Hevea brasiliensis 5-hydroxynitrile lyase

Hasslacher, M., Schall, M., Hayn, M. et al. (1997) High-level intracellular expression of hydroxynitrile lyase from the tropical rubber tree Hevea brasiliensis in microbial hosts. Protein Expression and Purification, 11, 61-71. 

Zuegg, J., Gruber, K., Gugganig, M. et al. (1999) Three-dimensional structures of enzyme-substrate complexes of the hydroxynitrile lyase from Hevea brasiliensis. Protein Science A Publication of the Protein Society, 8, 1990-2000. 

Wagner, U.G., Hasslacher, M., Griengl, H. et al. (1996) Mechanism of cyanogenesis the crystal structure of hydroxynitrile lyase from Hevea brasiliensis. Structure (London, England 1993), 4, 811-822. 

Veum, L., Hanefeld, U. and Pierre, A. (2004) The first encapsulation of hydroxynitrile lyase from Hevea brasiliensis in a sol-gel matrix. Tetrahedron, 60, 10419-10425. 

Purkarthofer, T., Gruber, K., Gruber-Khadjawi, M,et al. (2006) A biocatalytic Henry reaction - the hydroxynitrile lyase from Hevea brasiliensis also catalyzes nitroaldol reactions. Angewandte Chemie (International Edition in English), 45, 3454-3456. 

Johnson, D.V. and Griengl, H. (1997) The chemoenzymatic synthesis of (5)-13-hydroxyoctadeca-(9Z,ll )-dienoic acid using the hydroxynitrile lyase from Hevea brasiliensis. Tetrahedron, S3, 617-624. 

Another notable new advance is the development of hydroxynitrile lyases for the synthesis of enantiomerically active aromatic and aliphatic cyanohydrins. For instance, an S-specific hydroxynitrile lyase has been obtained from Hevea brasiliensis and the resulting fS)-cyanohydrin can be used to obtain both hydroxy acids and aminoalcohols. 

The highest reported heterologous production of a protein in yeast comes from the work by Hasslacher et al. (1997). The enzyme hydroxynitrile lyase (Hnl) from the tropical rubber tree Hevea brasiliensis was reported to produce levels of 22g/L intracellularly in P. pastoris. In the same study, S. cerevisiae and E. coli were tested in parallel experiments but were not competitive. Levels of proteins produced by yeasts have more typically been in the range of 1-15 g/L (Schmidt, 2004). A list of reported yields for expression of proteins in P. pastoris is provided by Cregg at http / faculty.kgi.edu/cregg/index.htm... 

Scheme 4.107 Enzymatic cyanohydrin formation catalyzed by HNL [429] (MTBE methyl-tert-butyl ether (R)-PaHNL (R)-hydroxynitrile lyase originating from Primus amygdalus (S)-HbHNL (S)-hydroxynitrile lyase originating from Hevea brasiliensis).

Selmar, D., Lieberei, R., Biehl, B. and Conn, E.E. (1989) a-Hydroxynitrile lyase in Hevea brasiliensis and its significance for rapid cyanogenesis. Physiol. Plantarum, 75, 97-101. 

These two methods involve the potential release of toxic HCN during the reaction and a cleaner as well as a safer method is to use acetone cyanohydrin 128 as a cyanide transfer reagent. The enzyme from the Brazilian rubber tree Hevea brasiliensis, called a hydroxynitrile lyase, catalyses cyanohydrin formation from aliphatic as well as aromatic aldehydes.40... 

Abbreviations HNL, hydroxynitrile lyase P.a., Prunus amygdalus L.u., Linum usitatissimum H.b., Hevea brasiliensis, M. e Manihot esculents. 

The chemoenzymatic synthesis of (13S)-hydroxy-18 2(9Z,ll ) was achieved in nine steps starting from (2 )-octenal. Of importance was the enzymatic conversion of (2 )-octenal to the (5)-cyanohydrin [5] with (S)-hydroxynitrile lyase cloned from Hevea brasiliensis (41). 13- Hydroxy-lO-oxo-18 1(11 ) was synthesized via a Kno-evenagel-type reaction of Isopropyl 11 - phenylsulfinyl-10-oxoundecanoate with hep-tanal to form y-hydroxyenone functionality together with carbon chain elongation (42). The regiospecific oxidation of a number of substituted unsaturated fatty esters with /7-benzoquinone in the presence of palladium(II) chloride under concomitant ultrasonic irradiation was reported. For example, methyl 9-hydroxy-18 1 (12Z) furnished methyl 9-hydroxy-12-keto-18 0 exclusively (43). 

Johnson, D.V., and H. Griengl, The Chemoenzymatic S5mthesis of (5)-13-Hydroxyoctadeca-(9Z, 1 l )-dienoic Acid Using the Hydroxynitrile Lyase from Hevea brasiliensis, Tetrahedron 53 617-624 (1997). 

On the other hand, (5)-hydroxynitrile lyases [1541-1544] were found in Sorghum bicolor [1545] (millet), Hevea brasiliensis [1546, 1547] (mbber tree). 

The (5)-hydroxynitrile lyase from Hevea brasiliensis has been made available in sufficient quantities by cloning and overexpressirai to allow industrial-scale applications [1563]. It should be noted that also a,p-unsaturated aliphatic aldehydes were transformed into the corresponding cyanohydrins in a clean reaction. No formation of saturated p-cyano aldehydes through Michael-type addition of hydrogen cyanide across the C=C double bond occurred. The latter is a common side reaction using traditional methodology. 

Bauer, M., Griengl, H., and Steiner, W. (1999) Parameters influencing stability and activity of a S-hydroxynitrile lyase from Hevea brasiliensis in two-phase systems. Enzyme Microb. Technol., 24, 514-522. 

Historically, enzyme catalysis has played a highly prominent role, with the first enzyme-catalyzed asymmetric addition of HCN to aldehydes dating back to 1908 [167]. A wide range of both aromatic and aliphatic ketones are suitable substrates and produce cyanohydrins of high optical purity. The most readily available and hence most commonly employed enzyme for asymmetric cyanohydrin formation is (R)-hydroxynitrile lyase isolated from almonds. Recent cloning and over-expression techniques have also made a number of (S)-hydroxynitrile lyases available for organic synthesis [164, 165]. This was utilized in Griengl s synthesis of coriolic acid (255), a natural product that displays calcium ionophoric activity and acts as a prostacyclin mimic (Scheme 2.32) [168]. Thus, an (S)-hydroxynitrile lyase was cloned from rubber trees (Hevea brasiliensis), overexpressed in Pichia pastoris, and used to provide cyanohydrin 254 in 99 % ee. 

S-Hydroxynitrile Lyases from Manihot esculenta and from Hevea brasiliensis... 

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