Bioprocess Enzymes

Proteases

Achieving the intact N-terminus in Biophrama products by use of recombinant proteases

Unique Features of Paras offerings

Product Offer from Paras Biopharmaceuticals Finland OY

Recombinant Enterokinase Unique Features

Product Offer from Paras Biopharmaceuticlas Finland OY

RapidTEVTM Unique Features

Recombinant Enterokinase

What is Protein Purification

Protein Purification is a serious of processes intended to isloate one or more proteins from a complex structure. It is vital for the characterisation of the function, structure and intercations of the protein of interest.

One effective techinque is the tagging of proteins to engineer an antigen peptide tag onto a protein, and then purify the protein through a column. Once Finished, the tag can be cleaved from the protein by a protease.

What is Protein Tags?

Protein tags are peptide sequences genetically grafted onto a recombinant protein which are often removabale by chemical agents or by enzymatic means. One such example are affinity tags which are appended to proteins so that they can be purified from their crude biological source using an affinity Technique.

Enterokinase can be used to cleave a fusion protein can taining a C-terminal affinity tag to produce a target protein following protein purification.

Enterokinase is a type 2 transmembrane serine protease produced by cells in the duodenum which are involved in the body's digestive system. It is responsible for the initial activation of pancreatic proteolytic proenzymes taht catalyses the conversion of trypsinogen to trypsin. Enterokinase's specificity makes it an ideal tool for biotechnological and biochemical removal of fusion proteins and tags.

  • As a human enzyme, enterokinase is 1019 amino acids in length containing 14 disulfide bond, 18 potential N-gly-cosylation sites and a N-myristoyl lipdation as a the N-terminus.
  • Specifically cleaves the bond between Lys23 and lle24 in human trypsin-1 and the equivalent peptide bond in other trypsin family members.
  • Enterokinase's catalytic light chain is 235 amino acids in length and contrains 4 disulfide bonds.
  • Enterokinase deficiency is alife threatening instestinal malabsorption disorder.

Molecular weight of enterokinase varies fro, 82-140kD (113kD PROTEIN CORE) AND 35-62kD for its disulfide - linked heavy chain and light-chain, respectively, depending on the organism.

Action

  • Recombinant Enterokinase cuts the following sequence
  • Asp-Asp-Asp-Asp-Lys-X-X-X-X

  • It has minimal requirements for specific amino acids in the P1 and P4 positions.
  • Natural substrates have a propensity for Gly or Set at P3' and/or P4' to ensure regular secondary structure formation does not limit accessiblity to cleavage site.
  • This independence makes enterokinase ideal for the removal of fusion proteins and/or tags with the subsequent generation of an authentic N-terminus.
  • Many campanies monitor activity by following the cleavage of a protein. The fusion partners used vary widely.
  • The assay conditions employed also vary widely and so, the units of the activity are often defined differently.
  • Paras uses one of the most credible industry standard method for recording the cativity of the enzyme.


PRODUCT SUMMARY

As a type 2 transmembrane serine protease, enterokinase is ideally sited for the biotechnological and biochemical removal of fusion proteins and cleavage of tags after purification. Produced by cells, enterokinase is responsible for inital activation of pancreatic proenzymes that catalyses the conversion of trypsinogen to trypsin.This means enterokinase is a very important enzyme in the biopharma and nutrition industry.

  • BRAND NAME:Enterokinase but also know as Enteropeptidase
  • ACTIVE INGREDIENT:Recombinant Enterokinase
  • DESCRIPTION:Type 2 transmembrane serine protease
  • TARGET GROUP:Nutrition industry and for the removal of fusion proteins and tags.
  • PRODUCT PATENT EXPIRY:NO patents exist.


RapidTEV® Protease

What is Protein Purification

Protein Purification is a serious of processes intended to isloate one or more proteins from a complex structure. It is vital for the characterisation of the function, structure and intercations of the protein of interest.

One effective techinque is the tagging of proteins to engineer an antigen peptide tag onto a protein, and then purify the protein through a column. Once Finished, the tag can be cleaved from the protein by a protease.

What is Protein Tags?

Protein tags are peptide sequences genetically grafted onto a recombinant protein which are often removabale by chemical agents or by enzymatic means. One such example are affinity tags which are appended to proteins so that they can be purified from their crude biological source using an affinity Technique.

TEV Protease can be used to cleave tags from recombinant fusion proteins that contain a TEV recognition site through a one step affinity removal of his-tagged TEV after cleavage.

As a member of the cysteine-like family of proteases, RapidTEV Protease is a improved version of the Tobacco ETCH Virus(TEV) protease enzymes. RapidTEV Protease has been engineered to possess enhanced activity,improved stability and site specificity. RapidTEV Protease specificity makes it an ideal tool for biotechnological and biochemical removal of fusion proteins and tags.

  • RapidTEV Protease facilitates high specificity cleavage between Gln and Gly (or Ser) of the seven amino acid recognition sequence.
  • RapidTEV Protease is extremely useful for removing affinity tags from fusion proteins under target proteins friendly conditions.
  • The enzyme is engineered for resistance against autolysis and improved catalytic activity and performance.
  • Due to the presence of a 6X-His at the N-Terminus, RapidTEV Protease can be easily removed after cleavage reaction by affinity chromatography.

With an active pH range between 6.0 and 8.5, 99% cleavage is often is often achieved with Rapid TEV Protease within 1-2 hours at its optimum conditions (pH 7.0 and 30ºC)

Action

  • Recombinant RapidTEV Protease cuts the following sequence
  • Glu-Asn-Leu-Tyr-phe-Gln-Gly/Ser(ENLYFQ(G/S)

  • RapidTEV® Protease's specificity makes it an ideal tool within the biotechnology industry and R&D for the the biochemical removal of fusion proteins and tags.
  • Due to its sequence specificity, TEV Protease is more strigent and specific than Xa and trombin.
  • Some metal Ions, for example Zn,have been reported to inhibit and activity of the enzyme at concentrations above 5mM.
  • Most suitable temperature range is 4-30ºC.
  • Howevwer, TEV Protease is reported to be 2-3 fold less active at 40ºC than at 20ºC.
  • Storage is recommended at-20ºC.
  • Resistant to many widely used cysteine and serine protease inhibitors.
  • RapidTEV® Protease is active in a wide range of different buffers.


PRODUCT SUMMARY

RapidTEV® Protease is a site-specific protease that has been engineered for enhanced acitivity and greater stability. Due to its hight specificity it is frequently used for the cleavage of fusion proteins and the removal of tags from recombinant proteins in vitro and in vivo. RapidTEV® Protease is available in varoius unit sizes.

  • PRODUCT NAME: RapidTEV® Protease
  • ACTIVE INGREDIENT:Recombinant TEV protease
  • SOURCE:E,coli
  • PURITY:95% BY SDS page
  • UNIQUE PROPERTIES:Engineered for greater stability (pH and °C)

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Paras Biopharmaceuticals
Finland Oy
,

Kiviharjunlenkki 10, OULU,
FI-90220 Finland

Dr Ashesh Kumar
E: kumar.ashesh@parasbiopharma.com
P: +358 (0) 442709462

Dr Mark Jackson
E: mark.jackson@parasbiopharma.com
P:+358 (0) 442905993

Skype: Paras.Finland