Heterologous Expression of Thermogutta terrifontis Endo-Xanthanase in Penicillium verruculosum, Isolation and Primary Characterization of the Enzyme

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Yury A. Denisenko^1,a*, Olga G. Korotkova¹, Ivan N. Zorov^1,2, Alexandra M. Rozhkova¹, Margarita V. Semenova¹, Alexandr G. Elcheninov¹, Ilya V. Kublanov¹, and Arkady P. Sinitsyn^1,2

¹Federal Research Center “Fundamentals of Fundamental Biotechnology”, Russian Academy of Sciences, 119071 Moscow, Russia

²Department of Chemical Enzymology, Faculty of Chemistry, Lomonosov Moscow State University, 119991 Moscow, Russia

^* To whom correspondence should be addressed.

Received November 10, 2020; Revised December 29, 2020; Accepted December 29, 2020

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Heterologous endo-xanthanase (EX) from the thermophilic planktomycete Thermogutta terrifontis strain was obtained using Penicillium verruculosum 537 (ΔniaD) expression system with the cellobiohydrolase 1 gene promoter. Homogeneous EX with a molecular weight of 23.7 kDa (pI 6.5) was isolated using liquid chromatography methods. This xanthan degrading enzyme also possesses the enzymatic activity towards CM-cellulose, β-glucan, curdlan, lichenan, laminarin, galactomannan, xyloglucan but not towards p-nitrophenyl derivatives of β-D-glucose, mannose and cellobiose. The temperature and pH optima of EX were 55°C and 4.0, respectively; the enzyme exhibited 90% of its maximum activity in the temperature range 50-60°C and pH 3-5.

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KEY WORDS: endo-xanthanase, Thermogutta terrifontis, Penicillium verruculosum, heterologous expression, xanthan destruction

DOI: 10.1134/S000629792104009X

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