PROGRAM

ENZYME ENGINEERING XV

October 10-15, 1999
Kona Surf Hotel
78-128 Ehukai Street
Kailua-Kona, Hawaii 96740
Phone: 1-808-322-3411
Fax: 1-808-322-3245

Conference Co-Chairs:
Dr. David Anton
DuPont
Professor Frances H. Arnold
California Institute of Technology
Professor Robert Kelly
North Carolina State University


United Engineering Foundation Inc.

Three Park Avenue, 27th Floor
New York, NY 10016-5902
T: 1-212-591-7836 F: 1-212-591-7441
E-mail: engfnd@aol.com Website: www.engnfnd.org

Poster Session I
Poster Session II
Poster Session III

Sunday, October 10, 1999

16:00 - 19:00 Registration

18:00 - 19:30 Dinner

19:30 - 19:45 Opening Remarks
Conference Co-Chair: Frances Arnold
Engineering Foundation Liaison: Allen Laskin

19:45 - 20:45 PLENARY LECTURE
Designs for Fribrous Proteins
David Eisenberg
University of California, Los Angeles, CA, USA

21:00 - 22:00 Opening Reception

Monday, October 11, 1999

07:00 - 08:15 Breakfast

08:30 - 12:00 SESSION I: BIODIVERSITY/NEW ENZYME DISCOVERY
Session Co-Chairs: Gao Xiang Li and Douglas Clark

08:30 - 09:00 Amino-Acid Metabolizing Enzymes from Hyperthermophilic
Microorganisms
Michael W. Adams
University of Georgia, Athens, GA, USA

09:00 - 09:30 Harvesting Biomolecules from the Tree of Life
Eric J. Mathur
Diversa Corporation, San Diego, CA, USA

09:30 - 10:00 Industrial Bioreactors as a Source for Novel Genes and Pathways
Vasantha Nagarajan
Dupont, Wilmington, DE, USA

10:00 - 10:30 Coffee Break

10:30 - 11:00 Bridged Bimetallic Enzymes: Why Two Heads are Better
Than One
Gregory Petsko
Brandeis University, Waltham, MA, USA

11:00 - 11:30 New Functions from Old Scaffolds: How Nature Re-engineers
Enzymes for New Functions
Patsy Babbitt
University of California, San Francisco, CA, USA

11:30 - 12:00 Discussion

12:00 -14:00 Lunch

14:00 - 16:30 Ad hoc Sessions and/or free time

17:00 - 18:30 SESSION II: HIGHLIGHTS IN ENZYME DESIGN
Session Chair: John Holbrook

17:00- 17:30 Towards the Automated Design of Protein Catalysts
Steve Mayo
California Institute of Technology, Pasadena, CA, USA

17:30 - 18:00 Enzyme Assembly
Stephen J. Benkovic
Pennsylvania State University, University Park, PA, USA

18:00 - 18:30 Molecular Breeding of Genes, Pathways and Organisms by DNA Shuffling
Willem P.C. Stemmer
Maxygen Inc., Redwood City, CA, USA

18:30 - 20:00 POSTER SESSION I
(Poster Presenters will be present to answer questions)

20:00 - 21:30 Dinner

21:30 - 22:30 Social Hour (in poster area)

Tuesday, October 12, 1999

07:00 - 08:15 Breakfast

08:30 - 12:00 SESSION III: ENZYME ENGINEERING/DIRECTED EVOLUTION
Session Co-Chairs: Joseph Affholter and Michael Danson

08:30 - 09:00 Directed Evolution of Enzymes Using THERMUS THERMOPHILUS
and E. COLI as Hosts: Thermal Stability and Activity
Akahiko Yamagishi
Tokyo University, Tokyo, JAPAN

09:00 - 09:30 Enzyme and Pathway Engineering for Gene Therapy
Margaret E. Black
Washington State University, Pullman, WA, USA

09:30 - 10:00 Isolation of Novel Enzymes from Large Libraries Displayed
on the Surface of Bacteria and Screened by FACS
George Georgiou
University of Texas-Austin, Austin, TX, USA

10:00 - 10:30 Coffee Break

10:30 - 11:00 Tailoring of Fatty Acid Desaturase and Hydroxylase Enzymes
John Shanklin
Brookhaven National Laboratory, Upton, NY, USA

11:00 - 11:30 Engineering the Reaction Mechanism of the Sulfolobus
Solfataricus Beta-Glycosidase: Construction of a New Thermophilic Glycosynthase
Marco Moracci
Institute of Protein Biochemistry and Enzymology Naples, ITALY

11:30 - 12:00 Discussion

12:00 -14:00 Lunch

14:00 - 16:45 Ad hoc Sessions and/or free time

17:30 - 18:30 SESSION IV: NEW BIOTRANSFORMATIONS
Session Chair: Maria Kula

17:00 - 17:30 Production of Both Enantiomers of Chiral Amines Using (S)-
Transaminase and its Unique Property of Enantioselective Substrate Inhibition
Byung-Gee Kim
Seoul National University, Seoul, KOREA

17:30 - 18:00 (Chemo)enzymatic Synthesis of Nucleotide-Activated Sugars and
Their Application in Glycobiotechnology
Lothar Elling
Institute of Enzyme Technology, Jülich, GERMANY

18:00 - 18:30 Engineerng Enzymes for Glycotechnology
Pamela Sears
Scripps Research Institute, La Jolla, CA, USA

18:30 - 20:00 POSTER SESSION II
(Poster Presenters will be present to answer questions)

20:00 - 21:30 Dinner

21:30 - 22:30 Social Hour (in poster area)

Wednesday, October 13, 1999

07:00 - 08:15 Breakfast

08:30 - 12:00 SESSION V: OXIDATION BIOTRANSFORMATIONS
Session Chair: Jonathan Dordick

08:30 - 09:00 Rational Design of a Semi-Synthetic Peroxidase
Roger Sheldon
Delft University of Technology, Delft, THE NETHERLANDS

09:00 - 09:30 Ancient Bio-Oxidation Processes as the Basis for New Biocatalytic Systems
Bernard Witholdt
ETH, Zurich, SWITZERLAND

09:30 - 10:00 Steps Towards a P450 Monooxygenase-Based Enzyme Reactor
Rolf Schmid
University of Stuttgart, Stuttgart, GERMANY

10:00 - 10:30 Coffee Break

10:30 - 11:00 Reaction Engineering for Redox Enzyme Applications
Vince Vilker
NIST, Gaithersburg, MD, USA

11:00 - 12:00 Discussion

12:00 - 14:00 Lunch

14:00 - 16:45 Ad hoc Sessions and/or free time

17:00 - 18:30 SESSION VI: ADVANCES IN ENZYME REACTION ENGINEERING
Session Chair: Christian Wandrey

17:00 - 17:30 Enzyme Processing Technology for the Production of a Vitamin C
Intermediate
Richard LaDuca
Genencor International, Palo Alto, CA, USA

17:30 - 18:00 Process Selection for the Aldolase-Catalized Synthesis of
N-Acetyl-D-Neuraminic Acid
John M. Woodley
University of College London, London, UNITED KINGDOM

18:00 - 18:30 Gas/Solid Biocatalysis: Fundamentals and Applications
Marie Dominique Legoy
University of Rochelle, FRANCE

18:30 - 20:00 POSTER SESSION III
(Poster Presenters will be present to answer questions)

20:00 - 21:30 Dinner

21:30 - 22:30 Social Hour (in poster area)

Thursday, October 14, 1999

07:00 - 08:15 Breakfast

08:30 - 12:00 SESSION VII: COMBINATORIAL BIOCATALYSIS/PATHWAY
ENGINEERING
Session Chair: Veronique Larreta Garde and TBA

08:30 - 09:00 Gene Expression Tools for Metabolic Engineering of Bacteria
Jay D. Keasling
University of California, Berkeley, CA, USA

09:00 - 09:30 Pathway Engineering for Polyunsaturated Fatty Acid
Production by an Arachidonic Acid-Producing Fungus
Mortierella Alpina 1S-4
Sakayu Shimizu
Kyoto University, Kyoto, JAPAN

09:30 - 10:00 Role of Catabolic Threonine Dehydratase on Pathway Engineering
Tony Sinskey
Massachusetts Institute of Technology, Cambridge, MA, USA

10:00 - 10:30 Coffee Break

10:30 - 11:00 Biocatalytic Synthesis of Combinatorial Libraries
Joseph Rich
Enzymed, Inc.

11:00 - 11:30 The Mitomycin Biosynthetic Gene Cluster: New Directions in
Combinatorial Biology
David Sherman
University of Minnesota, USA

11:30 - 12:00 Discussion

12:00 -13:30 Lunch

13:30 - 15:45 Ad hoc Sessions and/or free time

16:00 - 17:00 SESSION VIII: NEW DEVELOPMENTS
Session Chair: Harvey Blanch

16:00 - 16:30 What Limits in Vitro Protein Biosynthesis? State of the Art in
Protein Bioreactor Performance
Martin Siemann
University of Stuttgart, Stuttgart, GERMANY

16:30 - 17:00 Protein Splicing: Mechanism and Applications
Francine Perler
New Ennglad Biolabs

17:00 - 18:30 SESSION IX: ENZYME ENGINEERING AWARD LECTURE

19:15 - 22:00 Luau and Social Hour

Friday, October 15, 1999

07:00 - 09:00 Breakfast and Conference Adjournment

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Poster Presentations - Enzyme Engineering Conference

Co-Chairs: Jeffrey Moore and Wilfred Chen

Day 1
Monday, October 11, 1999
6:30 p.m. - 8:00 p.m.


Biodiversity / New Enzyme Discovery, Highlights in Enzyme Design, New Developments:

1 Masson, Jean-Michel
Screening for New Enzymes with Phenomics

2 Junying, Wang
A new microbial acid amylase.

3 Wu, Jin
The production, isolation and characterization of ß-mannanase secreted from Nocardioform Actinomycetes.

4 Liu, Yang
Cloning and sequence analyzing of trehalose synthase from bacterium isolated from near the spring of Changbai mountain.

5 Ying, Lin
Extracellular glutaminase from marine bacteria.

6 Yamamura, Shohei
A cold-active protease from Pseudomonas fluorescens strain 3A08.

7 Hasan, Quamrul
Extracellular proteinases from psychrotrophs: Keratinase.

8 Morita, Yasutaka
Cold-active protease and pyruvate dehydrogenase-like protein genes from psychrotrophic bacteria.

9 Tamiya, Eiichi
Purification and Characterization of Cold-Active L-Glutamate Dehydrogenase Independent of NAD(P) and Oxygen.

10 Zhang, Jinhong
Investigation of enantioselective hydrolysis of chiral ester by lipases.

11 Zhu, Xianjun
Cloning and high expression of urate oxidase from Candida utilis.

12 Shtemenko, N.
Activation of glucuseoxidase-peroxidase in vitro system with binuclear rhenium-dicarboxilates.

13 Jiang, Tai Jiao,
Expression and characterization of active horseradish peroxidase in Pichia pastoris.

14 Liu, Jungi
Chemoenzymatic synthesis of epothilones A and B.

15 Hua, Zi-Chun
Molecular imprinting of human urokinase in polysiloxane polymer.

16 Ishikawa, Kazuhiko
Thermostable Aminopeptidase from Pyrococcus horikoshii.

17 Clark, Douglas S.
Tailoring the Activity and Stability of Hyperthermophilic Enzymes.

18 Chaudhuri, Julian B.
Biocatalysis with Extremophilic Enzymes.

19 van den Burg, Bertus
Engineering a Protease Enzyme to Resist Boiling.

20 de Kreij, Arno
Rational Design - A Second Youth - Engineering of Thermolysin Like Proteases.

21 Li, Xinhui
Zinc-mediated thermal stabilization of carboxypeptidase A.

22 Kermasha, S.
Characterization of tyrosinase biocatalysis in chloroform medium, using high-performance liquid chromatography and capillary electrophoresis.

23 Kamiya, Noriho
A Novel Cytochrome B562 Mutant with Enhanced Peroxidase Activity.

24 Pohl, Martina
Tailoring enzymes for the application in biotransformation.

25 Koeller, Kathryn
Utility of glycosyltransferases in the chemoenzymatic synthesis of complex glucoconjugates.

26 Zhang, Hong Ying
Design and synthesis of a new signal peptide for secretory expression of µ-aspartyl dipeptidase.

27 Yingjiu, Zhang
Studies on the changes of conformations and activity of polygalacturonase in alcohol solution.

28 Koivula, Anu
Protein engineering of Trichoderma reesei cellobioydorlase CEL7A catalytic domain.

29 Monsan, Pierre
Amylosucrase: A Glucosyltransferase of the Alpha-Amylase Super Family Useful for Oligo- and Poly- Amylose Synthesis.

30 Willemot, René-Marc
Dextransucrases and Amylosucrases are Microbial Glucosyltransferases Acting on Sucrose but Belonging to the a-Amylase Super Family.

31 Bott, Richard R.
Quantitative Structure Activity Relationships in Subtilisin BPN' and Protein Engineered Variants having Altered Specificity and Proteolytic Activity.

32 Han, Xiao-Qing
The Kinetics of Proteolysis Cannot Fit the Steady State Kinetic Model.

33 Khan, Golam Faruque
Comparative Study of Proteolytic Specificity of Several Commercial Proteases.

34 Elliott, R. Jeffery
Development of a Novel Peptide Ligase.

35 Resnick, Sol M.
In vitro ATP Regeneration from Polyphosphate and AMP by Polyphosphate and AMP Phosphotransferase and Adenylate Kinase.

36 Maksimenko, A.V.
Experimental antioxidant biotherapy for protection of vessel wall by superoxide dismutase and catalase derivatives.

37 Manco, Giuseppe
Functional and structural properties of esterases belonging to the hormone sensitive lipase (HSL)-like group of the esterase-lipase family.

38 Gong, Z-Y
Reshuffle and redesign of L-aspartase Domains.

39 Kong, Xiang-Duo
Study of the amino acid residues at the active site of µ-aspartyl dipeptidase.

40 Kong, Xiang-Duo
Constructing a new type of ribozyme.

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Poster Presentations: Day 2
Tuesday, October 12, 1999
6:30 p.m. - 8:00 p.m.

Enzyme Engineering, Combinatorial Biocatalysis, Oxidative Biotransformations, New Developments

1 Nakano, Hideo
In vitro Methods for the Generation of Protein Libraries using PCR and Cell-Free Protein Synthesis.

2 Ostermeier, Marc
Using Incremental Truncation to Create Hybrid Enzymes Libraries.

3 May, Oliver
Inversion of Enantioselectivity by Directed Evolution.

4 Urabe, Itaru
Nonadditivity of mutational effects and its application to efficient directed evolution.

5 Nagamune, Teruyuki
Molecular Design of Artificial Antibody and Its Application to Immunoassay.

6 Wang, Jin-Shan
Directed evolution of human granulocyte colony stimulating factor by error-prone PCR and DNA shuffling.

7 Wang, Li-Juan
Construction of the active single subunit of ECSOD by directed evolution.

8 Wong Chi-Huey
Altering the Substrate Specificity and Catalytic Efficiency of 2-Keto-3-Deoxy-6-Phosphogluconate (Kdpg) Aldolase by Directed Evolution.

9 Oriel, Patrick
Greatly Enhanced Expression of a Thermostable Amylase from a Single Alteration in Primary Sequence.

10 Ford, Clark
Directed Evolution of Glucoamylase to Increase Thermostability.

11 Sundaram, P.V.
Improved properties of a variety of chemically modified enzymes.

12 Ahsan, Mohammad Mainul
Construction and Comparative Characterization of Two Family 10 Chimeric Xylanases.

13 Lee, Seung-Goo
L-DOPA selective tyrosine phenol-lyase engineered by error-prone PCR.

14 Chen, Wilfred
High efficiency biodegradation of organophosphates using genetically modified organophosphorus hydrolase.

15 Zhao, Huimin
Directed Evolution of a Dehalogenase for Industrial Scale Biocatalysis.

16 Showell, Michael S.
Engineering Detergent Proteases.

17 Liu, Yi-Ming
Directed evolution of µ-aspartyl dipeptidase for synthesis of the precursors of dipeptide sweenteners.

18 Kumar, Manoj
Biocatalytic production of ascorbic acid.

19 Hsu, Amy K.H.
Pathway engineering of E. coli and E. blattae for glycerol hyper-producing strains.

20 Nakamura, Charles E.
Biological production of 1,3-propanediol.

21 Chou, C. Perry
Genetic strategies to enhance penicillin acylase production in E. coli.

22 Kim, Geun-Joong
Evolution of a Novel Bifunctional D-Hydantoinase/ N-Carbamoylase Fusion Enzyme for Functional Expression with Enhanced Stability.

23 Hanson, Ronald L.
Enzymatic Synthesis of L-6-Hydroxynorleucine.

24 Cowan, Don
S. Solfataricus b-D-glycosidase-catalyzed synthesis of glucoconjugates in organic solvents.

25 Bentzien, Jorg
PDA: Protein Design Automation - A Computational Prescreen for Improving Enzymatic Activity.

26 Joo, Hyun
Directed evolution of peroxide-mediated cytochrome P450 hydrolxylation to enhance activity and alter regioselectivity.

27 Liu, Jianguo
Chemiluminescent sensor for enzymatic determination of creatinine content.

28 Jung, Heung-Chae
Bacteria cell surface display of enzymes and its application as a platform technology for direct selection of improved enzyme variants.

29 Tanaka, Atsuo
Construction of Arming Yeasts Responsible to the Change of Environmental Conditions.

30 Wood, Keith V.
Using Directed Evolution to Reveal the Molecular Mechanism of Color Variation in Beetle Luciferases.

31 Satoh, Ikuo
Flow Injection Amperometry Using a Laccase Column.

32 Akkara, Joseph A.
Enzyme-Catalyzed Synthesis of Polyaromatic Compounds with Multifunctional Groups for Industrial Applications.

33 Uchida, Akira
Microbial Regiospecific Hydroxylation of Pyridine Related Compounds.

34 Burton, Stephanie G.
Biotransformation Of Phenolics Using Oxidative Enzymes In Membrane Bioreactors.

35 Doig, Steven D.
Use of Recombinant Whole Cells as Biocatalysts in Organic Syntheses Stereo-Specific Baeyer-Villiger Oxidation of Cyclic Ketones.

36 Schmid, Andreas
Enantiospecific Preparation of Epoxides using Cell Free Bacterial Styrenemonooxygenase.

37 Kito, Mitsuaki
Microbial Production of a Useful Sulfoxide Compound.

38 Haltrich, Dietmar
Continuous Enzymatic Regeneration of Quinone Electron Acceptors -- Production of Lactobionic Acid by Enzyme Technology as an Example.

39 Nidetzky, Bernd
Groel-mediated enzyme stabilization: Glucose-fructose oxidoreductase from Zymononas mobilis as example.

40 Amanullah, A.
Rational Selection of a Microbial Host for the Indene Bioconversion Based on Multi-Staining Flow Cytometry.

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Poster Presentations: Day 3
Wednesday, October 13, 1999
6:30 p.m. - 8:00 p.m.

Advances in Reaction Engineering, New Biotransformations

1 Hansson, Therese
Enzymatic Synthesis of Oligosaccharides from Lactose using Wildtype and Mutants of Thermostable Beta-Glycosidases.

2 Zhang, Xue-Zhong
Enzymatic synthesis of small peptide in organic media.

3 Yoo, Young Je.
Enzyme Reactions in PEG Media.

4 Yoshida, Toyokazu
Carbon Dioxide Fixation by Reversible Pyrrole-2-Carboxylate Decarboxylase.

5 Radke, Clayton J.
Hydroxynitrile Lyase Activity at the Organic Solvent/Water Interface.

6 Miyawaki, Osato
Effect of Water Activity and Aqueous Solution Structure on Enzyme Kinetics of Alcohol Dehydrogenase, Lysozyme, and Beta-Galactosidase.

7 Clark, Douglas S.
Dramatic Enhancement of Enzyme Activity in Organic Solvents: Exploring the Mechanism of Salt Activation.

8 Andersson, Mats
Increased Almond Glucosidase Activity in Low Water Activity Media by Substrate and Medium Engineering.

9 Pham, Laura J.
Lipase modification of triglycerides and its effect on the foaming properties of coconut and non-lauric oils.

10 Marty, Alain
Lipase-Catalyzed Synthesis Reactions: How to Choose the Good Solvent?

11 Zaidi, Ahmed
Enzymatic Esterification of Long-Chain Fatty Acids using Nylon-Immobilized Lipase.

12 Chopineau, Joël
Enzyme-mediated formation of vesicles from mixed micelles.

13 Li, Gaoxiang
Enzymatic diagnosis of serum triglyceride by a coupled enzymes system.

14 Persson, Mattias
Lipase Catalysed Fatty Acid Exchange in Digalactosyldiacylglycerol.

15 Kunugi, Shigeru
Ring-Opening Bulk Polymerization of Lactones by using Commercial Lipases.

16 Combes, Didier
Effect of Temperature and Pressure on Lipase Stability.

17 Straathof, Adrie J. J.
Manipulation of Multiple Enzymatic Reaction and Phase Equilibria.

18 Larreta-Garde, Veronique M.
Enzyme catalyzed phase transitions: Protease against 'gel solase' activity.

19 Bommarius, Andreas S.
Novel Method for Measuring Biocatalyst Operating Stability.

20 Yamuna, A.
Chemical modification of trypsin with free and polyethyleneimine dervatives of disaccharides.

21 Russell, Alan
Biocatalytic chemical defense.

22 Novick, Scott J.
Protein-Based Composites for Reaction and Separation.

23 Kondo, Akihiko
Preparation of High Activity Whole Cell Biocatalysts Prepared by Permeabilization.

24 Jongejan, J. A.
Molecular and Thermodynamic Modeling of Enzyme Enantioselectivity.

25 Thomas, Katie C.
Rapid, Rational Selection of Cofactor Recycling Options.

26 Nidetzky, Bernd
Fungal trehalose phosphorylase and its use in the production of trehalose from sucrose or starch.

27 Shi, Yi-Feng
pH and Polyatomitic Alcohol Effect on Activity and Selectivity of Penicillin Acylase-Catalyzed Kinetically Synthesis of Cephalexin.

28 Na, Shu min
Enzymatic synthesis of serine by serine hydroxymethytransferase.

29 Lortie, Robert
Determination of Optimal Reaction Conditions for the Enzymatic Synthesis of Sorbitol Monoester in Organic Medium Under Reduced Pressure

30 Syldatk, Christoph
A Cascade of Immobilized Enzymes for the Production of Natural and Unnatural L-Amino Acids from D,L-5-Monosubstituted Hydantoins.

31 Payson, R.
Purification, cDNA Cloning, and Improvement of Activity by Directed Evolution of a Novel Ketoreductase from Zygosaccharomyces rouxii.

32 Kou, Xiufen
Mono- and diester of polyethylene glycol400 laurate synthesized with immobilized lipase.

33 Kula, Maria-Regina
Development of a Large Scale Production Process for Rec. LeuDH from Bacillus cereus.

34 Burton, Stephanie G.
Production and Application of Bacterial Hydantoin-Hydrolysing Enzymes for Synthesis of Enantiomerically Pure Amino Acids and Related Products.

35 Sakka, Kazuo
Cloning and Expression of the agaA Gene Encoding a-galactosidase from Clostridium josiu.

36 Li, Lin
Studies on transglutaminase from Streptovertivillium fradioverticillatum.

37 Nicolini, C.
Immobilization Techniques for Biocatalysis.

38 Rattray, Fergal P.
The Influence of Lactococcal Proteinase Activity, Diversity and Specificity of Bitter Flavor Development in Cheese.

39 Kermasham S.
Biocatalysis of immobilized chlorophyllase in a ternary micellar system.

40 Liu, Xingjuan
Removal of phenol from waste water by soluble tyrosinase.

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