Covalent Binding of an NAD Analogue to Liver Alcohol Dehydrogenase Resulting in an Enzyme‐Coenzyme Complex not Requiring Exogenous Coenzyme for Activity

Månsson, Mats-Olle; Larsson, Per‐Olof; Mosbach, Klaus

doi:10.1111/j.1432-1033.1978.tb12328.x

Cited by 65 publications

(8 citation statements)

References 28 publications

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“…Scheme 2 shows the chemical routes involved in this modification procedure. It is believed that the cofactor modifications were achieved through the amino group on the adenosine moiety as reported previously in literature (Buckmann et al, 1981;Mansson et al, 1978). The immobilized catalysts were washed extensively with both buffer solution and DI water to a point that no free NADH and enzyme in the washing solutions were detectable through either HPLC or activity analyses.…”

Section: Resultsmentioning

confidence: 97%

Enabling multienzyme biocatalysis using nanoporous materials

El‐Zahab

Jia

Wang

2004

Biotech & Bioengineering

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Multistep reactions catalyzed by a covalently immobilized enzyme-cofactor-enzyme system were achieved. Lactate dehydrogenase (LDH), glucose dehydrogenase (GDH), and cofactor NADH were incorporated into two porous silica glass supports. One of the glass supports had pores of 30 nm in diameter, while the other was of 100-nm pore size. Effective shuttling of the covalently bound NADH between LDH and GDH was achieved, such that regeneration cycles of NADH/NAD(+) were observed. The glass of 30-nm pore size afforded enzyme activities that were about twice those observed for the glass of 100-nm pore size, indicating the former provided better enzyme-cofactor integration. The effect of the size of spacers was also examined. The use of longer spacers increased the reaction rates by approximately 18 times as compared to those achieved with glutaraldehyde linkage. It appeared that the concave configuration of the nanopores played an important role in enabling the multistep reactions. The same multienzyme system immobilized on nonporous polystyrene particles of 500-nm diameter was only approximately 2% active as the glass-supported system. It is believed that the nanoporous structure of the glass supports enhances the molecular interactions among the immobilized enzymes and cofactor, thus improving the catalytic efficiency of the system.

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Section: Resultsmentioning

confidence: 97%

Enabling multienzyme biocatalysis using nanoporous materials

El‐Zahab

Jia

Wang

2004

Biotech & Bioengineering

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“…The same happened in the case presented in [102]. Using an NAD + analogue prepared like in [103] for binding to HLADH through a spacer, the authors obtained an enzyme-cofactor complex which was immobilized on a glassy carbon electrode. Coenzyme regeneration was not possible due to coenzyme decomposition at the surface of the electrode.…”

Section: Nad(p)(h) Derivativesmentioning

confidence: 79%

Redox Enzymes Used in Chiral Syntheses Coupled to Coenzyme Regeneration

Leonida¹

2001

CMC

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Due to their ability to perform reactions in a stereo- and regiospecific manner, while functioning under mild conditions of temperature and pH, enzymes are increasingly used for chiral synthesis in the pharmaceutical industry, in medicinal chemistry, agriculture, cosmetic and food industry. The oxidoreductases as catalysts require a coenzyme (cofactor) which functions as an electron carrier. If used in stoichiometric amounts the cost of coenzymes makes synthetic applications of redox enzymes prohibitively expensive for industrial applications. The present review updates previous discussions about the objectives of cofactor regeneration as a requirement for the applicability of enzymatic redox reactions, and about the strategies customarily used to this end. Different types of chiral syntheses coupled successfully to coenzyme regeneration (amino acid synthesis, lactone synthesis, synthetic reactions involving alcohols, hydroxy acid and steroid synthesis, deracemization) are then discussed. New catalysts, cross-linked enzyme crystals, prepared from redox enzyme are presented in a small paragraph together with some of their applications. Special attention is given to whole cells used in this type of synthesis and their advantages and disadvantages are presented in one paragraph. Finally, different reactors, within which were conducted chiral syntheses catalyzed by oxireductases and coupled to cofactor regeneration, are briefly discussed.

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“…1, in which cinnamoyl alcohol is oxidized to aldehyde and NAD(P)+ reduced to NAD(P)H. In the next step, the NAD(P)+ is regenerated by oxidation and the octylaldehyde reduced to octanol. No coenzyme was added, but the coenzyme coprecipitated with the enzyme was functioning in the enzyme active site and was continuously recycled as in the system with NAD' covalently bound to horse liver alcohol dehydrogenase (Minsson et al, 1978;Minsson and Mosbach, 1987). The reaction mixture was investigated after a recycling assay with cinnamoyl alcohol and octyl aldehyde in acetonitrilehuffer and it was noted that neither NAD(P)' nor enzyme had been solubilized from the precipitate during the assay.…”

Section: Resultsmentioning

confidence: 99%

Horse Liver Alcohol Dehydrogenase can Accept NADP⁺ as Coenzyme in High Concentrations of Acetonitrile

Johansson

Mosbach

Månsson

1995

European Journal of Biochemistry

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The coenzyme specificity of horse liver alcohol dehydrogenase assayed in mixtures of acetonitrile and buffer, 0.01 M Tris/HCI pH 7.4, was investigated. The enzyme could accept NADP' as coenzyme if it was first bio-imprinted with NADP', i.e. precipitated from an aqueous buffer with 1-propanol in the presence of NADP', dried, and then assayed in a system with less than 10% buffer. When assayed in a system with 25 % buffer, no activity with NADP' as coenzyme was observed. The activity was measured with a coenzyme regeneration assay with cinnamoyl alcohol and octylaldehyde as substrates. Other methods to prepare a binary complex of horse liver alcohol dehydrogenase and NADP', i.e. if the enzyme was immobilized on silica and NADP' added afterwards or if it was deposited on Celite together with NADP', failed to show any bio-imprinting effect. The activity of horse liver alcohol dehydrogenase bioimprinted with NADP' and assayed in acetonitrilehuffer mixtures with less than 10% buffer showed the same or higher activity than an enzyme preparation prepared by bio-imprinting with NAD'. The hypothesis is that the conformation of the active site of horse liver alcohol dehydrogenase is modified to one that is complementary to the ligand present during precipitation and drying. This is possible because of the restricted motility of the enzyme conformation in high concentrations of organic solvent. In the presence of more than 10% buffer the mobility increases in such a way that the imposed conformation with activity towards NADP' disappears.

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Covalent Binding of an NAD Analogue to Liver Alcohol Dehydrogenase Resulting in an Enzyme‐Coenzyme Complex not Requiring Exogenous Coenzyme for Activity

Cited by 65 publications

References 28 publications

Enabling multienzyme biocatalysis using nanoporous materials

Enabling multienzyme biocatalysis using nanoporous materials

Redox Enzymes Used in Chiral Syntheses Coupled to Coenzyme Regeneration

Horse Liver Alcohol Dehydrogenase can Accept NADP⁺ as Coenzyme in High Concentrations of Acetonitrile

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Covalent Binding of an NAD Analogue to Liver Alcohol Dehydrogenase Resulting in an Enzyme‐Coenzyme Complex not Requiring Exogenous Coenzyme for Activity

Cited by 65 publications

References 28 publications

Enabling multienzyme biocatalysis using nanoporous materials

Enabling multienzyme biocatalysis using nanoporous materials

Redox Enzymes Used in Chiral Syntheses Coupled to Coenzyme Regeneration

Horse Liver Alcohol Dehydrogenase can Accept NADP+ as Coenzyme in High Concentrations of Acetonitrile

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Horse Liver Alcohol Dehydrogenase can Accept NADP⁺ as Coenzyme in High Concentrations of Acetonitrile