escherichia coli strain XLOLR and Tuner (DE3) pLacl from vector pET Blue-1, method optimization
gene PBANKA_1106900, genetic structure, sequence comparisons
gene PF3D7_0507300, genetic structure, sequence comparisons
gene prtS8A, DNA and amino acid sequence determination and analysis, fusion to the Trichoderma reesei cbh1 (cel7A) promoter and recombinant expression in Trichoderma reesei
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gene SBT1.9 or At5g67090, recombinant expression of soluble mature enzyme AtSBT1.9 fused to maltose-binding protein lacking the signal peptide in Escherichia coli BL21(DE3)
gene ThSS45, DNA and amino acid sequence determination and analysis, phylogenetic tree, quantitative expression analysis, recombinant expression in in Escherichia coli strain BL21
gene vpr, recombinant expression of extracellular enzyme in Escherichia coli strain XL10-Gold from pET 11a vector, high-level expression of the cold adapted subtilase, VPR, utilizing the rhamnose titratable T7 system of Lemo21 in presence of Ca2+ ions (best at about 100 mM) and rhamnose (best at about 0.076 mM) resulting in a dramatic increase of soluble protein compared to the other systems. Method evaluation, overview. In the absence of rhamnose, toxic effects of the proteinase are clearly observed, as cultures grow slower and end up at lower cell density. Addition of Ca2+ increases the yield of soluble protein. The calcium present in the growth medium is not only aiding in correct folding and stabilization of VPR but is also increasing the resistance of Lemo21 against the toxic effects caused by enzyme VPR
in vivo formation and stability of engineered disulfide bonds in subtilisin
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inactive mutants of subtilisin BPNÂ’ expressed in Bacillus subtilis. Expression of prodomain mutant proR9 in Escherichia coli strain BL21(DE3)
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overexpressed in Escherichia coli as a His-tagged fucion protein
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overexpression of pro-S255A by pET25b in Escherichia coli BL21-Codon-Plus(DE3)
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pET25b derivatives for overproduction of Pro-G56W, Pro-G56E, G56W-propeptide, and G56E-propeptide. G56W-propeptide overproduced in soluble form, while G56E-propeptide overproduced in insoluble form. S324A-subtilisin overproduced in inclusion bodies
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prosubtilisin JB1 cloned into the GST fusion protein expression vector pGEX-4T-1 and transformed into Escherichia coli DH5alphaMCR
prosubtilisn, DELTAloop-subtilisin, DELTACa2-Pro-S324A, and DELTACa3-Pro-S324A cloned into pET25b vector and overexpressed in Escherichia coli in inclusion bodies
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recombinant expression of C-terminally His6- and FLAG-tagged wild-type and mutant enzymes in Escherichia coli
recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
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recombinant expression of mutant enzyme sin Bacillus subtilis strain DB104, subcloning in Escherichia coli strain DH5alpha
recombinant expression of the codon-optimized gene encoding enzyme subtilisin QK in Pichia pastoris strain GS115, the thrombolytic activity of QK reaches 112000 IU (urokinase unit) per ml and the specific activity is 14,679 IU/mg under optimal culture and fermentation conditions, method optimization, detailed overview
replacement of the Bacillus subtilis subtilisin structural gene with an in vitro-derived deletion mutant
subtilisin 7150, mutagenized subtilisin with enhanced thermal stability
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subtilisin BPN' cloned, sequenced and expressed at high levels from its natural promoter sequences in Bacillus subtilis
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subtilisin Carlsberg from Bacillus licheniformis expressed in Bacillus subtilis
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subtilisin E produced in Escherichia coli
subtilisin gene from Bacillus subtilis PTCC 1023 is cloned into the vector pET-15b and expressed in Escherichia coli strain BL21
subtilisin variants expressed in Escherichia coli
the gene encoding the 1689 derivative without a putative N-terminal signal sequence, Pro-subtilisin-like serine protease (Met + Ala1-Gly640), ligated into pET25b vector, mutant Pro-S359A overexpressed in Escherichia coli BL21-CodonPlus(DE3)-RIL cells
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vector pET22b-AprB2 encoded residues 1-474 of the AprB2 precursor with a C-terminal His6 transformed into Escherichia coli DH5alpha. Expression construct transformed into Escherichia coli strain Rosetta-Gami(DE3)pLysS
vector pET22b-AprV2 encoded residues 1-474 of the AprV2 precursor with a C-terminal His6 tag transformed into Escherichia coli strain Rosetta-Gami(DE3)pLysS
vector pET22b-BprB encoded residues 1-476 of BprB with a C-terminal His6 tag transformed into Escherichia coli strain Rosetta-Gami(DE3)pLysS
vector pET22b-BprV encoded residues 1-476 of BprV with a C-terminal His6 tag and transformed into Escherichia coli strain Rosetta-Gami(DE3)pLysS
vectors pRSET-acpII, pRSET-acpII-tr (truncated AcpII Met1-Arg675), pRSET-acpII-C (catalytic domain of AcpII) and pRSET-acpII-C-DELTAPA (AcpII-C with the protease-associated domain deleted) expressed in Escherichia coli BL21(DE3) pLysS