NL1005028C2 - Controlling enzyme reactions in polar solvent - Google Patents

Controlling enzyme reactions in polar solvent Download PDF

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Publication number
NL1005028C2
NL1005028C2 NL1005028A NL1005028A NL1005028C2 NL 1005028 C2 NL1005028 C2 NL 1005028C2 NL 1005028 A NL1005028 A NL 1005028A NL 1005028 A NL1005028 A NL 1005028A NL 1005028 C2 NL1005028 C2 NL 1005028C2
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Prior art keywords
polar solvent
enzyme reactions
enzyme
electromagnetic radiation
controlling enzyme
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NL1005028A
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Dutch (nl)
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Franciscus Matheus Everaerts
Theodorus Petrus Eug Verheggen
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Franciscus Matheus Everaerts
Theodorus Petrus Eugenius Mari
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Priority to NL1005028A priority Critical patent/NL1005028C2/en
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Publication of NL1005028C2 publication Critical patent/NL1005028C2/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J19/08—Processes employing the direct application of electric or wave energy, or particle radiation; Apparatus therefor
    • B01J19/12—Processes employing the direct application of electric or wave energy, or particle radiation; Apparatus therefor employing electromagnetic waves
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • Toxicology (AREA)
  • Electromagnetism (AREA)
  • Immunology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biomedical Technology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

Controlling enzyme reactions comprises conducting the reaction in a polar solvent which is subjected to electromagnetic radiation of 5\*107 to 1\* 1010 Hz.

Description

Werkwijze voor het controleren van een enzymreactieMethod for controlling an enzyme reaction

De onderhavige uitvinding heeft betrekking op een werkwijze voor het controleren van een enzymreactie in een polair oplosmiddel.The present invention relates to a method for controlling an enzyme reaction in a polar solvent.

In het vak wordt de snelheid van een enzymreactie 5 gecontroleerd, meestal versneld, door het toevoegen van co- factoren, zoals calcium- of magnesiumionen, of het wijzigen, meestal verhogen, van de temperatuur.In the art, the rate of an enzyme reaction is controlled, usually accelerated, by adding co-factors, such as calcium or magnesium ions, or changing, usually increasing, the temperature.

De onderhavige uitvinding beoogt het aantal wijzen waarop een enzymreactie kan worden gecontroleerd uit te brei-10 den.The present invention aims to expand the number of ways in which an enzyme reaction can be controlled.

Daartoe wordt de werkwijze volgens de uitvinding gekenmerkt doordat dat de enzymreactie wordt uitgevoerd in een met elektromagnetische straling behandeld polair oplosmiddel, waarbij de elektromagnetische straling een frequentie 15 heeft in het bereik van 5*107 tot 1010 Hz.To this end, the method according to the invention is characterized in that the enzyme reaction is carried out in a polar solvent treated with electromagnetic radiation, the electromagnetic radiation having a frequency in the range of 5 * 107 to 1010 Hz.

Verrassenderwijs is gebleken dat de enzymatische activiteit van een enzym kan worden gewijzigd door het gebruik van elektromagnetische straling in het voornoemde bereik. Deze verandering is niet het gevolg van een verhoging 20 van de temperatuur van het polaire oplosmiddel. Zonder aan deze theorie gebonden te zijn, wordt gemeend dat elektromagnetische straling binnen het genoemde frequentiebereik in staat is de structuur van het polaire oplosmiddel te wijzigen. Het is in de stand van de techniek bekend dat vloeistof-25 moleculen zich niet louter chaotisch bewegen, doch dat in een vloeistof enige mate van structuur voorkomt, zoals verschillende soorten clusters. Volgens aanvraagsters kan de verdeling van verschillende in de vloeistof optredende structuren worden beïnvloed door gebruik te maken van elektromagnetische 30 straling in het genoemde frequentiebereik, en kan daarmee de beschikbaarheid van het enzymsubstraat en de enzymactiviteit worden beïnvloed.Surprisingly, it has been found that the enzymatic activity of an enzyme can be modified using electromagnetic radiation in the aforementioned range. This change is not due to an increase in the temperature of the polar solvent. Without being bound by this theory, it is believed that electromagnetic radiation within the said frequency range is capable of altering the structure of the polar solvent. It is known in the art that liquid molecules do not move purely chaotically, but that a degree of structure occurs in a liquid, such as different types of clusters. According to the applicants, the distribution of different structures occurring in the liquid can be influenced by using electromagnetic radiation in the said frequency range, and can thereby influence the availability of the enzyme substrate and the enzyme activity.

Bij voorkeur wordt de enzymreactie uitgevoerd in een waterig oplosmiddel als het polaire oplosmiddel.Preferably, the enzyme reaction is performed in an aqueous solvent as the polar solvent.

1005028 21005028 2

Gebleken is dat water, in het bijzonder in de vorm van een buffer, een geschikt oplosmiddel is waarin de enzym-activiteit kan worden beïnvloed.It has been found that water, in particular in the form of a buffer, is a suitable solvent in which the enzyme activity can be influenced.

Aangezien verschillende enzymen verschillend op de 5 behandeling met elektromagnetische straling reageren, is het mogelijk om bij gebruik van meer dan een enzym de enzymacti-viteiten van de enzymen beter op elkaar af te stemmen.Since different enzymes react differently to the treatment with electromagnetic radiation, it is possible to better attune the enzyme activities of the enzymes to each other when using more than one enzyme.

De uitvinding zal thans worden toegelicht aan de hand van het volgende, niet-beperkende voorbeeld.The invention will now be illustrated by the following non-limiting example.

10 Voorbeeld10 Example

Het effect van bestralen op de door D-lactaatdehy-drogenase (D-LDH) gekatalyseerde reactie van D-lactaat met het co-enzym NAD+ tot pyruvaat, NADH en H* werd onderzocht onder gebruikmaking van een commercieel verkrijgbaar testpak-15 ket (Boehringer Mannheim, Duitsland). Het tijdens de door D-LDH gekatalyseerde evenwichtsreactie gevormde pyruvaat wordt, om D-lactaat volledig te laten reageren, met behulp van een tweede enzym (glutamaat-pyruvaat-transaminase (GPT)) en glutamaat omgezet.The effect of irradiation on the D-lactate dehydrogenase (D-LDH) catalyzed reaction of D-lactate with the coenzyme NAD + to pyruvate, NADH and H * was investigated using a commercially available test package (Boehringer Mannheim, Germany). The pyruvate formed during the equilibrium reaction catalyzed by D-LDH is reacted with a second enzyme (glutamate-pyruvate transaminase (GPT)) and glutamate to completely react D-lactate.

20 De helft van de buisjes met D-LDH- en GTP-enzymop- lossingen uit het testpakket werd onder koelen met lucht gedurende 24 uur bij ca. 90 MHz bestraald bij een vermogen van ca. 3 Watt. (De gebruikte inrichting is beschreven door Everaerts F.M. et al in J. Chromatog. Libr., deel 6,: Isota-25 chophoresis, theory, instrumentation and applications, Elsevier Scientific Publishing Company, Amsterdam (1976). De enige modificatie was dat de elektronische schakeling was voorzien van een deklaag van goud om aantasting ervan door gevormd ozon te voorkomen). Voor het oplossen van de sub-30 straten werd 50 ml gedeïoniseerd water waaraan 2,2 mg/ml NaCl was toegevoegd gedurende 7 dagen zoals hierboven beschreven behandeld. In tabel I is met een asterisk aangegeven welke oplossingen zijn bestraald.Half of the tubes with D-LDH and GTP enzyme solutions from the test package were irradiated under cooling with air for 24 hours at about 90 MHz at a power of about 3 watts. (The device used is described by Everaerts FM et al in J. Chromatog. Libr., Vol. 6, Isota-25 chophoresis, theory, instrumentation and applications, Elsevier Scientific Publishing Company, Amsterdam (1976). The only modification was that electronic circuit was coated with gold to prevent deterioration by ozone formed). To dissolve the substrates, 50 ml of deionized water to which 2.2 mg / ml NaCl was added over 7 days was treated as described above. In table I an asterisk indicates which solutions have been irradiated.

1005028 31005028 3

TABEL ITABLE I

Voorbeeld ControleExample Control

Bufferopl. pH 10 5 14,7 mg/ml L-glutaminezuur (testpakket) 1 ml 1 ml NAD-lyofilisaat1 35 mg/ml 0,2 ml* 0,2 ml GPT ca. 1100 IU 0,02 ml* 0,02 mlBufferopl. pH 10 5 14.7 mg / ml L-glutamic acid (test package) 1 ml 1 ml NAD lyophilisate1 35 mg / ml 0.2 ml * 0.2 ml GPT approx. 1100 IU 0.02 ml * 0.02 ml

Li-lactaat1 0,186 mg/ml 0,1 ml* 0,1 ml 10 Gedeïoniseerd water met 2,2 mg/ml NaCl 0,9 ml* 0,9 ml D-LDH1 ca. 3800 IU 0,02 ml* 0,02 ml 1 gedeïoniseerd water met 2,2 mg/ml NaCl 15 De omzetting van NAD+ in NADH werd spectrofotometrisch bij 340 nm gevolgd. Hieruit bleek dat de omzetting van lactaat in bestraald water versneld verloopt.Li-lactate1 0.186 mg / ml 0.1 ml * 0.1 ml 10 Deionized water with 2.2 mg / ml NaCl 0.9 ml * 0.9 ml D-LDH1 approx. 3800 IU 0.02 ml * 0, 02 ml 1 deionized water with 2.2 mg / ml NaCl 15 The conversion of NAD + to NADH was monitored spectrophotometrically at 340 nm. This showed that the conversion of lactate to irradiated water is accelerated.

10050281005028

Claims (2)

1. Werkwijze voor het controleren van een enzymre-actie in een polair oplosmiddel, met het kenmerk, dat de enzymreactie wordt uitgevoerd in een met elektromagnetische straling behandeld polair oplosmiddel, waarbij de elektromag- 5 netische straling een frequentie heeft in het bereik van 5*107 tot 1010 Hz.Method for controlling an enzyme reaction in a polar solvent, characterized in that the enzyme reaction is carried out in a polar solvent treated with electromagnetic radiation, the electromagnetic radiation having a frequency in the range of 5 * 107 to 1010 Hz. 2. Werkwijze volgens conclusie 1, met het kenmerk, dat de enzymreactie wordt uitgevoerd in een waterig oplosmiddel als het polaire oplosmiddel. 1005028A method according to claim 1, characterized in that the enzyme reaction is carried out in an aqueous solvent as the polar solvent. 1005028
NL1005028A 1997-01-17 1997-01-17 Controlling enzyme reactions in polar solvent NL1005028C2 (en)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
NL6904685A (en) * 1968-03-27 1969-09-30
FR2039250A1 (en) * 1969-04-17 1971-01-15 Gray Ind Inc
DE4142244A1 (en) * 1991-12-20 1993-06-24 Tramm Werner Sabine Dipl Biol Acceleration of enzymatic processes - by exposing the enzyme substrate combination to alternating electric field of fixed or variable frequency
WO1993021344A1 (en) * 1992-04-16 1993-10-28 Pioneer Hi-Bred International, Inc. Microwave modification of biological macromolecules
NL9300976A (en) * 1993-06-07 1995-01-02 Inst Voor Agrotech Onderzoek Method for altering the kinetics of a reaction involving at least one macromolecule, the macromolecule being treated with electromagnetic energy, use of said method for optimizing processes involving the macromolecule, and a macromolecule whose reactivity has been altered

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
NL6904685A (en) * 1968-03-27 1969-09-30
FR2039250A1 (en) * 1969-04-17 1971-01-15 Gray Ind Inc
DE4142244A1 (en) * 1991-12-20 1993-06-24 Tramm Werner Sabine Dipl Biol Acceleration of enzymatic processes - by exposing the enzyme substrate combination to alternating electric field of fixed or variable frequency
WO1993021344A1 (en) * 1992-04-16 1993-10-28 Pioneer Hi-Bred International, Inc. Microwave modification of biological macromolecules
NL9300976A (en) * 1993-06-07 1995-01-02 Inst Voor Agrotech Onderzoek Method for altering the kinetics of a reaction involving at least one macromolecule, the macromolecule being treated with electromagnetic energy, use of said method for optimizing processes involving the macromolecule, and a macromolecule whose reactivity has been altered

Non-Patent Citations (16)

* Cited by examiner, † Cited by third party
Title
BELKHODE, M. L. ET AL: "Thermal and athermal effects of 2.8 GHz microwaves on three human serum enzymes", J. MICROWAVE POWER (1974), 9(1), 23-9 CODEN: JLMPAB, XP002043635 *
CHEMICAL ABSTRACTS, vol. 103, no. 17, 28 October 1985, Columbus, Ohio, US; abstract no. 137719, POPESCU, T. ET AL: "Interaction of a high frequency electromagnetic field with lactate dehydrogenase activity" XP002043639 *
CHEMICAL ABSTRACTS, vol. 107, no. 9, 31 August 1987, Columbus, Ohio, US; abstract no. 76370, BABA, TORU ET AL: "Development of snack foods produced from sweet potatoes. IV. Effects of microwave irradiation on amylolytic activities in dehydrated sweet potato flour" XP002043638 *
CHEMICAL ABSTRACTS, vol. 108, no. 25, 20 June 1988, Columbus, Ohio, US; abstract no. 218275, GILDERSLEEVE, R. P. ET AL: "Serum enzymes in hemorrhaged Japanese quail after microwave irradiation during embryogeny" XP002043637 *
CHEMICAL ABSTRACTS, vol. 81, no. 9, 2 September 1974, Columbus, Ohio, US; abstract no. 46101, KIIREND, ENE ET AL: "Effect of radio frequency and microwave electromagnetic fields on the enzyme -substrate reaction in aqueous solutions and organic solvents" XP002043643 *
CHEMICAL ABSTRACTS, vol. 89, no. 23, 4 December 1978, Columbus, Ohio, US; abstract no. 193160, BINI, M. ET AL: "Analysis of the effects of microwave energy on enzymic activity of lactate dehydrogenase (LDH)" XP002043642 *
CHEMICAL ABSTRACTS, vol. 96, no. 25, 21 June 1982, Columbus, Ohio, US; abstract no. 213320, HALL, C. A. ET AL: "Interaction of microwave radiation with turkey sperm" XP002043641 *
CHEMICAL ABSTRACTS, vol. 99, no. 21, 21 November 1983, Columbus, Ohio, US; abstract no. 172069, ZHU, LINGMEI ET AL: "Effect of microwave irradiation on spermatozoa lactate dehydrogenase -X in adult human testes" XP002043640 *
COMP. BIOCHEM. PHYSIOL., A: COMP. PHYSIOL. (1988), 89A(4), 531-4 CODEN: CBPAB5;ISSN: 0300-9629 *
EESTI NSV TEAD. AKAD. TOIM., BIOL. (1974), 23(2), 117-82 CODEN: ETATAW *
J. MICROWAVE POWER (1978), 13(1), 95-9 CODEN: JLMPAB;ISSN: 0022-2739 *
NIPPON SHOKUHIN KOGYO GAKKAISHI (1987), 34(3), 178-84 CODEN: NSKGAX;ISSN: 0369-5727 *
RADIAT. ENVIRON. BIOPHYS. (1982), 20(2), 145-52 CODEN: REBPAT;ISSN: 0301-634X *
S.KERMASHA ET AL.: "Comparison of microwave,conventional and combination heat treatments on wheat germ lipase activity", INTERNATIONAL JOURNAL OF FOOD SCIENCE AND TECHNOLOGY, vol. 28, no. 6, 1993, pages 617 - 623, XP002043636 *
SICHUAN YIXUEYUAN XUEBAO (1983), 14(3), 252-5 CODEN: CIYPDA;ISSN: 0253-4290 *
STUD. CERCET. BIOCHIM. (1985), 28(1), 60-7 CODEN: SCBIA5;ISSN: 0049-2396 *

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