# US 5,350,686 A · Microwave acceleration of enzyme-catalyzed modification of macromolecules

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- Number: US 5,350,686 A
- Document ID: US5350686A
- Status: Granted
- Kind: A
- Application number: 07/869,604
- Filed: 1992-04-16
- Granted: 1994-09-27
- Earliest claimed: 1992-04-16
- Assignees: Pioneer Hi-Bred International, Inc.
- Inventors: Anil K. Jhingan
- Web page: https://patentgrep.com/patent/US5350686A

## CPC

- C12N 9/22: Ribonucleases {[RNase]; Deoxyribonucleases [DNase]}
- C12N 13/00: Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
- C12Q 1/00: Measuring or testing processes involving enzymes, nucleic acids or microorganisms (measuring or testing apparatus with condition measuring or sensing means, e.g. colony counters, C12M1/34); Compositions therefor; Processes of preparing such compositions
- C12Q 1/68: involving nucleic acids
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

A method of enhancing the enzyme-catalyzed modification of macromolecules comprising exposure of the enzyme and its macromolecule substrate to microwaves is disclosed. A frequency of from 1800 to 3000 MHz and a power of from 50 to 100 watts is preferred. The procedure is rapid, efficient, and suitable for many enzyme-catalyzed reactions. The method can be used for restriction endonuclease digestion of DNA samples and facilitates one of the time-consuming procedures of nucleic acid research. The method is also adaptable for automated procedures in biological research.

## Claims

### Claim 1

A method for enzymatic modification of a biological macromolecule comprising the step of exposing a composition comprising the biological macromolecule and an enzyme for which the macromolecule is a substrate to microwaves at a frequency of from 1800 to 3000 MHz and a power of from 50 watts to 1000 watts to accelerate the activity of the enzyme.

### Claim 2 (depends on 1)

A method according to claim 1 wherein the biological macromolecule is DNA or RNA.

### Claim 3 (depends on 2)

A method according to claim 2 wherein the biological macromolecule is DNA and the enzyme is a DNA restriction endonuclease or a mixture thereof.

### Claim 4 (depends on 1)

A method according to claim 1 wherein the total irradiation time is from 1 to 35 minutes.

### Claim 5 (depends on 4)

A method according to claim 4 wherein the composition is irradiated for intervals of from 1 to 60 seconds.

### Claim 6 (depends on 4)

A method according to claim 4 wherein the composition is intermittently irradiated by exposure to microwave radiation for intervals of from 1 to 60 seconds, the intervals of exposure to microwave radiation being separated by intervals of non-exposure to microwave radiation of from 1 to 60 seconds.

### Claim 7 (depends on 6)

A method according to claim 6 wherein the irradiation intervals are about 45 seconds and the non-irradiation intervals are from 1 to 2 seconds.

### Claim 8 (depends on 5)

A method according to claim 5 wherein the microwave radiation is supplied by a magnetron having a duty cycle during the irradiation intervals of from 10% to 80%.

### Claim 9 (depends on 8)

A method according to claim 8 wherein the magnetron has an output power of from 650 to 900 watts.

### Claim 10 (depends on 3)

A method according to claim 3, wherein the total irradiation time is from 1 to 35 minutes.

### Claim 11 (depends on 10)

A method according to claim 10, wherein the said composition is irradiated for intervals of from 1 to 60 seconds.

### Claim 12 (depends on 10)

A method according to claim 10, wherein the composition is intermittently irradiated by exposure to microwave radiation for intervals of from 1 to 60 seconds, the intervals of exposure to microwave radiation being separated by intervals of non-exposure to microwave radiation of from 1 to 60 seconds.

### Claim 13 (depends on 12)

A method according to claim 12 wherein the irradiation intervals are about 45 seconds and the non-irradiation intervals are from 1 to 2 seconds.

### Claim 14 (depends on 11)

A method according to claim 11 wherein the microwave radiation is supplied by a magnetron having a duty cycle during the irradiation intervals of from 10% to 80%.

### Claim 15 (depends on 14)

A method according to claim 14 wherein the magnetron has an output power of from 650 to 900 watts.