EC Number   |
Recommended Name |
Application |
|---|
   3.4.14.1 | dipeptidyl-peptidase I |
synthesis |
specific and efficient method for complete removal of polyhistidine purification tags from the N-termini of target proteins |
| 3.4.14.11 | Xaa-Pro dipeptidyl-peptidase |
synthesis |
comparison of the effects of cell disruption methods on the activity of PepX. The optimized values of high-pressure homogenization are one cycle at 130 MPa providing activity of 114.47 mU/ml, while sonication gives an activity of 145.09 mU/ml at 28 min with 91% power and three cycles |
| 3.4.14.12 | Xaa-Xaa-Pro tripeptidyl-peptidase |
synthesis |
strategy for expression and onestep-purification of the prolyl tripeptidyl peptidase |
 3.4.16.4 | serine-type D-Ala-D-Ala carboxypeptidase |
synthesis |
highest yield of protein expression and purification from 12 mg of protein per liter of culture at 3 l bioreactor-scale is achieved in Streptomyces venezuelae ATCC 10595, a fast growing Streptomyces susceptible to glycopeptides. The addition of His6-tag at the N-terminus of the protein abolishes its biological activity either in vitro or in vivo assays. The addition of His6-tag at the C-terminus of the protein leads to a functional protein |
| 3.4.16.5 | carboxypeptidase C |
synthesis |
synthesis of CPY in Pichia pastoris as procarboxypeptidase Y with a yield of about 605 mg/l in shake-flasks after 168 h induction with 1 % (v/v) methanol. This precursor is cleaved by endogenous proteinases of Pichia pastoris and released into the fermentation broth as active carboxypeptidase Y within 2 weeks at 10°C. The recombinant enzyme is optimally active at 30°C and pH 6.0, with an optimal activity of about 305 U/mg |
| 3.4.17.1 | carboxypeptidase A |
synthesis |
enzymatic modification of human haemoglobin, useful for functional studies |
| 3.4.17.1 | carboxypeptidase A |
synthesis |
immobilization for synthetic use |
| 3.4.17.1 | carboxypeptidase A |
synthesis |
useful tools for peptide biosynthesis in non-conventional media, considered |
| 3.4.17.1 | carboxypeptidase A |
synthesis |
procedure for the production of CPA in the cytosol of Escherichia coli that yields approximately 0.5 mg of pure enzyme per liter of cell culture. The expression strategy maintains the proCPA zymogen in a soluble state by fusing it to the C-terminus of maltose-binding protein while simultaneously overproducing the protein disulfide isomerase DsbC in the cytosol from a separate plasmid. The yield of active and properly oxidized CPA is highest when coexpressed with DsbC in BL21(DE3) cells that do not also contain mutations in the trxB and gor genes. Most of the active CPA is generated after cell lysis and amylose affinity purification of the MBP-proCPA fusion protein, during the time that the partially purified protein is held overnight at 4°C prior to activation with thermolysin |
| 3.4.17.B1 | Sulfolobus solfataricus carboxypeptidase |
synthesis |
nanobioconjugate of the enzyme immobilized on silica-coated magnetic nanoparticles exhibits enhanced stability in aqueous media at room temperature as well as in different organic solvents. The improved stability in ethanol paves the way to possible applications of the immobilized enzyme, in particular as a biocatalyst for the synthesis of N-blocked amino acids. Another potential application might be amino acid racemate resolution, a critical and expensive step in chemical synthesis |
| 3.4.17.2 | carboxypeptidase B |
synthesis |
production of human insulin |
| 3.4.17.19 | Carboxypeptidase Taq |
synthesis |
expression of enzyme gene optimized for the Pichia pastoris codon usage and secretory expression. System is applicable for large-scale preparation of enzyme |
| 3.4.19.1 | acylaminoacyl-peptidase |
synthesis |
the enzyme is a very useful tool for the synthesis and modification of peptides. The stabilized Sepharose-coupled form of the enzyme is used to couple a carboxy-methylated N-formyl amino acid or N-acetyl amino acid to a short pre-existing peptide |
| 3.4.19.12 | ubiquitinyl hydrolase 1 |
synthesis |
used as molecular scissors for releasing a peptide or protein product |
| 3.4.19.16 | glucosinolate gamma-glutamyl hydrolase |
synthesis |
transient expression in Nicotiana benthamiana results in increased benzylglucosinolate levels that are accompanied by a high accumulation of the last intermediate in benzylglucosinolate synthesis, desulfobenzylglucosinolate, and a derivative thereof. Coexpression of adenosine 5'-phosphosulfate kinase APK2 reduces the accumulation of desulfobenzylglucosinolate and its derivative by more than 98% and increases benzylglucosinolate accumulation 16fold |
| 3.4.21.1 | chymotrypsin |
synthesis |
reaction catalyzer in apolar organic solvents |
| 3.4.21.1 | chymotrypsin |
synthesis |
enzymes are often used in organic solvents for catalyzing organic synthesis. Two enzyme preparations, EPRP (enzyme precipitated and rinsed with n-propanol) and PCMC (protein coated microcrystals) show much higher activities than lyophilized powders in such systems. Both preparations involve precipitation by an organic solvent |
| 3.4.21.1 | chymotrypsin |
synthesis |
construction of three different molecular weight multimodal temperature-responsive chymotrypsin-poly(sulfobetaine methacrylamide)-block-poly(N-isopropylacrylamide) protein-polymer conjugates that respond structurally to both low and high temperature. In the block copolymer grown from the surface of the enzyme, upper critical solution temperature phase transition is dependent on the chain length of the polymers in the conjugates, whereas lower critical solution temperature phase transition is independent of molecular weight. Each protein conjugate shows temperature dependent changes in substrate affinity and productivity when assayed from 0 to 40°C. The conjugates show higher stability to harsh conditions, including temperature, low pH, and protease degradation. The modified enzyme is active for over 8 h in the presence of a stomach protease at pH 1.0 |
| 3.4.21.1 | chymotrypsin |
synthesis |
use of immobilized bovine alpha-chymotrypsin in a biotransformation process applying an aqueous micellar two-phase system for separation and recycling of the enzyme immobilisates. The thermoresponsive surfactant, Eumulgin ES, has no influence on the activity up to a concentration of 10%, while surfactants Tween 20, Triton X 114, Triton X 100 lead to an activity reduction. For Triton X-114, the activity rises to its previous value after washing or a buffer exchange influence. Optimization of the aqueous micellar two-phase system composition via a Design of Experiments approach allows for reuse of the immobilisates for hemoglobin digestion over eleven cycles in an Eumulgin ES aqueous micellar two-phase system |
| 3.4.21.4 | trypsin |
synthesis |
synthesis of benzoyl-Arg leucinamide by stabilized trypsin |
| 3.4.21.4 | trypsin |
synthesis |
production of human insulin-threonine-ester, which can be simply converted into human insulin by hydrolysis with subsequent purification steps |
| 3.4.21.4 | trypsin |
synthesis |
production of mono/di-arg insulin |
| 3.4.21.4 | trypsin |
synthesis |
attachment of O-carboxymethyl-poly-beta-cyclodextrin with molecular weight of 13000 Da to surface of enzyme. resulting neoglycoenzyme retains high proteolytic and esterolytic activity, optimum temperature is increased by 10°C, enzyme is more resistant to thermal inactivation and to denaturation by sodiumdodecyl sulfate |
| 3.4.21.4 | trypsin |
synthesis |
covalent immobilization of enzyme onto poly(methyl methacrylate)-co-(ethyl acrylate)-co-(acrylic acid) latex particles. Immobilized enzyme shows higher optimal temperature and pH-value than free form. Immobilized enzyme exhibits higher KM-value than free form and better chemical and thermal stability, it maintains 63% of initial activity after reusing ten times |
| 3.4.21.4 | trypsin |
synthesis |
immobilisation of enzyme on Fe3O4 nanoparticles. The immobilized enzyme is slightly morestable than the free enzyme at 45°C. 55% of activity of immobilized trypsin remains at 44°C after 2 h and 90% after 120 days storage. 40% of its initial activity remains after eight times of successive reuse |
| 3.4.21.5 | thrombin |
synthesis |
production of recombinant human prethrombin-2 in mouse myeloma cells, activation by recombinant ecarin and purification by affinity chromatography. Yield is about 70%, product is indistinguishable from plasma-derived enzyme |
| 3.4.21.5 | thrombin |
synthesis |
preparation of thrombin from human plasma. Isolation of prothrombin is followed by activation to thrombin and further purification, process is suitable for large-scale production with a high degree of virus safety |
| 3.4.21.5 | thrombin |
synthesis |
use of thrombin as enhancer in polymerase chain reaction. Presence of bovine thrombin is exceptionally effective at preventing the formation of primer dimers and enhancing the formation of the desired polymerase chain reaction products. The PCR enhancement effects of thrombin apply to low-copy synthetic single-stranded DNAs, synthetic ssDNA pools, human genomic DNA, or hepatitis B virus genomic DNA. Thrombin is also able to effectively relieve PCR inhibition by nanomaterial inhibitors such as gold nanoparticles and graphene oxide. Compared with bovine serum albumin, thrombin is more effective and requires concentrations 18-178 times less than that of serum albumin to achieve a similar level of PCR enhancement |
| 3.4.21.6 | coagulation factor Xa |
synthesis |
enzyme is a target for design of specific inhibitors |
| 3.4.21.9 | enteropeptidase |
synthesis |
the enzyme may be useful in amino acid sequence studies for the production of large fragents. The enzyme may also be useful in DNA-recombinant studies in releasing the desired polypeptide chain from neighboring sequences |
| 3.4.21.9 | enteropeptidase |
synthesis |
the cleavage immediately after the carboxyl-terminal residue of the (Asp)4-Lys recognition sequence allows regeneration of native amino-terminal residues of recombinant proteins, e.g. removal of the thioredoxin and polyhistidine fusion partners from proteins of intrest |
| 3.4.21.9 | enteropeptidase |
synthesis |
the enzyme can be used for cleavage of fusion proteins due to its high specific activity |
| 3.4.21.9 | enteropeptidase |
synthesis |
useful tool for in vitro cleavage of fusion proteins |
| 3.4.21.9 | enteropeptidase |
synthesis |
gene engineering studies on processing fusion proteins |
| 3.4.21.9 | enteropeptidase |
synthesis |
tool protease in the research and production of gene engineering |
| 3.4.21.9 | enteropeptidase |
synthesis |
a huge number of therapeutic proteins such as antibodies, coagulation factors, growth hormones or vaccines are produced as fusion proteins. To obtain the therapeutic protein in its monomeric, active form, the fusion partner has to be removed either by chemical or enzymatic cleavage. Enterokinase is a very attractive tool for the in vitro cleavage of fusion proteins |
| 3.4.21.19 | glutamyl endopeptidase |
synthesis |
the enzyme could be useful for commercial preparation of casein phosphopeptides |
| 3.4.21.19 | glutamyl endopeptidase |
synthesis |
enzyme can be used as catalyst for peptide synthesis in hydrophilic organic solvents with low water content, e.g. acetonitrile, overview |
| 3.4.21.19 | glutamyl endopeptidase |
synthesis |
increased accumulation of recombinant enzyme in Bacillus subtilis by growth in presence of casein or gelatin. During stationary growth phase, enzyme production is stimulated by Ca2+, Mn2+, and Co2+, and inhibited by Zn2+, Fe2+, and Cu2+ |
| 3.4.21.19 | glutamyl endopeptidase |
synthesis |
increased accumulation of recombinant enzyme in Bacillus subtilis during stationary growth phase by growth in presence of phosphate or ammonium ions, and in presence of gelatin or casein. During sporulation, enzyme production is stimulated by Ca2+, Mn2+, and Co2+, and inhibited by Zn2+, Fe2+, and Cu2+. Glucose is not inhibitory to enzyme production during stationary growth |
| 3.4.21.19 | glutamyl endopeptidase |
synthesis |
recombinant expression of enzyme as insoluble inclusion bodies, solubilization in 6 M guanidine-HCl in presence of reducing agent and renaturation by fast frequent dilution method. Highest yield of refolded protein at pH 8.4, 4°C. Renaturation is accompanied by gradual splitting of K12-E13 and T47-E48 bonds resulting in a 26 kDa protein that is converted to 25 kDa mature protein by limited proteolysis trypsin or subtilisin. Complete cleavage of N-terminal pro-peptide is necessary for final packing and activation of enzyme |
| 3.4.21.38 | coagulation factor XIIa |
synthesis |
the enzyme is used for preparation of factor Xia and plasma kallikrein |
| 3.4.21.48 | cerevisin |
synthesis |
high-level production of human mechano-growth factor in Saccharomyces cerevisiae is interfered by proteinase B. Expression of mechano-growth factor in a proteinase B deletion mutant results in a fivefold increase of yield |
| 3.4.21.48 | cerevisin |
synthesis |
much stricter experimental protocols than those routinely used are necessary to prevent the artefactual interpretation of protein levels in strains or conditions that increase proteinase B activity. Stability of some proteins, such as Slt2p or Chs4p, but not others, is severely compromised in the rim101 mutant due to the upregulation of the PRB1 gene. Proteolytic degradation during protein processing can be almost completely prevented by an overdose of subtilisin-like protease inhibitors, such as PMSF, or by avoiding cell freezing |
| 3.4.21.50 | lysyl endopeptidase |
synthesis |
use of the immobilized enzyme in the enzyme-assisted semisynthesis of human insulin |
| 3.4.21.62 | Subtilisin |
synthesis |
C221 subtilisin is catalytically wounded to the point that will barely hydrolyze peptide bonds but turn to be quite reactive with certain activated ester substrates, therefore a useful tool for catalyzing synthetic reactions |
| 3.4.21.62 | Subtilisin |
synthesis |
subtilisin BPN' is an important industrial enzyme |
| 3.4.21.62 | Subtilisin |
synthesis |
production of (R)-(2-methylpropyl)butanedioic acid 4-ethyl ester |
| 3.4.21.62 | Subtilisin |
synthesis |
production of (R)-2-benzyl-3-[[1-methyl-1-((morpholin-4-yl)-carbonyl)ehtyl]sulfonyl]propionic acid-ethyl ester, which was prepared as an intermediate for the renin inhibitors ciprokiren and remikiren |
| 3.4.21.62 | Subtilisin |
synthesis |
production of (S)-2-benzyl-3-(tert-butylsulfonyl)propionic acid, which was prepared as an intermediate for the renin inhibitor remikiren |
| 3.4.21.62 | Subtilisin |
synthesis |
production of phenylalanine, which is used as an intermediate for the synthesis of aspartame |
| 3.4.21.62 | Subtilisin |
synthesis |
conjugation of enzyme with comb-shaped poly-(ethylen glycol) and solubilization in ionic liquids without adding water. Enzyme exhibits higher transesterification activity in solution of [Eminm][Tf2N] than in toluene. No enzymic activity in DMSO, THF, or acetonitrile |
| 3.4.21.62 | Subtilisin |
synthesis |
preparation of liposomes containing the enzyme and modulation of membrane permeability by cholate results in a nano-bioreactor system with higher apparent substrate selectivity than free enzyme due to different permeation of membrane by the substrates |
| 3.4.21.62 | Subtilisin |
synthesis |
synthesis of benzyloxycarbonyl-L-Asp-L-Ser-NH2 with benzyloxycarbonyl-L-Asp methyl ester as acyl donor and serine amide as the nucleophile. Optimum conditions are pH 10.0, 35°C, in acetonitrile/Na2CO3-NaHCO3 buffer system, 85:15, with a dipeptide yield of 75.5% |
| 3.4.21.62 | Subtilisin |
synthesis |
the engineered enzyme displays synthetically useful enantioselectivity for most of the secondary alcohols tested. The enantioselectivity of ISCBLS is in particular good to high for m- or p-substituted 1-phenylethanols. The dynamic kinetic resolutions of these secondary alcohols by the combination of ISCBLS and a ruthenium-based racemization catalyst provide the products of (S)-configuration with good results (80-94% yield, 90-99% enantiomeric excess). Enzyme CBLS is of great use as the enantiocomplementary counterpart of (R)-selective lipase for the dynamic kinetic resolution of secondary alcohols |
| 3.4.21.62 | Subtilisin |
synthesis |
the enzyme subtilisin Carlsberg (SC) is industrially applied or synthetic organic chemistry applications, it has been used to enrich the S enantiomer of a racemic alcohol mixture by dynamic kinetic resolution (DKR) systems by combining SC with a ruthenium complex in organic solvent |
| 3.4.21.64 | peptidase K |
synthesis |
chemoenzymatic synthesis of oligo(L-phenylalanine) by the enzyme as a green and clean chemical reaction compared to organic synthesis |
| 3.4.21.79 | granzyme B |
synthesis |
expression system for the production of high yields of enzymatic and biologically active human grB by transfection of HEK-293 with grB. The HEK-293 host cells are protected from apoptotic cell death by fusing an inactivation site coupled to a (His)6 tag to the gene sequence of GrB. Inactive grB which is actively released from HEK-293 cells by insertion of a Igkappa leader sequence is purified on a nickel column utilizing the (His)6 tag. After enterokinase digestion and heparin affinity chromatography, high yields of enzymatic and biologically active human grB are obtained |
| 3.4.21.82 | Glutamyl endopeptidase II |
synthesis |
peptide coupling catalyzed by proteasesis an alternative to chemical solution and solid phase peptide synthesis |
| 3.4.21.91 | Flavivirin |
synthesis |
expression and purification of a natural form of DENV protease containing the full-length NS2B protein and the protease domain of NS3. The protease is expressed in Escherichia coli and purified in detergent micelles necessary for its folding. This purified protein is active in detergent micelles such as lyso-myristoyl phosphatidylcholine |
| 3.4.21.96 | Lactocepin |
synthesis |
optimisation of enzymatic hydrolysis of beta-casein to produce the angiotensin-I-converting enzyme (ACE) inhibitory peptides. Under optimal conditions (enzyme-to-substrate ([E]/[S]) ratio (w/w) of 0.132 and pH of 8.00 at 38.8°C), the ACE inhibitory activity of hydrolysates is 72.06% and the total peptides is 11.75 mg/ml. The resulting hydrolysates have higher thermal stability than beta-casein and show an increase in the free sulfhydryl content compared with raw beta-casein |
| 3.4.22.2 | papain |
synthesis |
use as catalyst in asymetric synthesis of acyl derivatives and in peptide synthesis |
| 3.4.22.2 | papain |
synthesis |
chemical modification of papain using different anhydrides of 1,2,4-benzenetricarboxylic and pyromellitic acids and immobilization on cotton fabric results in immobilized papain with optimum pH shifted from 6.0 to 9.0. Compared with immobilized native papain, the thermal stability and the resistance to alkali and washing detergent of immobilized modified enzyme are improved considerably. When the concentration of detergent is 20 mg/ml, the activity of the immobilized pyromellitic papain retains about 40% of its original activity, whereas the native papain is almost inhibited |
| 3.4.22.2 | papain |
synthesis |
enzymatic hydrolysis of casein to produce free amino acids by papain in a two-phase system, which is composed of n-propanol, NaCl and water. In this system, the top phase contains more n-propanol and the bottom phase contains more NaCl and water. Papain and casein are mainly distributed in bottom phase, and free aromatic amino acids tyrosine, tryptophan and phenylalanine produced by enzymatic hydrolysis aere mainly in top phase. When the two-phase system consists of 44% n-propanol, 60 g/l NaCl, 0.15 g/l papain and 13 g/l casein at 55°C and pH 5.6, the transformation yield is 99.5% |
| 3.4.22.2 | papain |
synthesis |
Fab antibody fragment production and purification by papain digestion of an intact monoclonal antibody. After digestion, separation of the Fab fragment from the Fc fragment and residual intact antibody is achieved using protein A affinity chromatography. The Fab fragments are of high quality suitable to produce diffraction quality crystals suitable for X-ray crystallographic analysis |
| 3.4.22.2 | papain |
synthesis |
immobilization of papain on Sepharose 6B in the presence of different concentrations of cysteine affect the enzyme activity depending on cysteine concentration. The maximum specific activity is observed when papain was immobilized with 200 mM cysteine. The immobilization process results in significant enhancement of stability to temperature and extreme pH. After immobilization, the optimum temperature of papain activity increases by 20 degrees from 60 to 80°C and its optimum pH activity shifts from 6.5 to 8.0. Catalytic efficiency and specific activity of the immobilized enzyme do not significantly change after immobilization |
| 3.4.22.3 | ficain |
synthesis |
kinetically controlled formation of peptide bonds by coupling the ester substrates benzyloxycarbonyl-Ala methyl ester and benzyloxycarbonyl-Gly methyl ester with L-Ala, D-Ala, L-Gln, D-Gln and L-Cys(acetamidomethyl) respectively |
| 3.4.22.3 | ficain |
synthesis |
digestion of murine monoclonal IgG to obtain immunoreactive bivalent antibody fragments |
| 3.4.22.8 | clostripain |
synthesis |
the enzyme can be used for the high yielding synthesis of a variety of peptide bond using L-Arg and, to a lesser degree, L-Lys as substrates with L-amino acid amides or D-amino acid amides |
| 3.4.22.8 | clostripain |
synthesis |
efficient biocatalyst for the synthesis of peptide isosteres |
| 3.4.22.8 | clostripain |
synthesis |
recombinant clostripain might prove useful in the production of insulin from the proinsulin fusion protein |
| 3.4.22.14 | actinidain |
synthesis |
the enzyme can be used efficiently for hydrolysis of collagen and isolation of different cell populations from various solid tissues |
| 3.4.22.68 | Ulp1 peptidase |
synthesis |
comparison of Ulp1 protease in active upon expression as inclusion bodies and soluble protein. Fusion of the N-terminal selfassembling peptide GFIL8 to Ulp1 increases production of active inclusion bodies in Escherichia coli. Attachment of the N-terminal cellulose-binding module facilitates the immobilization on regenerated amorphous cellulose with a binding capacity up to about 235 mg protein per gram of cellulose. The immobilized soluble Ulp1 maintains about 42% initial cleavage activity with repetitive use, whereas the aggregated Ulp1 loses its cleavage capacity after cleaving the protein substrate once. Crosslinking of inclusion bodies using glutaraldehyde inactivates Ulp1 |
| 3.4.22.68 | Ulp1 peptidase |
synthesis |
immobilization of Ulp1 as a tool for cleavage of the SUMO tag of recombinant proteins. Ulp1 immobilized on N-hydroxysuccinimide-activated Sepharose maintains 95% substrate-cleavage ability and significantly enhanced pH and thermal stability. The immobilized Ulp1 can tolerate 15% (v/v) DMSO and 20% (v/v) ethanol. It can be reused for more than 15 batch reactions with 90% activity retention |
| 3.4.22.68 | Ulp1 peptidase |
synthesis |
optimization of expression of the catalytic domain. Optimization of cultivation conditions at shake flask results in Ulp1 expression of 195 mg/l in TB medium. Ni-NTA affinity purification of Ulp1 using 0.1% Triton X-100, 0.01mM DTT, 0.02mM EDTA and 1% glycerol leads to a purity of about 95% with a recovery yield of 80% and specific activity of 398600 U/mg. The protease cleaves the SUMO tag even at 1:10,000 enzyme to substrate ratio The in vivo cleavage of SUMO tag via coexpression strategy also results in more than 80% cleavage of SUMO fusion protein |
| 3.4.22.70 | sortase A |
synthesis |
in vitro applications of sortase A to protein conjugation. Application of recombinant Staphylococcus aureus sortase A to attach a tagged model protein substrate (green fluorescent protein) to polystyrene beads chemically modified with either alkylamine or the in vivo sortase A ligand, Gly-Gly-Gly, on their surfaces. Sortase A can be used to sequence-specifically ligate eGFP to amino-terminated poly(ethylene glycol) and to generate protein oligomers and cyclized monomers using suitably tagged eGFP An alkylamine can substitute for the natural Gly3 substrate, which suggests the possibility of using the enzyme in materials applications. The highly specific and mild sortase A-catalyzed reaction, based on small recognition tags unlikely to interfere with protein expression represents a useful addition to the protein immobilization and modification tool kit |
| 3.4.22.70 | sortase A |
synthesis |
semienzymatic cyclization of disulfide-rich peptides using sortase A, overview |
| 3.4.22.70 | sortase A |
synthesis |
SrtA works as a versatile tool in protein engineering |
| 3.4.22.70 | sortase A |
synthesis |
the bacterial transpeptidase sortase A is a well-established tool in protein chemistry and catalyzes the chemoselective ligation of peptides and proteins |
| 3.4.23.1 | pepsin A |
synthesis |
comparison of hydrolysis kinetics of hemoglobin after immobilization of enzyme on aluminium oxide and on 2-ethanolamine-O-phosphate-modified acidic alumina. Modified alumina results in comparatively less adsorption of peptides and complete adsorption of heme |
| 3.4.23.4 | chymosin |
synthesis |
use of chymosin to cleave a pro-chymosin derived fusion tag releasing native target proteins. After modification of the pro-chymosin fusion tag chymosin can remove this tag at more neutral pH 6.2, less prone to compromise the integrity of target proteins. Chymosin produces intact native target protein both at the level of small and large-scale preparations |
| 3.4.23.41 | yapsin 1 |
synthesis |
single disruption of either PEP4 gene or yapsin family member YPS1 gene leads to reduced degradation of recombinant human serum albumin and human parathyroid hormone upon expression in Pichia pastoris. In a PEP4YPS1 double disruptant, more than 80% of the human serum albumin and human parathyroid hormone secreted by the remains intact, after 72 h of incubation, in comparison to only 30% with the wild-type strain |
| 3.4.23.43 | prepilin peptidase |
synthesis |
scaled-up synthesis of PilD, followed by solubilization in dodecyl-beta-D-maltoside and chromatography, leads to a pure enzyme that retains its known biochemical activities |
| 3.4.23.52 | preflagellin peptidase |
synthesis |
scaled-up synthesis of PilD, followed by solubilization in dodecyl-beta-D-maltoside and chromatography, leads to a pure enzyme that retains its known biochemical activities |
| 3.4.24.7 | interstitial collagenase |
synthesis |
improved method for high-level expression of soluble human MMP-1 catalytic domain in Escherichia coli |
| 3.4.24.B7 | matrix metalloproteinase-26 |
synthesis |
use of Brevibacillus choshinensis as the host system for the soluble and active expression of MMP26. The enzyme is secreted in soluble form in the supernatant of cell culture medium. The yields of purified proform of MMP26 and catalytic form of MMP-26 are 12 and 18 mg/L, respectively, with high purity and homogeneity. Both pro- and catalytic form show gelatin zymography activity and the purified catalytic form has high enzymatic activity against DQ-gelatin substrate. Expression using several widely used expression vectors in Escheriochia coli cells results in insoluble expressions or soluble expressions with little catalytic activity |
| 3.4.24.20 | peptidyl-Lys metalloendopeptidase |
synthesis |
isolation of blocked N-terminal peptides |
| 3.4.24.24 | gelatinase A |
synthesis |
application of MMP-2 from sea cucumber (Stichopus japonicas) to prepare bioactive collagen hydrolysate |
| 3.4.24.26 | pseudolysin |
synthesis |
the Pseudomonas aeruginosa elastase, produced by Pichia pastoris, is a promising biocatalyst for peptide synthesis in organic solvents |
| 3.4.24.26 | pseudolysin |
synthesis |
the enzyme can be applied to obtain bioactive soluble peptides from eggshell-membrane. Potential applicability of its peptides as functional food and cosmetic additives |
| 3.4.24.B26 | myroilysin |
synthesis |
optimization of the culture conditions for protease production, best conditions are (w/v) 1% corn powder, 4% wheat bran, 2% bean powder, 0.4% Na2HPO4, 0.03% KH2PO4, 0.1% CaCl2, and artificial seawater, with the initial pH of 8.0 and 1% inoculation amount, 15°C culture temperature. Under the optimized conditions, the protease activity reaches 1137 U/ml, i.e., 174% of that before optimization. The protease activity in small scale fermentations reaches 1546.4 U/ml after parameter optimization |
| 3.4.24.27 | thermolysin |
synthesis |
production of L-alpha-aspartame, which is used as a low-calorie sweetener in food, including soft drinks, table-top sweeteners, dairy products, instant mixes, dressings, jams, confectionary, toppings and in pharmaceuticals |
| 3.4.24.27 | thermolysin |
synthesis |
hydrolysis and condensation reactions of peptides catalyzed by enzyme can be reversibly controlled by on/off ultrasound irradiation depending on its frequency region |
| 3.4.24.27 | thermolysin |
synthesis |
immobilized enzyme catalyzes the formation of beta-cyclodextrin esters using vinyl esters of butyrate, decanoate and laurate, as acyl donors in dimethylsulfoxide. Esterification occurs exclusively at the glucose C-2 position. Enzyme also catalyzes the synthesis of alpha-, beta-, gamma- and maltosyl-beta-cyclodextrin esters with vinyl laurate as the acyl donor in dimethylsulfoxide and dimethylformamide |
| 3.4.24.27 | thermolysin |
synthesis |
immobilized enzyme catalyzes the transesterification of vinyl laurate to several sucrose-containing tri- and tetrasaccharides. Preferred position of acylation is the 2-OH group of the alpha-D-glucopyranose moiety linked 1 to 2 to the beta-D-fructofuranose unit |
| 3.4.24.27 | thermolysin |
synthesis |
introduction of ionizing residues into the active site of enzyme as a means of modifying its pH-activity profile |
| 3.4.24.27 | thermolysin |
synthesis |
the enzyme is used for synthesis of N-carbobenzyloxy L-Asp-L-Phe methyl ester, a precursor of the artificial sweetener aspartam |
| 3.4.24.27 | thermolysin |
synthesis |
thermolysin is industrially used for the synthesis of N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester, a precursor of an artificial sweetener aspartame, from N-carbobenzoxy-L-aspartic acid and L-phenylalanine methyl ester |
| 3.4.24.27 | thermolysin |
synthesis |
the enzyme can be used for production of caseicin A, an antimicrobial active peptide, from alpha-casein, for potential improvement of the safety of infant milk formula using milk-derived bioactive peptides |
| 3.4.24.27 | thermolysin |
synthesis |
using the enzyme to catalyze the condensation of the chiral aspartame-precursor, carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester, from the protected amino acid substrates carbobenzoxy-L-aspartic acid and L-phenylalanine methyl ester in large scale production. Analysis of the protease mediated peptide synthesisof a precursor of the artificial sweetener aspartame, a multiton peptide synthesis catalyzed by the enzyme thermolysin. X-ray structures of thermolysin in complex with aspartame substrates separately, and after protease mediated peptide synthesis in a crystal, rationalize the reaction's substrate preferences and reveal an unexpected form of substrate inhibition that explains its sluggishness. Structure guided optimization of this and other PMPS reactions could expand the economic viability of commercial peptides beyond current high-potency, low-volume therapeutics, with substantial green chemistry advantages |
