PMID- 20951671
OWN - NLM
STAT- MEDLINE
DCOM- 20110322
LR  - 20171116
IS  - 1096-0309 (Electronic)
IS  - 0003-2697 (Linking)
VI  - 409
IP  - 1
DP  - 2011 Feb 1
TI  - Visible wavelength spectrophotometric assays of L-aspartate and D-aspartate using 
      hyperthermophilic enzyme systems.
PG  - 1-6
LID - 10.1016/j.ab.2010.10.016 [doi]
AB  - Methods with which to simply and rapidly assay L-aspartate (L-Asp) and 
      D-aspartate (D-Asp) would be highly useful for physiological research and for 
      nutritional and clinical analyses. Levels of L- and D-Asp in food and cell 
      extracts are currently determined using high-performance liquid chromatography. 
      However, this method is time-consuming and expensive. Here we describe a simple 
      and specific method for using an L-aspartate dehydrogenase (L-AspDH) system to 
      colorimetrically assay L-Asp and a system of three hyperthermophilic 
      enzymes--aspartate racemase (AspR), L-AspDH, and L-aspartate oxidase (L-AO)--to 
      assay D-Asp. In the former, the reaction rate of nicotinamide adenine 
      dinucleotide (NAD(+))-dependent L-AspDH was measured based on increases in the 
      absorbance at 438 nm, reflecting formation of formazan from water-soluble 
      tetrazolium-1 (WST-1), using 1-methoxy-5-methylphenazinum methyl sulfate (mPMS) 
      as a redox mediator. In the latter, D-Asp was measured after first removing L-Asp 
      in the sample solution with L-AO. The remaining D-Asp was then changed to L-Asp 
      using racemase, and the newly formed L-Asp was assayed calorimetrically using 
      NAD(+)-dependent aspartate dehydrogenase as described above. This method enables 
      simple and rapid spectrophotometric determination of 1 to 100 muM L- and D-Asp in 
      the assay systems. In addition, methods were applicable to the L- and D-Asp 
      determinations in some living cells and foods.
CI  - Copyright (c) 2010 Elsevier Inc. All rights reserved.
FAU - Mutaguchi, Yuta
AU  - Mutaguchi Y
AD  - Microbial Genetic Division, Institute of Genetic Resources, Faculty of 
      Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, 
      Japan.
FAU - Ohmori, Taketo
AU  - Ohmori T
FAU - Sakuraba, Haruhiko
AU  - Sakuraba H
FAU - Yoneda, Kazunari
AU  - Yoneda K
FAU - Doi, Katsumi
AU  - Doi K
FAU - Ohshima, Toshihisa
AU  - Ohshima T
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20101015
PL  - United States
TA  - Anal Biochem
JT  - Analytical biochemistry
JID - 0370535
RN  - 0 (Escherichia coli Proteins)
RN  - 0U46U6E8UK (NAD)
RN  - 299-11-6 (Methylphenazonium Methosulfate)
RN  - 30KYC7MIAI (Aspartic Acid)
RN  - 4SR0Q8YD1X (D-Aspartic Acid)
RN  - 65162-13-2 (1-methoxy-5-methylphenazinium methyl sulfate)
RN  - EC 1.4.- (Amino Acid Oxidoreductases)
RN  - EC 1.4.1.- (aspartate dehydrogenase)
RN  - EC 1.4.3.16 (L-aspartate oxidase, E coli)
RN  - EC 5.1.1.- (Amino Acid Isomerases)
RN  - EC 5.1.1.13 (aspartate racemase)
RN  - Q40Q9N063P (Acetic Acid)
SB  - IM
MH  - Acetic Acid/chemistry
MH  - Amino Acid Isomerases/metabolism
MH  - Amino Acid Oxidoreductases/metabolism
MH  - Animals
MH  - Aspartic Acid/*analysis
MH  - D-Aspartic Acid/*analysis
MH  - Escherichia coli Proteins
MH  - Isomerism
MH  - Liver/chemistry
MH  - Methylphenazonium Methosulfate/analogs & derivatives/chemistry
MH  - Mice
MH  - NAD/chemistry
MH  - Oxidation-Reduction
MH  - Spectrophotometry/*methods
MH  - Swine
EDAT- 2010/10/19 06:00
MHDA- 2011/03/23 06:00
CRDT- 2010/10/19 06:00
PHST- 2010/03/11 00:00 [received]
PHST- 2010/10/12 00:00 [revised]
PHST- 2010/10/12 00:00 [accepted]
PHST- 2010/10/19 06:00 [entrez]
PHST- 2010/10/19 06:00 [pubmed]
PHST- 2011/03/23 06:00 [medline]
AID - S0003-2697(10)00659-7 [pii]
AID - 10.1016/j.ab.2010.10.016 [doi]
PST - ppublish
SO  - Anal Biochem. 2011 Feb 1;409(1):1-6. doi: 10.1016/j.ab.2010.10.016. Epub 2010 Oct 
      15.