Final Program

RECOMBINANT DNA BIOTECHNOLOGY:
FOCUS ON METABOLIC ENGINEERING

October 6-11, 1996
Ferncroft Conference Resort
50 Ferncroft Road
Danvers, Massachusetts 01923
1-508-777-2500 - Fax: 1-508-750-7959

Conference Chair
Professor Gregory Stephanopoulos
Massachusetts Institute of Technology
Department of Chemical Engineering

Conference Co-Chairs
Dr. Barry Buckland
Chemical Engineering R&D
Merck Sharp & Dohme Research Labs

Dr. David Anton
DuPont Central R&D
Bioprocess Development

Engineering Foundation Conferences
345 East 47th Street, New York, N.Y. 10017
1-212-705-7836; Fax 1-212-705-7441; E-mail: engfnd@aol.com
World Wide Web: http://www/engfnd.org

Program Committee
Prof. Spiro Agathos, Catholic University of Louvain - Belgium
Prof. James Bailey, Institute of Biotechnology - Switzerland
Dr. Roger Beachy, Scripps Research Institute - USA
Dr. Rick Benson, Cargill - USA
Prof. Douglas Cameron, University of Wisconsin - USA
Dr. Thomas Chattaway, ORSAN - France
Prof. Peter Dunnill, University College London - UK
Prof. Jean Marc Engasser, Institut National Polytechnique de Lorraine - France
Dr. David Fell, Oxford Brookes University - UK (invited)
Dr. S. Heijnen, Delft University of Technology - Netherlands
Dr. Fred Heineken, National Science Foundation - USA
Dr. Ganesh Kishore, Monsanto - USA
Dr. Hiroshi Kuriyama, National Institute of Bioscience & Human Technology - Japan
Prof. James Liao, Texas A&M University - USA
Prof. Basil Macris, National Technical University of Athens - Greece
Prof. E.T. Papoutsakis, Northwestern University - USA
Prof. Dr.-Ing. Matthias Reuss, Universitat Stuttgart - Germany
Prof. Herman Sahm, Institute fur Biotechnologie - Germany
Prof. Carlos Sola, Unitat d'Enginyeria Quimica - Spain
Prof. Hans Westerhoff, Free University - Netherlands
Prof. John Villadsen, Technical University of Denmark - Denmark

ACKNOWLEDGEMENTS
The Chair, Co-Chairs and Organizing Committee would like to acknowledge the
following organizations and thank them for their support of the conference:
National Science Foundation
Whittaker Foundation
Asahi Chemical Industry Co., Ltd.
Merck Research Laboratories
Schering-Plough Inc.
Yamanounchi Pharmaceutical

Sunday, October 6, 1996

3:00 p.m. - 6:00 p.m. Registration (Lobby)

6:00 p.m. - 7:30 p.m. Dinner (Ferncroft East/Center)

7:45 p.m. - 8:15 p.m. Opening Remarks
Dr. Gregory Stephanopoulos, Chair
Dr. Barry Buckland, Co-chair
Dr. David Anton, Co-chair
Allen Laskin, Engineering Foundation Liaison

8:15 p.m. - 9:15 p.m. Keynote Lecture:
Dr. Gerald Fink
Director, Whitehead Institute, MIT
"Fungal biology in the post-genome era"

9:15 p.m. - 11:00 p.m. Reception (Living Room)

NOTE: All technical sessions will be in the Grand Tara I & II
Poster sessions will be in the King Henry Suite

Monday, October 7, 1996

7:00 a.m. - 8:30 a.m. Breakfast (Grand Tara III & IV)

8:30 a.m. - 12:00 noon SESSION V: METABOLIC ENGINEERING
APPLICATIONS TO FUELS & CHEMICALS
Chairs: Prof. Herman Sahm
Biotechnology Institute
KFA, Julich, Germany
&
Dr. Thomas Erpicum
Roquette Freres, France

Speakers:
Prof. Loni Ingram
Department of Microbiology
University of Florida, Gainesville, Florida
"Metabolic engineering of bacteria for ethanol production"

Dr. Alan Berry
Genencor International, Palo Alto, California
"Metabolic engineering of E. coli for improved production of
aromatic compounds"

Coffee Break (Tara Foyer)

Prof. George Bennett
Department of Biochemistry and Cell Biology
Rice University, Houston, Texas
"Genetic manipulation of acetate and butyrate formation pathways in
Clostridium acetobutylicum"

Dr. L. Eggeling
Institute of Biotechnology
KFA, Julich, Germany
"Fluxes resolved and fluxes increased: A detailed view on
intracellular fluxes to obtain superior amino acid producers of
Corynebacterium glutamicum"

Dr. Merja Penttila, et al.
VTT Biotechnology, Finland
"Engineering Saccharomyces cerevisiae for xylose utilization"

12:00 noon - 1:30 p.m. Lunch (Grand Tara III & IV)

1:30 p.m. - 4:00 p.m. Ad hoc sessions/Free Time

4:00 p.m. - 6:00 p.m. WORKSHOP: Selection of Optimal Host-Cell Platform
for Metabolic Engineering
Chair: Dr. Scott Power
Genencor International

6:00 p.m. - 7:30 p.m. Dinner (Grand Tara III & IV)

7:30 p.m. - 10:00 p.m. SESSION II: MATHEMATICAL & COMPUTATIONAL TOOLS
OF METABOLIC ENGINEERING

Chairs: Dr. David Fell
Biology & Molecular Sciences
Oxford Brookes University, United Kingdom
&
Dr. K. Konstantinov
Bayer Co., Berkeley, California

(Coffee self-service at rear of meeting room)

Speakers:
Prof. James C. Liao
Department of Chemical Engineering
Texas A & M University
"Dynamic metabolic control analysis"

Troy Simpson and Gregory Stephanopoulos
Department of Chemical Engineering
Massachusetts Institute of Technology
"Flux determination and flux amplification in complex metabolic
networks"

V. Hatzimanikatis and J.E. Bailey
Institute of Biotechnology
ETH, Zurich
"Mathematical and computational tools for constructive metabolic
engineering"

Prof. Kazuyuki Shimizu
Department of Biochemical Engineering & Sciences
Kyushu Institute of Technology, Japan
"An integrated metabolic system engineering approach based on
metabolic signal flow diagram and cellular energetics"

P. Vanrolleghem and J.J. Heijnen
University of Gent and Delft University of Technology
"A structured approach for selection among candidate metabolic
models and estimation of unknown stoichiometric coefficients"

10:00 p.m. - 11:00 p.m. Social Hour (Living Room)

Tuesday, October 8, 1996

7:00 a.m. - 8:15 a.m. Breakfast (Grand Tara III & IV)

8:30 a.m. - 10:00 a.m. SESSION III: METABOLIC ENGINEERING APPLICATIONS
TO NEW MATERIALS
Chairs: Dr. Ashwin Madia
Cargill Corporation
&
Dr. Robert Wetegrove
Nalco Chemicals

Speakers:
Prof. A.J. Sinskey
Department of Biology
Massachusetts Institute of Technology
"Application of metabolic engineering principles to the
manipulation of polymer structure and function"

Prof. Dan Urry
University of Alabama School of Medicine at Birmingham
"Engineering protein-based machines to emulate key steps of
metabolism"

10:00a.m. - 10:30 a.m. Coffee Break (Tara Foyer)

10:30 a.m. - 12:30 p.m. SESSION IV: METABOLIC ENGINEERING APPLICATIONS
TO THE MANUFACTURING OF CHIRAL PHARMACEUTICALS
Chairs: Dr. Michel Chartrain
Merck & Co.
&
Dr. Gregg Whited
Genencor International

Speakers:
Dr. Paul Reider
VP Process Research
Merck Research Labs
"Opportunities for biocatalysis in the pharmaceutical industry:
Biology as a chemical tool"

Prof. C. Richard Huchinson
University of Wisconsin
"Engineering Antibiotic Producers for the Production of Novel
Metabolites as Potential Drugs"

Dr. Neil Bruce
Institute of Biotechnology
University of Cambridge, United Kingdom
"Engineering recombinant strains of Escherichia coli for the
efficient production of "semisynthetic opiate drugs"

12:00 noon - 1:30 p.m. Lunch (Grand Tara III & IV)
Dessert and Coffee will be served at the Poster Session

1:30 p.m. - 3:30 p.m. Poster Session I (King Henry Suite)
Chair: Dr. Ramesh Nair, DuPont Co.

3:30 p.m. - 6:00 p.m. Ad hoc sessions/Free Time

6:00 p.m. - 7:30 p.m. Dinner (Grand Tara III & IV)

7:30 p.m. - 10:00 p.m. SESSION V: APPLICATIONS OF METABOLIC ENGINEERING
TO PLANTS
Chairs: Dr. Phillip Lessard
Department of Biology
Massachusetts Institute of Technology
&
Dr. Bill Hitz
DuPont Co.

Speakers:
Prof. Jackie Shanks
Department of Chemical Engineering
Rice University, Houston, Texas
"Quantification of fluxes in the indole alkaloid pathway of
Catharanthus roseus: Implications for metabolic engineering"

Dr. Bill Hitz
DuPont Co.
"Genetic modification to alter the saturated to unsaturated fatty
acid ratio in the seed lipid of canola"

Dr. Ken Gruys
Ceregen, Inc.
Chesterfield, MO
"Metabolic engineering of offseed plants for production of
biodegradable plastics"

Dr. M.M. Burrell
Advanced Technologies
Cambridge, UK
"The use of transgenic technolgies to alter carbohydrate
metabolisms"

10:00 p.m. - 11:00 p.m. Social Hour (Living Room)

Wednesday, October 9, 1996

7:00 a.m. - 8:30 a.m. Breakfast (Grand Tara III)

8:30 a.m. - 12:30 p.m. SESSION VI: CONCEPTS OF METABOLIC ENGINEERING
Chairs: Prof. John Villadsen
Department of Biotechnology
Technical University of Denmark
&
Prof. Hiroshi Kuriyama
National Institute of Biosciences and Human Technology
Japan

Speakers:
Prof. Arnold L. Demain
Department of Biology
MIT
"The evolution of metabolic engineering"

Dr. David Fell
Biological & Molecular Sciences
Oxford Brookes University, United Kingdom
"Increasing the flux in metabolic pathways: A metabolic control
analysis perspective"

Coffee Break (Tara Foyer)

Prof. Bernhard O. Palsson
Department of Bioengineering
University of California San Diego, USA
"How should we approach the "Engineering" of metabolic function"

Dr. Jens Nielsen
Department of Biotechology
Denmark Technical University, Denmark
"Metabolic Engineering: concepts and applications"

Prof. Martin Yarmush
Harvard Medical School, USA
"The potential of metabolic engineering in studying human disease:
A new window into organ and tissue function"

12:30 p.m. - 2:00 p.m. Lunch (Grand Tara III)

2:00 p.m. - 4:00 p.m. WORKSHOP: Defining the Field of Metabolic
Engineering
Chair: A.J. Sinskey
Department of Biology
Massachusetts Institute of Technology, Massachusetts

4:00 p.m. - 6:00 p.m. Ad hoc sessions/Free Time

6:00 p.m. - 7:30 p.m. Dinner (Grand Tara III)

7:30 p.m. - 10:30 p.m. SESSION VII: EXPERIMENTAL TOOLS OF METABOLIC
ENGINEERING
Chairs: Prof. Doug Cameron
Department of Chemical Engineering
University of Wisconsin, Madison, Wisconsin
&
Dr. Max Follettie
Genetics Institute, Cambridge, Massachusetts

(Coffee self-service at rear of room)

Speakers:
Prof. Matthias Reuss
Institute for Biotechnology
University of Stuttgart, Germany
"Dynamic aspects of metabolic control analysis based on
measurements of intracellulars metabolites during transient
conditions"

Dr. Max Follettie
Genetics Institute, Cambridge, Massachusetts
"Transcript imaging using oligonucleotide arrays"

Prof. S. Shioya
Department of Biotechnology
Osaka University, Osaka, Japan
"On-line recognition and operation of the lysine fermentation
process based on a metabolic flux model"

Dr. Peter R. Jensen
Department of Microbiology
Technical University of Denmark, Lyngby, Denmark
"Artificial promoters for metabolic engineering of microorganisms"

Prof. Nancy DaSilva
Department of Chemistry & Biochemical Engineering
University of California, Irvine, California
"Regulated cloned gene integration in yeast: Improved methods for
metabolic engineering"

Dr. A. Matsuyama
Kikkoman Co., R & D Division, Chiba, Japan
"Novel production system for useful compounds through multi-step
enzyme reactions by sleeper vectors"

10:00 p.m. - 11:00 p.m. Social Hour (Living Room)

Thursday, October 10, 1996

7:00 a.m. - 8:15 a.m. Breakfast (Grand Tara III)

8:30 a.m. - 12:00 noon SESSION VIII: CELLULAR AND GENETIC
REGULATORY NETWORKS & METABOLIC ENGINEERING
Chair: Prof. B. Magasanik
Department of Biology
Massachusetts Institute of Technology, Massachusetts

Speakers:
Prof. Milton Saier
Department of Biology
University of California San Diego, LaJolla, California
"Mechanisms of carbon metabolic control in gram- negative vs. gram
positive bacteria"

Prof. Chris Kaiser
Department of Biology
Massachusetts Institute of Technology, Massachusetts
"Nitrogen regulation of protein localization in yeast"

Coffee Break (Tara Foyer)

Dr. Harley McAdams
Department of Developmental Biology
Stanford University, Stanford, California
"Genetic regulatory networks: Stochastic mechanisms affect timing
and regulatory outcomes"

Dr. Tom Rapoport
Harvard Medical School, Boston, Massachusetts
"Regulation of protein transport in mammalian cells"

12:00 noon - 1:30 p.m. Lunch (Grand Tara III)

Dessert and Coffee will be served at the Poster Session

1:30 p.m. - 3:00 p.m. Poster Session II (King Henry Suite)

Chair: Dr. Ramesh Nair, DuPont Co.

3:00 p.m. - 7:00 p.m. Ad hoc sessions/Free Time

7:00 p.m. - 9:00 p.m. Conference Banquet - New England Clam Bake (Grand Tara
III)

Hosted Bar until 11:00 p.m.

Dinner Lecture:
Prof. Boris Magasanik
Massachusetts Institute of Technology
"Genomics, Cell Physiology and Metabolic Engineering"

Friday, October 11, 1996

7:00 a.m. - 8:30 a.m. Breakfast (Grand Tara III & IV)

8:30 a.m. - 11:00 a.m. SESSION IX: APPLICATIONS OF METABOLIC
ENGINEERING TO PROTEIN TRAFFICKING AND CELL
CULTURE
Chairs: Dr. John Birch
Director of Development
Celltech, Slough, United Kingdom
&
Dr. David Robinson
Manager Fermentation R & D
Schering-Plough, New Jersey

Speakers:
Prof. Dhinakar Kompala
Department of Chemical Engineering
University of Colorado, Boulder, Colorado
"Engineering CHO cells to enhance foreign gene expression
selectively from the inducible MMTV promoter"

Prof. Nigel Jenkins
School of Applied Sciences
DeMontfort University, Leicester, United Kingdom
"Genetic engineering of glycosyltransferases in animal cells"

Coffee Break (Tara Foyer)

Prof. Michael Betenbaugh
Department of Chemical Engineering
Johns Hopkins University, Baltimore, Maryland
"Protein trafficking in insect cells: Multiple bottlenecks in the
assembly, secretion and glycosylation pathway"

Dr. David K. Robinson
Schering-Plough Research Institute, Union, NJ
"No pipe dream: Industrial applications of metabolic engineering in
cell culture"

11:00 a.m. - 12:30 p.m. SESSION X: ENVIRONMENTAL APPLICATIONS OF
METABOLIC ENGINEERING
Chairs: Prof. Spiro Agathos
Unit of Bioengineering
Catholic University of Louvain, Belgium
&
Prof. Murray Moo Young
Department of Chemical Engineering
University of Waterloo, Canada

Speakers:
Prof. Jay Keasling
Department of Chemical Engineering
University of California Berkeley, California
"Metabolic engineering for bioremediation of inorganic pollutants"

Prof. Hak-Sung Kim
Department of Biological Sciences
KAIST, Taejon, Korea
"Redesigning a metabolic pathway of Pseudomonas putida for the
treatment of aromatic hydrocarbons"

Prof. P. Dunnill & R. Sheridan
Advanced Centre for Biochemical Engineering
U.C. London, United Kingdom
"Controlled engineering of the tol meta- cleavage pathway"

12:30 p.m. - 2:00 p.m. Lunch (Grand Tara III & IV) and Adjournment

POSTERS

SESSION I: Tuesday, 8 October 1996, 1:30 - 3:30 pm

(King Henry Suite - Dessert and coffee will be served)

1. Min Zhang, Kristine Deanda, Christina Eddy, Ali Mohagheghi, Mark
Finkelstein, and Stephen Picataggio
Metabolic Engineering of Pentose Metabolism Pathways in Zymomonas Mobilis
for Ethanol Production

2. Fabien Barbirato, Andre Bories, and Philippe Soucaille
Regulation of the fermentation of glycerol to 1,3-propanediol by
Exnterobacter agglomerans in relation to the accumulation of
3-hydroxypropionaldehyde

3. Deborah A. Rathbone and Neil C. Bruce
The Improvement of Morphine Transformation Efficiency by the Use of a
(3-17)b- Hydroxysteroid Dehydrogenase

4. Pedro Mendes and Douglas B. Kell
A User-Oriented Simulation System for the Improvement of Biotechnological
Processes through Rational Metabolic Engineering

5. Fernando Valle, Elizabeth Ponce, Noemi Flores, and Francisco Bolivar
Alterations in Carbon Flow in Escherichia coli Strains Affected in the
Glucose Transport System and the Pyruvate Kinases or Phosphoglucose
Isomerase Genes

6. John H. Woods, David A. Fell, Stefan Schuster and Claus Hilgetag
Elementary Modes for Metabolic Network Stoichiometry Analysis

7. Yoshimasa Saito, Yoshinori Ishii, Hiromi Hayashi, Koji Yoshikawa, Yuji
Noguchi, Shinsuko Soeda, Mineo Niwa, Masaru Yoshida, Junji Hosoda and
Kyoichi Shimomura
Cloning of Genes for L-sorbose and L-sorbosone Dehydrogenases from
Gluconobacter oxydans and the Application to Microbial Process Engineering
for the Production of 2-keto-gulonate, a Direct Precursor of L-Ascorbic Acid

8. Jon P. Dean and George A. Dervakos
Redesigning Metabolic Networks Using Mathematical Programming

9. Sang Yup Lee, Young Lee and Jong Il Choi
Polyhydroxyalkanoate Synthesis by Metabolically Engineered Escherichia coli
Strains

10. Ramesh V. Nair, Edward M. Green, George N. Bennett, and Eleftherios T.
Papoutsakis
Strain improvement of and deregulated butanol and acetone production by
Clostridium acetobutylicum ATCC 824 via inactivation of a repressor protein
gene

11. Michael L. Hoffman, Frank A. Skraly, Anita Shaw and Douglas C. Cameron
Metabolic Engineering of 1,3-propanediol Production in Saccharomyces
cerevisiae

12. Anurag Khetan, Li-Hong Malmberg, Yun Seung Kyung, David Sherman and
Wei-Shou Hu
Identification of a-ketoglutarate as a Limiting Cofactor in the Biosynthesis
of Cephamycin C in Streptomyces clavuligerus Using Kinetic Simulation of the
Pathway Dynamics

13. Jae-Gu Pan, Heung Chae Jung, Dong Eun Chang, Soo An Shin, Chan Kyu Park,
Joon Shick Rhee
Metabolic Significance of pta-ackA Pathway in Escherichia coli and Acetate
Flux Modification for the Overproduction of Indigenous (D-lactate) and
Non-indigenous Metabolites (L-lactate, poly-b-hydroxybutyrate)

14. Hee-Sung Park, Jang-Young Lee, Moon-Hee Sung and Hak-Sung Kim
Production of L-DOPA (3,4-Dihydroxyphenyl-L-alanine) from benzene by using a
hybrid pathway

15. J. Varner and D. Ramkrishna
Application of Cybernetic Models to Metabolic Engineering

16. N. Kuchm, J.L. Goergen and J.M. Engasser
METAFLUX: A New Procedure for Intracellular Fluxes Evaluation

17. R.P. Desai and E.T. Papoutsakis
A Computational Method for Pathway Flux Analysis and Its Use in the Analysis
of Clostridium acetobutylicum Fermentations

18. Elizabeth Bull Daae, Andrew P. Ison and Peter Dunnill
Modelling of Carbohydrate Flux in Streptomyces lividans

19. S. Delaunay, M.F. Baucher, J.M. Engasser, A. Guyonvarch, and J.L. Goergen
Efficient promoter identification and PEP Carboxylase activity amplification
in
Corynebacterium glutamicum producing glutamate

20. Saburo Komatsubara and Naoki Sakurai
Fermentative production of D-Biotin by a recombinant strain of Serratia
marcesens

21. Norihiko Misawa
Metabolic engineering for the production of carotenoids in Escherichia coli
and other microorganisms

22. Mikhael Ponomarenko, Julia Ponomarenko, Alexander Kel, and Nikolay
Kolchanov
Signals and anti-signals of the td intron homing site

23. Mikhael Ponomarenko, Ludmila Savinkova, Alexander Kel, and Nikolay
Kolchanov
Simulation of the DNA sequences for protein-binding sites

24. Igor Titov
Transport asymmetry in inhomogeneous membrane with traps

25. Esperanza Troyano, Sam-Pin Lee, Oliver Peoples, Tae-Rak Kim, ChoKyan Rha,
and Anthony J. Sinskey
Cloning, sequencing, and characterization of an acetyl-transferase gene
involved in the exopolysaccharide biosynthesis of Zoolgloea ramigera 115

26. K.D. Snell, S.A. Hogan, S.J. Sim, J. Stubbe, C. Rha, and A.J. Sinskey
Polyhydroxybutyrate biosynthesis in recombinant Escherichia coli: molecular
weight control via in vivo synthase activity modulation

27. Fusao Tomita, and Atsushi Yokota
Change in metabolic flux in an F1-ATPase-defective mutant of Escherichia
coli W1485LIP2

28. G. Peter van Walsum and Lee R. Lynd
Allocation of ATP to synthesis of cells and hydrolytic enzymes in cellulotic
fermentative microorganisms: bioenergetics, kinestics, and bioprocessing

SESSION II: Thursday, 10 October 1996, 1:30 - 3:30 pm

(King Henry Suite - Dessert and coffee will be served)

29. Lee J. Sweetlove, T. ap Rees, and Michael M. Burrell
Starch metabolism in transgenic potato tubers (Solanum tuberosum 1.) with
increased adenosinediphosphoglucose pyrophosphorylase

30. Stelios Andreadis and Bernhard Palsson
Intracellular Stability of Retroviral Vectors: A Rate Limiting Factor in
Retrovirus- Mediated Gene Transfer

31. Simon Thomas, Peter J.F. Mooney, Michael M. Burrell and David A. Fell
Metabolic Control Analysis of Glycolytic Flux Control in Transgenic Potatoes
Overexpressing Phosphofructokinase in Tuber Tissue

32. Pau Ferrer, Ivan Diers, Juan A. Asenjo and Barbara A. Andrews
Yeast Cell Permeabilizing b-1,3-Glucanases: A Tool for the Integration of
Downstream Processes and Metabolic Engineering Applications to Yeast

33. Claudia Schmidt-Dannert, M. Luisa Rùa and Rolf D. Schmidt
Overexpression of a Thermophilic Lipase of Bacillus thermocatenulatus in E.
Coli, Large-scale Production and Purification

34. Atsuhiko Shinmyo, Takeshi Shoji, Kazuya Yoshida, and Masami Sekine
Metabolic engineering of cultured tobacco cells

35. Diethard Mattanovich, Walter Kramer and Christine Lüttich
Rational Design of an Improved Induction Scheme for Recombinant Escherichia
coli

36. J.J. Cairo , C. Paredes, C.R. Bebbington, C. Solà and F. Gòdia
Effect of the Expression of Glutamine Synthetase Gene on the Intracellular
Fluxes of a Hybridoma Cell Line

37. Nikolaos A. Katerelos and Peter W. Goodenough
Protein Engineering of a Plant Cysteine Proteinase as a Contribution to
Deciphering the Catalytic Mechanism of this Family of Enzymes

38. P. Tavernier, L. Besson, J.C. Portais, J. Courtois, B. Courtois and J.N.
Barbotin
In Vivo 13C-NMR Study of Polymer Synthesis in Two Rhizobium Meliloti Strains

39. Dirk E. Martens, Hendrik P.J. Bonarius, Cornelius D. de Gooijer and
Johannes Tramper
Cell-Cycle and Metabolic-Flux Analysis of Hybridoma Cells

40. Trygve Brautaset, Steffen Pettersen and Svien Valla
Correlation Between Kinetic Constants In Vitro and Biological Effects In
Vivo of Enzyme-mutants created by Site-directed Mutagenesis of
phosphoglucomutase from the bacterium Acetobacter xylinum

41. Frank W.R. Chaplen, William E. Fahl and Douglas C. Cameron
Improving the Properties of Animal Cells Used in Industrial Cell Culture:
Reducing Methylglyoxal Accumulation in Chinese Hamster Ovary Cells Grown in
Culture

42. Henrik Møllgaard and Bjørn Eggert Christensen
Mycotoxins: Quality Control of Fermentations with Aspergillus Oryzae

43. Dimitris G. Hatzinikolaou, Peter Stougaard, Dimitris Kekos and Basil J.
Macris
Metabolic Engineering of Apsergillus Niger: The Effect of Calcium Carbonate
on Glucose Oxidase Production

44. Kyongbum Lee, Francois Berthiaume, Gregory Stephanopoulos, and Martin L.
Yarmush
The effect of Interleukin-1b and Interleukin-6 on mitochondrial energy
metabolism in cultured hepatocytes

45. Alan J. Fischman, Edward A. Carter, Hongbing Hsu, Nathaniel M. Alpert,
Steven Weise, Martin L. Yarmush, and Ronald G. Tompkins
Measurement of Regional Blood Flow and Energy Metabolism in Burn Injury by
PET

46. Hendrik P.J. Bonarius, Ahmet Oezemre, Georg Schmid and Johannes Tramper
Metabolic Flux Analysis of Mammalian Cell Using Mass Balances and Their
Validation Using 13CO2 Mass Spectrometry and 13C-NMR Spectrometry

47. Karsten Schmidt, Achim Marx, Albert A. de Graaf, Wolfgang Wiecher,
Hermann Sahm, Jens Nielsen and John Villadsen
13C Tracer Experiments and Metabolic Balancing for Flux Analysis: Comparing
Two Approaches

48. Tomoya Iemura, Keisuke Furuichi, Takeshi Omasa, Yoshio Katakua, and
Ken-ichi Suga
Analysis of antibody production based on energy metabolism in hybridoma

49. Peter Ruhdal Jensen and Karin Hammer
Artificial promoters for metabolic engineering of microorganisms

50. Vasudevan Namasivayam
Treatment of petroleum refinery oil sludge contaminated soil through
bioaugmentation

51. Brian D. Follstad, Robert Balcarcel, Gregg Nyberg, A.J. Sinskey, D.I.C.
Wang, and Gregory Stephanopoulos
Metabolic fluxes, culture physiology, and product quality inc hemostat
cultures of
Chinese Hamster Ovary cells producing g-IFN

52. Erik N. Whiteley, Tsu-An Hsu, and Michael J. Betenbaugh
Modeling the assembly, aggregation, and chaperoning of Immunoglobulin G
production in insect cells

53. L. Mathieu, C. Rollin, F. Wojcik, and N.D. Lindley
Interruption of PEP carboxylase and the effect on glutamic acid production
in Corynebacterium melassecola ATCC 17965