Pepsin immobilized by covalent fixation to hydroxyalkyl methacrylate gels: Preparation and characterization

Valentová, Olga; Turková, J.; Lapka, R; Zima, Jan; Čoupek, J.

doi:10.1016/0005-2744(75)90021-2

Cited by 24 publications

(5 citation statements)

References 13 publications

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“…Pepsin has been immobilized on a large number of supports by methods including covalent binding, crosslinking or copolymerization and it has been shown that immobilized pepsin was able to hydrolyse many proteins such as albumin (Hirano et al, 1979), serum albumin (Grubhofer and Schleith, 1954;Vretblad and Axen, 1971), haemoglobin (Goldstein, 1973;Valentova et al, 1975;AbdeI-Hay et al, 1980), casein (Puvanakrishnan and Bose, 1984;Findlay et al, 1986) or immunoglobulins (Tomono et al, 1981). However those studies were generally carried out in batch reactions and the stability of the enzyme was limited to the possibility of repeated uses in a short length of time or to the verification of enzyme activity after a long storage at low temperature.…”

Section: Introductionmentioning

confidence: 99%

Stabilization of pepsin on duolite for the continuous hydrolysis of bovine haemoglobin at pH2 and 40�C

Sannier¹,

Piot²,

Guillochon³

et al. 1993

Biotechnol Tech

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Several methods of pepsin immobilization have been applied in order to achieve the continuous hydrolysis of a 2,5% haemoglobin solution at pH 2 and 40°C. Methods using glutaraldehyde were unsuccesful because of the unstability of the derived enzyme at low pH. Pepsin covalently bounded to a Duolite amine resin by a carbodiimide showed a half life of 15 days during the hydrolysis of haemoglobin in a column reactor, No enzyme activity was detected in the hydrolysates. No accumulation of haem in the column was noticed which could have limited long term studies by plugging the system. Modulation of the degree of hydrolysis was also performed by changing the feeding flow rate of the reactor.

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Section: Introductionmentioning

confidence: 99%

Stabilization of pepsin on duolite for the continuous hydrolysis of bovine haemoglobin at pH2 and 40�C

Sannier¹,

Piot²,

Guillochon³

et al. 1993

Biotechnol Tech

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“…The EDC was added dropwise to the gel slurry with continuous stirring, and the reaction was allowed to continue for 1 h. The reaction was stopped with sequential washes of (1) 6 L of H20 (pH 4.5), (2) 6 L of 50 mM NaCl (pH 4.5), (3) 6 L of H20 (pH 4.5), and (4) 1 L of 0.15 M sodium acetate (pH 5.0). The amount of pepsin coupled to the AH-Sepharose gel was then estimated by a hemoglobin digestion assay (Valentova et al, 1975) or by direct titration with known quantities of A. suum pepsin inhibitor. One gram of suction-filtered affinity gel bound approximately 1500 units of inhibitor.…”

Section: Methodsmentioning

confidence: 99%

Primary structure of the major pepsin inhibitor from the intestinal parasitic nematode Ascaris suum

Martzen¹,

McMullen²,

Smith³

et al. 1990

Biochemistry

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The major pepsin inhibitor from Ascaris suum was isolated by affinity chromatography and chromatofocusing. Its amino acid sequence was determined by automated Edman degradation of peptide fragments. Peptides were produced by chemical and enzymatic cleavage of pyridylethylated protein and were purified by reverse-phase high-performance liquid chromatography. The inhibitor consists of 149 residues with the following sequence: QFLFSMSTGP10FICTVKDNQV20FVANLPWTML30EGDDIQVGKE40 FAARVEDCTN50VKHDMAPTCT60KPPPFCGPQD70MKMFNFVGCS80VLGNKLFIDQ90KYVRDLTAK D100 HAEVQTFREK110IAAFEEQQEN120QPPSSGMPHG130AVPAGGLSPP140PPPSFCTVQ149. It has a molecular weight of 16,396. All cysteines are engaged as disulfide bonds: Cys(13)-Cys(59), Cys(48)-Cys(66), and Cys(79)-Cys(146). The protein is probably composed of two domains connected by a short hydrophobic region. This is the first aspartyl protease inhibitor of animal origin that has been sequenced. The sequence has no significant homology with any other known protein.

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“…These results are consistent with the results of Valentova. 38 In addition to covalent cross-linking with glutaraldehyde, a few pepsin molecules may be physically adsorbed in the pores of the support. 39,40 Therefore, the initial concentration of pepsin for the immobilization process was set at 35 mg mL −1 for further experiments.…”

Section: Characterization Of Immobilized Pepsinmentioning

confidence: 99%

Preparation of immobilized pepsin for extraction of collagen from bovine hide

Duan

Cheng

2022

RSC Adv.

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Pepsin immobilized by covalent fixation to hydroxyalkyl methacrylate gels: Preparation and characterization

Cited by 24 publications

References 13 publications

Stabilization of pepsin on duolite for the continuous hydrolysis of bovine haemoglobin at pH2 and 40�C

Stabilization of pepsin on duolite for the continuous hydrolysis of bovine haemoglobin at pH2 and 40�C

Primary structure of the major pepsin inhibitor from the intestinal parasitic nematode Ascaris suum

Preparation of immobilized pepsin for extraction of collagen from bovine hide

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