S-selective hydroxynitrile lyase from a plant Baliospermum montanum: Molecular characterization of recombinant enzyme

Dadashipour, Mohammad; Yamazaki, Misako; Momonoi, Kazumi; Tamura, Ken’ichirou; Fuhshuku, Ken‐ichi; Kanase, Yurina; Uchimura, Etsuzoh; Guan, Kang; Asano, Yasuhisa

doi:10.1016/j.jbiotec.2011.02.004

Cited by 42 publications

(35 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We excluded two other hydroxynitrile lyases in the α/β-hydrolase superfamily from this comparison: Bm HNL from Baliospermum montanum (a medicinal shrub from India) [15] and At HNL from Arabidopsis thaliana , [16] (see supporting information). The amino acid sequences of these HNL are 57% and 47%, respectively, identical to Hb HNL, but neither plant contains cyanogenic glycosides, so the enzymes’ natural function is unknown.…”

Section: Resultsmentioning

confidence: 99%

Increasing the Reaction Rate of Hydroxynitrile Lyase fromHevea brasiliensistoward Mandelonitrile by Copying Active Site Residues from an Esterase that Accepts Aromatic Esters

2014

View full text Add to dashboard Cite

The natural substrate of hydroxynitrile lyase from rubber tree (HbHNL, Hevea brasiliensis) is acetone cyanohydrin, but synthetic applications usually involve aromatic cyanohydrins such as mandelonitrile. To increase the activity of HbHNL toward this unnatural substrate, we replaced active site residues in HbHNL with the corresponding ones from esterase SABP2 (salicylic acid binding protein 2). Although this enzyme catalyzes a different reaction (hydrolysis of esters), its natural substrate (methyl salicylate) contains an aromatic ring. Three of the eleven single-amino-acid-substitution variants of HbHNL reacted faster with mandelonitrile. The best was HbHNL-L121Y, with a 4.2-fold higher kcat and high enantioselectivity. Site-saturation mutagenesis at position 121 identified three other improved variants. We hypothesize that the smaller active site orients the aromatic substrate more productively.

show abstract

Section: Resultsmentioning

confidence: 99%

Increasing the Reaction Rate of Hydroxynitrile Lyase fromHevea brasiliensistoward Mandelonitrile by Copying Active Site Residues from an Esterase that Accepts Aromatic Esters

2014

View full text Add to dashboard Cite

show abstract

“…Because millipedes exude defensive secretions of aromatic compounds, such as mandelonitrile and benzaldehyde (19), millipede HNLs seem likely to have substrate specificity toward aromatic compounds. Thus, we sought to synthesize cyanohydrins using ChuaHNL in a citrate buffer, using the aromatic aldehydes in our chemical library (26,27) as starting materials, and evaluated the chiral configuration and ee, which are important indices for fine chemical syntheses. ChuaHNL catalyzed the synthesis of cyanohydrins from monosubstituted, disubstituted, biphenyl, heteroaromatic, and bicyclic aldehydes with an almost exclusive R configuration and an ee range of 7.5-90% (Table S2).…”

Section: Substrate Specificity and Kinetic Studies In The Synthesis Omentioning

confidence: 99%

Discovery and molecular and biocatalytic properties of hydroxynitrile lyase from an invasive millipede,Chamberlinius hualienensis

Dadashipour

Ishida

Yamamoto

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

100

View full text Add to dashboard Cite

Hydroxynitrile lyase (HNL) catalyzes the degradation of cyanohydrins and causes the release of hydrogen cyanide (cyanogenesis). HNL can enantioselectively produce cyanohydrins, which are valuable building blocks for the synthesis of fine chemicals and pharmaceuticals, and is used as an important biocatalyst in industrial biotechnology. Currently, HNLs are isolated from plants and bacteria. Because industrial biotechnology requires more efficient and stable enzymes for sustainable development, we must continuously explore other potential enzyme sources for the desired HNLs. Despite the abundance of cyanogenic millipedes in the world, there has been no precise study of the HNLs from these arthropods. Here we report the isolation of HNL from the cyanide-emitting invasive millipede Chamberlinius hualienensis, along with its molecular properties and application in biocatalysis. The purified enzyme displays a very high specific activity in the synthesis of mandelonitrile. It is a glycosylated homodimer protein and shows no apparent sequence identity or homology with proteins in the known databases. It shows biocatalytic activity for the condensation of various aromatic aldehydes with potassium cyanide to produce cyanohydrins and has high stability over a wide range of temperatures and pH values. It catalyzes the synthesis of (R)-mandelonitrile from benzaldehyde with a 99% enantiomeric excess, without using any organic solvents. Arthropod fauna comprise 80% of terrestrial animals. We propose that these animals can be valuable resources for exploring not only HNLs but also diverse, efficient, and stable biocatalysts in industrial biotechnology.millipede hydroxynitrile lyase | bioresource exploration | biocatalysis | white biotechnology | arthropod

show abstract

“…S-selective HNL (S-HNL) activity was found in a homogenate of the leaves of Baliospermum montanum, while R-selective HNLs (R-HNLs) were detected in the leaves and seeds of Passiflora edulis, and the seeds of Eriobotrya japonica, Prunus mume, Prunus persica, Chaenomles sinensis and Sorbus aucuparia. The HNLs from E. japonica (EjHNL) (Ueatrongchit et al, 2008(Ueatrongchit et al, , 2009, P. edulis (PeHNL) (Ueatrongchit et al, 2010), P. mume (PmHNL) (Fukuta et al, 2011;Nanda et al, 2005Nanda et al, , 2006, and B. montanum (BmHNL) (Dadashipour et al, 2011) have been purified to homogeneity and their enzymatic properties characterized. PmHNL shows excellent potential for the asymmetric synthesis of cyanohydrins in organic solvents.…”

Section: Introductionmentioning

confidence: 99%

Synthesis of (R)-β-nitro alcohols catalyzed by R-selective hydroxynitrile lyase from Arabidopsis thaliana in the aqueous–organic biphasic system

Fuhshuku

Asano

2011

Journal of Biotechnology

Self Cite

View full text Add to dashboard Cite

S-selective hydroxynitrile lyase from a plant Baliospermum montanum: Molecular characterization of recombinant enzyme

Cited by 42 publications

References 46 publications

Increasing the Reaction Rate of Hydroxynitrile Lyase fromHevea brasiliensistoward Mandelonitrile by Copying Active Site Residues from an Esterase that Accepts Aromatic Esters

Increasing the Reaction Rate of Hydroxynitrile Lyase fromHevea brasiliensistoward Mandelonitrile by Copying Active Site Residues from an Esterase that Accepts Aromatic Esters

Discovery and molecular and biocatalytic properties of hydroxynitrile lyase from an invasive millipede,Chamberlinius hualienensis

Synthesis of (R)-β-nitro alcohols catalyzed by R-selective hydroxynitrile lyase from Arabidopsis thaliana in the aqueous–organic biphasic system

Contact Info

Product

Resources

About