US Government As Represented by the Secretary of the Army

University of Massachusetts

The Administrators of the Tulane Educational Fund

Joseph A Akkara

David L Kaplan

Ferdinando F Bruno

Madhu S Ayyagari

Jonathan S Dordick

Sukant K Tripathy

Vijay Thomas John

Kenneth A Marx

Francisco J Aranda

Devulapalli V Rao

Jayant Kumar

Lynne Ann Samuelson

Zhongping Chen

Joby Joseph

Braja K Mandal

Arthur M Kaplan

Rajiv Pande

Madhu S R Ayyagari

Sanjay Kamtekar

Joseph F Roach

Gary L McPherson

Desai Narayana Rao

Masato Nakashima

Protease enzyme from and Bacillus sp. Catalyzes the acylation of organic solvent-insoluble macromolecules in isooctane solution containing vinyl esters of fatty acids, lactones or lactides as acyl donors. The reaction occurs only when the enzyme is solubilized via ion-pairing with the anionic surfactant dioctylsulfosuccinate, sodium salt (AOT). Enzyme based acylation was demonstrated in macromolecules such as silk proteins. These macromolecules are reactive either as cryogenically milled powder suspended in the organic solvent or as a thin film deposited onto ZnSe slides. This selective acylation approach represents the first attempt at using enzymes to modify organic-insoluble macromolecules in nonaqueous media.

Enzyme-catalyzed modifications of macromolecules in organic solvents

A process for large-scale, low cost, batch or continuous production of polyphenols using enzyme-mediated reactions and methods for recycling non-consumed reactants.

Large-scale production of polyphenols or polyaromatic amines using enzyme-mediated reactions

A process of controlling the molecular weight and dispersity of poly(p-ethylphenol) and poly(m-cresol) synthesized enzymatically by varying the composition of the reaction medium. Polymers with low dispersities and molecular weights from 1000 to 3000 are synthesized in reversed micelles and biphasic systems. In comparison, reactions in bulk solvents resulted in a narrow range of molecular weights (281 to 675 with poly(p-ethylphenol) in a DMF/water system and 1,400 to 25,000 with poly(m-cresol) in an ethanol/water system). Poly(p-ethylphenol) was functionalized at hydroxyl positions with palmitoyl, cinnamoyl, and biotin groups.

Process to control the molecular weight and polydispersity of substituted polyphenols and polyaromatic amines by enzymatic synthesis in organic solvents, microemulsions, and biphasic systems

protease catalyzes the acylation of organic solvent-insoluble polysaccharides in isooctane solution containing vinyl esters of fatty acids as acyl donor. The reaction occurs only when the enzyme is solubilized via ion-pairing with the anionic surfactant dioctyl sulfosuccinate, sodium salt (AOT). Enzyme based acylation was demonstrated with amylose, cyclodextrins, cellulose, cellulose derivatives, and other polysaccharides such as chitosan, pullulan, and maltodextrose. These polysaccharides are reactive either as a cryogenically milled powder suspended in the organic solvent or as a thin film deposited onto ZnSe slides. For chitosan, &agr;-cyclodextrin, and hydroxyethyl cellulose (HEC), the enzymatic crosslinking reaction occurs using adipic acid divinyl ester (C6DVE). HEC forms a compound that gels in solvents such as ethyl alcohol and dimethyl sulfone oxide (DMSO). Electron spectroscopy chemical analysis (ESCA) of the first 100 &angst; of the amylose thin film amylose indicates that the acylated surface had a degree of substitution of 0.9&plusmn;0.1 acyl chains per glucose moiety and this corresponded well to the expected regioselectivity of subtilisin catalysis on glucose-containing compounds.,H-NMR studies indicated that only the C-6 hydroxyl groups of the glucose moiety were acylated with amylose and &ggr;-cyclodextrin. However, &bgr;-cyclodextrin, and &agr;-cyclodextrin were modified at secondary alcohols and at all three alcohols, respectively. This approach represents the first attempt at using enzymes to modify organic solvent-insoluble polymers in nonaqueous media.

Transesterification of insoluble polysaccharides

Protease enzyme from,and Bacillus sp. Catalyzes the acylation of organic solvent-insoluble macromolecules in isooctane solution containing vinyl esters of fatty acids, lactones or lactides as acyl donors. The reaction occurs only when the enzyme is solubilized via ion-pairing with the anionic surfactant dioctylsulfosuccinate, sodium salt (AOT). Enzyme based acylation was demonstrated in macromolecules such as silk proteins. These macromolecules are reactive either as cryogenically milled powder suspended in the organic solvent or as a thin film deposited onto ZnSe slides. This selective acylation approach represents the first attempt at using enzymes to modify organic-insoluble macromolecules in nonaqueous media.

Polyaromatic compound made by polymerizing one or more aromatic monomers, least one of the one or more aromatic monomers having the following structure: ##STR1## wherein X is selected from the group consisting of OH and NH.sub.2, Y is selected from the group consisting of a halogen, a halogen-containing group, a boron-containing group, a phosphorus-containing group, a carboxyl group, a hydroxyacid group, a carboxyacid group, an amino group, an amino acid group, an amide group, an ester group, a hydroxyl group and a sulfonic acid group, R is an alkyl, phenyl, naphthyl, hydroxyphenyl, hydroxy naphthyl or hydroxy (.alpha., .beta., .gamma.) alkyl group, and n is an integer from 0 to 12. The (R).sub.n --Y group is preferably either ortho or para to X and more preferably is para to X. The polyaromatic compound may be a homopolymer consisting of one species of aromatic monomer of formula (I), a copolymer consisting of two or more species of aromatic monomers of formula (I), or a copolymer comprising one or more species of aromatic monomer of formula (I) and one or more species of an aromatic monomer not of formula (I). Preferably, the one or more aromatic monomers are reacted with a stoichiometric amount of hydrogen peroxide in the presence of horseradish peroxidase, the reaction taking place in water or in a mixture of water and one or more polar solvents. To obtain a substantial yield, the pH of the reaction medium is preferably around or below the pK.sub.a of the functional group of the monomer(s) being polymerized.

Polyaromatic compounds and method for their production

Process to control the molecular weight and polydispersity of

Bacillus subtilis protease catalyzes the acylation of organic solvent-insoluble polysaccharides in isooctane solution containing vinyl esters of fatty acids as acyl donor. The reaction occurs only when the enzyme is solubilized via ion-pairing with the anionic surfactant dioctyl sulfosuccinate, sodium salt (AOT). Enzyme based acylation was demonstrated with amylose, cyclodextrins, cellulose, cellulose derivatives, and other polysaccharides such as chitosan, pullulan, and maltodextrose. These polysaccharides are reactive either as a cryogenically milled powder suspended in the organic solvent or as a thin film deposited onto ZnSe slides. For chitosan, .alpha.-cyclodextrin, and hydroxyethyl cellulose (HEC), the enzymatic crosslinking reaction occurs using adipic acid divinyl ester (C6DVE). HEC forms a compound that gels in solvents such as ethyl alcohol and dimethyl sulfone oxide (DMSO). Electron spectroscopy chemical analysis (ESCA) of the first 100 .ANG. of the amylose thin film amylose indicates that the acylated surface had a degree of substitution of 0.9.+-.0.1 acyl chains per glucose moiety and this corresponded well to the expected regioselectivity of subtilisin catalysis on glucose-containing compounds. .sup.1 H-NMR studies indicated that only the C-6 hydroxyl groups of the glucose moiety were acylated with amylose and .gamma.-cyclodextrin. However, .beta.-cyclodextrin, and .alpha.-cyclodextrin were modified at secondary alcohols and at all three alcohols, respectively. This approach represents the first attempt at using enzymes to modify organic solvent-insoluble polymers in nonaqueous media.

A process is described for the enzyme catalyzed synthesis of polymeric microspheres from monomers such as ethylphenol, naphthols and hydroxypyrenes in reverse micelles in the presence of a detergent. Polymeric microspheres synthesized are porous with nonuniform densities. Polymers prepared from naphthols and hydroxypyrenes are fluorescent with electroluminescent properties. A number of materials such as enzymes, drugs and inorganic materials such as iron oxide and cadmium sulfide are encapsulated in the hollow polymeric microspheres synthesized. These encapsulations are carried out during or after the syntheses of the polymeric microspheres. Functional properties of the encapsulated materials such as enzyme, magnetic and nonlinear optical properties are demonstrated in the polymeric composites.

Polymer encapsulation and polymer microsphere composites

A method is described for a simple, fast and efficient synthesis of homopolymers and copolymers by the enzymatic ring opening polymerization of lactones and lactides. The enzyme used is an ion paired protease. The advantage of this enzymatic system is in using small amount of enzyme per monomer and lower reaction time. Homopolymers and copolymers are synthesized with molecular weights between 1000 and 4600 daltons, and dispersity as low as 1.1. The monomer conversion after 4 days, for reactions catalyzed by protease S, has reached 100%. Different initiators are used to control the rate and degree of polymerization. Synthesis of block copolymers with defined block size and crystallinity are described in this invention. These biodegradable and bioerodable polyesters and copolyesters with controlled molecular weight, dispersity and crystallinity have applications in medical, drug, cosmetic and food industries.

Enzyme-catalyzed synthesis of macromolecules in organic solvents

The invention relates to systems and methods for optical Fourier processing and logic operations based on the discovery that the photoinduced anisotropy of photochromic materials such as bacteriorhodopsin, organic fulgides, azo and fluorescent dyes, phycobiliproteins, rhodopsins, and their analogs, is dependent on the intensity of a polarized actinic beam that illuminates the material and the intensity profile of one or more input beams. This intensity dependence can be used to implement a simple, real-time, self-adaptive optical processing, i.e., spatial filtering, system for Fourier processing of optical input images. This optical processing system can be used to process a wide variety of optical input images, from projected still images to live motion picture images.

Optical fourier processing

A method of detecting the presence of a substance being monitored in a medium, selected from the group of substances including organophosphorus compounds and the metal ions Zn, Be and Bi, including the steps of: providing a 1,2-dioxetane phenyl phosphate compound; providing a phosphatase that catalytically degrades the 1,2-dioxetane phenyl phosphate compound to produce light, the catalytic activity of the phosphatase toward 1,2-dioxetane phenyl phosphate compound being altered by the substance being monitored; exposing the 1,2-dioxetane phenyl phosphate compound and the phosphatase together to a medium which may contain the substance being monitored; detecting light produced after the exposing step; and determining, from the detected light, the presence and concentration in the medium of the substance being monitored.

Chemiluminescence-based method for rapid and sensitive in-situ detection

All-optical devices, e.g., optical switches and modulators, and logic gates such as optical AND and OR gates, that include photochromic materials having first and second stable states, such as bacteriorhodopsin, organic fulgides, azo and fluorescent dyes, phycobiliproteins, rhodopsins, and their analogs, irradiated in a four-wave mixing geometry, are described. These devices can be used in a wide variety of systems, such as, e.g., optical signal processors and optical computers.

Growing community of inventors

Holliston, MA, United States of America

Joseph A Akkara