Cloning of the scytalidium thermophilum bifunctional catalase / phenol oxidase gene and expression in aspergillus sojae

Erçin, Hatice Özlem
Scytalidium thermophilum is a thermophilic fungus with an important role in the composting process of mushroom cultivation. An extracellular phenol oxidase of Scytalidium thermophilum (STEP) with novel features was previously studied in our laboratory. This enzyme later turned out to be a catalase having phenol oxidase activity. The aim of this study was to clone Scytalidium thermophilum bifunctional catalase/phenol oxidase encoding gene and express the gene in Aspergillus sojae for future site directed mutagenesis studies. Scytalidium thermophilum catalase gene was first cloned into E. coli XL1 Blue MRF’ and then heterologously expressed in Aspergillus sojae ATCC11906. For that aim, the catalase gene was amplified using specific primers, excluding the signal and pro-peptide coding regions and amplified fragment was then cloned into E.coli XL1 Blue MRF’ and sequenced. It was observed that the cloned gene, named as catpo, was 10 amino acids different from the amino acid sequence of the S.thermophilum catalase gene formerly cloned by Novo Nordisk. The catpo gene encoding a mature protein of 681 amino acids was then ligated onto expression vector pAN52-4 and the recombinant plasmid was transformed into Aspergillus sojae ATCC11906. Heterologous expression was observed under the control of the glyceraldehydes 3-phosphate dehydrogenese promoter of Aspergillus nidulans and the secretion signal of the glucoamylase gene of Aspergillus niger. Catalase activity of the transformants reached at a level of 13206 U/g at the end of the fourth day of cultivation. However, this is still lower than the catalase activity of the gene donor strain of Scytalidium thermophilum.


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An identification method, based on the total cholesterol level determination for identifying whether archaeological food residues have animal or vegetable origin, has been developed. The method has been applied for the identification of various archaeological food samples excavated in Turkey belonging to the 800, 3000 and 4000 Bc. periods.
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Many plants produce allelopathic chemicals, such as stilbenes, to inhibit pathogenic fungi. The degradation of allelopathic compounds by bacteria associated with the plants would limit their effectiveness, but little is known about the extent of biodegradation or the bacteria involved. Screening of tissues and rhizosphere of peanut (Arachis hypogaea) plants revealed substantial enrichment of bacteria able to grow on resveratrol and pterostilbene, the most common stilbenes produced by the plants. Investigati...
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The identification of foodborne microorganisms and their endospores in food products are important for food safety. The present work compares Bacillus (Bacillas licheniformis, Bacillus circulans and Bacillus silbtilis) and Micrococcus (Micrococcus luteus) species with Fourier transform infrared (FTIR) spectroscopy. Our results show that there are several characteristic peaks belonging to both the Micrococcus and Bacillus Species Which Call be used for the identification of these foodborne bacteria and their...
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Gels prepared from whey proteins can be used for controlled release of nutrients or active ingredients in food systems. The objective of this study was to characterize the water uptake by these hydrophilic gels to aid in the design of release systems. Whey protein isolate (WPI) gels (17% w/w protein) of different aspect ratios were submersed in aqueous solution at pH 7.0. Modeling of mass uptake is presented in terms of Case I (Fickian diffusion) and Case II (kinetic) models. Due to the extent of swelling, ...
Molecular methods for identification of Lactobacillus delbrueckii subsp bulgaricus and Streptococcus thermophilus using methionine biosynthesis and 16S rRNA genes
Cebeci, Aysun; Gültekin, Güzin Candan (Cambridge University Press (CUP), 2008-11-01)
Yoghurt and starter culture producers are still searching strains of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus to produce healthier yogurt with longer shelf life, better texture, taste and quality. However, selective identification of Lb. delbrueckii subsp. bulgaricus and Strep. thermophilus from a mixed population using microbiological and biochemical methods is difficult, time consuming and may not be accurate. In this study, a quick, sensitive and accurate method is propo...
Citation Formats
H. Ö. Erçin, “Cloning of the scytalidium thermophilum bifunctional catalase / phenol oxidase gene and expression in aspergillus sojae,” M.S. - Master of Science, Middle East Technical University, 2008.