Article

The effects of zeolite and silica gel on synthesis of amylisobutyrate catalyzed by lipase from Candida rugosa

Hemijska industrija (Impact Factor: 0.56). 01/2008; 62(2). DOI: 10.2298/HEMIND0802064S
Source: DOAJ

ABSTRACT Lipase-catalyzed synthesis of esters has been widely investigated due to numerous advantages in comparison with the conventional chemical process catalyzed with concentrated acids since use of chemical catalysts leads to several problems. The most important disadvantages are corrosion of equipment, hazards of handling of the corrosive acids that are not reused, loss of conversion, yield and selectivity. The activity of water in reaction mixture seems to be one of the crucial factors affecting yield of enzymatic synthesis, since the excess of water promotes hydrolysis of the ester. Therefore, a significant part of related studies was focused on attempts to increase the yield of esters by efficient control of water activity. In our experimental work an attempt was made to regulate the activity of water by addition of water adsorbents in reaction mixture for synthesis of amyl isobutyrate. Two adsorbents were used: silica gel and molecular sieves (zeolites). The use of water adsorbents was optimized with respect to timing of adsorbent addition and pretreatment of adsorbent. The pretreatment of adsorbent with substrate did not increase the yield of ester. It was found that the timing of adsorbent addition is crucial factor. The addition in early phases of reaction leads to the decrease in ester yield compared to reaction without adsorbents. On the other hand, when the adsorbent was added in the late stage of reaction it exhibited beneficial influence on the yield of ester. The increase of yield was higher with silica gel (10%) than with molecular sieves (3.5%). These discrepancies can be ascribed to higher water adsorbing capacity of silica gel in systems with higher water content. Superior behavior of silica gel in ester synthesis in conjunction with simple process of regeneration recommends the use of this adsorbent for increasing the yield of lipase-catalyzed ester synthesis.

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Available from: D. Ž. Mijin, Dec 15, 2013
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    • "Lipaze spadaju među najznačajnije industrijske enzime, koji se uspešno primenjuju u prehrambenoj , farmaceutskoj i industriji detergenata, proizvodnji biodizela, itd. [3] [4]. Danas interesovanje za lipazama , prvenstveno mikrobnog porekla, raste zbog njihove izuzetne hemoselektivnosti, stereoselektivnosti i regioselektivnosti [5]. "
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    ABSTRACT: Yeast Candida antarctica produces two lipase forms, which are widely used as catalysts in variety of organic reactions, many of which are applied on a large scale. In this work, production of two forms of lipase from C. antarctica DSM 70725 (CAL A and CAL B) was monitored during seven days of cultivation in the optimal medium using different electrophoretic and zymographic techniques. According to electrophoresis after silver staining, C. antarctica lipase A (molecular mass 45 kDa) was produced starting from the second day of cultivation. C. antarctica lipase B (CAL B) was also produced starting from the second day, but protein was present in the fermentation broth predominantly as dimer (molecular weight 66 kDa), while presence of monomeric form of CAL B (molecular weight of 33 kDa) was observed starting from the fourth day of cultivation. Both types of zymograms (based on hydrolysis and synthesis reactions) were used for detection of lipase activity in the fermentation broth. C. antarctica lipase A showed activity only in hydrolytic zymogram, when alpha-naphtyl butyrate was used as substrate. In the same zymogram, with alpha-naphtyl acetate as substrate no CAL A activity was detected. Similarly, CAL A showed no activity in synthesis based zymograms towards oleic acid and octanol as substrates, indicating that CAL A is not active towards very short- or long-chain substrates. As opposite of CAL A, both monomeric and dimeric form of CAL B were detected in the all zymograms, suggesting that CAL B is active towards wide range of substrates, regardless to the chain length. Thus, zymogram based on hydrolysis of alpha-naphtyl butyrate represents a simple method for monitoring the production of two forms of lipase from C. antarctica, that greatly differ in their characteristics.
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