Joseph J Defrank

US Patent:

6469145, Oct 22, 2002

Filed:

Jun 26, 2000

Appl. No.:

09/603450

Inventors:

Vipin K. Rastogi - Bel Air MD
Tu-Chen Cheng - Timonium MD
Joseph J. DeFrank - Bel Air MD

Assignee:

The United States of America as represented by the Secretary of the Army - Washington DC

International Classification:

C08H 100

US Classification:

530412, 530350, 530369, 435 701, 435 711, 435 697, 435174, 4352523, 4353201, 424 941, 536 234

Abstract:

Novel and improved processes for isolating organophosphorus hydrolase enzyme from an aqueous solution and obtaining substantially purified enzyme at high yield are provided, as well as compositions, including storage stable lyophilyzed organophosphorus hydrolase enzyme compositions, that are prepared by the provided methods. The organophosphorus hydrolase enzyme is purified by contacting an aqueous solution of cell free bacterial proteins with a strong cation exchange resin, the aqueous solution comprising soluble organophosphorus hydrolase enzyme, washing the strong cation exchange resin with a washing buffer to remove unbound proteins from the strong cation exchange resin, eluting proteins that remain bound to the strong cation exchange resin by washing the resin with an eluting buffer comprising salt in a concentration that starts at about zero and is raised during the eluting process to about 0. 5M, and detecting and collecting eluate comprising a protein having organophosphorus hydrolase enzyme activity. All processing steps are conducted at a pH that is less than the isoelectric point of the organophosphorus hydrolase enzyme.

US Patent:

6599733, Jul 29, 2003

Filed:

Apr 27, 2001

Appl. No.:

09/844829

Inventors:

Ilona J. Fry - Edgewood MD
Joseph J. DeFrank - Bel Air MD
James P. Earley - South Bend IN

Assignee:

The United States of America as represented by the Secretary of the Army - Washington DC

International Classification:

C12N 120

US Classification:

4352524, 435 42, 435244, 4352625, 424 933

Abstract:

A biodegradation process for the organophosphonate product of Sarin (O-isopropyl methylphosphonofluoridate) hydrolysis, i. e. , isopropylmethylphosphonate (IMPA). This process provides a feasible biodegradation demilitarization alternative to Sarin incineration. Public opposition of nerve agent incineration is widespread, and alternative methods are sought to help the U. S. Army meet the 2007 demilitarization deadline imposed by the Chemical Weapons Convention. This process uses a two-step approach to IMPA biodegradation. In the first step, a concentrated IMPA solution is used as the sole nutritional carbon and phosphorus source for microbial cultures. The second step involves diluting the culture and adding an inexpensive carbon source to encourage bacterial phosphate assimilation. The biodegradation typically involves a consortium of microorganisms comprising GB21, GB2GA, GB2CS, GB272, Aureobacterium sp. GB2 and three bacterial isolates belonging to the same species GB23, GB272, and GB292.

US Patent:

7001758, Feb 21, 2006

Filed:

May 12, 2003

Appl. No.:

10/436342

Inventors:

Ilona J. Fry - Edgewood MD, US
Joseph J. DeFrank - Bel Air MD, US
James P. Earley - South Bend IN, US

Assignee:

The United States of America as represented by the Secretary of the Army - Washington DC

International Classification:

C12N 1/20
B09B 3/00

US Classification:

4352625, 435 42, 4352524, 435244, 424 933

Abstract:

A biodegradation process for the organophosphonate product of Sarin (O-isopropyl methylphosphonofluoridate) hydrolysis, i. e. , isopropylmethylphosphonate (IMPA). This process provides a feasible biodegradation demilitarization alternative to Sarin incineration. Public opposition of nerve agent incineration is widespread, and alternative methods are sought to help the U. S. Army meet the 2007 demilitarization deadline imposed by the Chemical Weapons Convention. This process uses a two-step approach to IMPA biodegradation. In the first step, a concentrated IMPA solution is used as the sole nutritional carbon and phosphorus source for microbial cultures. The second step involves diluting the culture and adding an inexpensive carbon source to encourage bacterial phosphate assimilation. The biodegradation typically involves a consortium of microorganisms comprising GB21, GB2GA, GB2CS, GB272, sp. GB2 and three bacterial isolates belonging to the same species GB23, GB272, and GB292.

US Patent:

7229819, Jun 12, 2007

Filed:

Oct 27, 2003

Appl. No.:

10/694206

Inventors:

Tu-Chen Cheng - Timonium MD, US
Joseph J. DeFrank - Bel Air MD, US
Steven P. Harvey - Fallston MD, US
Vipin K. Rastogi - Bel Air MD, US

Assignee:

The United States of America as represented by the Secretary of the Army - Washington DC

International Classification:

A62D 3/00
C02F 3/34
D06M 16/00

US Classification:

435264, 435262, 4352625

Abstract:

The present invention relates to a near-universal non-corrosive, non-toxic, environmentally safe and user friendly decontaminant capable of detoxifying organophosphorus (OP)-based G-type, V-type neurotoxic chemical warfare, sulfur-mustard, and related OP based hazardous industrial materials in a dry powder form. The decontaminant contains OPH enzyme, OPAA enzyme, DFPase enzyme, dehalogenase enzyme, quaternary ammonium salt, a pH control reagent, a fire-fighting agent, and a foaming agent. The decontaminant is mixed with available water for use.

US Patent:

7704724, Apr 27, 2010

Filed:

Jan 26, 2006

Appl. No.:

11/345679

Inventors:

Ilona J. Fry - Edgewood MD, US
Joseph J. DeFrank - Bel Air MD, US

Assignee:

The United States of America as represented by the Secretary of the Army - Washington DC

International Classification:

C12N 15/55
C12N 1/21
C12N 9/80

US Classification:

4352523, 4353201, 435228, 536 232

Abstract:

Compositions and methods for catalytic buffering of enzymatic decontamination reactions are provided. Enzymatic decontamination of organophosphorus or organohalogen compounds generates acidic reaction products that precipitously reduce the pH of the medium, thus impairing activity of the decontaminating enzymes. Catalytic buffering, that is, the use of an enzyme to produce ions from a substrate to modulate pH, can provide effective pH control. The compositions provided here include urease enzymes with mutations in the alpha subunit of the urease holoenzyme. These mutant ureases maintain urease activity in the presence of fluoride ions, which are organophosphorus and organohalogen hydrolysis products that otherwise inhibit urease activity. The fluoride-resistant ureases act as effective catalytic buffers during organofluorophosphorus hydrolysis reactions. Methods for using the fluoride-resistant ureases in enzymatic decontamination are also provided.

US Patent:

7723558, May 25, 2010

Filed:

Jun 5, 2007

Appl. No.:

11/758431

Inventors:

Tu-Chen Cheng - Timonium MD, US
Joseph J. DeFrank - Bel Air MD, US
Steven P. Harvey - Fallston MD, US
Vipin K. Rastogi - Bel Air MD, US

Assignee:

The United States of America as represented by the Secretary of the Army - Washington DC

International Classification:

A62D 3/34

US Classification:

588408, 588316, 588405

Abstract:

The present invention relates to a near-universal non-corrosive, non-toxic, environmentally safe and user friendly decontaminant capable of detoxifying organophosphorus (OP)-based G-type, V-type neurotoxic chemical warfare, sulfur-mustard, and related OP based hazardous industrial materials in a dry powder form. The decontaminant contains OPH enzyme, OPAA enzyme, DFPase enzyme, dehalogenase enzyme, quaternary ammonium salt, a pH control reagent, a fire-fighting agent, and a foaming agent. The decontaminant is mixed with available water for use.

US Patent:

8357335, Jan 22, 2013

Filed:

Nov 9, 2005

Appl. No.:

11/272422

Inventors:

Steven P. Harvey - Fallston MD, US
Joseph J. DeFrank - Bel Air MD, US

Assignee:

The United States of America as Represented by the Secretary of the Army - Washington DC

International Classification:

G01N 33/00

US Classification:

422401, 422 50, 422561, 436104, 436165, 4352625, 588405

Abstract:

An assay for screening potential hydrolysis enhancing agents capable of facilitating the hydrolysis of a substantially water insoluble halogenated compound such as mustard gas (HD) in an aqueous reaction mixture is disclosed. The assay includes at least one chamber adapted for receiving and retaining the substantially water insoluble mustard gas compound and a potential hydrolysis agent in an aqueous reaction mixture, and a pH indicating agent adapted to produce a visible color change corresponding to the amount of the substantially water insoluble mustard gas compound hydrolyzed in the aqueous reaction mixture wherein the rate of hydrolysis can be established by measuring the change in the detectable signal over time.

US Patent:

60543108, Apr 25, 2000

Filed:

Nov 18, 1998

Appl. No.:

9/195113

Inventors:

Michael H. Kim - Ellicott City MD
Joseph J. DeFrank - Bel Air MD

Assignee:

The United States of America as represented by the Secretary of the Army - Washington DC

International Classification:

C07C 102

US Classification:

4352524

Abstract:

A process for biodegradation of an amine compound by contacting the amine mpound with a consortium of microorganisms effective for consuming carbon and nitrogen components of the amine compound under aerobic conditions, wherein the enzymatically degraded the amine compound forms an ammonia residue, nitrifying the ammonia residue under aerobic conditions, wherein the ammonia residue forms nitrite and nitrate residues and denitrifying the compound with the addition of a supplementary carbon source under anoxic conditions. The amine compound may be DS2 or similar amine structures. The microorganisms include Bacillus circulans, the genera Nitrosomonas, and the genera Nitrobacter. The process is a continuous-fed process in a bioreactor. A composition of microorganisms comprising Bacillus circulans, the genera Nitrosomonas, the genera Nitrobacter, and facultative heterotrophic denitrifiers effective to degrade DS2 also is disclosed.