Kinetics and native and modified liver alcohol dehydrogenase with coenzyme analogs: isomerization of enzyme-nicotinamide adenine dinucleotide complex

We have studied the binding of 1 ,lo-phenanthroline to specifically active-site cobalt(I1)-substituted horse-liver alcohol dehydrogenase [Co(II)-LADH]. The dissociation constant is a factor of 6500 smaller than in the native enzyme. Spectral evidence is given which shows that 1 ,lo-phenanthroline does not remove the catalytic Co(I1) ion and that binding of 1 ,I 0-phenanthroline renders the catalytic metal ion pentacoordinate. The maximum limiting rate constant for the association of 1,lO-phenanthroline to Co(I1)-LADH is about 60 s -' . This is about a third of the value (1 69 s-') determined for native horse-liver alcohol dehydrogenase, Zn(1I)LADH [Frolich et al. (1978) Arch. Biochem. Biuphys. 189,471 -4801. For cadmium(l1)-substituted horse-liver alcohol dehydrogenase, [Cd(II)LADH] the maximum limiting rate constant for association of 1 ,lo-phenanthroline increased to 590 s-'. These findings demonstrate that the rate-limiting step is strongly dependent on the chemical nature of the catalytic metal ion and its immediate environment. 1,lO-Phenanthroline is shown to bind to the Co(I1)-LADH . NAD' complex in the open conformation. The maximum limiting rate constant remains unchanged in the presence of NAD'. The data have been used to derive a kinetic scheme for the formation of ternary complexes including NAD' that involves a slow intermediary step.

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“…NAD' has been proposed by Plapp et al [28]. In chemically modified LADH this step became rate-limiting for the oxidation of ethanol by …”

Section: Lo-phenanthroline As Monitor and Effector Of The Open Confmentioning

confidence: 98%

The binding of 1,10‐phenanthroline to specifically active‐site cobalt(II)‐substituted horse‐liver alcohol dehydrogenase

Helander

Lindner

Brade

et al. 1988

European Journal of Biochemistry

202

140

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“…1 b), and BNA is a typical biomimetic nicotinamide coenzyme. Few wild-type redox enzymes have been reported to have promiscuous activities on NMN, including liver alcohol dehydrogenase [111] and glutamic dehydrogenase [112] . Scott and coworkers have engineered Pyrococcus furiosus alcohol dehydrogenase to act on NMN, but the enzyme activity remains very low [113] .…”

Section: Biomimetic Coenzyme Engineeringmentioning

confidence: 99%

Protein engineering of oxidoreductases utilizing nicotinamide-based coenzymes, with applications in synthetic biology

You

Huang

Wei

et al. 2017

Synthetic and Systems Biotechnology

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Two natural nicotinamide-based coenzymes (NAD and NADP) are indispensably required by the vast majority of oxidoreductases for catabolism and anabolism, respectively. Most NAD(P)-dependent oxidoreductases prefer one coenzyme as an electron acceptor or donor to the other depending on their different metabolic roles. This coenzyme preference associated with coenzyme imbalance presents some challenges for the construction of high-efficiency in vivo and in vitro synthetic biology pathways. Changing the coenzyme preference of NAD(P)-dependent oxidoreductases is an important area of protein engineering, which is closely related to product-oriented synthetic biology projects. This review focuses on the methodology of nicotinamide-based coenzyme engineering, with its application in improving product yields and decreasing production costs. Biomimetic nicotinamide-containing coenzymes have been proposed to replace natural coenzymes because they are more stable and less costly than natural coenzymes. Recent advances in the switching of coenzyme preference from natural to biomimetic coenzymes are also covered in this review. Engineering coenzyme preferences from natural to biomimetic coenzymes has become an important direction for coenzyme engineering, especially for in vitro synthetic pathways and in vivo bioorthogonal redox pathways.

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“…5). A few wild-type redox enzymes function using NMN, including liver alcohol dehydrogenase [88] and glutamic dehydrogenase [89]. Recently, Scott et al have shown that engineered P. furiosus alcohol dehydrogenase can work on NMN [90].…”

Section: Redox Enzyme Engineeringmentioning

confidence: 99%

Biomanufacturing by in vitro biosystems containing complex enzyme mixtures

You

Zhang

2017

Process Biochemistry

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Microbial fermentation is the current predominant biomanufacturing platform. However, it suffers from low production yields, slow reaction rates, and scaling-up challenges. In vitro enzymatic biosystems are emerging to expand the traditional biotechnological mode by utilizing more than three enzymes for manufacturing the desired product from cheap substrate. In the past few years, numerous proofs of the concept of in vitro biosystems containing complex enzyme mixtures from different groups worldwide have inspired the development of these platforms for biomanufacturing, these biosystems show advantages such as near-theoretical product yields, faster reaction rates, reduced interference from toxic compounds, and unprecedented level of engineering. In this review, several examples of in vitro systems are presented to illustrate these advantages and possible solutions to overcome the remaining challenges are discussed. The continuing decrease in enzyme cost and improvements in enzyme engineering techniques will make in vitro biosystems a comparable biomanufacturing platform for microbial fermentation in the near future.

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Kinetics and native and modified liver alcohol dehydrogenase with coenzyme analogs: isomerization of enzyme-nicotinamide adenine dinucleotide complex

Cited by 26 publications

References 45 publications

The binding of 1,10‐phenanthroline to specifically active‐site cobalt(II)‐substituted horse‐liver alcohol dehydrogenase

The binding of 1,10‐phenanthroline to specifically active‐site cobalt(II)‐substituted horse‐liver alcohol dehydrogenase

Protein engineering of oxidoreductases utilizing nicotinamide-based coenzymes, with applications in synthetic biology

Biomanufacturing by in vitro biosystems containing complex enzyme mixtures

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