Alcaligenes sp. ECU0401 has been isolated from soil samples with high nitrilase activity against glycolonitrile using the enrichment culture technique. The preferred carbon/nitrogen sources and metal ions were sodium acetate, a composite of peptone and yeast extract, and Cu(2+), respectively. Glycolic acid was obtained in a yield of 96.5% after 14 h of biotransformation from a total of 200 mM glycolonitrile in the mode of sequential addition during the cultivation of Alcaligenes sp. ECU0401 in a 5-L jar fermenter. Fifty micromolars of glycolonitrile could be hydrolyzed in a yield of 94.1% by resting cells after 36 h. The microbial nitrilase system could hydrolyze various nitriles with high activity, and no amidase activity and glycolic acid were observed in hydrolyzing glycolamide. It significantly exhibited high enantioselectivity in the hydrolysis of mandelonitrile and 2-chloromandelonitrile (>99.9% e.e.( p )). Efficient biocatalyst recycling was achieved as a result of immobilization in glutaraldehyde/polyethylenimine cross-linked carrageenan with immobilized cells exhibiting a biocatalyst productivity of 1,042.2 g glycolic acid per gram dry cell weight after 29 batch recycles.

A new strategy for the construction of optically active 5'-CF3 spiro[pyrrolidin-3,2'-oxindole] was described. A series of unprecedented 1,3-dipoles were obtained by condensation of CF3CH2NH2 with isatins. The 1,3-dipolar cycloaddition reactions of these ketimines with enals gave the products bearing four contiguous stereogenic centers in excellent yields, diastereoselectivities and enantioselectivities.

A Cinchona alkaloid-derived squaramidecatalyzed asymmetric cycloaddition of trfluoromethyl-containing azomethine ylides with b-nitroalkenes was realized under mild conditions.Aserieso fb iologically important 5'-trifluoromethyl-spiro[pyrrolidin-3,2'-oxindoles] was synthesizede fficiently by this process in excellent yields,e nantioseletivities and diastereoselectivities.

A catalytic asymmetric method for the synthesis of 2-CF3 chromanes has been described. Generally, the squaramide-catalyzed cascade reaction of 2-hydroxychalcones with β-CF3-nitroalkenes gave the CF3-containing heterocyclic compounds bearing three contiguous stereogenic centers in excellent yields, diastereoselectivities, and enantioselectivities.