Microbial Production of l-Amino Acids

194 Pages·2003·3.47 MB·English

by  Masato Ikeda (auth.)

79
Advances in Biochemical
Engineering / Biotechnology
Managing Editor: T. Scheper
Editorial Board:
W. Babel. H. W. Blanch. I. Endo. S.-O. Enfors
A. Fiechter • M. Hoare • B. Mattiasson • H. Sahm
K. Schiigerl • G. Stephanopoulos • U. yon Stockar
G. T. Tsao. J. ViUadsen • C. Wandrey • J.-J. Zhong
Springer
Berlin
Heidelberg
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Hong Kong
London
Milan
Paris
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Microbial Production
of L-Amino-Acids
Volume Editors:
Robert Faurie • Jiirgen Thommel
With contributions by
B. Bathe, V. G. Debabov, S. Huebner, M. Ikeda, E. Kimura,
A. Marx, B. M6ckel, U. Mueller, W. Pfefferle
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Dr. Jtirgen Thommel
AMINO GmbH
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E-mail: [email protected]
Editorial Board
Prof. Dr. W. Babel Prof. Dr. H.W. Blanch
Section of Environmental Microbiology Department of Chemical Engineering
Leipzig-HalleG mbH University of California
Permoserstrafle 15 Berkely,C A 94720-9989,U SA
04318 Leipzig,G ermany E-mail: [email protected]
E-mail: [email protected]
Prof. Dr. I. Endo Prof. Dr. S.-O. Enfors
Faculty of Agriculture Department of Biochemistry and
Dept. of Bioproductive Science Biotechnology
Laboratory of Applied Microbiology Royal Institute of Technology
Utsunomiya University Teknikringen 34,
Mine-cho 350, Utsunomiya-shi 100 44 Stockholm, Sweden
Tochigi 321-8505, Japan E-mail: [email protected]
E-maik [email protected]
Prof. Dr. A. Fiechter Prof. Dr. M. Hoare
Institute of Biotechnology Department of Biochemical Engineering
Eidgen6ssische Technische Hochschule University College London
ETH-H6nggerberg Torrington Place
8093 Ziirich, Switzerland London, WCIE 71E,U K
E-maik [email protected] E-mail: [email protected]
VI Editorial Board
Prof. Dr. B. Mattiasson Prof. Dr. H. Sahm
Department of Biotechnology Institute of Biotechnolgy
Chemical Center, Lund University Forschungszentrum Jiilich GmbH
P.O. Box 124, 221 00 Lund, Sweden 52425 Jfilich, Germany
E-maih [email protected] E-maih [email protected]
Prof. Dr. K. Schfigerl Prof. Dr. G. Stephanopoulos
Institute of Technical Chemistry Department of Chemical Engineering
University of Hannover Massachusetts Institute of Technology
Callinstrat~e 3 Cambridge, MA 02139-4307, USA
30167 Hannover, Germany E-maih [email protected]
E-maih [email protected]
Prof. Dr. U. von Stockar Prof. Dr. G. T. Tsao
Laboratoire de G4nie Chimique et Director
Biologique (LGCB) Lab. of Renewable Resources Eng.
D6partment de Chimie A. A. Potter Eng. Center
Swiss Federal Institute Purdue University
of Technology Lausanne West Lafayette, IN 47907, USA
1015 Lausanne, Switzerland E-maih [email protected]
E-maih [email protected]
Prof. Dr. J. Villadsen Prof. Dr. C. Wandrey
Center for Process of Biotechnology Institute of Biotechnology
Technical University of Denmark Forschungszentrum ]iilich GmbH
Building 223 52425 ]filich, Germany
2800 Lyngby, Denmark E-rnaih c. [email protected]
E-maih [email protected]
Prof. Dr. J.-J. Zhong
State Key Laboratory
of Bioreactor Engineering
East China University of Science
and Technology
130 Meilong Road, Shanghai 200237, China
E-maih [email protected]
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Preface
It was in 1908, when Prof. Kinue Ikeda, grandfather of Dr. T. Ikeda, a Japanese
chemist discovered glutamic acid as a flavoring component in seaweed ("Kon-
bu") and soy hydrolysates. After neutralization with caustic soda, he isolated
Mono Sodium Glutamate (MSG), a flavor enhancer that represented the initial
product for the company Ajinomoto. The company's name is synonymous with
its first product and MSG was setting the stage for large scale production of
amino acids. MSG is still today's most important amino acid and flavor
enhancer.
This was not only the birth of the first industrial scale amino acid production
from natural raw material hydrolysates but also the fundamental process for the
isolation of other amino acids. Several proteins derived from soy, corn or wheat
contain only approximately 10-20% glutamic acid and glutamine whereas the
residues contain many other amino acids, such as the essential ones (leucine,
isoleucine, valine, threonine, methionine, lysine, phenylalanine and tryptophan)
and the semi essential ones (arginine, cyst(e)ine, histidine and tyrosine). It is,
however, very difficult to separate and purify all these components from the
crude initial hydrolysate and therefore not surprising that the market price for
those amino acids was comparably high.
In order to meet the growing demand of MSG worldwide which was especially
used in instant food and developing seasoning blends and to reduce production
costs, the German companies Degussa AG/Frankfurt and the Amino GmbH/
Frellstedt produced the MSG from vinasses or from beet sugar molasses. This
was done by alkaline hydrolysis of the pyrrolidonic carboxylic acid (PCA)
extracted in large amounts from these cheap vegetable by-products.
The real breakthrough was however achieved in the late 1960s and early 1970s
when Kyowa Hakko and Ajinomoto applied a completely new technology and
thus circumvented the expensive technology of protein hydrolysis and subse-
quent by-products recovery as a source of amino acids using highly sophisticat-
ed separation technologies.
It was the birth of modern biotechnology when two Japanese scientists,
Dr. Kinoshita and Dr. Udaka, at the research labs of Kyowa Hakko Kogyo dis-
covered in 1957 that a certain strain of the Gram-positive soil bacterium
"Corynebacterium" was able to excrete significant amounts of glutamic acid
under certain conditions. This corynebacterium was exploited for large scale
fermentation of glutamic acid for MSG production and the world market prices
dropped dramatically from over US$ 8 to below US$ 2. Today MSG represents
X Preface
the commercially dominating amino acid with an annual production rate of
more than 800,000 tons worldwide.
The application of fermentation technology did not remain limited to MSG
production. By natural screening accompanied by shot gun mutagenesis many
Corynebacterum derived strains and also Gram-negative bacteria such as E. coti
were discovered and proven to be suitable for amino acid breeding. However, the
upcoming efforts of modern molecular biology offered new sophisticated tools
such as the revolutionary enzyme engineering technology in the late 1970s.
Enzyme engineering represents the fundamental tool for modern metabolic
engineering. These techniques made it possible to modify individual key
enzymes of the amino acid anabolism by site-directed mutagenesis in order to
optimize their turn over and/or by knocking out feedback inhibition.
The next milestone was the introduction of Polymerase Chain Reaction
(PCR) combined with high throughput sequencing that made it possible to deci-
pher complete organism genomes. The complete 3 million base pair genome of
Corynebacterium glutamicurn is known today and on the way to being commer-
cially exploited by companies like BASF and Degussa AG.
What is the commercial interest behind this research in the field of amino
acid biotechnology.~
Besides the use of MSG and some other amino acids as flavoring substances,
the largest market for amino acids is the feed market showing outstanding
growth rates.While the market for amino acids in general is said to double every
decade, the market for L-lysine being the most important and best selling bio-
technological produced feed amino acid demonstrated a tremendous growth
by a factor of twenty during the last two decades. L-Lysine has reached the
large output of the chemically produced D,z-methionine with approximately
400,000 tons per year.
The significance of these amino acids on the world market is derived from
their physiological essentiality and that their limited content in many tradition-
al animal feed sources such as soy and corn.
This fact is usually visualized by the so called "barrel model" to demonstrate
the nutritive value of certain feedstuffs. This means that the value of the total
proteins present is limited by the intrinsic content of the essential amino acid
present at the lowest level. Methionine, e.g., is the value limiting Amino Acid in
soy protein. By fortifying proteins by addition of only small amounts of the lim-
iting amino acid the total efficacy of the individual feed product increases dra-
matically. The addition of only one part of D,z-methionine allows the substitu-
tion of 15 times the amount of soy at identical biological value and therefore to
reduce cost of pig growing for dramatically.
D,z-Methionine and ~.-lysine represent the dominating feed amino acids. In a
ranking of limiting Amino Acids and in dependence of the feeding raw material
used, L-threonine, L-tryptophan and the so called "Branched Chain Amino
Acids" (BCAA) e-isoleucine, z-valine and z-leucine are the next amino acids to
follow.
z-Threonine has been marketed for several years in the feed market
and seems to be the new star with an annual production rate of more than
15,000 tons.