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Chapter 30 : Tools for Enzyme Discovery
Author:
Yasuhisa Asano
Content Type: Reference
Publication Year:
2010
Category: Applied and Industrial Microbiology
Book DOI:
10.1128/9781555816827
Chapter DOI:
10.1128/9781555816827.ch30
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Abstract:
This chapter discusses recent remarkable examples in the screening for new enzymes, biocatalysis, and enzyme evolution. A section discusses screening for enzymes by enrichment culture technique from stock cultures, followed by enzyme purification and characterization, coupled with gene cloning. Examples include the discovery of nitrile hydratase and industrial production amides, aldoxime dehydratase and nitrile synthesis, ω-laurolactam hydrolase for enzymatic transcrystallization, and hydroxynitrile lyase from plants. These traditional approaches often yield primary information that can be industrialized successfully. Enrichment culture is a technique to isolate microorganisms having special growth characteristics. Acrylamide synthesis is now the largest enzyme-catalyzed organic synthesis in the world. Metagenomic approaches include sequence-based screening after shotgun large-scale sequencing of environmental DNA, enzyme activity-based screening of expressed libraries of environmental DNA, and a recent methodology called substrate-induced gene expression screening approaches (SIGEX). Reductive amination reaction of α-keto acid to form d-amino acid was made possible with this new enzyme. D-Isoleucine, D-glutamic acid, D-phenylalanine, and D-4-chlorophenylalanine were the substrates on which the wild-type enzyme was completely nonactive. The directed evolution of enzymes has proved to be a very powerful technique, as can be seen in the development of the new enzyme D-amino acid dehydrogenase, a thermostable DCase, and industrial production of nucleic acids with umami flavor by the evolved acid phosphatase.
Citation:
Asano Y.
2010.
Tools for Enzyme Discovery,
p 441-452.
In
Baltz R, Demain A, Davies J, Bull A, Junker B, Katz L, Lynd L, Masurekar P, Reeves C, Zhao H (ed),
Manual of Industrial Microbiology and Biotechnology, Third Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555816827.ch30
Figures
Tools for Enzyme Discovery
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Citation:
Asano Y.
2010.
Tools for Enzyme Discovery,
p 441-452.
In
Baltz R, Demain A, Davies J, Bull A, Junker B, Katz L, Lynd L, Masurekar P, Reeves C, Zhao H (ed),
Manual of Industrial Microbiology and Biotechnology, Third Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555816827.ch30
/content/10.1128/9781555816827.ch30.ch30fig01
FIGURE 1
Discovery of industrial enzymes, biocatalysis, and enzyme evolution. These various steps in discovery and development are cited by their respective letters throughout this chapter.
10.1128/9781555816827/f0442-01_thmb.gif
10.1128/9781555816827/f0442-01.gif
FIGURE 1
Discovery of industrial enzymes, biocatalysis, and enzyme evolution. These various steps in discovery and development are cited by their respective letters throughout this chapter.
Citation:
Asano Y.
2010.
Tools for Enzyme Discovery,
p 441-452.
In
Baltz R, Demain A, Davies J, Bull A, Junker B, Katz L, Lynd L, Masurekar P, Reeves C, Zhao H (ed),
Manual of Industrial Microbiology and Biotechnology, Third Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555816827.ch30
/content/10.1128/9781555816827.ch30.ch30fig02
FIGURE 2
Plant and microbial aldoxime-nitrile pathway.
10.1128/9781555816827/f0442-02_thmb.gif
10.1128/9781555816827/f0442-02.gif
FIGURE 2
Plant and microbial aldoxime-nitrile pathway.
Citation:
Asano Y.
2010.
Tools for Enzyme Discovery,
p 441-452.
In
Baltz R, Demain A, Davies J, Bull A, Junker B, Katz L, Lynd L, Masurekar P, Reeves C, Zhao H (ed),
Manual of Industrial Microbiology and Biotechnology, Third Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555816827.ch30
/content/10.1128/9781555816827.ch30.ch30fig03
FIGURE 3
Synthesis of 12-aminolauric acid from ω-laurolactam.
10.1128/9781555816827/f0443-01_thmb.gif
10.1128/9781555816827/f0443-01.gif
FIGURE 3
Synthesis of 12-aminolauric acid from ω-laurolactam.
Citation:
Asano Y.
2010.
Tools for Enzyme Discovery,
p 441-452.
In
Baltz R, Demain A, Davies J, Bull A, Junker B, Katz L, Lynd L, Masurekar P, Reeves C, Zhao H (ed),
Manual of Industrial Microbiology and Biotechnology, Third Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555816827.ch30
/content/10.1128/9781555816827.ch30.ch30fig04
FIGURE 4
Reactions catalyzed by nitrilases.
10.1128/9781555816827/f0444-01_thmb.gif
10.1128/9781555816827/f0444-01.gif
FIGURE 4
Reactions catalyzed by nitrilases.
Citation:
Asano Y.
2010.
Tools for Enzyme Discovery,
p 441-452.
In
Baltz R, Demain A, Davies J, Bull A, Junker B, Katz L, Lynd L, Masurekar P, Reeves C, Zhao H (ed),
Manual of Industrial Microbiology and Biotechnology, Third Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555816827.ch30
/content/10.1128/9781555816827.ch30.ch30fig05
FIGURE 5
Enzyme-catalyzed peptide synthesis. (a) A peptidase from E. brevis catalyzing ami-nolysis reaction. (b) LAL.
10.1128/9781555816827/f0445-01_thmb.gif
10.1128/9781555816827/f0445-01.gif
FIGURE 5
Enzyme-catalyzed peptide synthesis. (a) A peptidase from E. brevis catalyzing ami-nolysis reaction. (b) LAL.
Citation:
Asano Y.
2010.
Tools for Enzyme Discovery,
p 441-452.
In
Baltz R, Demain A, Davies J, Bull A, Junker B, Katz L, Lynd L, Masurekar P, Reeves C, Zhao H (ed),
Manual of Industrial Microbiology and Biotechnology, Third Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555816827.ch30
/content/10.1128/9781555816827.ch30.ch30fig06
FIGURE 6
Biosynthesis of menaquinone in microorganisms.
10.1128/9781555816827/f0446-01_thmb.gif
10.1128/9781555816827/f0446-01.gif
FIGURE 6
Biosynthesis of menaquinone in microorganisms.
Citation:
Asano Y.
2010.
Tools for Enzyme Discovery,
p 441-452.
In
Baltz R, Demain A, Davies J, Bull A, Junker B, Katz L, Lynd L, Masurekar P, Reeves C, Zhao H (ed),
Manual of Industrial Microbiology and Biotechnology, Third Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555816827.ch30
/content/10.1128/9781555816827.ch30.ch30fig07
FIGURE 7
Industrial dynamic kinetic resolution of hydantoin by hydantoinase and DCase to produce p-hydroxy-d-phenylglycine.
10.1128/9781555816827/f0447-01_thmb.gif
10.1128/9781555816827/f0447-01.gif
FIGURE 7
Industrial dynamic kinetic resolution of hydantoin by hydantoinase and DCase to produce p-hydroxy-d-phenylglycine.
Citation:
Asano Y.
2010.
Tools for Enzyme Discovery,
p 441-452.
In
Baltz R, Demain A, Davies J, Bull A, Junker B, Katz L, Lynd L, Masurekar P, Reeves C, Zhao H (ed),
Manual of Industrial Microbiology and Biotechnology, Third Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555816827.ch30
/content/10.1128/9781555816827.ch30.ch30fig08
FIGURE 8
Enzymatic synthesis of d-amino acids.
10.1128/9781555816827/f0448-01_thmb.gif
10.1128/9781555816827/f0448-01.gif
FIGURE 8
Enzymatic synthesis of d-amino acids.
Citation:
Asano Y.
2010.
Tools for Enzyme Discovery,
p 441-452.
In
Baltz R, Demain A, Davies J, Bull A, Junker B, Katz L, Lynd L, Masurekar P, Reeves C, Zhao H (ed),
Manual of Industrial Microbiology and Biotechnology, Third Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555816827.ch30
/content/10.1128/9781555816827.ch30.ch30fig09
FIGURE 9
Acid phosphatase-catalyzed 5′-monophosphorylation of nucleoside with PPi as a phosphorylating agent.
10.1128/9781555816827/f0448-02_thmb.gif
10.1128/9781555816827/f0448-02.gif
FIGURE 9
Acid phosphatase-catalyzed 5′-monophosphorylation of nucleoside with PPi as a phosphorylating agent.
Citation:
Asano Y.
2010.
Tools for Enzyme Discovery,
p 441-452.
In
Baltz R, Demain A, Davies J, Bull A, Junker B, Katz L, Lynd L, Masurekar P, Reeves C, Zhao H (ed),
Manual of Industrial Microbiology and Biotechnology, Third Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555816827.ch30
/content/10.1128/9781555816827.ch30.ch30fig010
FIGURE 10
Directed evolution of acid phosphatase as an industrial biocatalyst to produce 5′-IMP and 5′-GMP. ○, wild type; •, mutant.
10.1128/9781555816827/f0449-01_thmb.gif
10.1128/9781555816827/f0449-01.gif
FIGURE 10
Directed evolution of acid phosphatase as an industrial biocatalyst to produce 5′-IMP and 5′-GMP. ○, wild type; •, mutant.
Citation:
Asano Y.
2010.
Tools for Enzyme Discovery,
p 441-452.
In
Baltz R, Demain A, Davies J, Bull A, Junker B, Katz L, Lynd L, Masurekar P, Reeves C, Zhao H (ed),
Manual of Industrial Microbiology and Biotechnology, Third Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555816827.ch30
/content/10.1128/9781555816827.ch30.ch30fig011
FIGURE 11
Dynamic kinetic resolution of amino acid amide.
10.1128/9781555816827/f0449-02_thmb.gif
10.1128/9781555816827/f0449-02.gif
FIGURE 11
Dynamic kinetic resolution of amino acid amide.
Citation:
Asano Y.
2010.
Tools for Enzyme Discovery,
p 441-452.
In
Baltz R, Demain A, Davies J, Bull A, Junker B, Katz L, Lynd L, Masurekar P, Reeves C, Zhao H (ed),
Manual of Industrial Microbiology and Biotechnology, Third Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555816827.ch30
/content/10.1128/9781555816827.ch30.ch30fig012
FIGURE 12
Dynamic kinetic resolution of alanine amide with d-aminopeptidase and ACL racemase. •, l-alanine;
, d-alanine amide;
, d-alanine.
, d-alanine amide;
, d-alanine.10.1128/9781555816827/f0450-01_thmb.gif
10.1128/9781555816827/f0450-01.gif
FIGURE 12
Dynamic kinetic resolution of alanine amide with d-aminopeptidase and ACL racemase. •, l-alanine;
, d-alanine amide;
, d-alanine.
Citation:
Asano Y.
2010.
Tools for Enzyme Discovery,
p 441-452.
In
Baltz R, Demain A, Davies J, Bull A, Junker B, Katz L, Lynd L, Masurekar P, Reeves C, Zhao H (ed),
Manual of Industrial Microbiology and Biotechnology, Third Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555816827.ch30
References
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