166
combinations of mutations that would provide the introduction of desired functions
into the enzymes (Yoshikuni and Keasling 2007; Yoshikuni et al. 2006).
Less stability and inferior activity towards unusual substrates were considered as
the weaknesses of biocatalysts; these issues can be solved by efficient utilization of
various protein engineering methods. Previously, in order to compensate for less
activity, large amounts of protein were used, which caused emulsions that hindered
reaction and reduced the yield. Highly active enzymes solve this problem because
emulsions do not form using smaller amounts of protein. Modified enzymes with
extended shelf life, stability in organic solvents, and good activity should promote
biocatalysis to spread further into industrial laboratories. Recent advancement in
protein engineering has attained the equivalent of converting mouse proteins into
human proteins. The amino acid sequences of similar proteins in mice and human
typically differ by approximately 13% (Consortium 2002). Modern advanced protein engineering methods create similar changes in converting a wild-type enzyme
into an enzyme suitable for chemical process applications.
In recent years, the catalytic properties of the enzymes have enhanced quantitatively by factors of thousands to millions, and the engineered enzymes have the
capability to act in unusually harsh conditions. In order to contribute more expeditious advancement in the field of protein engineering, excellent knowledge of how
protein structure influences protein properties and a crucial evaluation of the many
protein engineering approaches are needed.
References
Adamczak MS, Krishna H (2004) Strategies for improving enzymes for efficient biocatalysis.
Food Technol Biotechnol 42:251–264
BCC Research (2017) Global markets for enzymes in industrial applications. BCC Research
BIO030J
Becker S, Hobenreich H, Vogel A, Knorr J, Wilhelm S, Rosenau F, Jaeger KE, Reetz MT, Kolmar H
(2008) Single-cell high-throughput screening to identify enantioselective hydrolytic enzymes.
Angew Chem Int Ed Eng 47:5085–5088
Behe MJ, Snoke DW (2004) Simulating evolution by gene duplication of protein features that
require multiple amino acid residues. Protein Sci 13:2651–2664
Ben Ali M, Khemakhem B, Robert X, Haser R, Bejar S (2006) Thermostability enhancement and
change in starch hydrolysis profile of the maltohexaose-forming amylase of Bacillus stearothermophilus US100 strain. Biochem J 394:51–56
Bloom JD, Arnold FH (2009) In the light of directed evolution: pathways of adaptive protein evolution. Proc Natl Acad Sci U S A 106(Suppl 1):9995–10000
Bloom JD, Labthavikul ST, Otey CR, Arnold FH (2006) Protein stability promotes evolvability.
Proc Natl Acad Sci U S A 103:5869–5874
Bornscheuer UT, Huisman GW, Kazlauskas RJ, Lutz S, Moore JC, Robins K (2012) Engineering
the third wave of biocatalysis. Nature 485:185–194
Bradshaw RA, Purton M (2012) Proteins: form and function. Elsevier Science Publishers, St.
Louis
Busto MD, Meza V, Ortega N, Perez-Mateos M (2007) Immobilization of naringinase from
Aspergillus niger CECT 2088 in poly(vinyl alcohol) cryogels for the debittering of juices.
Food Chem 104:1177–1182
S.M. Basheer and S. Chellappan
combinations of mutations that would provide the introduction of desired functions
into the enzymes (Yoshikuni and Keasling 2007; Yoshikuni et al. 2006).
Less stability and inferior activity towards unusual substrates were considered as
the weaknesses of biocatalysts; these issues can be solved by efficient utilization of
various protein engineering methods. Previously, in order to compensate for less
activity, large amounts of protein were used, which caused emulsions that hindered
reaction and reduced the yield. Highly active enzymes solve this problem because
emulsions do not form using smaller amounts of protein. Modified enzymes with
extended shelf life, stability in organic solvents, and good activity should promote
biocatalysis to spread further into industrial laboratories. Recent advancement in
protein engineering has attained the equivalent of converting mouse proteins into
human proteins. The amino acid sequences of similar proteins in mice and human
typically differ by approximately 13% (Consortium 2002). Modern advanced protein engineering methods create similar changes in converting a wild-type enzyme
into an enzyme suitable for chemical process applications.
In recent years, the catalytic properties of the enzymes have enhanced quantitatively by factors of thousands to millions, and the engineered enzymes have the
capability to act in unusually harsh conditions. In order to contribute more expeditious advancement in the field of protein engineering, excellent knowledge of how
protein structure influences protein properties and a crucial evaluation of the many
protein engineering approaches are needed.
References
Adamczak MS, Krishna H (2004) Strategies for improving enzymes for efficient biocatalysis.
Food Technol Biotechnol 42:251–264
BCC Research (2017) Global markets for enzymes in industrial applications. BCC Research
BIO030J
Becker S, Hobenreich H, Vogel A, Knorr J, Wilhelm S, Rosenau F, Jaeger KE, Reetz MT, Kolmar H
(2008) Single-cell high-throughput screening to identify enantioselective hydrolytic enzymes.
Angew Chem Int Ed Eng 47:5085–5088
Behe MJ, Snoke DW (2004) Simulating evolution by gene duplication of protein features that
require multiple amino acid residues. Protein Sci 13:2651–2664
Ben Ali M, Khemakhem B, Robert X, Haser R, Bejar S (2006) Thermostability enhancement and
change in starch hydrolysis profile of the maltohexaose-forming amylase of Bacillus stearothermophilus US100 strain. Biochem J 394:51–56
Bloom JD, Arnold FH (2009) In the light of directed evolution: pathways of adaptive protein evolution. Proc Natl Acad Sci U S A 106(Suppl 1):9995–10000
Bloom JD, Labthavikul ST, Otey CR, Arnold FH (2006) Protein stability promotes evolvability.
Proc Natl Acad Sci U S A 103:5869–5874
Bornscheuer UT, Huisman GW, Kazlauskas RJ, Lutz S, Moore JC, Robins K (2012) Engineering
the third wave of biocatalysis. Nature 485:185–194
Bradshaw RA, Purton M (2012) Proteins: form and function. Elsevier Science Publishers, St.
Louis
Busto MD, Meza V, Ortega N, Perez-Mateos M (2007) Immobilization of naringinase from
Aspergillus niger CECT 2088 in poly(vinyl alcohol) cryogels for the debittering of juices.
Food Chem 104:1177–1182
S.M. Basheer and S. Chellappan