Laboratory of Biotechnology of Enzymes

Arkady P. Sinitsyn
Professor, Dr.Sci. (Chemistry)
Head of Laboratory
INBI, build. 2, room 321
Телефон +7 (495) 939-59-66
E-Mail apsinitsyn@gmail.com


Keywords:

carbohydrase enzymes, filamentous fungi, biochemical properties of enzymes, genetic engineering, the expression of heterologous and homologous genes, the industrial enzyme, products of microbiological synthesis

Main achievements:

The main activity of the laboratory is the creation of technological bases of efficient conversion of renewable biomass to sugars and further into products with high added value – amino acids, organic solvents, carboxylic acids, bifunctional and multifunctional molecules for future use in production processes of chemical industry.

The optimization of the qualitative and quantitative composition of enzyme complexes for effective production of simple sugars from polysaccharide substrates of plant origin.
The study of biochemical and physico-chemical properties of the enzymes secreted by fungi Trichoderma reesei, Chrysosporium lucknowense, Thelavia terrestris, Penicillium canescens, Penicillium verruculosum, etc. with the aim of finding enzymes with a new, unique properties.
The use of fungal host-vector systems (Penicillium spp., Aspergillus sp.) for homologous and heterologous gene expression.

Rational design of protein molecules to improve operational and technological properties of enzymes.
Creation of new strains-producers of carbohysrases and other enzymes for commercial production.
Enzyme application in food, feed, textile, pulp and paper and other types of industry.

Since 2006 the laboratory was prepared 9 PhD, 10 graduate students in the specialties “chemistry” and “biotechnology”. Laboratory staff prepared more than 100 scientific publications, over 10 patents. The laboratory completed about 25 projects in the field of biotechnology and industrial enzymology, including 6 international projects. Scientific and technological bases of biotechnology processes for many industries were developed. On the basis of the strains and technologies, developed in the laboratory, the first in post-Soviet Russia a modern industrial Fermentation Plant (Agroferment) for the production of industrial enzymes was built up. The technological processes for environmentally friendly and energy efficient bioconversion of cellulose-containing non-food raw materials into simple sugars was developed; a Pilot Plant for production of simple sugars from non-food cellulose-containing raw material with a capacity of 300 tons/year was created.


Selected publications:

  1. Dotsenko A.S., Dotsenko G.S., Senko O.V., Stepanov N.A., Lyagin I.V., Efremenko E.N.,  Gusakov A.V., Zorov I.N., Rubtsova E.A. Complex effect of lignocellulosic biomass pretreatment with 1-butyl-3-methylimidazolium chloride ionic liquid on various aspects of ethanol and fumaric acid production by immobilized cells within SSF, Bioresource Technology, 2018, v.250, p.429-438
  2. Доценко A.С., Гусаков А.В., Рожкова А.М., Волков П.В., Синицын А.П., Ферментативный гидролиз целлюлозы смесями мутантных форм целлюлаз Penicillium verruculosum, Вестн.Моск.Ун-та. Серия 2. Химия, 2018, т.59, №2, с.138-143
  3. Рожкова А.М., Мерзлов Д.А., Баширова А.В., Зоров И.Н., Короткова О.Г., Шашков И.А., Синицын А.П. Новый ферментный препарат для снижения вязкости цельнозерновых экстрактов ржи, Вестн.Моск.Ун-та. Серия 2. Химия, 2018, т.59, №2, с.132-137
  4. Короткова О.Г., Рубцова Е.А., Шашков И.А., Волчок А.А., Кондратьева Е.Г., Синицына О.А., Рожкова А.М., Сатрудинов А.Д., Денисенко Ю.А., Синицын А.П. Сравнительный анализ состава и свойств кормовых ферментных препаратов, Катализ в промышленности, 2018, т.18, №4, с.72-78
  5. Dotsenko A., Gusakov A., Rozhkova A., Sinitsyna O., Sinitsyn A. Enzymatic hydrolysis of cellulosic materials using synthetic mixes of purified cellulases bioengineered at N-glycosylation sites, 3 Biotech, 2018, 8:396, http://doi.org/10.1007/s13205-018-1419-4
  6. Семёнова М.В., Волков П.В., Рожкова А.М., Зоров И.Н., Синицын А.П. Клонирование, выделение и изучение свойств новой гомологичной экзо-арабиназы гриба Penicillium canescens, Прикладная биохимия и микробиология, 2018, т.54, №4, с.375-384
  7. Volkov P.V., Gusakov A.V., Rubtsova E.A., Rozhkova A.M., Matys V.Yu.,  Nemashkalov V.A., Sinitsyn A.P. Properties of recombinant GH49 family dextranase heterologously expressed in two recipient strains of Penicillium species, 2019, Biochimie, v.157,  р.123-130, https://doi.org/10.1016/j.biochi.2018.11.010
  8. Denisenko Y., Rozhkova A., Zorov I., Bashirova A., Matys V., Nemashkalov V., Sinitsyn A., Gusakov A. Protein engineering of GH10 family xylanases for gaining a resistance to cereal proteinaceous inhibitors, 2019, Biocatalysis and Agricultural Biotechnology, https://doi.org/10.1016/j.bcab.2019.01.042 , v.17, p.690-695
  9. Semenova M.V., Gusakov A.V., Volkov P.V., Matys V.Yu., Nemashkalov V.A., Telitsin V.D., Rozhkova A.M., Sinitsyn A.P. Enhancement of the enzymatic cellulose saccharification by Penicillium verruculosum multienzyme cocktails containing homologously expressed lytic polysaccharide monooxygenase, Molecular Biology Reports, https://doi.org/10.1007%2Fs11033-019-04693-y
  10. Bashirova А., Pramanik S., Volkov P., Rozhkova A., Nemashkalov V., Zorov I., Gusakov A., Sinitsyn A., Schwaneberg U., Davari M. Disulfide bond engineering of an endoglucanase from Penicillium verruculosum to improve its thermostability,  International Journal of Molecular Science, 2019, v.20, 1602, doi.org/10.3390/ijms20071602
  11. Semenova M., Gusakov A., Telitsin V., Rozhkova A., Sinitsyn A. Purification and characterization of two forms of the homologously expressed lytic polysaccharide monooxygenase (PvLPMO9A) from Penicillium verruculosum, BBA-Protein and Proteomics, 2020, doi.org/10.1016/j.bbapap.2019.140297 1868(1),140297
  12. Contreras F., Pramanik S., Rozhkova A.M., Zorov I.N., Коrotkova O., Sinitsyn A.P., Schwaneberg U., Davari M.D. Engineering robust cellulases for tailored lignocellulosic degradation cocktails, Int.J.Mol.Sci., 2020, v.21, 1589, doi:103390/ijms21051589
  13. Dotsenko A.S., Dotsenko G.S., Rozhkova A.M., Zorov I.N., Sinitsyn A.P. Rational design and structure insights for thermostability improvement of Penicillium verruculosum Cel7A cellobiohydrolase, Biochimie, 176, 103-109
  14. Contreras F., Thiele M.J., Pramanik S., Rozhkova A., Dotsenko A., Zorov I., Sinitsyn A., Davari M.D., Schwaneberg U. KnowVolution of a GH5 Cellulase from Penicillium verruculosum to Improve Thermal Stability for Biomass Degradation, ACS Sustainable Chemistry and Engineering, 2020, https://dx.doi.org/10.1021/acssuschemeng.0c02465
  15. Karp S.G., Osipov D.O., Semenova M.V., Rozhkova A.M., Zorov I.N., Sinitsyna O.A., Soccol C.R., Sinitsyn A.P. Effect of novel Penicillium verruculosum enzyme preparations on the saccahrification of acid- and alkali-pretreated agro-industrial residues. Agronomy, 2020, 10, 1348, doi:10.3390/agronomy10091348
  16. Osipov D.O., Dotsenko G.S., Sinitsyna O.A., Kondratieva E.G., Zorov I.N., Shashkov I.A., Satrutdinov A.D., Sinitsyn A.P. Comparative study of the convertibility of agricultural residues and other cellulose-containing materials in hydrolysis with Penicillium verruculosum cellulase. Agronomy, 2020, 10, 1712; doi:10.3390/agronomy10111712
  17. Karp S.G., Rozhkova A.M., Semenova M.V., Osipov D.O., de Pauli S.T.Z., Sinitsyna O.A., Zorov I.N., de Souza Vandenberghe L.P., Soccol C.R., Sinitsyn A.P., Enzyme cocktails from Penicillium verruculosum, Penicillium canescens and Aspergillus spp. for the effective saccharification of agro-industrial wastes, Bioresource Technology, 2021, 124888, doi.org/10.1016/j.biortech.2021.124888
  18. Sinelnikov I.G., Siedhoff N.E., Chulkin A.M., Zorov I.N., Schwaneberg U., Davari M.D., Sinitsyna O.A., Shcherbakova L.A., Sinitsyn A.P., Rozhkova A.M. Expression and refolding of the plant chitinase from Drosera capensis for applications as a sustainable and integrated pest management, Frontiers in Bioengineering and Biotechnology, 2021, V.9, doi:10.3389/fbioe.2021.728501
  19. Pramanik S., Semenova M.V., Rozhkova A.M., Zorov I.N., Korotkova O.G., Sinitsyn A.P., Dvarai M.D. An engineered cellobiohydrolase I for sustainable degradation of lignocellulosic biomass, Biotechnology and Bioengineering, 2021, v.118, p.4014-4027, doi.org/10.1002/bit.27877
  20. Dotsenko A.G., Denisenko Yu.A., Rozhkova A.M., Zorov I.N., Shashkov I.A. Implementation of microfiltration in recycled usage of ionic liquid BmimCl and deep eutectic solvent ChCl/acetic acid for biomass conversion, Bioresource Technology Reports, 2022, 17, 100887, doi.org/10.1016/j.biteb.2021.100887
  21. Kislitsin V.Yu, Chulkin A.M., Zorov I.N., Denisenko Yu.A., Shashkov I.A., Sinitsyn A.P., Rozhkova A.M. The effect of cellobiohydrolase 1 gene knockout for composition and hydrolytic activity of the enzyme complex secreted by filamentous fungus Penicillium verruculosum, Bioresource Technoloogy Reports, 2022, 18, 101023