Application of nitrite reductase from Alcaligenes faecalis S-6 for nitrite measurement

Sasaki, Satoshi; Karube, Isao; Hirota, Nobuko; Arikawa, Yoshiko; Nishiyama, Makoto; Kukimoto, Mutsuko; Horinouchi, Sueharu; Beppu, Teruhiko

doi:10.1016/s0956-5663(97)00100-0

Cited by 35 publications

(17 citation statements)

References 11 publications

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“…The corresponding acid could be the nitrite (pK a ϭ 3.25), the active site aspartate (usual pK a range in proteins (31) is 2-5.5) that is known to hydrogen bond to the bound nitrite (14,16,18), or the active site His-255 (usual range for histidine is [5][6][7][8] Fig. 4D).…”

Section: Discussionmentioning

confidence: 99%

A Random-sequential Mechanism for Nitrite Binding and Active Site Reduction in Copper-containing Nitrite Reductase

Wijma

Jeuken

Verbeet

et al. 2006

Journal of Biological Chemistry

128

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The homotrimeric copper-containing nitrite reductase (NiR) contains one type-1 and one type-2 copper center per monomer. Electrons enter through the type-1 site and are shuttled to the type-2 site where nitrite is reduced to nitric oxide. To investigate the catalytic mechanism of NiR the effects of pH and nitrite on the turnover rate in the presence of three different electron donors at saturating concentrations were measured. The activity of NiR was also measured electrochemically by exploiting direct electron transfer to the enzyme immobilized on a graphite rotating disk electrode. In all cases, the steady-state kinetics fitted excellently to a randomsequential mechanism in which electron transfer from the type-1 to the type-2 site is rate-limiting. At low [NO 2 ؊ ] reduction of the type-2 site precedes nitrite binding, at high [NO 2 ؊ ] the reverse occurs.Below pH 6.5, the catalytic activity diminished at higher nitrite concentrations, in agreement with electron transfer being slower to the nitrite-bound type-2 site than to the water-bound type-2 site. Above pH 6.5, substrate activation is observed, in agreement with electron transfer to the nitrite-bound type-2 site being faster than electron transfer to the hydroxyl-bound type-2 site. To study the effect of slower electron transfer between the type-1 and type-2 site, NiR M150T was used. It has a type-1 site with a 125-mV higher midpoint potential and a 0.3-eV higher reorganization energy leading to an ϳ50-fold slower intramolecular electron transfer to the type-2 site. The results confirm that NiR employs a random-sequential mechanism.

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Section: Discussionmentioning

confidence: 99%

A Random-sequential Mechanism for Nitrite Binding and Active Site Reduction in Copper-containing Nitrite Reductase

Wijma

Jeuken

Verbeet

et al. 2006

Journal of Biological Chemistry

128

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“…An enzymatic reaction of nitrite reductase (NIR) from Alcaligenes faecalis S-6 was used by Sasaki et al to measure nitrite. Amperometric methods using NIR and the electron mediator 1-methoxy-5-methylphenazinium methylsulfate (1-methoxy PMS) have also been used [253]. Measurements using a batch-flow type system gave a lower detection limit of 0.01 mg L -1 .…”

Section: Acid Rain Sensorsmentioning

confidence: 99%

Current research activity in biosensors

Nakamura

Karube

2003

Analytical and Bioanalytical Chemistry

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261

129

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Biosensors consist of a molecular recognition element and a transducer. Since the first biosensor was developed by Updike and Hicks many biosensors and their associated techniques have been studied and developed. In this review current research activity on the fundamentals and applications of biosensors is summarized. In discussion of the former, molecular recognition elements, techniques and tools for biosensor construction, and basic biosensor devices are introduced. Coverage of the latter includes description of biosensors for environmental, food, and clinical fields. Chemical sensors developed in our laboratory for environmental monitoring are also described. This review mainly summarizes work performed by our group and by our colleagues, and refers to the main review articles summarizing each field of biosensor research in recent years.

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“…Examples include a liquid ion-exchange sensor (7) and a sensor based on an immobilized enzyme (35). However, as tools for online measurement of NO 2 Ϫ in wastewater or microscale analysis of NO 2 Ϫ in marine sediment, these types of sensors have distinct limitations.…”

Section: Nitrite (Nomentioning

confidence: 99%

Bacterium-Based NO ₂ ⁻ Biosensor for Environmental Applications

Nielsen

Larsen

Jetten

et al. 2004

Appl Environ Microbiol

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A sensitive NO 2؊ biosensor that is based on bacterial reduction of NO 2 ؊ to N 2 O and subsequent detection of the N 2 O by a built-in electrochemical N 2 O sensor was developed. Four different denitrifying organisms lacking NO 3 ؊ reductase activity were assessed for use in the biosensor. The relevant physiological aspects examined included denitrifying characteristics, growth rate, NO 2 ؊ tolerance, and temperature and salinity effects on the growth rate. Two organisms were successfully used in the biosensor. The preferred organism was Stenotrophomonas nitritireducens, which is an organism with a denitrifying pathway deficient in both NO 3 ؊ and N 2 O reductases. Alternatively Alcaligenes faecalis could be used when acetylene was added to inhibit its N 2 O reductase. The macroscale biosensors constructed exhibited a linear NO 2 ؊ response at concentrations up to 1 to 2 mM. The detection limit was around 1 M NO 2 ؊ , and the 90% response time was 0.5 to 3 min. The sensor signal was specific for NO 2 ؊ , and interference was observed only with NH 2 OH, NO, N 2 O, and H 2 S. The sensor signal was affected by changes in temperature and salinity, and calibration had to be performed in a system with a temperature and an ionic strength comparable to those of the medium analyzed. A broad range of water bodies could be analyzed with the biosensor, including freshwater systems, marine systems, and oxic-anoxic wastewaters. The NO 2 ؊ biosensor was successfully used for long-term online monitoring in wastewater. Microscale versions of the NO 2 ؊ biosensor were constructed and used to measure NO 2 ؊ profiles in marine sediment. Nitrite (NO 2Ϫ ) has a central position in the global nitrogen (N) cycle and is involved in many important biological N transformations. With an intermediate oxidation state, NO 2 Ϫ acts as an electron donor in nitrification and serves as an electron acceptor in denitrification, dissimilative nitrite reduction to ammonium, and anammox.In natural ecosystems NO 2 Ϫ is of interest because of its toxicity for microorganisms and higher organisms (20,37). NO 2 Ϫ concentrations in natural bulk water bodies are usually very low, and in freshwater systems the average worldwide NO 2 Ϫ concentration has been estimated to be about 1 g of N liter Ϫ1 (ϳ0.07 M NO 2 Ϫ ) (28). Recently, however, there have been several reports of NO 2 Ϫ accumulation to concentrations of 5 to 140 M in European eutrophic rivers and estuaries (3,8,15,19,44). Studies have indicated that NO 2 Ϫ accumulations can be related to imbalances in NO 2 Ϫ production and consumption rates for either aerobic sediment processes (nitrification) or anaerobic sediment processes (denitrification and dissimilatory nitrate reduction to ammonia). In marine systems NO 2 Ϫ concentrations are normally negligible, but concentrations of 0.3 to 2.5 M have been reported near the oxic-anoxic boundary of stratified marine water bodies (5, 22). The distribution of NO 2 Ϫ in sediments is largely uncharacterized, except for a few studies that have documented NO 2 Ϫ a...

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Application of nitrite reductase from Alcaligenes faecalis S-6 for nitrite measurement

Cited by 35 publications

References 11 publications

A Random-sequential Mechanism for Nitrite Binding and Active Site Reduction in Copper-containing Nitrite Reductase

A Random-sequential Mechanism for Nitrite Binding and Active Site Reduction in Copper-containing Nitrite Reductase

Current research activity in biosensors

Bacterium-Based NO ₂ ⁻ Biosensor for Environmental Applications

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Application of nitrite reductase from Alcaligenes faecalis S-6 for nitrite measurement

Cited by 35 publications

References 11 publications

A Random-sequential Mechanism for Nitrite Binding and Active Site Reduction in Copper-containing Nitrite Reductase

A Random-sequential Mechanism for Nitrite Binding and Active Site Reduction in Copper-containing Nitrite Reductase

Current research activity in biosensors

Bacterium-Based NO 2 − Biosensor for Environmental Applications

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Product

Resources

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Bacterium-Based NO ₂ ⁻ Biosensor for Environmental Applications