Enantioselective hydrolysis of a Schiff's base of D,L-phenylalanine ethyl ester in water-poor media via the reaction catalyzed with α-chymotrypsin immobilized on hydrophilic macroporous gel support

被引:26
作者
Belokon, YN
Kochetkov, KA
Plieva, FM
Ikonnikov, NS
Maleev, VI
Parmar, VS
Kumar, R
Lozinsky, VI
机构
[1] AN Nesmeyanov Organoelement Cpds Inst, Moscow 117813, Russia
[2] Univ Delhi, Dept Chem, Delhi 110007, India
关键词
biocatalysis in MeCN water-poor media; immobilized alpha-chymotrypsin; macroporous poly(vinyl alcohol) cryogel carrier; L-Phe; D-Phe;
D O I
10.1385/ABAB:88:1-3:097
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The application of immobilized alpha-chymotrypsin for the purpose of enantioselective hydrolysis of a Schiff's base of D,L-Phe-OEt (D,L-SBPH) in the mixed water-acetonitrile media with the different content of water is described. The immobilized biocatalyst was prepared by the chemical coupling of the enzyme to poly(vinyl alcohol) (PVA) cryogel-the macroporous hydrogel prepared by means of the freezing-thawing techniques. SBPH is water insoluble, and, therefore, acetonitrile (MeCN) with minor water additives was used as a solvent for the reaction of enantioselective hydrolysis of the racemic substrate. The process was conducted for 96-200 h, and L-Phe with the purity up to 98% e.e. precipitated in both the reaction medium and gel-carrier bulk. The product was recovered by washing the organo-insoluble sediment with aqueous ammonia. D-Phe with the purity up to 85% e.e. was recovered from the organic solution of D-ester after its acidic hydrolysis. The PVA-cryogel-attached enzyme was effective in SBPH hydrolysis in MeCN/water mixtures. The immobilized biocatalyst was active for more than 1 mo of application and could be successfully used after another 4 mo storage at +10 degrees C.
引用
收藏
页码:97 / 106
页数:10
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