EFFECT OF VARIOUS INHIBITORS ON TREHALASE ACTIVITY IN ASPERGILLUS NIGER

Deepti Chrungu

Abstract


The black-spored fungus Aspergillusniger produces trehalase, which catalyzes the conversion of trehalose to glucose. Conidia from 15-day old culture were used for trehalase extraction, purification and characterization. Enzyme activity was assayed by estimating glucose released using 3,5-Dinitrosalicylic acid. The enzyme was purified 46-fold using Diethylaminoethyl-cellulose column chromatography. Then this purified enzyme was used for inhibition studies using three known trehalase inhibitors, Trehazolin, Validamycin and Validoxylamine A. The results showed that Trehazolin couldn’t inhibit trehalase whereas Validamycin and Validoxylamine A showed strong inhibition.


Keywords


Aspergillusniger, Trehalase, Inhibition, Trehazolin, Validamycin, ValidoxylamineA.

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References


Alizadeh P &Klionsky D J, (1996) Purification and biochemical characterization of the ATH1 gene product, vacuolar acid trehalase, from Saccharomyces cerevisiae. FEBS Letters, 391, p. 273-278.

Bradford M M, (1976) Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, p. 248-254.

Carroll D, Pastuszak I, Edavana V, Pan Y &Elbein A, (2007). A novel trehalase from Mycobacterium smegmatis- purification, properties, requirements. FEBS Journal, 274 (7), p. 1701-1714.

Francois J &Parrou J L, (2001) Reserve carbohydrates metabolism in the yeast Saccharomyces cerevisiae. FEMS Microbiology Reviews, 25, p. 125-145.

Frison M, Parrou J L, Guillaumot D, Masquelier D, Francois J, Chaumont F &Batoko H, (2007). The Arabidopsis thaliana is a plasma membrane-bound enzyme with extracellular activity. FEBS Letters, 581, p. 4010-4016.

Jin K, Peng G, Liu Y& Xia Y, (2015) The acid trehalase, ATM1, contributes to the in vivo growth and virulence of the entomopathogenic fungus, Metarhiziumacridum. Fungal Genetics and Biology, 77, p. 61-67.

Kamei Y, Hasegawa Y, Niimi T, Yamashita O & Yaginuma T, (2011). Trehalase-2 protein contributes to trehalase activity enhanced by diapauses hormone in developing ovaries of the silkworm, Bombyxmori. Journal of Insect Physiology, 57, p. 608-613.

Kamiya T, Hirata K, Matsumoto S, Arai C, Yoshizane C, Kyono F, Ariyasu T, Hanaya T, Arai S & Okura T, (2004) Targeted disruption of the trehalase gene: determination of the digestion and absorption of trehalose in trehalase-deficient mice. Nutrition Research, 24, p. 185-196.

Kandror O, Bretschneider N, Kreydin E, Cavalieri D & Goldberg A L, (2004) Yeast adapt to near-freezing temperatures by STRE/Msn2, 4-dependent induction of trehalose synthesis and certain molecular chaperones. Molecular Cell, 13, p. 771-781.

Kord M A E, Youssef E, Ahmed H &Qaid E, (2013) Purification and characterization of trehalase from seeds of chickpea CicerarietinumL. Turkish Journal of Biology, 37, p. 661-669.

Lillie S H & Pringle J R, (1980) Reserve carbohydrate metabolism in Saccharomyces cerevisiae: responses to nutrient limitation. Journal of Bacteriology, 143,p. 1384-1394.

Miller G L, (1959). Use of Dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry, 3, p. 426-428.

Murata M, Nagai M, Takao M, Suzuki A, Sakai T &Terashita T, (2001). Purification and characterization of an extracellular acid trehalase from Lentinulaedodes. Mycoscience, 42, p. 479-482.

Nwabuisi C &Ologe F E, (2001) The fungal profile of otomycosis patients in Ilorin, Nigeria. Nigerian Medical Journal, 10, p. 124-126.

Parrou J, Jules M, Beltran G & Francois J, (2005) Acid trehalase in yeasts and filamentous fungi: Localization, regulation and physiological function. FEMS Yeast Research, 5, p. 503-511

Reguera M, Peleg Z &Blumwald E, (2012) Targeting metabolic pathways for genetic engineering abiotic stress-tolerance in crops. BiochimicaetBiophysicaActa, 1819, p. 186-194.

Saubolle M A, (2000) Fungal pneumonias. Seminars in Respiratory Infections,15, p. 162-177.

Shukla E , Thorat L J, Bimalendu B N &Gaikwad S M, (2015) Insect trehalase: Physiological significance and potential applications. Glycobiology, 25(4), p. 357-367.

Silva M C P, Terra W R & Ferreira C, (2004) The role of carboxyl, guanidine and imidazole groups in catalysis by a midguttrehalase purified from an insect larvae. Insect Biochemistry and Molecular Biology, 34, p. 1089-1099.

Simola M, Hanninen A L, Stranius S M &Makarow M, (2000) Trehalose is required for conformational repair of heat denatured proteins in the yeast endoplasmic reticulum but not for maintenance of membrane traffic functions after severe heat stress. Molecular Microbiology 37, p.42-53.

Wiederhold N P, Lewis R E &Kontoyiannis D P, (2003) Invasiveaspergillosis in patients with hematologic malignancies. Pharmacotherapy, 23, p. 1592-1610.


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