potassium cyanide, the enzyme shows a fivefold higher specific
activity compared with PaHNL (the already industrialized and
highly active almond enzyme), effectively saving materials and en-
ergy previously needed for the reaction. In HNL research, cycles of
enzyme identification, physicochemical property characterization,
and rational protein engineering have been repeated for the sus-
tainable development of industrial biotechnology for a century (38).
In this respect, the discovery of millipede HNL can serve as a
template for the isolation of promising and efficient enzymes and
the design of tailor-made enzymes by rational protein engineering,
which are needed to spur the industrial synthesis of fine chemicals
and pharmaceuticals in the next generation.
Nucleotide Sequence Accession Number. The nucleotide sequence data of
ChuaHNL cDNA have been deposited in the DNA Data Bank of Japan (ac-
cession no. LC004755).
the methods of millipede collection, enzyme assays, enzyme purification,
protein sequencing, detection of sugar molecules in the HNL, UV-Vis, and CD
spectra of the purified millipede HNL, cDNA cloning, recombinant gene
expression, tissue collection, RT-PCR, Western-blot analysis, immunohisto-
ACKNOWLEDGMENTS. We thank T. Tanabe, M. Kamakura, T. Sakaki,
M. Kameya, H. Komeda, Y. Takakura, and K. Isobe for supporting our
experiments and valuable discussions. This work was supported by the
Exploratory Research for Advanced Technology program of the Japan
Science and Technology Agency.
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