MANAGEMENT OF ENERGY EFFICIENT PRODUCTION OF INULINASE ASPERGILLUS AWAMORI 2250

Цель работы – повышение точности и надежности управления процессом производства биомассы аэробных микроорганизмов, снижение удельных энергозатрат и повышение выхода готовой культуры. Разработан алгоритм управления процессом производства биомассы аэробных микроорганизмов. В качестве объекта производства использован ферментный препарат инулиназа, полученный глубинным способом с использованием продуцента грибов Aspergillus awamori 2250. Процесс культивирования осуществлялся в вертикальном ферментере серии Biostat (Sartorius Stedim Biotech GmbH, Германия) с рабочим объемом 100 л, предназначенным для выращивания микроорганизмов глубинным способом. Для стабилизации температурных режимов культивирования в инокуляторе, непосредственном выращивании микроорганизмов в ферментере и охлаждении готовой культуры в приемных сборниках осуществляли подготовку теплой и холодной воды с использованием пароэжекторной холодильной машины, работающей в режиме теплового насоса. Алгоритм управления производства биомассы микроорганизмов Aspergillus awamori 2250 позволил обеспечить стабилизацию параметров в области оптимальных значений и увеличить выход готовой культуры; предотвратить выброс отработанных теплоносителей в окружающую среду; обеспечить существенную экономию теплоэнергетических затрат за счет рекуперации и утилизации отработанных энергоносителей в замкнутых термодинамических циклах по материальным и энергетическим потокам. The work purpose – increase in accuracy and reliability of management of process of production of biomass of aerobic microorganisms, decrease in specific energy consumption and increase in an exit of ready culture. Algorithm of process of production of biomass of aerobic microorganisms is developed. Process of cultivation was carried out in a vertical fermenter of Sartorius Stedim Biotech of the Biostat series with the working volume of 100 liters intended for cultivation of microorganisms. For stabilization of temperature conditions of cultivation in an inokulyator, direct cultivation of culture of microorganisms in a fermenter and cooling of ready culture in reception collections carried out preparation of warm and cold water with use of the paroezhektorny refrigerator working in the thermal pump mode. The control algorithm of production of biomass of microorganisms of Aspergillus awamori 2250 has allowed to provide stabilization of parameters in the field of optimum values and to increase an exit of ready culture; to prevent emission of the fulfilled heat carriers in the environment; to provide essential economy of heat power expenses due to recovery and utilization of the fulfilled energy carriers in the closed thermodynamic cycles on material and power streams.

UV-C Treatment Of Intermediates Of Biologicals To Inactivate Viruses Without Denaturing Desired Protein

Dedicated virus reduction steps implemented in the manufacturing process of biologicals, either isolated from human plasma or produced as recombinant proteins, are essential safety measures to assure that a potential virus contamination of the source material will not be transmitted to patients requiring these therapeutic proteins. Currently applied virus reduction steps as solvent/detergent treatment and virus filtration are very effective virus reduction methods with inherent method-dependent gaps regarding the reduction capacity for a very wide range of viruses of diverse physico-chemical characteristics: solvent/detergent treatment does not inactivate non-enveloped viruses and, depending on the pore size of the virus filter, small viruses are not removed when the desired protein is large and has to pass the filter. Therefore, another virus inactivation method was studied which is considered especially effective for small viruses: UV-C treatment using the UVivatec system provided by Sartorius Stedim Biotech GmbH, Göttingen, Germany. Experiments were performed to study the impact of UV-C treatment on the integrity of proteins employing fibrinogen as an example for a large protein and on the inactivation capacity for poliovirus (a small non-enveloped virus). The integrity of fibrinogen was assessed by comparing the untreated fibrinogen with the UV-C treated fibrinogen using HPLC, Clauss assay and thromboelastometry. Virus inactivation was studied in a bioassay using a sensitive cell culture infectivity assay employing a cynomolgus cell line. The results show that UV-C treatment inactivates viruses and modifies fibrinogen in a dose dependent manner; the monomer, dimer and polymer peak in the fibrinogen preparation studied changed from approx. 75% to 60%, 17% to 25% and 8% to 13%, respectively, at a UV-C intensity of 400 J/m² demonstrated by HPLC measurement. In order to protect fibrinogen from modifications, the antioxidant glutathione was added to the fibrinogen preparation. At an UV-C intensity of approx. 300 J/m², sufficient to effectively inactivate viruses studied, a modification of fibrinogen was not any longer detectable. Disclosures: Groener: CSL Behring: Employment.