Covalent Chromatography

Batista‐Viera, Francisco; Rydén, Lars; Carlsson, Jan

doi:10.1002/9780470939932.ch8

Cited by 4 publications

(1 citation statement)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The bond can be broken by reaction with some suitable agent, such as dithiothreitol (DTT) under mild circumtainance (Guisan, 2013). Additionally, because the reactivity of the thiol groups was modulated via pH alteration, the activity yield of the methods involving disulfide bond method was usually provided when the appropriate thiol-reactive adsorbent with high specificity was applied (Carlsson et al, 1998).…”

Section: Formation Of Disulfide Bondsmentioning

confidence: 99%

Study the Best Support for Hydrolysis of Tiger Grass

Sayamnikorn

View full text Add to dashboard Cite

Bioethanol, one of the outstanding biofuels, is not only a renewable bio-based resource, but also interesting to research for improving better properties of the enzyme needed in the hydrolysis process. The purpose of this work is to study how to reuse the enzyme used in the hydrolysis step for bioethanol production by immobilizing Trichoderma reesei (T.reesei) on various silica supports, namely, SBA-15, TUD-1, and MCM-48. The amount of the adsorbed enzymes was determined by UV-visible spectrophotometry. Among those studied supports, SBA-15 showed 100% enzymatic adsorption on the support owing to its larger pore diameter of 6.14 nm, which is large enough to accommodate T.reesei enzyme molecules inside the pore channel. Various parameters, viz. temperature, pH, time, and amount of the support for optimizing the immobilized enzyme were investigated. The immobilized T.reesei on SBA-15 support was characterized by N2 adsorption-desorption. The amount of monomeric sugar after the hydrolysis process was measured by high performance liquid chromatography (HPLC).

show abstract

Section: Formation Of Disulfide Bondsmentioning

confidence: 99%

Study the Best Support for Hydrolysis of Tiger Grass

Sayamnikorn

View full text Add to dashboard Cite

show abstract

Immobilization of Enzymes: A Literature Survey

Brena

González-Pombo

Batista‐Viera

2013

Methods in Molecular Biology

283

155

View full text Add to dashboard Cite

The term immobilized enzymes refers to "enzymes physically confined or localized in a certain defined region of space with retention of their catalytic activities, and which can be used repeatedly and continuously." Immobilized enzymes are currently the subject of considerable interest because of their advantages over soluble enzymes. In addition to their use in industrial processes, the immobilization techniques are the basis for making a number of biotechnology products with application in diagnostics, bioaffinity chromatography, and biosensors. At the beginning, only immobilized single enzymes were used, after 1970s more complex systems including two-enzyme reactions with cofactor regeneration and living cells were developed. The enzymes can be attached to the support by interactions ranging from reversible physical adsorption and ionic linkages to stable covalent bonds. Although the choice of the most appropriate immobilization technique depends on the nature of the enzyme and the carrier, in the last years the immobilization technology has increasingly become a matter of rational design. As a consequence of enzyme immobilization, some properties such as catalytic activity or thermal stability become altered. These effects have been demonstrated and exploited. The concept of stabilization has been an important driving force for immobilizing enzymes. Moreover, true stabilization at the molecular level has been demonstrated, e.g., proteins immobilized through multipoint covalent binding.

show abstract

Reversible Covalent Immobilization of Enzymes via Disulfide Bonds

Ovsejevi

Manta

Batista‐Viera

2013

Methods in Molecular Biology

View full text Add to dashboard Cite

This enzyme immobilization approach involves the formation of disulfide (-S-S-) bonds with the support. Thus, enzymes bearing exposed nonessential thiol (SH) groups can be immobilized onto thiol-reactive supports provided with reactive disulfides or disulfide oxides under mild conditions. The great potential advantage of this approach is the reversibility of the bonds formed between the activated solid phase and the thiol-enzyme, because the bound protein can be released with an excess of a low-molecular-weight thiol (e.g., dithiothreitol [DTT]). This is of particular interest when the enzyme degrades much faster than the adsorbent, which can be reloaded afterwards. The possibility of reusing the polymeric support after inactivation of the enzyme may be of interest for the practical use of immobilized enzymes in large-scale processes in industry, where their use has often been hampered by the high cost of the support material. Disulfide oxides (thiolsulfinate or thiolsulfonate groups) can be introduced onto a wide variety of support materials with different degrees of porosity and with different mechanical resistances. Procedures are given for the preparation of thiol-activated solid phases and the covalent attachment of thiol-enzymes to the support material via disulfide bonds. The possibility of reusing the polymeric support is also shown.

show abstract

Covalent Chromatography

Cited by 4 publications

References 44 publications

Study the Best Support for Hydrolysis of Tiger Grass

Study the Best Support for Hydrolysis of Tiger Grass

Immobilization of Enzymes: A Literature Survey

Reversible Covalent Immobilization of Enzymes via Disulfide Bonds

Contact Info

Product

Resources

About