Abstract
The rumen anaerobic fungusPiromonas communis, unlike the rumen anaerobic fungiNeocallimastix frontalis andNeocallimastix patriciarum, produced extracellular α-(4-O-methyl)-d-glucuronidase when grown in cultures containing filter-paper, barley straw, birchwood xylan or birchwood sawdust as carbon source. The highest concentration of enzyme was produced in cultures containing birchwood sawdust. The aldobiouronic acidO-α-(4-O-methyl-d-glucopyran-osyluronic acid)-(1 → 2)-d-xylopyranose (MeGlcAXyl) was the best substrate of those tested: the aldotriouronic acidO-α-(4-O-methyl-d-glucopyranosyluronic acid (1 → 2)-O-\-d-xylopyranosyl-(1 → 4)-d-xylopyranose (MeGlcAXyl2) and the aldotetraouronic acidO-α-(4-O-methyl-d-glucopyranosyluronic acid)-(1 → 2)-O-\-d-xylopyranosyl-(1 → 4)-O-\-d-xylopyranosyl-(1 → 4)-d-xylopyranose (MeGlcAXyl3) were also attacked but the rate fell as the degree of polymerisation increased. When the same substituted xylooligosaccharides were reduced to the corresponding alditols the enzyme activity disappeared. Similarly,p-nitrophenyl-α-d-glucuronide was not a substrate. Remarkably, the relative rates of attack shown by the α-(4-O-methyl)-d-glucuronidase on the aldouronic acids and on xylans extracted from birchwood, oat spelts and oat straw differed according to the carbon source used to produce the enzyme. The α-(4-O-methyl)-d-glucuronidase had a pH optimum of 5.5 and a temperature optimum of 50°C. On gel filtration the enzyme was shown to be associated with proteins covering the range 100–300 kDa, but a major peak of activity in the column effluent appeared to have a molecular mass of 103 kDa.
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References
Bucht B, Eriksson KE (1969) Extracellular enzyme system utilized by the rot fungusStereum sanguinolentum for the breakdown of cellulose. Arch Biochem Biophys 129:416–420
Garcia-Campayo V, Wood TM (1993) Purification and characterisation of a \-d-xylosidase from the anaerobic rumen fungusNeocallimastix frontalis. Carbohydr Res 242:229–245
Garcia-Campayo V, McCrae SI, Wood TM (1993) Hydrolysis of oligosaccharides of the \-(1 → 4)-linkedd-xylose series by endo-(1 → 4)-\-d-xylanase from the anaerobic rumen fungusNeocallimastix frontalis. World J Microbiol Biotechnol 10:64–68
Hébraud M, Fèvre M (1988) Characterization of glycoside and polysaccharide hydrolases secreted by the rumen anaerobic fungiNeocallimastix frontalis, Sphaeromonas communis andPiromonas communis. J Gen Microbiol 134:1123–1129
Hébraud M, Fèvre M (1990) Purification and characterisation of an extracellular \-xylosidase from the rumen anaerobic fungusNeocallimastix frontalis. FEMS Microbiol Lett 72:11–16
Johnson KG, Silva MC, Mackenzie CR, Schneider H, Fontana JD (1989) Microbial degradation of hemicellulosic materials. Appl Biochem Biotechnol 20/21:245–258
Khandake KM, Vithayathil PJ, Murthy SK (1989) Purification and characterisation of an α-d-glucuronidase from a thermophilic fungusThermoascus aurantiacus Arch Biochem Biophys 274:511–517
Korte H (1991) Reinigung und Charakterisierung einer α-Glucuronidase ausAgaricus Hbisporus (lge) Sing. und Untersuchungen an substiuiernten Xylooligomeren. PhD thesis, University of Hamburg
Levy GA, Marsh CA (1959) Preparation and properties of glucuronidases. Adv Carbohydr Chem 14:428–440
Lowe SE, Theodorou MK, Trinci APJ, Hespell RB (1985) Growth of anaerobic fungi on defined medium and semi-defined medium lacking rumen fluid. J Gen Microbiol 31:2225–2229
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Marsh CA (1958) Synthesis of aryl glycosiduronic acids. Biochem J 68:617–621
Mueller-Harvey L, Hartley RD, Harris PJ, Curzon EH (1986) Linkage ofp-coumaroyl and feruloyl groups to cell wall polysaccharides of barley straw. Carbohydr Res 148:71–85
Mountford DO (1987) The rumen anaerobic fungi FEMS Microbiol Rev 46:401–408
Nelson N (1952) A photometric adaptation of the Somogyi method for the determination of glucose. J Biol Chem 19:19–23
Orpin CG (1988) Genetic approaches to the improvement of lignocellulose degradation in the rumen In: Aubert JP, Beguin P, Millet J (eds) Biochemistry and genetics of cellulose degradation. Academic Press, London, pp 171–180
Puls J, Poutanen K (1989) Mechanism of enzymatic hydrolysis of hemicelluloses (xylan) and procedures for the determination of the enzyme activities involved In: Coughlan MP (ed) Enzyme systems for lignocellulose degradation. Elsevier Applied Science, Barking, UK, pp 151–166
Selvendran RR, Stevens BJH, O'Neill MA (1985) Developments in the isolation and analysis of cell walls from edible plants. In: Brett CT, Hillman JR (eds) Biochemistry of plant cell walls, Cambridge University Press, Cambridge, UK, pp 39–78
Scott J (1979) Colorimetric determination of hexuronic acids in plant materials. Anal Chem 51:936–941
Siika-aho M, Tenkanen M, Buchert J, Puls J, Viikari L (1994) An α-glucuronidase fromTrichoderma reesei RUT C-30. Enzyme Microb Technol 16:813–819
Smith MM, Hartley RD (1983) Occurrence and nature of ferulic acid substitution of cell wall polysaccharides in graminaceous plants. Carbohydr Res 118:65–80
Stephen AM (1983) Other plant polysaccharides In: Aspinall (ed) The polysaccharides, Vol 2. Academic Press, Orlando, Fla, pp 196–286
Sundberg M, Poutanen K (1991) Purification and properties of two acetylxylan esterases ofTrichoderma reesei. Biotechnol Appl Biochem 13:1–11
Uchida H, Nanri T, Kawabata Y, Kusakabe I, Murakami K (1992) Purification and characterisation of intracellularAspergillus niger 5–16. Biosci Biotechnol Biochem 56:1608–1615
Whistler RL, BeMiller JN (1963) Holocellulose from annual plants by chlorous acid delignification. In: Whistler RL (ed) Methods in carbohydrate chemistry. III. Academic Press, New York, pp 21–22
Williams AG, Orpin CG (1987) Glycoside hydrolase enzymes present in the zoospore and vegetative growth stages of the rumen fungiNeocallimastix patriciarum, Piromonas communis, and an unidentified isolated, grown on a range of carbohydrates. Can J Microbiol 33:427–434
Wilson CA, Wood TM (1992) The anaerobic fungusNeocallimastix frontalis: isolation and properties of a cellulosome-type enzyme fraction with the capacity to solubilize hydrogen bond-ordered cellulose. Appl Microbiol Biotechnol 37:125–129
Wood TM, Wilson CA (1995) Studies on the capacity of the cellulase of the anaerobic rumen fungusP. communis P to degrade hydrogen bond-ordered cellulose. Appl Biochem Biotechnol (in press)
Wood TM, Wilson CA, McCrae SI, Joblin KN (1986) A highly active extracellular cellulase from the anaerobic rumen fungusNeocallimastix frontalis. FEMS Microbiol Lett 34:37–40
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Wood, T.M., Wilson, C.A. α-(4-O-Methyl)-d-glucuronidase activity produced by the rumen anaerobic fungusPiromonas communis: A study of selected properties. Appl Microbiol Biotechnol 43, 893–900 (1995). https://doi.org/10.1007/BF02431925
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DOI: https://doi.org/10.1007/BF02431925