Physico-chemical properties and substrate specificity of α-(1→3)- d -glucan degrading recombinant mutanase from Trichoderma harzianum expressed in Penicillium verruculosum

Physico-chemical properties and substrate specificity of α-(1→3)- d -glucan degrading recombinant mutanase from Trichoderma harzianum expressed in Penicillium verruculosum Full article

Journal

Applied and Environmental Microbiology
ISSN: 0099-2240 , E-ISSN: 1070-6291

Output data

Year: 2025, Volume: 91, Number: 2, Article number : e00226-24, Pages count : DOI: 10.1128/aem.00226-24

Authors

Sinitsyna Olga A. ¹ , Volkov Pavel V. ² , Zorov Ivan N. ^1,2 , Rozhkova Alexandra M. ² , Emshanov Oleg V. ³ , Romanova Yulia M. ⁴ , Komarova Bozhena S. ⁵ , Novikova Natalia S. ⁵ , Nifantiev Nikolay E. ⁵ , Sinitsyn Arkady P. ^1,2

Affiliations

1	Department of Chemistry, M.V. Lomonosov Moscow State University, Moscow, Russia
2	Federal Research Centre «Fundamentals of Biotechnology» of the Russian Academy of Sciences
3	LLC BFR Laboratories, Moscow, Russia
4	The National Research Center for Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya of the Ministry of Health of the Russian Federation, Moscow, Russia
5	Laboratory of Glycoconjugate Chemistry, N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia

The gene mutAW encoding Trichoderma harzianum fungus mutanase (MutA, GH71 family, α-1,3-glucanase, EC 3.2.1.59) was cloned and heterologously expressed by the highly productive Penicillium verruculosum fungus. P. verruculosum MutA strain secreted crude enzyme preparations with the recombinant MutA content of 40% of the total secreted protein, and the specific activity increased 150 folds compared to that of enzyme preparation obtained by the host strain. Homogeneous MutA had molecular mass of 70 kDa and displayed maximum of the activity on mutan at pH 5.0 and 50°C, with Km and kcat being 1.0 g/L and 30 s−1, respectively. At 40-50°C, the MutA was stable for at least 3 h. Glucose was the main product of long-term mutan hydrolysis. HPLC analysis of hydrolysis product of oligo-α-(1→3)-D-glucosides bearing UV-detectable N-trans-cinnamoyl residue in the aglycon clearly indicated that MutA has an endo-processive hydrolytic mode of action. It was demonstrated that MutA can destroy the polysaccharide matrix of both gram-positive and gram-negative pathogenic bacteria biofilms.

Sinitsyna O.A. , Volkov P.V. , Zorov I.N. , Rozhkova A.M. , Emshanov O.V. , Romanova Y.M. , Komarova B.S. , Novikova N.S. , Nifantiev N.E. , Sinitsyn A.P.
Physico-chemical properties and substrate specificity of α-(1→3)- d -glucan degrading recombinant mutanase from Trichoderma harzianum expressed in Penicillium verruculosum
Applied and Environmental Microbiology. 2025. V.91. N2. e00226-24 . DOI: 10.1128/aem.00226-24 WOS Scopus OpenAlex

≡ Web of science:	WOS:001404815200001
≡ Scopus:	2-s2.0-85218455196
≡ OpenAlex:	W4406740990