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Penicillium purpurogenum produces a highly stable endo-β-(1,4)-galactanase

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Abstract

The polysaccharides of galactose present in the pectin of the plant cell wall are degraded by endo-β-1,4-galactanases. The filamentous fungus Penicillium purpurogenum, which grows on a number of natural carbon sources, among them sugar beet pulp which contains pectin, has a gene (ppgal1) coding an endo-β-1,4-galactanase (PpGAL1). This enzyme was expressed heterologously in Pichia pastoris. It has a molecular mass of 38 kDa, a pH optimum of 4–4.5, and an optimal temperature of 60 °C. It is 100 % stable for up to 24 h at pH 4–4.5 and 40 °C. These stability properties, which exceed those from other endo-β-1,4-galactanases reported to date, make it particularly suitable for industrial processes requiring acidic conditions and temperatures up to 40 °C. PpGAL1 is, therefore, a potentially effective tool in the food industry and in other biotechnological applications.

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Acknowledgments

This work has been supported by grants from FONDECYT (1130180), Universidad Andrés Bello (DI-478-14/R and DI-31-12/R)

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  1. Departamento de Ciencias Biológicas, Universidad Andrés Bello, Santiago, Chile

    Vasni Zavaleta & Jaime Eyzaguirre

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  1. Vasni Zavaleta
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Correspondence to Jaime Eyzaguirre.

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Fig. S1

Analysis of the ppgal1 promoter. The GARE element (5′ TCCNCCAAT 3′) at -693 is highlighted in green. CREA binding sites at -353, -396, -503, -779, -784, -839 are shown in yellow. G underlined indicates a nucleotide shared by two CREA binding sites. (DOC 30 kb)

Fig. S2

Identification of the GARE element in fungal promoters. The GARE element (5′ TCCNCCAAT 3′), in green, is found in the promoters of the endo-β-1,4-galactanase gen of A. aculeatus (-335), A. niger (-299), E. nidulans (-195), P. chrysogenum (-346) and T. stipitatus (-154). Purple background indicates a nucleotide not conserved in GARE. (DOC 36 kb)

Fig. S3

Model of the molecular surface of the PpGAL1. a. Side view of the model. b. Front view of the model. The acid/base residue (Glu156) is indicated in yellow and the nucleophilic residue (Glu 267) in green. The image was obtained by means of the Swiss-PDB Viewer program. (DOCX 202 kb)

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Zavaleta, V., Eyzaguirre, J. Penicillium purpurogenum produces a highly stable endo-β-(1,4)-galactanase. Appl Biochem Biotechnol 180, 1313–1327 (2016). https://doi.org/10.1007/s12010-016-2169-6

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