Management Development

Spiros Kambourakis

Dr. Spiros Kambourakis, Ph.D.

**** ***** ******, **** ****, San Diego, CA, 92101

Cell: 626-***-****

Email: *****************@*****.***

CAREER SUMMARY AND OBJECTIVE

I have over 10 years of “hands on” industrial experience in

the areas of biocatalysis, organic chemistry, molecular

biology, protein expression optimization in bacteria and

yeasts, fermentation development, and protein and whole

cell engineering. Much of this work has focused on the

development and optimization of biocatalytic routes for the

synthesis of industrial and high value chemicals.

Since I have mostly worked in start-up and intermediate size companies (less than two hundred

employees), I understand and thrive in the fast-pace, multitasking and results driven environment

of such organizations. For example, as one of the first employees of BioCatalytics (Pasadena,

CA) I played a key role in the company’s growth from five to over thirty employees over a seven

year period. Biocatalytics grew organically, only relying on product sales (mainly enzymes), with

no external investment capital. Using bioinformatics and gene mining techniques, I expanded

BioCatalytics’ product catalogue (and corresponding sales) from a few enzymes to more than

two hundred. BioCatalytics’ success was recognized by its acquisition by Codexis in 2007. I

retained my group leader position at Codexis, where I created new enzyme panels using directed

evolution techniques. In February of 2009, I joined a start-up company in San Diego (Strategic

Enzyme Applications) as the director of biology and biotransformation.

My current objective is to find a position that utilizes my

scientific and management skills. My preferred position

would be in scientific management as a director or group

leader. I would also be happy to work in project

management or product development, where I can utilize my

technical background to communicate and coordinate

science and business development.

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Spiros Kambourakis

SKILLS AND EXPERIENCE

Management skills

1. Lab Director: Directed and supervised the research activities of a group of scientists

(PhD, MS, and research technicians).

2. Project research management: Attended project management courses and participated in

business meetings for new product development and strategic research planning.

3. Technical writing: Paper, proposal and patent writing experience.

4. Data and other presentations: Presented data at conferences, and business meetings

(internally and with clients).

Scientific technical skills

5. Organic chemistry: Synthesis, purification, structure determination and chiral analysis of

organic molecules.

6. Molecular biology: Expert in cloning techniques and cDNA/gDNA library construction.

7. Microbial engineering: Construction of biocatalysts by altering the host strain (of E. coli

and Pseudomonas) and by modifying or creating new enzymatic pathways. Used

recombinant cells in the synthesis of chemicals from glucose or other precursors.

8. Protein Expression. Expert in recombinant protein expression techniques in bacteria

such as E. coli and Pseudomonas. Experience with expression in yeasts (Pichia and K.

lactis).

9. Fermentation: Growth of bacteria in 1L to 15L fed batch fermentors for the purpose of

large-scale protein production, or for the synthesis of organic chemicals.

10. Enzymology and protein purification: Enzyme purification and determination of kinetic

parameters of enzymatic reactions. Experience in the expression of membrane bound

proteins, such as the P450s.

11. Enzyme and whole cell immobilization: Experience in immobilization of proteins and

whole cells into different polymers (natural and synthetic)

12. Enzyme development and panel construction. Mutagenesis, library construction and

screening: Experience in DNA shuffling, error-prone PCR, as well as other random and

structure-guided mutagenesis techniques.

13. Gene discovery: Cloned novel genes using bioinformatics and genome mining

techniques.

14. Biocatalytic reaction process development and optimization: Optimized cell free or

whole cell enzymatic reactions for pilot scale synthesis of chemical intermediates.

15. New product development: Discovered over 200 new enzymes which were marketed as

enzyme kits (KREDs EREDs, etc) by Biocatalytics and Codexis.

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Spiros Kambourakis

COMPREHENSIVE PROFESSIONAL EXPERIENCE

Director of biology; Strategic Enzyme Applications (SEA), San

Diego, CA, 2009- Current position

As one of the first employees of SEA (operations began in

February 2009), I was hired to build the scientific team and

organize the research laboratories. This group was fully

operational and produced high quality results within the

first three months. These results exceeded the first quarter

expectations of our partners, Monsanto Inc.

The group’s responsibilities included:

• Cloning, screening and mutagenesis of new enzymes

• Expression optimization in various bacteria (E. coli and

Pseudomonas)

• Modeling and bioinformatic analysis to support

mutagenesis and enzyme evolution

• Whole cell metabolic engineering: creation of robust

biocatalysts for use in whole cell bio-transformations

(using both E. coli and Pseudomonas strains)

• Fermentation development (fed-batch and continuous)

for the production of enzymes and for use as

biocatalysts.

• Enzyme and whole cell immobilization

• Reaction optimization

As part of the scientific management of SEA, I have designed, planned and written research

proposals for potential clients, including full-time employee (FTE) requirements and project

management charts.

Manager Panel development; Codexis Inc 2007-2008 (Pasadena,

CA)

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Spiros Kambourakis

After the acquisition of BioCatalytics by Codexis in July

2007, I managed a group of scientists whose main focus was

the development of enzyme panels. Enzyme panels

(pioneered by Codexis Inc.) are created by structure-

designed mutagenesis and shuffling techniques to produce a

library of enzymes with diverse properties (wide substrate

range, improved stability, and good expression in bacterial

hosts). Group responsibilities included:

• Molecular biology techniques for targeted and random

mutagenesis, as well as shuffling techniques that

randomly recombined beneficial mutations

• Development and optimizitaion of high throughput

techniques for growing cells in plates and for their

screening using colorometric, UV, GC and HPLC

methods.

• Data analysis using data management software (LIMS).

In order to keep up with the scientific progress of the group,

I commonly used data management software (LIMS) for

analyzing data and library screening results. In addition to

technical support, I was responsible for the overall project

planning, setting of timelines, identifying and utilizing

resources, and facilitating communications between the

scientific teams in Pasadena and the Codexis headquarters

in Redwood city, CA. I also frequently communicated with

other groups within the company, particularly the

fermentation development and manufacturing group, in

order to select the best clones for manufacturing, or to

improve industrial strains.

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Spiros Kambourakis

As part of the Codexis Pasadena management team, I

attended quarterly meetings with executive management and

business development groups in order to evaluate current

and future research directions for new product

development..

Senior scientist; BioCatalytics Inc.: 2000-2007 (Pasadena, CA)

During my tenure at BioCatalytics, Inc., I was heavily involved in the development and

expansion of the company’s existing technology and product platform. Some representative

examples of research performed include the following;

• Discovery and development of over 200 new enzymes products.

• Construction and high throughput screening (HTS) of mutant and c-DNA

libraries.

• Development and optimization of chemo-enzymatic processes for the synthesis

of chiral intermediates.

• Expression optimization of various proteins including mammalian membrane-

bound enzymes, such as P450s in various bacterial and eukaryotic hosts.

• Design and execution of the large scale production of enzymes expressed in E.

coli using 15L fed-batch fermentations.

• Partial purification, lyophilization or immobilization of the enzymes to various

polymers.

• Measurement, characterization and comparison of kinetic parameters (Km,

Vmax) for different enzymatic reaction processes.

• Responsible for the planning, overlooking, directing and execution of

contracted research projects that were assigned to my group.

• Writing of patents, papers and research grants.

PhD; Michigan State University: 1994-2000 (Adviser: Prof John

W. Frost)

My doctorate thesis research involved development of both chemical and biological catalysts for

the synthesis of value-added hydroaromatic organic compounds. I accomplished the following

projects in this thesis:

• Using genetic engineering techniques, a number of whole cell biocatalysts capable of

synthesizing a number of hydroaromatic compounds from glucose were constructed.

• Optimized fed-batch fermentation conditions for growth and biotransformation

• Developed of a general high-yielding reaction for the oxidation of α-hydroxy carbonyl

functionality using catalytic amounts of Cu+2 and Zn+2 ions.

MS; University of Crete, Greece: 1992-1994(Adviser: Prof Michael Orfanopoulos)

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Spiros Kambourakis

During my masters thesis I was among the first to examine oxidations of organic olefins by

singlet oxygen that was generated after photosensitization of molecular oxygen by low

concentrations of buckminsterfullerenes (C60 and C70) in polar and nonpolar solvents. I identified

the reaction mechanism as Norris Type II by synthesizing a large number of deuterium-labeled

and unlabeled olefins.

EDUCATION

1994-2000 Ph.D., Dept. of Chemistry, Michigan State University, MI

“Synthesis of Value-added Chemicals from Glucose Using Chemical and Microbial Catalysis:

Gallic Acid, Protocatechuic Acid Pyrogallol and Catechol.” Adviser Prof John W. Frost

1992-1994 M.S. Dept. of Chemistry University of Crete, Heraklion, Crete, Greece

“Catalytic Photooxidations of Olefins Using C 60 and C70 as Photosensitizers”

Adviser Prof. Michael Orfanopoulos

1988-1992 B.S. Dept. of Chemistry University of Crete, Heraklion, Crete Greece

PUBLICATIONS

1. “A Diversified Library of Bacterial and Fungal Bifunctional Cytochrome P450 Enzymes

for Drug Metabolite Synthesis”

Roland Weis, Margit Winkler, Matthias Schittmayer, Spiros Kambourakis, Mandy Vink, J.

David Rozzell, Anton Glieder Adv. Synth Catal 2009, 351, 2140

2. “Stereoselective chemoenzymatic synthesis of sitophilate: a natural pheromone”

Dimitris Kalaitzakis, Spiros Kambourakis, J. David Rozzell, Ioulia Smonou Tetrahedron:

Asymmetry, 2007, 18(20), 2148.

3. “Stereoselective formation of a-substituted-b-hydroxy ketones and 1,3-diols using isolated

ketoreductases.”

Dimitris Kalaitzakis, J. David Rozzell, Ioulia Smonou and Spiros Kambourakis* Adv. Synth.

Catal 2006, 348, 1958

4. “A two step chemoenzymatic synthesis of the natural pheromone -Sitophilure utilizing an

isolated, NADPH-dependent ketoreductase.”

Dimitris Kalaitzakis, J. David Rozzell, Ioulia Smonou and Spiros Kambourakis* Eur. J.

Org. Chem 2006, 2309

5. “A recombinant ketoreductase tool-box. Assessing the substrate selectivity and stereo-

selectivity toward the reduction of β-ketoesters”

Dunming Zhu, D; Chandrani Mukherjee, J. David Rozzell, Spiros Kambourakis and Ling

Hua Tetrahedron 2006, 62, 901

6. “Ketoreductases: Stereoselective Catalysts for the Facile Synthesis of Chiral Alcohols”

Iwona A Kaluzna, J. David Rozzell, and Spiros Kambourakis* Tetrahedron:Asymmetry.

2005, 16, 3682.

7. “Highly stereoselective reductions of α -alkyl 1,3 diketones and α -alkyl β -ketoesters

catalyzed by isolated NADPH-dependent ketoreductases”

Dimitris Kalaitzakis, J. David Rozzell, Ioulia Smonou and Spiros Kambourakis* Org.

Lett 2005, 7, 4799

8. “Ketoreductases in the Synthesis of Valuable Chiral Intermediates: Application in the

Synthesis of α -hydroxy β -amino and β -hydroxy γ -amino acids”

Spiros Kambourakis* and J. David Rozzell Tetrahedron, 2004, 60, 663

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Spiros Kambourakis

9. “Chemo-enzymatic Method for the Synthesis of Statine, Phenylstatine and Analogues”

Spiros Kambourakis* and J. David Rozzell, Adv. Synth. Catal. 2003, 345(6+7), 699.

10. “Synthesis of Gallic Acid and Pyrogallol from Glucose: Replacing Natural Product Isolation

with Microbial Catalysis”

Spiros Kambourakis, Karen M Draths, John W. Frost J. Am. Chem. Soc. 2000, 122, 9042.

11. “Synthesis of Gallic Acid: Cu+2-Mediated Oxidation of 3-Dehydroshikimic acid”

Spiros Kambourakis, John W. Frost J. Org. Chem. 2000, 65, 6904

12. “Reaction of Dehydroshikimic Acid With Molecular Oxygen and Hydrogen Peroxide:

Products, Mechanism, and Associated Antioxidant Activity”

Jack Richman, Yu-Chen Chang, Spiros Kambourakis, K.M. Draths, Erick Almy, Kristi D.

Snell, Gale M. Strasburg and John W. Frost J. Am. Chem. Soc. 1996, 118, 11587.

13. “Generation and Trapping of Singlet Oxygen During Strong Illumintation of Photosystem II

core complex”

R. K. Misha, N. P. Mishra, Spiros Kambourakis, Michael Orfanopoulos Plant Science 1996,

115, 151.

16. “Chemical Evidence of Singlet Oxygen Production from C60 and C70 in Aqueous and Other

Polar Media”

Spiros Kambourakis, Michael Orfanopoulos Tetrahedron Lett. 1995, 36, 435.

17. “Fullrene C60 and C70 Photosensitized Oxygenation of Olefins”

Spiros Kambourakis, Michael Orfanopoulos Tetrahedron Lett. 1994, 35, 1945.

BOOK CHAPTERS

“3-Dehydroshikimic acid: a building block for chemical synthesis from renewable feedstocks”

Karen M. Draths; Spiros Kambourakis; Kai Li; John W. Frost. ACS Synp. Ser. 2001, 784

“Chemicals and materials from renewable resources” pg. 133.

“Biosynthesis of drug metabolites” Wenying Li, J. David Rozzell, Spiros Kambourakis, Martin

Mayhew in “Biocatalysis for the Pharmaceutical Industry” (2009) pp 183-211, 2009, Publisher:

(John Wiley&Sons (Asia), Singapore, Singapore)

PATENTS

1. Spiros Kambourakis and David Rozzell “Methods for producing hydroxy amino acids

and derivatives thereof” US Pat 6,833,471 B2, December 21, 2004

2. Spiros Kambourakis and David Rozzell “Hydroxy amino acids” US 7,081,535 B2 July 25

2006

3. Anton Glieder, Matthias Schittmayer, Spiros Kambourakis, Simone Zach “Methods for

redox reaction using an old yellow enzyme” US Patent Appl US2009/0117613A1 May 7

2009

Two more patents are recently prepared for submission (with SEA and Codexis)

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Spiros Kambourakis

RESEARCH PROPOSALS

1. “Synthesis of libraries of statine analogues” Phase I SBIR (Small Business Innovation

Research grant). $100,000 awarded for 6 months (August 2003-January 2004). Prime

Investigator and author: Spiros Kambourakis

2. “Construction of whole cell biocatalysts for ketone reduction” Phase II SBIR $850,000

awarded for 2 years. (Start date April 2006). Co-authored with: Jon Stweart, J David

Rozzell and Spiros Kambourakis

3. “Cloning and overexpression of novel alkene reductases” Phase I SBIR Submitted July

2006, $100,000 awarded for 6 months (May 2007-Octomber 2007). Prime Investigator

and author: Spiros Kambourakis

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