US3679139A - Process for the disrupting of biological material - Google Patents
Process for the disrupting of biological material Download PDFInfo
- Publication number
- US3679139A US3679139A US37536A US3679139DA US3679139A US 3679139 A US3679139 A US 3679139A US 37536 A US37536 A US 37536A US 3679139D A US3679139D A US 3679139DA US 3679139 A US3679139 A US 3679139A
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- US
- United States
- Prior art keywords
- biological material
- pressure
- pressure vessel
- disrupting
- disruption
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B02—CRUSHING, PULVERISING, OR DISINTEGRATING; PREPARATORY TREATMENT OF GRAIN FOR MILLING
- B02C—CRUSHING, PULVERISING, OR DISINTEGRATING IN GENERAL; MILLING GRAIN
- B02C19/00—Other disintegrating devices or methods
- B02C19/0056—Other disintegrating devices or methods specially adapted for specific materials not otherwise provided for
Definitions
- the present invention relates to a process for the disruption of biological material and to a device for this process.
- the device according to the invention comprises vin combination a pressure vessel provided with inlet and outlet means, pressure regulating means connecting the pressure vessel with a container of a compressed gaseous medium and means for determining the pressure established in the pressure vessel.
- the invention relates to a process of disrupting biological material and to a device for effecting-such process.
- the device according to the invention is quite inexpensive and simple.
- the disruption of the biological material can be effected under accurately controlled and reproducible conditions, and it can be effected under any desired gaseous atmosphere.
- the process according to the invention comprises introducing the biological material into a suitable pressure vessel provided with inlet and outlet means, establishing above the material a desired. gaseous atmosphere and applying a controlled predetermined pressure by connecting the pressure vessel via pressure regulating means with a high-pressure gas container containing the desired gas, and permitting the biological material to flow out of the pressure vessel through a release valve. If necessary, the process may be repeated.
- the device according to the present invention shown schematically in the enclosed drawing, comprises a pressure vessel (1), provided with a closure (2), an outlet valve (3), an outlet (4), an inlet conduit for the gaseous medium (5), an inlet valve (6), a pressure regulating and pressure indicating valve (7) which is connected via a conduit (8) with a high-pressure gas container (9).
- the pressure vessel (1) is charged with the biological material, the atmosphere above the sample is flushed with the gaseous medium (if this is to be other than air), so as to establish a desired gaseous atmosphere above the biological material, the cap (2) is closed tightly while the valve (6) is in the closed position, the pressure bottle (9) is opened, the pressure is adjusted by means of the valve (7), the valve 6) is opened so as to establish in the vessel (1) the desired pressure, and subsequently the valve (3) is opened so as to gradually eject the biological material through the outlet (4).
- the gaseous medium if this is to be other than air
- the pressure regulating valve 7 makes it possible to establish a desired gaseous atmosphere of predetermined pres- I exemplified for a number of biological systems, without being restricted-thereto.
- EXAMPLE 1 A sample of 20 ml of 50 percent w/v Escherichia Coli in phosphate bufier was introduced into a pressure cell and an air pressure of 1,500 p.s.i. was applied; The release was effected'at a, rate of 3 to 4 drops per second. After one passage, a very. viscous suspension was obtaineddue to the liberation of DNA from the disrupted cells. After a second passage through the pressure cell, in a similar manner, a breakage of 50 percent of the cells was obtained.
- a protein content of 48mg/ml and a nucleic acids content of 14 mg/ml was obtained. This was determined by optical measurements of a diluted sample.
- EXAMPLE 2 A breakage of 50 percent of the bacterium Rhodospirillum rubrum. was obtained by passage through the device according to the invention under a pressure of 1,500 p.s.i. of nitrogen. This was effected in a manner similar to that of Example 1.
- the biological activities after disruption by the device according to the invention were superior as compared with the disruption by sonication or by passage through the French press, as determined by trans-hydrogenase activity.
- EXAMPLE 3 The pressure cell according to the invention was charged EXAMPLE 4 Human red blood cells l-2-percent in saline) were almost 100 percent broken by passage through the pressure cell according to the invention under a pressure of 500 p.s.i. of air.
- a process of disrupting biological material which comprises introducing the biological material into a suitable pres sure vessel provided with inlet and outlet means, establishing above the biological material a desired gaseous atmosphere, applying a predetermined pressure on the sample by connecting the pressure vessel with a compressed source of said gas, and gradually ejecting the biological material from the pressure vessel through a release valve. 5
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- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The present invention relates to a process for the disruption of biological material and to a device for this process. The device according to the invention comprises in combination a pressure vessel provided with inlet and outlet means, pressure regulating means connecting the pressure vessel with a container of a compressed gaseous medium and means for determining the pressure established in the pressure vessel.
Description
United States Patent Schneyour et a1.
[5 PROCESS FOR THE DISRUPIING OF BIOLOGICAL MATERIAL [72] lnventors: Amir Schneyour, 12 a Shalem St., Ramat- Gan; Mordhay Avron, 9 Neve Weizmann, Rehovot, both of Israel [22] Filed: May 15, 1970 [21] Appl.No.: 37,536
[30] Foreign Application Priority Data May 21, 1969 lsrael ..32,268
[52] U.S. Cl. ...241/2 [51] Int. CL... ...-.B02e 19/12 [58] Field olSearch... ..241/1,2, 18,30
[56] References Cited UNITED STATES PATENTS 1,578,609 3/1926 Mason ..241/1 X [451 July 25, 1972 7/1969 Edebe..., ...241/1 1,002,990 9/191 1 Herendeen ..241/2 2,928,614 3/1960 Emanuel et a1. ..241/1 X 3,309,032 3/1967 Fitz et al. ....241/1 X 2,318,693 5/1943 Joyce et al.... ,.241/l 3,165,266 1/1965 -B1um et 241/1 3,556,414 1/1971 Eberly,.lr ..241/2 Primary Examiner-Granville Y. Custer, Jr. Attorney-Ostrolenk, Faber, Gerb & Solfen [5 7] ABSTRACT The present invention relates to a process for the disruption of biological material and to a device for this process. The device according to the invention comprises vin combination a pressure vessel provided with inlet and outlet means, pressure regulating means connecting the pressure vessel with a container of a compressed gaseous medium and means for determining the pressure established in the pressure vessel.
4 Clairm, 1 Drawing Figure BACKGROUND OF THE INVENTION 1. Field of the Invention The invention relates to a process of disrupting biological material and to a device for effecting-such process.
2. Description of the Prior Art It is frequently necessary to disrupt biological materiahsuch as bacteria, algae, blood cells, chloroplasts, mitochondria and the like under controlled. and reproducible conditions. In a disruption process, the cell walls are ruptured by first pressurizing the'cells so'that an equilibrium across the cell walls is attained, andthereafter, suddenly lowering the external pres-- sure, resulting in the cells bursting open due tothe sudden decompression and expansion of the gas. In the past, rather inconvenient devices were used for this purpose. Amongst these, there may be mentioned the French. press and the Hughes press, used in many laboratories. Both devices are rather inconvenient and it is rather laborious to effect the disruption of the biological material. With the known devices it is hard to maintain a substantially constant pressure, and it is difficult to carry out the process in a reproducible manner so SUMMARY OF THE PRESENT INVENTION The present invention provides a simple and convenient device for accomplishing the foregoing.
The device according to the invention is quite inexpensive and simple. The disruption of the biological material can be effected under accurately controlled and reproducible conditions, and it can be effected under any desired gaseous atmosphere.
The process according to the invention comprises introducing the biological material into a suitable pressure vessel provided with inlet and outlet means, establishing above the material a desired. gaseous atmosphere and applying a controlled predetermined pressure by connecting the pressure vessel via pressure regulating means with a high-pressure gas container containing the desired gas, and permitting the biological material to flow out of the pressure vessel through a release valve. If necessary, the process may be repeated.
BRIEF DESCRIPTION OF THE DRAWING The invention is described by way of example only with reference to the enclosed schematical drawing, which is a schematic side view of a device according to the invention.
DESCRIPTION OF THE PREFERRED EMBODIMENT The device according to the present invention, shown schematically in the enclosed drawing, comprises a pressure vessel (1), provided with a closure (2), an outlet valve (3), an outlet (4), an inlet conduit for the gaseous medium (5), an inlet valve (6), a pressure regulating and pressure indicating valve (7) which is connected via a conduit (8) with a high-pressure gas container (9).
For carrying out the process of disrupting biological material according to the present invention, the pressure vessel (1) is charged with the biological material, the atmosphere above the sample is flushed with the gaseous medium (if this is to be other than air), so as to establish a desired gaseous atmosphere above the biological material, the cap (2) is closed tightly while the valve (6) is in the closed position, the pressure bottle (9) is opened, the pressure is adjusted by means of the valve (7), the valve 6) is opened so as to establish in the vessel (1) the desired pressure, and subsequently the valve (3) is opened so as to gradually eject the biological material through the outlet (4).
The pressure regulating valve 7 makes it possible to establish a desired gaseous atmosphere of predetermined pres- I exemplified for a number of biological systems, without being restricted-thereto.
EXAMPLE 1 A sample of 20 ml of 50 percent w/v Escherichia Coli in phosphate bufier was introduced into a pressure cell and an air pressure of 1,500 p.s.i. was applied; The release was effected'at a, rate of 3 to 4 drops per second. After one passage, a very. viscous suspension was obtaineddue to the liberation of DNA from the disrupted cells. After a second passage through the pressure cell, in a similar manner, a breakage of 50 percent of the cells was obtained.
In the resulting sample, a protein content of 48mg/ml and a nucleic acids content of 14 mg/ml was obtained. This was determined by optical measurements of a diluted sample.
With a French press, a disruption of percent of the cells can be obtained at a pressure of 15,000 p.s.i., but the biological activity of the material is diminished by such high pressures.
EXAMPLE 2 A breakage of 50 percent of the bacterium Rhodospirillum rubrum. was obtained by passage through the device according to the invention under a pressure of 1,500 p.s.i. of nitrogen. This was effected in a manner similar to that of Example 1.
By sonication, a breakage of 100 percent was obtained; both of these being determined by the release of bacteric chlorophyll.
The biological activities after disruption by the device according to the invention were superior as compared with the disruption by sonication or by passage through the French press, as determined by trans-hydrogenase activity.
Also in the case of Euglena (Example 3), the activity, based on determination of phosphorylation activity, was superior in the case of disruption by the process according to the invention as compared with sonication.
EXAMPLE 3 The pressure cell according to the invention was charged EXAMPLE 4 Human red blood cells l-2-percent in saline) were almost 100 percent broken by passage through the pressure cell according to the invention under a pressure of 500 p.s.i. of air.
Comparative runs were made with other compressed gases and'also under increased pressures. In the latter case, the disruption of subcellular structures also occurred.
We claim:
1. A process of disrupting biological material, which comprises introducing the biological material into a suitable pres sure vessel provided with inlet and outlet means, establishing above the biological material a desired gaseous atmosphere, applying a predetermined pressure on the sample by connecting the pressure vessel with a compressed source of said gas, and gradually ejecting the biological material from the pressure vessel through a release valve. 5
2. A process as claimed in claim 1, wherein the applied gas is air, oxygen, nitrogen, argon or helium.
3. A process as claimed in claim 1, wherein the passage through the pressure cell is repeated.
4. A process as claimed in claim 1, wherein the biological material is provided in the form of a suspension in a suitable suspension medium so as to obtain a liquid suspension of desired viscosity.
* *IK l
Claims (4)
1. A process of disrupting biological material, which comprises introducing the biological material into a suitable pressure vessel provided with inlet and outlet means, establishing above the biological material a desired gaseous atmosphere, applying A predetermined pressure on the sample by connecting the pressure vessel with a compressed source of said gas, and gradually ejecting the biological material from the pressure vessel through a release valve.
2. A process as claimed in claim 1, wherein the applied gas is air, oxygen, nitrogen, argon or helium.
3. A process as claimed in claim 1, wherein the passage through the pressure cell is repeated.
4. A process as claimed in claim 1, wherein the biological material is provided in the form of a suspension in a suitable suspension medium so as to obtain a liquid suspension of desired viscosity.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| IL32268A IL32268A (en) | 1969-05-21 | 1969-05-21 | Process and device for disrupting biological material |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| US3679139A true US3679139A (en) | 1972-07-25 |
Family
ID=11044949
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| US37536A Expired - Lifetime US3679139A (en) | 1969-05-21 | 1970-05-15 | Process for the disrupting of biological material |
Country Status (2)
| Country | Link |
|---|---|
| US (1) | US3679139A (en) |
| IL (1) | IL32268A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3848812A (en) * | 1973-02-26 | 1974-11-19 | Scp Exploatering Ab | Process for extracting protein from microorganisms |
| US4746071A (en) * | 1985-03-19 | 1988-05-24 | Kohlensaeurewerk Deutschland Gmbh | Process for cracking blossom pollen |
| US5620730A (en) * | 1993-09-09 | 1997-04-15 | Van Noort; Gerard | Method of enhancing shelf-stability of an edible biological product |
| US6405948B1 (en) * | 1997-07-18 | 2002-06-18 | Pulsewave Llc | Liberating intracellular matter from biological material |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US1002990A (en) * | 1910-06-20 | 1911-09-12 | Charles Herendeen | Process of producing flour or meal. |
| US1578609A (en) * | 1924-09-24 | 1926-03-30 | William H Mason | Process and apparatus for disintegration of wood and the like |
| US2318693A (en) * | 1939-11-29 | 1943-05-11 | Gen Motors Corp | Method of shredding rubber |
| US2928614A (en) * | 1958-04-16 | 1960-03-15 | Emanuel | Hydraulic tissue homogenizer |
| US3165266A (en) * | 1962-02-07 | 1965-01-12 | Sorvall Inc Ivan | Cell fractionator apparatus and method |
| US3309032A (en) * | 1964-03-23 | 1967-03-14 | Sorvall Inc Ivan | Cell fractionator apparatus |
| US3458139A (en) * | 1965-09-17 | 1969-07-29 | Biox Ab | Method and arrangement for disintegrating biological material under high pressure |
| US3556414A (en) * | 1968-02-28 | 1971-01-19 | United States Banknote Corp | Method and apparatus for disrupting cells |
-
1969
- 1969-05-21 IL IL32268A patent/IL32268A/en unknown
-
1970
- 1970-05-15 US US37536A patent/US3679139A/en not_active Expired - Lifetime
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US1002990A (en) * | 1910-06-20 | 1911-09-12 | Charles Herendeen | Process of producing flour or meal. |
| US1578609A (en) * | 1924-09-24 | 1926-03-30 | William H Mason | Process and apparatus for disintegration of wood and the like |
| US2318693A (en) * | 1939-11-29 | 1943-05-11 | Gen Motors Corp | Method of shredding rubber |
| US2928614A (en) * | 1958-04-16 | 1960-03-15 | Emanuel | Hydraulic tissue homogenizer |
| US3165266A (en) * | 1962-02-07 | 1965-01-12 | Sorvall Inc Ivan | Cell fractionator apparatus and method |
| US3309032A (en) * | 1964-03-23 | 1967-03-14 | Sorvall Inc Ivan | Cell fractionator apparatus |
| US3458139A (en) * | 1965-09-17 | 1969-07-29 | Biox Ab | Method and arrangement for disintegrating biological material under high pressure |
| US3556414A (en) * | 1968-02-28 | 1971-01-19 | United States Banknote Corp | Method and apparatus for disrupting cells |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3848812A (en) * | 1973-02-26 | 1974-11-19 | Scp Exploatering Ab | Process for extracting protein from microorganisms |
| US4746071A (en) * | 1985-03-19 | 1988-05-24 | Kohlensaeurewerk Deutschland Gmbh | Process for cracking blossom pollen |
| US5620730A (en) * | 1993-09-09 | 1997-04-15 | Van Noort; Gerard | Method of enhancing shelf-stability of an edible biological product |
| US6405948B1 (en) * | 1997-07-18 | 2002-06-18 | Pulsewave Llc | Liberating intracellular matter from biological material |
Also Published As
| Publication number | Publication date |
|---|---|
| IL32268A0 (en) | 1969-07-30 |
| IL32268A (en) | 1973-03-30 |
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