Crude enzyme preparation from Candida

Enzyme Preparation. A crude enzyme mixture was prepared from the fermentate of Candida rugosa (American Type Culture Collection, ATCC No. 14830. The rugosa was revived and cultivated in YM Agar slants at 24°C for one week. The growth on one YM Agar slant was transferred aseptically to a two-liter flask containing 400 ml sterilized fermentation medium consisting of 2.0% defatted soyflour,... 

Several crude lipase preparations are available from the yeasts Candida lipolytica, C. antarctica (CAL), and C. rugosa (CRL, syn. C. cylindracea). The latter enzyme, the three-dimentional structure of which has been resolved by X-ray analysis [333], has been frequently used for the resolution of esters of secondary alcohols [401-406] and, to a lesser extent, for the resolution of a-substituted carboxylates [407,408]. The CRL preparations from several commercial sources which contain up to 16% of protein [409] differ to some extent in their activity but their selectivity is very similar [410]. As CRL is able to accommodate relatively bulky esters in its active site, it is the lipase of choice for the selective hydrolysis of esters of cyclic secondary alcohols. To illustrate this point, some representative examples are given below. 

The most common limitation for the enantioselective synthetic methods is poor enantio-selectivity. Crude Candida rugosa (CRL, formerly known as C. cylindracea) commercial lipase, one of the most used enzymes for the resolution of racemates, only exhibits poor to moderate enantiodiscrimination for the resolution of some racemic 2-arylpropionic acids and their derivatives (see Table lA). This might be due to the presence of other proteinaceous impurities (hydrolases and proteases with MW < 62 kDa) in the crude preparation [118], which may display opposite or poor enantioselectivity. A similar situation has also been described for crude lipase from pig pancreas [119]. 

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