ß-Glucosidases from white rot fungus Phlebia radiata - purification and properties.

Monika A. Prendecka ,  Dorota Ścibior ,  Jerzy Rogalski 

Abstract

ß-Glucosidases are heterogenous group of enzymes, widely distributed from microorganisms to vertebrates, which hydrolyze O-glycosidic bonds at the terminal, nonreducing end of carbohydrates. ß-Glucosidases can be divided into three groups with respect to their ability to hydrolyze different substrates: enzymes that exhibit high specificity towards aryl-ß-D-glucosides, other ß-glucosidases preferentially hydrolyse cellobiose and cellooligosaccharides and are also called cellobiases and with broad-specificity, cleaving both types of substrates to the same extent [Decker et al., 2000].


ß-Glucosidases have potentially board application. The use of ß-glucosidase in the wine industry seems to be very interesting. It can hydrolyse the monoterpen glycosides which naturally occur in wines and are responsible for determination of aroma enhancement [Williams et al., 1982; Shosheyov et al., 1988]. Moreover ß-glucosidase can find application in process of bioconwersion of phenolic compounds from cranberries pomace [Zheng and Shetty, 2000]. Free fenolics and phenolic acids can be used as natural food ingradients as antioxidants [Bocco et al., 1998], nutraceuticals [Shetty, 1997]. Cranberry pomace contains many phenolic compounds which occur in bound forms of fenolic acids conjugated with glucose or other sugars. And this is why enzymatic hydrolysis of phenolic glycosides by for example ß-glucosidase appears to be an attractive mean of increasing the free fenolic acids in wine or fruit juice to enrich taste, aroma and potentially increasing nutraceutical value [Gueguen et al., 1997].


The purpose of this work was to isolate, purify and characterize the

ß-glucosidase and aryl-ß-glucosidase from culture filtrates of P. radiata. Phlebia radiata was found to secrete three ß-glucosidases when it was grown in liquid culture on various carbon sources. It was found that all isoforms ware optimally active at 50ºC and pH 5,0. Highly purified isoforms of ß-glucosidase (BGS) and aryl-ß-glucosidase (aBGSm, aBGS) ware obtained by means of the column chromatography on DEAE-Sepharose, SP-Sepharose and chromatofocussing on Polybuffer Exchanger PBE-94. All these forms were purified with purification fold over 80-times and their physico-chemical and kinetic characteristics were determined.

Key words: Phlebia radiata, b-glucosidase, aryl-b-glucosidase, purification, biodegradation

References:

Decker, C.H., Visser, J., Schreier, P. (2000) Appl Microbiol Biotechnol 55: 157-163.

Williams, P.J., Strauss, C.R., Wilson, B. Massy Weststapp R.A. (1982) Phytochemistry 21: 2013-20.

Shosheyov, O., Bravdo, B.A., Ikan,R. (1988) J Agric Food Chem 38: 1387-90.

Zheng, Z., and Shetty, K. (2000) J Agric Food Chem 48: 895-900.

Bocco, A., Cuvelier, M.E., Richard, H., Berset, C (1998) J Agric Food Chem 46: 2123-2129.

Shetty, K. (1997) Asia Pacific J Clin Nutr 6: 162-171.

Gueguen, Y., Chemardin, P., Pien, S., Arnaud, A., Galzy, P. (1997) J Biotechnol 55: 151-156.

 

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Presentation: Poster at Zjazd Polskiego Towarzystwa Biochemicznego, Sympozjum G, by Monika A. Prendecka
See On-line Journal of Zjazd Polskiego Towarzystwa Biochemicznego

Submitted: 2007-04-30 13:52
Revised:   2009-06-07 00:44