The synthesis of chiral cyanohydrins by oxynitrilases

doi:10.1016/S0167-7799(00)01452-9

Trends in Biotechnology

Volume 18, Issue 6, 1 June 2000, Pages 252-256

https://doi.org/10.1016/S0167-7799(00)01452-9 Get rights and content

Abstract

Enantiomerically pure cyanohydrins are important synthetic intermediates for pharmaceuticals and agrochemicals. They are produced by enzyme-catalysed synthesis using oxynitrilases. Sufficient quantities of enzyme are available via cheap natural sources and there have been recent advances in overexpression production of cyanohydrins on an industrial scale.

Section snippets

Oxynitrilases: their occurrence, properties, structure and availability

Hnls are naturally widespread enzymes: ∼3000 plant species and a small number of other organisms are known to possess these enzymes14, 15, 16. Within cyanogenic plants, the cyanohydrins of several aldehydes and ketones (e.g. benzaldehyde, acetone and 2-butanone) exist as glycosides or lipids. When a plant cell is damaged, Hnl comes into contact with the cyanohydrins, which results in cleavage and liberation of hydrocyanic acid. Thus, a form of plant defence is activated. Another natural

Molecular cloning and characterization

In general, the successful commercial application of enzymes depends on their availability in sufficient quantities at a reasonable cost, and on the ability to adapt the enzymes to the needs of the industrial production process. Recombinant DNA techniques enable efficient enzyme production, provide rapid access to molecular data on primary structure and facilitate the establishment of enzyme-engineering strategies. The genes for various Hnls have already been cloned and characterized12, 28, 30,

Recombinant production

There are several expression systems available for the recombinant production of Hnls12, 13, 28, 29, 30, 31. Those for the enzyme from H. brasiliensis were developed for large-scale production using recombinant DNA technology. This Hnl can be expressed efficiently in various microbial hosts. A strong, inducible tac-promoter-based expression system results in high-level expression in Escherichia coli, but E. coli is not a good environment for Hnl synthesis because the Hnl is mostly present in an

(R)- and (S)-selective Hnls, available enzymes and substrate acceptance

As shown in Fig. 1a, the addition of hydrogen cyanide to the carbonyl group of an aldehyde or ketone is a reversible reaction, leading to an equilibrium that, as a rule, favours cyanohydrin formation. The use of Hnls opens up the unique possibility of convenient synthetic access to enantiopure cyanohydrins of both (R)- and (S)- configurations in preparative and industrial quantities. P. amygdalus Hnl [for (R)-cyanohydrins] and the M. esculenta and H. brasiliensis Hnls [for (S)-cyanohydrins] all

Outlook

Over the past few years, tremendous progress has been achieved in the enzyme-catalysed cyanohydrin reaction. Not only has the broad applicability of this reaction been established but also enzymes for industrial application have been developed. This method will be used in the future to obtain enantiomerically pure chiral intermediates, and this method will become increasingly important.

Uncited references

22, 54, 55

Acknowledgements

We thank D. Johnson for his help in preparing the final version of the manuscript.

References (57)

M. Hasslacher
Molecular cloning of the full-length cDNA of (S)-hydroxynitrile lyase from Hevea brasiliensis: functional expression in Escherichia coli and Saccharomyces cerevisiae and identification of an active site residue
J. Biol. Chem.
(1996)
M.S. Jorns
Mechanism of catalysis by the flavoenzyme oxynitrilase
J. Biol. Chem.
(1979)
J. Brussee
Synthesis of optically active silyl protected cyanohydrins
Tetrahedron
(1990)
T.T. Huuhtanen et al.
Enzyme-catalysed synthesis of optically active aliphatic cyanohydrins
Tetrahedron Asymmetry
(1992)
F. Effenberger
Enzyme-catalysed synthesis of (R)-ketone cyanohydrins and their hydrolysis to (R)-α-hydroxy-α-methyl carboxylic acids
Tetrahedron Lett.
(1991)
F. Effenberger et al.
Stereoselective synthesis of thienyl and furyl analogues of ephedrine
Tetrahedron Asymmetry
(1997)
M. Schmidt
Preparation of optically active cyanohydrins using the (S)-hydroxynitrile lyase from Hevea brasiliensis
Tetrahedron
(1996)
H. Griengl
Enzyme catalysed formation of (S)-cyanohydrins derived from aldehydes and ketones in a biphasic solvent system
Tetrahedron
(1998)
U.G. Wagner
Mechanism of cyanogenesis: the crystal structure of hydroxynitrile lyase from Hevea brasiliensis
Structure
(1996)
J. Hughes
Purification, characterization, and cloning of α-hydroxynitrile lyase from cassava (Manihot esculenta Crantz)
Arch. Biochem. Biophys.
(1994)

K. Trummler et al.

Molecular cloning of acetone cyanohydrine lyase from flax (Linum usitatissimum): definition of a novel class of hydroxynitrile lyases

J. Biol. Chem.

(1997)

H. Breithaupt

Cloning and expression of (R)-hydroxynitrile lyase from Linum usitatissimum (flax)

J. Mol. Catal. B: Enzymatic

(1999)

E. Kiljunen et al.

Novel (R)-oxynitrilase sources for the synthesis of (R)-cyanohydrins in diisopropyl ether

Tetrahedron Asymmetry

(1997)

J. Hughes

Genomic organization and structure of α-hydroxynitrile lyase in cassava (Manihot esculenta Crantz)

Arch. Biochem. Biophys.

(1998)

C. Reimmann

Characterization of a rice gene induced by Pseudomonas syringae pv. syringae: requirement for the bacterial lemA gene function

Physiol. Mol. Plant Pathol.

(1995)

G. Boche

Electrophilic amination of acyl anion equivalents: mild oxidation of aldehyes to amides via O-(trimethylsilyl)- aldehyde cyanohydrin anions

Tetrahedron Lett.

(1982)

H. Wajant et al.

Identification of potential active-site residues in the hydroxynitrile lyase from Manihot esculenta by site-directed mutagenesis

J. Biol. Chem.

(1996)

D. Costes

Hydroxynitrile-lyase-catalysed synthesis of cyanohydrins in organic solvents: parameters influencing activity and enantiospecificity

Enzyme Microb. Technol.

(1999)

M. Bauer

Parameters influencing stability and activity of S-hydroxynitrile lyase from Hevea brasiliensis in two-phase systems

Enzyme Microb. Technol.

(1999)

J. Brussee

Bioorganic synthesis of optically active cyanohydrins and acyloins

Tetrahedron Lett.

(1988)

L. Rosenthaler

Durch Enzyme bewirkte asymmetrische Synthesen

Biochem. Z.

(1908)

V.K. Krieble et al.

The properties of oxynitrilase

J. Am. Chem. Soc.

(1921)

H. Albers et al.

Über die synthetisierende Wirkung des Emulsin

Biochem. Z.

(1932)

H. Albers et al.

Über die Kinetik der durch Emulsin beschleunigten Synthese des (+)-Mandelsäurenitrils

Biochem. Z.

(1934)

W. Becker

Cyanohydrin synthesis: purification and properties of oxynitrilase of bitter almonds

Biochem. Z.

(1963)

W. Becker et al.

Über das Flavinenzym d-Oxynitrilase

Biochem. Z.

(1966)

W. Becker

Stereospecific synthesis of d-hydroxy nitriles and optically active ethanolamines

Angew. Chem., Int. Ed. Engl.

(1965)

W. Becker et al.

Continuous synthesis of optically active α-hydroxynitriles

J. Am. Chem. Soc.

(1966)

Cited by (115)

Enzymes in dairy products
2021, Value-Addition in Food Products and Processing Through Enzyme Technology
The potential role of enzymes in the processing and production of food has cognizable pertinence. The dairy food manufacturing industries are considered as conventional exploiter of diverse enzymes. Enzyme symbolizes biological molecules and is defined as proteins that catalyze a particular reaction like brewing, milk processing, yogurt, cheese manufacturing, etc., intending to generate particular products. Various enzymes are existing in human bodies and are utilized to improve all chemical reactions to bolster life. With the advent of advanced-level technologies, numerous novels enzymes have been manufactured for their broad range applications with enhanced specificity which is still being explored. Predominantly, the growing concern of researchers to ameliorate the status of dairy food industries is toward interdisciplinary strategies of enzymes to assure rational and less expensive production processes by utilizing diverse novel enzymes. This chapter will acknowledge comprehensive information about various significant sources of enzymes and their utilization in particular production processes. Furthermore, the discussion holds strategies and approaches being used to improve the quality and quantity of dairy products by illustrating numerous examples. Finally, we will discuss in brief the future perspectives of enzymes to intensify the status of dairy food industries.
Nanoparticles decorated carbon nanotubes as novel matrix: A comparative study of influences of immobilization on the catalytic properties of Lens culinaris β-galactosidase (Lcβ-gal)
2020, International Journal of Biological Macromolecules
In the present study, Multiwalled carbon nanotubes (MWCNT) decorated with two different nanoparticles namely tungsten disulfide (WS₂) and tin oxide (SnO₂), nanocomposites (NCs) were synthesized via hydrothermal method. Spectroscopic studies showed that both synthesized NCs possess nearly same functional groups but MWCNT-SnO₂ NCs are rich in O-functional group. Microscopic studies revealed that both NCs have different morphological microstructure. Lens culinaris β-galactosidase (Lcβ-gal) was immobilized using glutaraldehyde cross-linker resulted in immobilization efficiency of 91.5% and 88% with MWCNT-WS₂ and MWCNT-SnO₂ NCs, respectively. Remarkable increase in rate of hydrolysis of whey lactose has been observed with both NCs i.e. Lcβ-gal immobilized MWCNT-WS₂ hydrolyzes the 97% whey lactose in 1.5 h while MWCNT-SnO₂ showed maximum 92% of whey hydrolysis in 2 h at optimum conditions. Both nanobiocatalyst could serve as a promising candidates for dairy industries and would offer a potential platform for enzyme based biosensor fabrication.
Catalytic applications of enzymes encapsulated in metal–organic frameworks
2019, Coordination Chemistry Reviews
Citation Excerpt :
To further expand enzyme applicability, biotechnology researchers have dedicated considerable effort to synthesizing designer biological macromolecules [10–13]. Currently, enzymes play a pivotal role in the food [14–16] and pharmaceutical [17–19] industries where substrate selectivity is of utmost importance. Intensive process optimization has even allowed enzymatic catalysis to contribute to the manufacture of biorenewables [20], polymers [21,22], fine chemicals [23], and bulk chemicals [24]; however, broader industrial utilization of enzymes is impeded by their poor stability in organic media, low recyclability, and dependence on coenzymes [25].
The global catalyst industry is valued at nearly 20 billion USD and demand continues to rise. Enzymes, biological macromolecules, are highly selective and efficient catalysts; however, their commercial implementation has been hindered due to their poor chemical and thermal stability and low reusability. Numerous strategies have been investigated to stabilize enzymes and improve their processability. Recently, metal–organic frameworks (MOFs) have been investigated as enzyme immobilization supports. Herein, we highlight the immobilization of enzymes in MOF cages and pores for catalysis applications and discuss the future of this enzyme encapsulation strategy.
Baliospermum montanum hydroxynitrile lyase catalyzed synthesis of chiral cyanohydrins in a biphasic solvent
2018, Biocatalysis and Agricultural Biotechnology
This study presents asymmetric synthesis of cyanohydrins in aqueous–organic biphasic system using crude Baliospermum montanum hydroxynitrile lyase (BmHNL). The effect of various biocatalytic parameters e.g. different organic solvents, ratio of organic solvents, substrate concentration (benzaldehyde and HCN), pH, and temperature were studied to achieve highest % conversion and enantiomeric excess of (S)-mandelonitrile. Among the organic solvents i.e. hexane, toluene, acetonitrile, tert-butyl methyl ether (TBME), di-isopropyl ether (DIPE) and n-butyl acetate studied, 10% n-butyl acetate showed best results. Optimal results were also found with 0.8 mM benzaldehyde, 10 min reaction time, pH 4.2 and substrate to KCN ratio of 1:125. Under optimal biocatalytic conditions more than a dozen of different aromatic aldehydes were converted into corresponding chiral (S)-cyanohydrins. This method also reports the synthesis of eight chiral (S)-cyanohydrins for the first time, not synthesized by any (S)-selective HNL earlier.
State-of-the-art protein engineering approaches using biological macromolecules: A review from immobilization to implementation view point
2018, International Journal of Biological Macromolecules
Citation Excerpt :
This wave of engineering enabled the use of biocatalysts for the biosynthesis of value-added pharmaceutical and fine chemicals. Notable examples are the lipase-driven development of a blood pressure drug (diltiazem), hydroxy nitrile-lyase-mediated biosynthesis of herbicides [27], synthesis of cholesterol-lowering statin drugs by carbonyl-reductase-catalyzed reaction [29], and nitrile-hydratase-catalyzed hydration of acrylonitrile to acrylamide for polymers [41]. In parallel to stabilizing features, the challenges now involved optimization of the biocatalyst towards unnatural substrates.
Over the past years, technological and scientific advances have proven biocatalysis as a sustainable alternative than traditional chemical catalysis including organo- or metallocatalysis. In this context, immobilization based approaches represent simple but effective routes for engineering enzyme catalysts with higher activities than wild-type derivatives. Many enzymes including oxidoreductases have been engineered by rational and directed evolution, to realize the catalytic activity, enantioselectivity, and stability attributes which are essential for their biotechnological exploitation. Induce yet stable activity in enzyme catalysis offer new insights and motivation to engineer efficient catalysts for practical and commercial purposes. It has now become possible to envisage substrate accessibility to the catalytic site of the enzyme by current computational capabilities that reduce the experimental work related to the enzyme selection, screening, and engineering. Herein, state-of-the-art protein engineering approaches for improving enzymatic activities including chemical modification, directed evolution, and rational design or their combination methods are discussed. The emphasis is also given to the applications of the resulting tailored catalysts ranging from fine chemicals to novel pharmaceutical compounds that use biocatalysts as a vital step.
Enzymatic C--C Bond Formation
2016, Organic Synthesis Using Biocatalysis
Carbon—carbon bond formation catalyzed by lyases and carboligases are finding increasing acceptance in chemical research and laboratory as well as large-scale production of stereo pure compounds. Carboligating enzymes being highly selective and efficient, offer a unique tool to perform asymmetric carbon—carbon bond formation in a sustainable, environmentally benign fashion. In this chapter, the most significant up to date carboligating enzymes, methodologies and procedures are described. This includes the catalytic addition of hydrogen cyanide by hydroxynitrile lyases, the umpolung aldol condensation by thiamin diphosphate (ThDP)-dependent enzymes, catalytic transfer of hydroxyacetyl unit by transketolases and aldol addition reactions of dihydroxyacetone phosphate (DHAP), 1-hydroxyalkan-2-ones (i.e., dihydroxyacetone, hydroxyacetone, and hydroxybutanone), pyruvate, glycine, acetaldehyde, and glycolaldehyde to a variety of electrophilic aldehyde structures.

View all citing articles on Scopus

View full text

Trends in Biotechnology

ReviewThe synthesis of chiral cyanohydrins by oxynitrilases

Abstract

Section snippets

Oxynitrilases: their occurrence, properties, structure and availability

Molecular cloning and characterization

Recombinant production

(R)- and (S)-selective Hnls, available enzymes and substrate acceptance

Outlook

Uncited references

Acknowledgements

J. Biol. Chem.

J. Biol. Chem.

Tetrahedron

Tetrahedron Asymmetry

Tetrahedron Lett.

Tetrahedron Asymmetry

Tetrahedron

Tetrahedron

Structure

Arch. Biochem. Biophys.

J. Biol. Chem.

J. Mol. Catal. B: Enzymatic

Tetrahedron Asymmetry

Arch. Biochem. Biophys.

Physiol. Mol. Plant Pathol.

Tetrahedron Lett.

J. Biol. Chem.

Enzyme Microb. Technol.

Enzyme Microb. Technol.

Tetrahedron Lett.

Durch Enzyme bewirkte asymmetrische Synthesen

Biochem. Z.

The properties of oxynitrilase

J. Am. Chem. Soc.

Über die synthetisierende Wirkung des Emulsin

Biochem. Z.

Über die Kinetik der durch Emulsin beschleunigten Synthese des (+)-Mandelsäurenitrils

Biochem. Z.

Cyanohydrin synthesis: purification and properties of oxynitrilase of bitter almonds

Biochem. Z.

Über das Flavinenzym d-Oxynitrilase

Biochem. Z.

Stereospecific synthesis of d-hydroxy nitriles and optically active ethanolamines

Angew. Chem., Int. Ed. Engl.

Continuous synthesis of optically active α-hydroxynitriles

J. Am. Chem. Soc.

Review
The synthesis of chiral cyanohydrins by oxynitrilases