BIOCATALYSIS AND BIOPROCESSING OF MACROMOLECULES:
A NATIONAL SCIENCE FOUNDATION INDUSTRY/UNIVERSITY COOPERATIVE RESEARCH CENTER

Biocatalysis encompasses a wide platform of chemistries that afford opportunities for innovative new products and processes. Hallmarks of biocatalysts are their ability to operate under mild conditions, with impressive selectivity, on a diverse range of natural and non-natural substrates. Immobilized enzymes can function over a broad range of conditions that are adaptable to current manufacturing equipment and processes. Rapid advances in biotechnology continues to decrease the time and resources required to engineer organisms to produce desired products in high titers as well as to engineer enzymes with increased thermal stability, efficiency and specificity. The Center works with its members to apply existing or develop new biocatalytic methods that solve short or long term product development challenges. To address a broad range of technological challenges, the Center assembled an impressive network of scientists with a broad range of experimental and analytical knowledge. Furthermore, members self direct research to ensure that they maximize their return on their investment.

POLYMER RESEARCH AT THE POLYTECHNIC UNIVERSITY:

Polytechnic Institute of New York University (NYU-POLY), fondly known to many as ‘Brooklyn Poly’, was the first to establish a graduate program in Polymer Science. The Polytechnic is steeped in a tradition of excellence in Polymer Research and has sent into the world many of today’s leaders in the field. Today, NYU-POLY is embracing a new frontier at the interface of Bioengineering, Macromolecules and the Biological sciences. Major transformations are underway at NYU-POLY to make the University a world leader in Biomacromolecular Science and Engineering.

VISION:

We stand at the threshold of a revolution in the way Industry will look towards biologically based materials and processes. The launching of new products is increasingly dependent on cradle-to-grave assessments. Significant additional costs will be imposed on industry for the disposal of toxic chemicals and by-products. These considerations will accelerate the rate-of-change in the way that chemistry and processes are designed and conducted. Completely new methods will emerge that reach well beyond incremental improvements to the discovery of new paradigms in synthesis and processing.

Biocatalysis, whether using isolated enzymes or whole cells, offers unique opportunities to meet the evolving needs of environmentally compatible processes. In reviewing the level of activity and knowledge in biocatalysis, we found a substantial effort in the development of low molar mass molecules but polymers have received much less attention. A need was expressed by industry for a cost-effective mechanism that would allow them to rapidly get up-to-speed in Biocatalysis and Bioprocessing, determine opportunities in the field, gain access to ideas and new technology, initiate projects of interest for which they did not have sufficient resources, and find business partners that compliment their expertise. To meet these needs, the NSF Center on Biocatalysis and Bioprocessing of Macromolecules (NSF-BBM) was established in April 2000.

WHY ENZYME-CATALYSIS?

The following describes how biocatalysts are being applied by the ‘Center’ to create new technologies for product synthesis, processing and utilization:

Control of Structure – Enzyme selectivity can be directed towards the synthesis of unique surfactants, peptides, monomers, macromers and polymers. Functionality that would otherwise be difficult to achieve without protection-deprotection steps can be attained by regioselective transformations.

Simplicity of Reactions Over an Ever-Widening Range of Conditions - An important feature of biocatalytic reactions is the growing range of reaction conditions in which it can be performed. These include bulk systems, organic solvents, biphasic conditions, emulsions, and in supercritical fluids.

Multistep Conversions in Aqueous Media: Whole-cell transformations convert low-cost materials to value-added products. Concepts of metabolic engineering used in combination with enzyme evolution provide powerful methods for the development of efficient microbial catalysts.

Green Chemistry - The use of environmentally compatible methods for synthesis and processing is good business. Enzymes allow reductions in processing temperatures, provide metal-free safe catalysts, and convert multiple-step processes to one-pot reactions. They are also made and degraded by natural processes.

Protein Engineering - Incorporated diversity can be retrieved through structural constraints, phylogenetic diversity, random mutations or immunological constraints. Analysis of systematically varied sets of sequences is being applied to improve key enzymes under study within the Center. This strategy has the advantage of requiring orders of magnitude fewer variants than directed evolution methods, while not being constrained by structurally interpretable changes.

RESEARCH AREAS:

We are exploring how the diverse chemistries, mild reaction conditions and selectivity of enzymes can provide unique opportunities to develop new or improve synthetic routes to molecules and materials of industrial importance. The following summarizes Center research activities:

• Lipases, cutinases, esterases, proteases, peroxidases, laccases and other enzymes families are being used as biocatalysts.
• Availability of proprietary enzymes.
• Enzyme Engineering
• Biobased functional lipids (e.g. w-hydroxy and w-carboxyfatty acids).
• Chemical polymerization of biobased monomers.
• New technologies for low-cost peptide synthesis using protease catalysis. Study of their potential use in metal binding, as macromers, self-assembly, and their biological properties.
• Biosurfactants for uses in cosmetics, antimicrobial agents and as templates for directed material assembly.
• Synthesize vinyl monomers by selective acrylation of natural building blocks (e.g. carbohydrates, amino acids and lipids).
• Controlling end group functionality (e.g. diol, thiol, and acrylate groups) of prepolymers.
• Enzyme-catalysis in microfluidic reactors.
• Develop mild ‘triggered’ enzyme-catalyzed crosslinking and degradation reactions.
• Silicone based materials.
• Innovative new bioresorbable polymers for drug delivery and tissue-engineering scaffolds.
• Biopolishing and surface modification.
• Enzyme immobilization to optimize its activity and recyclability.
• Fiber electrospinning.
• Design and synthesis of protein based materials.
• AFM and other spectroscopic methods to analyze the surface topology and structure.
• Thermal and mechanical properties of polymeric materials.

APPLICATIONS OF MATERIALS FROM BIOCATALYSIS:

Plastics, elastomers, adhesives, coatings, macromers, functional prepolymers, formulation ingredients (e.g. biosurfactants, peptides), polyurethanes, water-soluble polymers, surfactants, vinyl monomers, cosmetics, fine chemicals, nutraceuticals, agricultural materials, bioresorbable polymers, scaffolds for tissue engineering, protein therapeutics, modified peptides, functionalized surfaces.

CENTER STAFF:

The Center has assembled an impressive group of Faculty and Scientists from a wide-range of participating universities. Faculty, postdoctoral fellows and graduate students work in interdisciplinary teams. The following lists these participants, their affiliation and core research interests (in alphabetical order):

Dr. Kathryn Beers (micro-reactors and microfluidics) NIST; Professor Stephen Clarson (enzymatic reactions used by diatoms and sponges, (bio)silicon chemistry) University of Cincinnati; Dr. John Decator (High field NMR spectroscopy) Columbia University; Professor Yuying Gosser (protein modeling and crystallization) The City College of NY, CUNY; Professor George John (self assembly, biosurfactants, soft materials) City College at CUNY); Professor X.-P. Kong (protein modeling and crystallization) NYU Medical Center, NY, NY; Dr. William L’Amoreaux (director of the Advanced Imaging Facility that includes AFM, confocal microscopy, SEM, TEM at CUNY Staten Island); Professor Katja Loos, characterization of surfaces, block copolymer self-assembly) Univ. of Groningen; Professor Jin Kim Montclare (protein engineering, protein based materials, non-natural amino acids) Polytechnic University; Professor Fred Naider (peptide synthesis and characterization) CUNY Staten Island; Professor Miriam Rafailovich (Surface engineering and characterization: SUNY at Stony Brook, L.I., NY); Professor P. Somasundaran (Surface and interfacial properties of surfactants) Columbia University; Professor Maristella Scandola (Solid-state material properties, fiber electrospinning) University of Bologna, Italy); Professor Ruth Stark (solid state NMR, molecular structure and biomechanics of plant cuticles) The City College of NY, CUNY; Professor Iwao Teraoka (Polymer Physics and chromatography) Polytechnic University; Professor Benjamin Zhao (polymer crystallization, polymer nanocomposites, synchrotron X-ray scattering) SUNY Stony Brook.

WORKING TOGETHER: AN INDUSTRY-UNIVERSITY PARTNERSHIP

The difficult economy through which most of us are navigating have tightened budgets. Industrial sponsorship of academic research needs to be more tied to proprietary arrangements that have valuable deliverables addressing current needs. To this end industrial partners are offered the opportunity to leverage the cost of a small percentage of one researcher into the power of 10 professors and 25 researchers & students. In response to your needs, Research teams are assembled from the Center network and timelines are established to complete specific tasks. Members have the opportunity to become ‘true partners’ in the process of problem solving and discovery. Since different models fit different problems and organizations, Members can define their level of involvement in projects. This ranges from visits to our laboratories, regular teleconferences to quarterly progress reports. The following outlines the benefit of Center membership:

• Leverage the cost of a small percentage of one researcher into the power of 10 professors and 25 researchers & students.
• Annually renewable membership allowing you the flexibility to opt in or out as needed.
• Cost-effective access to a portfolio of research projects and a network of researchers with an extraordinary breadth of expertise.
• Use 60% of your contribution to support a specific research that is kept confidential from other members. Prioritize the use of the remaining 40 % the membership fee that is pooled with other member dollars to address research challenges of mutual interest to members.
• NSF Center status avoids the normal university overhead so that only 10% of funding goes to administrative overhead.
• Rapid information transfer through frequent reports, cost-effect tele- or video-conferencing.
• On-site learning of experimental methods.
• Annual review meeting where you can network with other industrial members.
• Get to know highly trained potential future employees.
• Access to results from other federally funded projects. Four million in external funds were available from 2005-2008.
• Access to highly specialized equipment.

Some benefits to members are more subtle but provide powerful arguments to management for participation. For example, Center research may save Members from investing in similar projects that, if performed in house, would be much more costly. Also, by participation, Members can rapidly target promising new technologies for in-house investment.

PATENTS, PUBLICATIONS, and INTELLECTUAL PROPERTY

Patents are actively pursued. As of September 2005, 14 patents have been granted (US 6,424,271; US 6,486,295; US 6,316,581; US 6,093,792; 5,378,807; US 5,440,007; US 5,439,985; WO9961482AI; US 05981743; US 05883199; WO 97071531AI; WO 09724387A1. Ten applications are currently active within the US Patent office.

INDUSTRIAL MEMBERS:

Since the Centers inception in April 2000, members have included BASF, DSM, Evonik, Novozymes, Genencor, W.R. Grace, Johnson & Johnson, Esteé Lauder, Cognis, Sherwin Williams, Grain Processing Company, EcoSynthetic’s, DNA 2.0 and Nalco.

PRESIDENTIAL HONOR: The Center was the recipient of the 2003 Presidential Green Chemistry Award winner in the academic category.

The award honors the Centers research on a broad range of research on lipase-catalyzed polyester syntheses.