Search results for: bioprocess biotechnology

Authors: Marcela dos P. G. Baltazar, Louise H. Gracioso, Luciana J. Gimenes, Bruno Karolski, Ingrid Avanzi, Elen A. Perpetuo

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In this work, bacterial strains were isolated from environmental samples from a copper mine and three of them presented potential for bioremediation of copper. All the strains were identified by mass spectrometry (MALDI-TOF-Biotyper) and grown in three diferent media supplemented with 100 ppm of copper chloride in flasks of 500mL and it was incubated at 28 °C and 180 rpm. Periodically, samples were taken and monitored for cellular growth and copper biosorption by spectrophotometer UV-Vis (600 nm) and Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES), respectively. At the end of exponential phase of cellular growth, the biomass was utilized to construct a correlation curve between absorbance and dry mass of the cells. Among the three isolates with potential for biorremediation, 1 strain exhibit capacity the most for bioremediation of effluents contaminated by copper being identified as Rhodococcus erythropolis.

Keywords: bioprocess, bioremediation, biosorption, copper

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Authors: Muhammad Zainuddin Arriafdi, Hamudah Hakimah Abdullah, Mohd Helmi Sani, Wan Azlina Ahmad, Muhd Nazrul Hisham Zainal Alam

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The biotechnology process development demands numerous experimental works. In laboratory environment, this is typically carried out using a shake flask platform. This paper presents the design and fabrication of a miniature bioreactor system as an alternative research tool for bioprocessing. The working volume of the reactor is 100 ml, and it is made of plastic. The main features of the reactor included stirring control, temperature control via the electrical heater, aeration strategy through a miniature air compressor, and online optical cell density (OD) sensing. All sensors and actuators integrated into the reactor was controlled using an Arduino microcontroller platform. In order to demonstrate the functionality of such miniature bioreactor concept, series of batch Saccharomyces cerevisiae fermentation experiments were performed under various glucose concentrations. Results attained from the fermentation experiments were utilized to solve the Monod equation constants, namely the saturation constant, Ks, and cells maximum growth rate, μmax as to further highlight the usefulness of the device. The mixing capacity of the reactor was also evaluated. It was found that the results attained from the miniature bioreactor prototype were comparable to results achieved using a shake flask. The unique features of the device as compared to shake flask platform is that the reactor mixing condition is much more comparable to a lab-scale bioreactor setup. The prototype is also integrated with an online OD sensor, and as such, no sampling was needed to monitor the progress of the reaction performed. Operating cost and medium consumption are also low and thus, making it much more economical to be utilized for biotechnology process development compared to lab-scale bioreactors.

Keywords: biotechnology, miniature bioreactor, research tools, Saccharomyces cerevisiae

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Authors: Francielo Vendruscolo

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This review summarizes the potential of starch agroindustrial residues as substrate for biohydrogen production. Types of potential starch agroindustrial residues, recent developments and bio-processing conditions for biohydrogen production will be discussed. Biohydrogen is a clean energy source with great potential to be an alternative fuel, because it releases energy explosively in heat engines or generates electricity in fuel cells producing water as only by-product. Anaerobic hydrogen fermentation or dark fermentation seems to be more favorable, since hydrogen is yielded at high rates and various organic waste enriched with carbohydrates as substrate result in low cost for hydrogen production. Abundant biomass from various industries could be source for biohydrogen production where combination of waste treatment and energy production would be an advantage. Carbohydrate-rich nitrogen-deficient solid wastes such as starch residues can be used for hydrogen production by using suitable bioprocess technologies. Alternatively, converting biomass into gaseous fuels, such as biohydrogen is possibly the most efficient way to use these agroindustrial residues.

Keywords: biofuel, dark fermentation, starch residues, food waste

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Authors: Sirous Eydivandi

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Diacylglycerol-O-acyltransferase (DGAT1) gene encodes diacylglycerol transferase enzyme that plays an important role in glycerol lipid metabolism. DGAT1 is considered to be the key enzyme in controlling the synthesis of triglycerides in adipocytes. This enzyme catalyzes the final step of triglyceride synthesis (transform triacylglycerol (DAG) into triacylglycerol (TAG). A total of 20 DGAT1 gene sequences and corresponding amino acids belonging to 4 species include cattle, goats, sheep and yaks were analyzed, and the differentiation within and among the species was also studied. The length of the DGAT1 gene varies greatly, from 1527 to 1785 bp, due to deletion, insertion, and stop codon mutation resulting in elongation. Observed genetic diversity was higher among species than within species, and Goat had more polymorphisms than any other species. Novel amino acid variation sites were detected within several species which might be used to illustrate the functional variation. Differentiation of the DGAT1 gene was obvious among species, and the clustering result was consistent with the taxonomy in the National Center for Biotechnology Information.

Keywords: DGAT1gene, bioinformatic, ruminnants, biotechnology information

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Authors: Ana L. Sánchez Regla, Igor A. Rivera González, María del Pilar Monserrat Pérez Hernández

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The ability of an organization to identify and get useful information from external sources, assimilate it, transform and apply to generate products or services with added value is called absorptive capacity. Absorptive capabilities contribute to have market opportunities to firms and get a leader position with respect to others competitors. The Bioprocess Development and Research Unit (UDIBI) is a Research and Development (R&D) laboratory that belongs to the National Polytechnic Institute (IPN), which is a higher education institute in Mexico. The UDIBI was created with the purpose of carrying out R and D activities for the Transferon®, a biopharmaceutical product developed and patented by IPN. The evolution of competence and scientific and technological platform made UDIBI expand its scope by providing technological services (preclínical studies and bio-compatibility evaluation) to the national pharmaceutical industry and biopharmaceutical industry. The relevance of this study is that those industries are classified as high scientific and technological intensity, and yet, after a review of the state of the art, there is only one study of absorption capabilities in biopharmaceutical industry with a similar scope to this research; in the case of Mexico, there is none. In addition to this, UDIBI belongs to a public university and its operation does not depend on the federal budget, but on the income generated by its external technological services. This fact represents a highly remarkable case in Mexico's public higher education context. This current doctoral research (2015-2019) is contextualized within a case study, its main objective is to identify and analyze the absorptive capabilities that characterise the UDIBI that allows it had become in a one of two third authorized laboratory by the sanitary authority in Mexico for developed bio-comparability studies to bio-pharmaceutical products. The development of this work in the field is divided into two phases. In a first phase, 15 interviews were conducted with the UDIBI personnel, covering management levels, heads of services, project leaders and laboratory personnel. These interviews were structured under a questionnaire, which was designed to integrate open questions and to a lesser extent, others, whose answers would be answered on a Likert-type rating scale. From the information obtained in this phase, a scientific article was made (in review and a proposal of presentation was submitted in different academic forums. A second stage will be made from the conduct of an ethnographic study within this organization under study that will last about 3 months. On the other hand, it is intended to carry out interviews with external actors around the UDIBI (suppliers, advisors, IPN officials, including contact with an academic specialized in absorption capacities to express their comments on this thesis. The inicial findings had shown two lines: i) exist institutional, technological and organizational management elements that encourage and/or limit the creation of absorption capacities in this scientific and technological laboratory and, ii) UDIBI has had created a set of multiple transfer technology of knowledge mechanisms which have had permitted to build a huge base of prior knowledge.

Keywords: absorptive capabilities, biopharmaceutical industry, high research and development intensity industries, knowledge management, transfer of knowledge

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Authors: Nelson Barrientos, Matthew J. Davies, Marco C. Marques, Darren N. Nesbeth, Gary J. Lye, Nicolas Szita

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Microbioreactor technology is making important advances towards its application in cell culture and bioprocess development. In particular, the technology promises flexible and controllable devices capable to perform parallelised experimentation at low cost. Currently, state of the art methods (e.g. optical sensors) allow the accurate monitoring of the microbioreactor operation. In addition, the laminar flow regime encountered in these devices allows more predictive fluid dynamics modelling, improving the control over the soluble, physical and mechanical environment of the cells. This work describes the development and characterisation of a novel microbioreactor cassette system (microbioreactor volume is 150 μL. The volumetric oxygen transfer coefficient (KLa) and mixing time have been characterised to be between 25 to 113 h-1 and 0.5 and 0.1 s, respectively. In addition, the Residence time distribution (RTD) analysis confirms that the reactor operates at well mixed conditions. Finally, Staphylococcus carnosus TM300 growth is demonstrated via batch culture experiments. Future work consists in expanding the optics of the microbioreactor design to include the monitoring of variables such as fluorescent protein expression, among others.

Keywords: microbioreactor, cell-culture, fermentation, microfluidics

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Authors: Elsayed Ahmed Elsayed, Azza Noor El-Deen, Mohamed A. Farid, Mohamed A. Wadaan

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Recently, a great attention has been paid to the use of dietary carbohydrates as prebiotic functional foods. Among the new commercially available products, fructooligosaccharides (FOS), which are microbial produced from sucrose, have attracted special interest due to their valuable properties and, thus, have a great economic potential for the sugar industrial branch. They are non-cariogenic sweeteners of low caloric value, as they are not hydrolyzed by the gastro-intestinal enzymes, promoting selectively the growth of the bifidobacteria in the colon, helping to eliminate the harmful microbial species to human and animal health and preventing colon cancer. FOS has been also found to reduce cholesterol, phospholipids and triglyceride levels in blood. FOS has been mainly produced by microbial fructosyltransferase (FTase) enzymes. The present work outlines bioprocess optimization for different cultivation parameters affecting the production of FTase by Penicillium aurantiogriseum AUMC 5605. The optimization involves both traditional as well as fractional factorial design approaches. Additionally, the production process will be compared under batch and fed-batch conditions. Finally, the optimized process conditions will be applied to 5-L stirred tank bioreactor cultivations.

Keywords: prebiotics, fructooligosaccharides, optimization, cultivation

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Authors: Sushil Pradhan

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Biotechnology contributes to sustainable development in several ways such as biofertilizer production, biopesticide production and management of environmental pollution, tissue culture and biodiversity conservation in vitro, in vivo and in situ, Insectivorous medicinal plant Drosera burmannii Vahl belongs to the Family-Droseraceae under Order-Caryophyllales, Dicotyledoneae, Angiospermeae which has 31 (thirty one) living genera and 194 species besides 7 (seven) extinct (fossil) genera. Locally it is known as “Patkanduri” in Odia. Its Hindi name is “Mukhajali” and its English name is “Sundew”. The earliest species of Drosera was first reported in 1753 by Carolous Linnaeus called Drosera indica L (Indian Sundew). The latest species of Drosera reported by Fleisch A, Robinson, AS, McPherson S, Heinrich V, Gironella E and Madulida D.A. (2011) is Drosera ultramafica from Malaysia. More than 50 % species of Drosera have been reported from Australia and next to Australia is South Africa. India harbours only 3 species such as D. indica L, Drosera burmannii Vahl and D. peltata L. From our Odisha only D. burmannii Vahl is being reported for the first time from the district of Subarnapur near Sonepur (Arjunpur Reserve Forest Area). Drosera plant is autotrophic but to supplement its Nitrogen (N2) requirement it adopts heterotrophic mode of nutrition (insectivorous/carnivorous) as well. The colour of plant in mostly red and about 20-30cm in height with beautiful pink or white pentamerous flowers. Plants grow luxuriantly during November to February in shady and moist places near small water bodies of running water stream. Medicinally it is a popular herb in the locality for the treatment of cold and cough in children in rainy season by the local Doctors (Kabiraj and Baidya). In the present field investigation an attempt has been made to understand the unique reproductive phase and life cycle of the plant thereby planning for its conservation and propagation through various techniques of tissue culture and biotechnology. More importantly besides morphological and anatomical studies, cytological investigation is being carried out to find out the number of chromosomes in the cell and its genomics as there is no such report as yet for Drosera burmannii Vahl. The ecological significance and biodiversity conservation of Drosera with special reference to energy, environmental and chemical engineering has been discussed in the research paper presentation.

Keywords: insectivorous, medicinal, drosera, biotechnology, chromosome, genome

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Authors: Reshu Chaudhary, R. S. Sengar

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To increase the agricultural production Indian farmer needs to aware of the latest technology i.e. precision farming to maximize the crop yield and minimize the input (fertilizer, pesticide etc.) through monitoring the environmental factors. Biotechnology and information technology have provided lots of opportunities for the development of agriculture. But, still we have to do much more for increasing our agricultural production in order to achieve the target growth of agriculture to secure food, to eliminate poverty and improve living style, to enhance agricultural exports and national income and to improve quality of agricultural products. Nanotechnology can be a great element to satisfy these requirements and to boost the multi-dimensional development of agriculture in order to fulfill the dream of Indian farmers. Nanotechnology is the most rapidly growing area of science and technology with its application in physical science, chemical science, life science, material science and earth science. Nanotechnology is a part of any nation’s future. Research in nanotechnology has extremely high potential to benefit society through application in agricultural sciences. Nanotechnology has greater potential to bring revolution in the agricultural sector.

Keywords: agriculture, biotechnology, crop yield, nanotechnology

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Authors: R. Hurtado-Ortiz, D. Clermont, M. Schüngel, C. Bizet, D. Smith, E. Stackebrandt

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Microbiological resources and their derivatives are the essential raw material for the advancement of human health, agro-food, food security, biotechnology, research and development in all life sciences. Microbial resources, and their genetic and metabolic products, are utilised in many areas such as production of healthy and functional food, identification of new antimicrobials against emerging and resistant pathogens, fighting agricultural disease, identifying novel energy sources on the basis of microbial biomass and screening for new active molecules for the bio-industries. The complexity of public collections, distribution and use of living biological material (not only living but also DNA, services, training, consultation, etc.) and service offer, demands the coordination and sharing of policies, processes and procedures. The Microbial Resource Research Infrastructure (MIRRI) is an initiative within the European Strategy Forum Infrastructures (ESFRI), bring together 16 partners including 13 European public microbial culture collections and biological resource centres (BRCs), supported by several European and non-European associated partners. The objective of MIRRI is to support innovation in microbiology by provision of a one-stop shop for well-characterized microbial resources and high quality services on a not-for-profit basis for biotechnology in support of microbiological research. In addition, MIRRI contributes to the structuring of microbial resources capacity both at the national and European levels. This will facilitate access to microorganisms for biotechnology for the enhancement of the bio-economy in Europe. MIRRI will overcome the fragmentation of access to current resources and services, develop harmonised strategies for delivery of associated information, ensure bio-security and other regulatory conditions to bring access and promote the uptake of these resources into European research. Data mining of the landscape of current information is needed to discover potential and drive innovation, to ensure the uptake of high quality microbial resources into research. MIRRI is in its Preparatory Phase focusing on governance and structure including technical, legal governance and financial issues. MIRRI will help the Biological Resources Centres to work more closely with policy makers, stakeholders, funders and researchers, to deliver resources and services needed for innovation.

Keywords: culture collections, microbiology, infrastructure, microbial resources, biotechnology

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Authors: H. Necef, A. Benayad

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Fungi are becoming more and more important in our life. Therefore, as a start for the symposium on filamentous fungi in biotechnology a short survey of the role of fungi in biotechnology. Salin soils occupy about 7% of land area; they are characterized by unsuitable physical conditions for the growth of living organisms. However, researches showed that some microorganisms especially fungi are able to grow and adapt to such extreme conditions; it is due to their ability to develop different physiological mechanisms in their adaptation. This is the first study on the physiological and biological characteristics of El-Beida marsh. Nine soil samples were taken at different points in two steps, the first was in winter (low temperature), and the second was in summer (high temperature). The physicochemical analyses of the soil were conducted, then the isolation process was applied using two methods, direct method and dilution method (10-1, 10-2, 10-3, 10-4). Different species of fungi were identified belong to 21 genera in addition to 3 yeast species, Aspergillus showed the highest proportion by 43%, then Penicillium by 20% then Alternaria by 7%, in addition to various genera in different proportions. As for the sampling periods, it was observed that the spread of fungi in winter was higher than in summer with the proportion 75.47% and 24.53% respectively. Some halotolerant fungi have a biotechnological importance especially if the salinity of the medium is necessary for the fermentation, and if the halotolerance genes of the fungus will define, this will open the research to study and improve this property for the industrial important micro-organisms.

Keywords: salinity, identification, aspergillus oryzae, halotolerance, fungi

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Authors: Mohsen Rezaei Aghdam

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Recombinant DNA technology has meaningfully augmented the conventional crop improvement and has a great possibility to contribution plant breeders to encounter the augmented food request foretold for the 21st century. Predictable changes in weather and its erraticism, chiefly extreme fevers and vicissitudes in rainfall are expected to brand crop upgrading even more vital for food manufacture. Tissue attitude has been downtrodden to create genetic erraticism from which harvest plants can be better, to improve the state of health of the recognized physical and to upsurge the number of wanted germplasms obtainable to the plant breeder. This appraisal delivers an impression of the chances obtainable by the integration of vegetable biotechnology into plant development efforts and increases some of the social subjects that need to be considered in their application. Public-private companies offer chances to catalyze new approaches and investment while accelerating integrated research and development and commercial supply chain-based solutions. Novel varieties derivative by encouraged mutatgenesis are used commonly: rice in Thailand. These paper combinations obtainable data about the influence of change breeding-derived crop changes around the world, traveler magnetism the possibility of mutation upbringing as a flexible and feasible approach appropriate to any crop if that suitable objectives and selection approaches are used.

Keywords: crop, improve, genetic, agricultural

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Authors: Eugene C. Lim

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This paper seeks to examine the extent to which ‘forum shopping’ is available to patent filers seeking protection of GMO (genetically modified organisms)-based inventions in Hong Kong. Under Hong Kong’s current re-registration system for standard patents, an inventor must first seek patent protection from one of three Designated Patent Offices (DPO) – those of the People’s Republic of China (PRC), the Europe Union (EU) (designating the UK), or the United Kingdom (UK). The ‘designated patent’ can then be re-registered by the successful patentee in Hong Kong. Interestingly, however, the EU and the PRC do not adopt a harmonized approach toward the patenting of GMOs, and there are discrepancies in their interpretation of the phrase ‘animal or plant variety’. In view of these divergences, the ability to effectively manage ‘conflict of law’ issues is an important priority for multinational biotechnology firms with a patent portfolio in the Greater China region. Generally speaking, both the EU and the PRC exclude ‘animal and plant varieties’ from the scope of patentable subject matter. However, in the EU, Article 4(2) of the Biotechnology Directive allows a genetically modified plant or animal to be patented if its ‘technical feasibility is not limited to a specific variety’. This principle has allowed for certain ‘transgenic’ mammals, such as the ‘Harvard Oncomouse’, to be the subject of a successful patent grant in the EU. There is no corresponding provision on ‘technical feasibility’ in the patent legislation of the PRC. Although the PRC has a sui generis system for protecting plant varieties, its patent legislation allows the patenting of non-biological methods for producing transgenic organisms, not the ‘organisms’ themselves. This might lead to a situation where an inventor can obtain patent protection in Hong Kong over transgenic life forms through the re-registration of a patent from a more ‘biotech-friendly’ DPO, even though the subject matter in question might not be patentable per se in the PRC. Through a comparative doctrinal analysis of legislative provisions, cases and court interpretations, this paper argues that differences in the protection afforded to GMOs do not generally prejudice the ability of global MNCs to obtain patent protection in Hong Kong. Corporations which are able to first obtain patents for GMO-based inventions in Europe can generally use their European patent as the basis for re-registration in Hong Kong, even if such protection might not be available in the PRC itself. However, the more restrictive approach to GMO-based patents adopted in the PRC would be more acutely felt by enterprises and inventors based in mainland China. The broader scope of protection offered to GMO-based patents in Europe might not be available in Hong Kong to mainland Chinese patentees under the current re-registration model for standard patents, unless they have the resources to apply for patent protection as well from another (European) DPO as the basis for re-registration.

Keywords: biotechnology, forum shopping, genetically modified organisms (GMOs), greater China region, patent portfolio

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Authors: Deepak Loura

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Production and productivity of several crops in the country continue to be adversely affected by biotic (e.g., Insect-pests and diseases) and abiotic (e.g., water temperature and salinity) stresses. Over-dependence on pesticides and other chemicals is economically non-viable for the resource-poor farmers of our country. Further, pesticides can potentially affect human and environmental safety. While traditional breeding techniques and proper- management strategies continue to play a vital role in crop improvement, we need to judiciously use biotechnology approaches for the development of genetically modified crops addressing critical problems in the improvement of crop plants for sustainable agriculture. Modern biotechnology can help to increase crop production, reduce farming costs, and improve food quality and the safety of the environment. Genetic engineering is a new technology which allows plant breeders to produce plants with new gene combinations by genetic transformation of crop plants for improvement of agronomic traits. Advances in recombinant DNA technology have made it possible to have genes between widely divergent species to develop genetically modified or genetically engineered plants. Plant genetic engineering provides the strength to harness useful genes and alleles from indigenous microorganisms to enrich the gene pool for developing genetically modified (GM) crops that will have inbuilt (inherent) resistance to insect pests, diseases, and abiotic stresses. Plant biotechnology has made significant contributions in the past 20 years in the development of genetically engineered or genetically modified crops with multiple benefits. A variety of traits have been introduced in genetically engineered crops which include (i) herbicide resistance. (ii) pest resistance, (iii) viral resistance, (iv) slow ripening of fruits and vegetables, (v) fungal and bacterial resistance, (vi) abiotic stress tolerance (drought, salinity, temperature, flooding, etc.). (vii) quality improvement (starch, protein, and oil), (viii) value addition (vitamins, micro, and macro elements), (ix) pharmaceutical and therapeutic proteins, and (x) edible vaccines, etc. Multiple genes in transgenic crops can be useful in developing durable disease resistance and a broad insect-control spectrum and could lead to potential cost-saving advantages for farmers. The development of transgenic to produce high-value pharmaceuticals and the edible vaccine is also under progress, which requires much more research and development work before commercially viable products will be available. In addition, molecular-aided selection (MAS) is now routinely used to enhance the speed and precision of plant breeding. Newer technologies need to be developed and deployed for enhancing and sustaining agricultural productivity. There is a need to optimize the use of biotechnology in conjunction with conventional technologies to achieve higher productivity with fewer resources. Therefore, genetic modification/ engineering of crop plants assumes greater importance, which demands the development and adoption of newer technology for the genetic improvement of crops for increasing crop productivity.

Keywords: biotechnology, plant genetic engineering, genetically modified, biotic, abiotic, disease resistance

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Authors: Tomasz Borowski, Dawid Sołoducha, Rafał Rakoczy, Marian Kordas

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The application of magnetic field is essential for a wide range of technologies or processes (i.e., magnetic hyperthermia, bioprocessing). From the practical point of view, bioprocess control is often limited to the regulation of temperature at constant values favourable to microbial growth. The main aim of this study is to determine the effect of various types of electromagnetic fields (i.e., static or alternating) on the heat transfer in a self-designed magnetically assisted reactor. The experimental set-up is equipped with a measuring instrument which controlled the temperature of the liquid inside the container and supervised the real-time acquisition of all the experimental data coming from the sensors. Temperature signals are also sampled from generator of magnetic field. The obtained temperature profiles were mathematically described and analyzed. The parameters characterizing the response to a step input of a first-order dynamic system were obtained and discussed. For example, the higher values of the time constant means slow signal (in this case, temperature) increase. After the period equal to about five-time constants, the sample temperature nearly reached the asymptotic value. This dynamical analysis allowed us to understand the heating effect under the action of various types of electromagnetic fields. Moreover, the proposed mathematical description can be used to compare the influence of different types of magnetic fields on heat transfer operations.

Keywords: heat transfer, magnetically assisted reactor, dynamical analysis, transient function

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Authors: Rosa Buxeda, Lorenzo Saliceti-Piazza, Rodolfo J. Romañach, Luis Ríos, Sandra L. Maldonado-Ramírez

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Biopharmaceuticals manufacturing is one of the major economic activities worldwide. Ninety-three percent of the workforce in a biomanufacturing environment concentrates in production-related areas. As a result, strategic collaborations between industry and academia are crucial to ensure the availability of knowledgeable workforce needed in an economic region to become competitive in biomanufacturing. In the past decade, our institution has been a key strategic partner with multinational biotechnology companies in supplying science and engineering graduates in the field of industrial biotechnology. Initiatives addressing all levels of the educational pipeline, from K-12 to college to continued education for company employees have been established along a ten-year span. The Amgen BioTalents Program was designed to provide undergraduate science and engineering students with training in biomanufacturing. The areas targeted by this educational program enhance their academic development, since these topics are not part of their traditional science and engineering curricula. The educational curriculum involved the process of producing a biomolecule from the genetic engineering of cells to the production of an especially targeted polypeptide, protein expression and purification, to quality control, and validation. This paper will report and describe the implementation details and outcomes of the first sessions of the program.

Keywords: biomanufacturing curriculum, interdisciplinary learning, workforce development, industry-academia partnering

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Authors: V. Shapaval, N. K. Afseth, D. Tzimorotas, A. Kohler

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Globally there is a dramatic increase in the demand for food, energy, materials and clean water since natural resources are limited. As a result, industries are looking for ways to reduce rest materials and to improve resource efficiency. Microorganisms have a high potential to be used as bio factories for the production of primary and secondary metabolites that represent high-value bio-products (enzymes, polyunsaturated fatty acids, bio-plastics, glucans, etc.). In order to find good microbial producers, to design suitable substrates from food rest materials and to optimize fermentation conditions, rapid analytical techniques for quantifying target bio products in microbial cells are needed. In the EU project FUST (R4SME, Fp7), we have developed a fully automated high-throughput FUST system based on micro-cultivation and FTIR spectroscopy that facilitates the screening of microorganisms, substrates and fermentation conditions for the optimization of the production of different high-value metabolites (single cell oils, bio plastics). The automated system allows the preparation of 100 samples per hour. Currently, The FUST system is in use for screening of filamentous fungi in order to find oleaginous strains with the ability to produce polyunsaturated fatty acids, and the optimization of cheap substrates, derived from food rest materials, and the optimization of fermentation conditions for the high yield of single cell oil.

Keywords: FTIR spectroscopy, FUST system, screening, biotechnology

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Authors: Shagufta Gulraiz, Abu Saeed Hashmi, Muhammad Mohsin Javed

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Lysine required for poultry feed is imported in Pakistan to fulfil the desired dietary needs. Present study was designed to produce maximum lysine by utilizing cheap sources to save the foreign exchange. To achieve the goal of lysine production through fermentation, large scale production of lysine was carried out in 7.5 L stirred glass vessel fermenter with wild and mutant Brevibacterium flavum (B. flavum) using all pre-optimized conditions. The identification of produced lysine was carried out by TLC and amino acid analyzer. Toxicity evaluation of produced lysine was performed before feeding to broiler chicks. During biological trial concentrated fermented broth having 8% lysine was used in poultry rations as a source of Lysine for test birds. Fermenter scale studies showed that the maximum lysine (20.8 g/L) was produced at 250 rpm, 1.5 vvm aeration, 6.0% inoculum under controlled pH conditions after 56 h of fermentation with wild culture but mutant (BFENU2) gave maximum yield of lysine 36.3 g/L under optimized condition after 48 h. Amino acid profiling showed 1.826% Lysine in fermented broth by wild B. flavum and 2.644% by mutant strain (BFENU2). Toxicity evaluation report showed that the produced lysine is safe for consumption by broilers. Biological evaluation results showed that produced lysine was equally good as commercial lysine in terms of weight gain, feed intake and feed conversion ratio. A cheap and practical bioprocess of Lysine production was concluded, that can be exploited commercially in Pakistan to save foreign exchange.

Keywords: lysine, fermentation, broiler chicks, biological evaluation

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Authors: Astorga-Trejo Rebeca, Fonseca-Peralta Héctor Manuel, Beltrán-Arredondo Laura Ivonne, Castro-Martínez Claudia

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The use of biomass as feedstock for the production of fuels and other chemicals of interest is an ever-growing accepted option in the way to the development of biorefinery complexes; in the Mexican state of Sinaloa, two million tons of residues from corn crops are produced every year, most of which can be converted to bioethanol and other products through biotechnological conversion using yeast and other microorganisms. Therefore, the objective of this work was to take advantage of corn stover and evaluate its potential as a substrate for the production of second-generation bioethanol (2G), enzymes, and xylitol. To produce bioethanol 2G, an acid-alkaline pretreatment was carried out prior to saccharification and fermentation. The microorganisms used for the production of enzymes, as well as for the production of xylitol, were isolated and characterized in our workgroup. Statistical analysis was performed using Design Expert version 11.0. The results showed that it is possible to obtain 2G bioethanol employing corn stover as a carbon source and Saccharomyces cerevisiae ItVer01 and Candida intermedia CBE002 with yields of 0.42 g and 0.31 g, respectively. It was also shown that C. intermedia has the ability to produce xylitol with a good yield (0.46 g/g). On the other hand, qualitative and quantitative studies showed that the native strains of Fusarium equiseti (0.4 IU/mL - xylanase), Bacillus velezensis (1.2 IU/mL – xylanase and 0.4 UI/mL - amylase) and Penicillium funiculosum (1.5 IU / mL - cellulases) have the capacity to produce xylanases, amylases or cellulases using corn stover as raw material. This study allowed us to demonstrate that it is possible to use corn stover as a carbon source, a low-cost raw material with high availability in our country, to obtain bioproducts of industrial interest, using processes that are more environmentally friendly and sustainable. It is necessary to continue the optimization of each bioprocess.

Keywords: biomass, corn stover, biorefinery, bioethanol 2G, enzymes, xylitol

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Authors: Mauricio Cruz, Andrés Díaz García, Martha Isabel Gómez, Juan Carlos Serrato Bermúdez

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The disadvantages of using natural substrates in SSF processes have been well recognized and mainly are associated to gradual decomposition of the substrate, formation of agglomerates and decrease of porosity bed generating limitations in the mass and heat transfer. Additionally, in several cases, materials with a high agricultural value such as sour milk, beets, rice, beans and corn have been used. Thus, the use of economic inert supports (natural or synthetic) in combination with a nutrient suspension for the production of biocontrol microorganisms is a good alternative in SSF processes, but requires further studies in the fields of modeling and optimization. Therefore, the aim of this work is to compare the performance of two inert supports, a synthetic (polyurethane foam) and a natural one (rice husk), identifying the factors that have the major effects on the productivity of T. asperellum Th204 and the maximum specific growth rate in a PROPHYTA L05® fixed bed bioreactor. For this, the six factors C:N ratio, temperature, inoculation rate, bed height, air moisture content and airflow were evaluated using a fractional design. The factors C:N and air flow were identified as significant on the productivity (expressed as conidia/dry substrate•h). The polyurethane foam showed higher maximum specific growth rate (0.1631 h-1) and productivities of 3.89 x107 conidia/dry substrate•h compared to rice husk (2.83x106) and natural substrate based on rice (8.87x106) used as control. Finally, a quadratic model was generated and validated, obtaining productivities higher than 3.0x107 conidia/dry substrate•h with air flow at 0.9 m3/h and C:N ratio at 18.1.

Keywords: bioprocess, scale up, fractional design, C:N ratio, air flow

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Authors: Ahmad Ali Shahid, M Shakil Shaukat

Abstract:

Agriculture is the backbone of economy of Pakistan and Cotton is the major agricultural export and supreme source of raw fiber for our textile industry. To combat against the developing resistance in the target insects and combating these challenges wholesomely, a novel combination of pyramided/stacked genes was conceptualized and later realized, through the means of biotechnology i.e., transformation of three genes namely, Cry1Ac, Cry2A, and EPSP synthase (glyphosate tolerant) genes in the locally cultivated cotton variety. The progenies of the transformed plants were successfully raised and screened under the tunnel conditions for two generations and the present study focused on the screening of plants which were confirmed for containing all of these three genes and their expressions. Initially, the screening was done through glyphosate spray assay and the plants which were healthy and showed no damage on leaves were selected after 07 days of spray. In the laboratory, the DNA of these plants were isolated and subjected to amplification of the three genes. Thus, seventeen out of twenty were confirmed positive for Cry1Ac gene and ten out of twenty were positive for Cry2A gene and all twenty were positive for presence of EPSP synthase gene. Then, the ten plant samples which were confirmed with presence of all three genes were subjected to expression analysis of these proteins through ELISA. The results showed that eight out of ten plants were actively expressing the three transgenes. Real-time PCR was also done to quantify the expression levels of the EPSP synthase gene. Finally, eight plants were confirmed for the presence and active expression of all three genes in T3 generation of the triple gene transformed cotton. These plants may be subjected to T4 generation to develop a new stable variety in due course of time.

Keywords: agriculture, cotton, transformation, cry genes, ELISA, PCR

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Authors: Andrea Schiffauerova, Amnah Alzeyoudi

Abstract:

This paper studies patterns of innovation within national constitutional context. Its objective is to examine national systems of innovation in biotechnology in six leading innovative countries: the US, Japan, Germany, the UK, France and Canada. The framework proposed for this purpose consists of specific factors considered critical for the development of national systems of innovation, which are industry size, innovative activities, area of specialization, industry structure, national policy, the level of government intervention, the stock of knowledge in universities and industries, knowledge transfer from universities to industry and country-specific conditions for start-ups. The paper then uses the framework to provide detailed cross-country comparisons while highlighting particular features of national institutional context which affect the creation and diffusion of scientific knowledge within the system. The study is primarily based on the extensive survey of literature and it is complemented by the quantitative analysis of the patent data extracted from the United States Patent and Trademark Office (USPTO). The empirical analysis provides numerous insights and greatly complements the data gained from the literature and other sources. The final cross-country comparative analysis identifies three patterns followed by the national innovation systems in the six countries. The proposed cross-country relative positioning analysis may help in drawing policy implications and strategies leading to the enhancement of national competitive advantage and innovation capabilities of nations.

Keywords: comparative analysis, framework, national systems of innovation, patent analysis, United States Patent and Trademark Office (USPTO)

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Authors: Odeyemi Ifeoluwayemi, Maha Joushua, Fulani Omoyeni

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In the search for sustainability, over time, architectural approaches to design have moved from just nature inspired design to the study of nature’s principles to produce effective designs that solve the issue of sustainability. Nature has established materials, shapes and processes that are effective right from a minor scale to an enormous scale. A branch of human knowledge that studies nature is called biology. Biology helps us to grasp and understand nature. Biomimicry is a new way of viewing and valuing nature, based not on what we can extract from the natural world but on what we can learn from it. Life has sustained on the earth for the last 3.85 billion years, and it is necessary for us to find out how life has been able to stay sustained for that long. The building must teach the society new ecological morals, thus, a better understanding of how nature works can usefully inspire architectural designs to resolve issues that have already been resolved by nature. This will not only help in creating a healthy environment but will also produce positive environmental impacts. Biomimetic Architecture connects and reproduces the ideologies found in nature in order to create built environment which benefit people and other living creatures as well as preserving it for the future. Understanding the bioprocess would lead to the establishment of ecological approaches that serve as a platform for creating a built environment that goes beyond sustaining current settings but also mimic nature’s regenerative ecosystem. This paper aims to explain these design methods under the name of biomimicry and biomimetic architecture by reviewing literature and research works done by examining these approaches classified as forms, processes and ecosystems. It is expected that this research will provide information that would, therefore, lead to the creation of buildings that are eco-friendly and provide greater comfort to the populaces.

Keywords: biomimetic architecture, biomimicry, ecological design, nature

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Authors: Avinash Vellore Sunder, Shikha Shah, Pramod P. Wangikar

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The enzymatic conversion of nitriles to carboxylic acids by nitrilases has gained significance in the green synthesis of several pharmaceutical precursors and fine chemicals. While nitrilases have been characterized from different sources, the industrial application requires the identification of nitrilases that possess higher substrate tolerance, wider specificity and better thermostability, along with the development of an efficient bioprocess for producing large amounts of nitrilase. To produce large amounts of nitrilase, we developed a fed-batch fermentation process on defined media for the high cell density cultivation of E. coli cells expressing the well-studied nitrilase from Alcaligenes fecalis. A DO-stat feeding approach was employed combined with an optimized post-induction strategy to achieve nitrilase titer of 2.5*105 U/l and 78 g/l dry cell weight. We also identified 16 novel nitrilase sequences from genome mining and analysis of substrate binding residues. The nitrilases were expressed in E. coli and their biocatalytic potential was evaluated on a panel of 22 industrially relevant nitrile substrates using high-throughput screening and HPLC analysis. Nine nitrilases were identified to exhibit high activity on structurally diverse nitriles including aliphatic and aromatic dinitriles, heterocyclic, -hydroxy and -keto nitriles. With fed-batch biotransformation, whole-cell Zobelia galactanivorans nitrilase achieved yields of 2.4 M nicotinic acid and 1.8 M isonicotinic acid from 3-cyanopyridine and 4-cyanopyridine respectively within 5 h, while Cupravidus necator nitrilase enantioselectively converted 740 mM mandelonitrile to (R)–mandelic acid. The nitrilase from Achromobacter insolitus could hydrolyze 542 mM iminodiacetonitrile in 1 h. The availability of highly active nitrilases along with bioprocesses for enzyme production expands the toolbox for industrial biocatalysis.

Keywords: biocatalysis, isonicotinic acid, iminodiacetic acid, mandelic acid, nitrilase

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Authors: Ekramul Hoque, Zinat Ara Eakut Zarin, Ershad Ali

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The growth and development of explants is governed by the effect of nutrient medium. Ammonium nitrate (NH4NO3) as a major salt of stock solution-1 for the preparation of tissue culture medium. But, it has several demerits on human civilization. It is use for the preparation of bomb and other destructive activities. Hence, it is totally ban in our country. A new chemical was identified as a substitute of ammonium nitrate. The concentrations of the other ingredients of major and minor salt were modified from the MS medium. The formulation of new medium is totally different from the MS nutrient composition. The most widely use MS medium composition was used as first check treatment and MS powder (Duchefa Biocheme, The Netherland) was used as second check treatment. The experiments were carried out at the Department of Biotechnology, Sher-e-Bangla Agricultural University, Dhaka, Bangladesh. Two potato varieties viz. Diamant and Asterix were used as experimental materials. The regeneration potentiality of potato onto new medium was best as compare with the two check treatments. The traits -node number, leaf number, shoot length, root lengths were highest in new medium. The plantlets were healthy, robust and strong as compare to plantlets regenerated from check treatments. Three subsequent sub-cultures were made in the new medium to observe the growth pattern of plantlet. It was also showed the best performance in all the parameter under studied. The regenerated plantlet produced good quality minituber under field condition. Hence, it is concluded that, a new plant tissue culture medium as discovered from the Department of Biotechnology, Sher-e-Bangla Agricultural University, Dhaka, Bangladesh under the leadership of Professor Dr. Md. Ekramul Hoque.

Keywords: new medium, potato, regeneration, ammonium nitrate

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Authors: John Onolame Unuofin, Uchechukuw U. Nwodo, Anthony I. Okoh

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Laccase is constantly gaining status as important biocatalyst in biotechnology. The illimitable potential of its industrial applications and the corresponding aggressive need for phenomenal volumes of extracellularly secreted laccases have called for its interminable production from sources which are able to meet this demand within a relatively short period of time, preferably bacteria. In response to this call, this study was designed to source for laccase-producing bacteria from different environmental matrices. Three sampling environments were chosen such as wastewater treatment plants, University of Fort Hare vicinity and the Hogback woodland, all within the Eastern Cape, South Africa. Samples such as effluents, sediments, leaf litters, degrading wood and rock scrapings were selectively enriched with some model aromatic compounds and were further screened qualitatively and quantitatively on five phenolic substrates ABTS (2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), Guaiacol, 1-Naphthol, Potassium Ferric Cyanide and Syringaldazine). Basis for selection was their ability to elicit a colour change on at least three of the above mentioned agar based assay substrates. The choice isolates were further identified based on 16S rRNA molecular identification techniques. 33 isolates were screened out of the 40 representative distinct colonies during the qualitative plate screens, while quantitative screens selected out 11 bacterial isolates. They were, based on molecular identification, desginated as members of the genera Pseudomonas, Stenotrophomonas and Citrobacter of the gammaproteobacteria and Bordetalla and Achromobacter of the betaproteobacteria respectively. We therefore conclude based on our outcomes that we may have isolated efficient laccase-producing bacteria, which might be of beneficial significance in catalysis and biotechnology.

Keywords: beta proteobacteria, catalysis, gammaproteobacteria, laccase

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Authors: Laura Mejias, Sandra Monteagudo, Oscar Martinez-Avila, Sergio Ponsa

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Bioplastics are gaining attention as potential substitutes of conventional fossil-derived plastics and new components of specialized applications in different industries. Besides, these constitute a sustainable alternative since they are biodegradable and can be obtained starting from renewable sources. Thus, agro-industrial wastes appear as potential substrates for bioplastics production using microorganisms, considering they are a suitable source for nutrients, low-cost, and available worldwide. Therefore, this approach contributes to the biorefinery and circular economy paradigm. The present study assesses the solid-state fermentation (SSF) technology for the co-synthesis of exopolysaccharides (EPS) and polyhydroxyalkanoates (PHA), two attractive biodegradable bioplastics, using the leftover of the brewery industry brewer's spent grain (BSG). After an initial screening of diverse PHA-producer bacteria, it was found that Burkholderia cepacia presented the highest EPS and PHA production potential via SSF of BSG. Thus, B. cepacia served to identify the most relevant aspects affecting the EPS+PHA co-synthesis at a lab-scale (100g). Since these are growth-dependent processes, they were monitored online through oxygen consumption using a dynamic respirometric system, but also quantifying the biomass production (gravimetric) and the obtained products (EtOH precipitation for EPS and solid-liquid extraction coupled with GC-FID for PHA). Results showed that B. cepacia has grown up to 81 mg per gram of dry BSG (gDM) at 30°C after 96 h, representing up to 618 times higher than the other tested strains' findings. Hence, the crude EPS production was 53 mg g-1DM (2% carbohydrates), but purity reached 98% after a dialysis purification step. Simultaneously, B. cepacia accumulated up to 36% (dry basis) of the produced biomass as PHA, mainly composed of polyhydroxybutyrate (P3HB). The maximum PHA production was reached after 48 h with 12.1 mg g⁻¹DM, representing threefold the levels previously reported using SSF. Moisture content and aeration strategy resulted in the most significant variables affecting the simultaneous production. Results show the potential of co-synthesis via SSF as an attractive alternative to enhance bioprocess feasibility for obtaining these bioplastics in residue-based systems.

Keywords: bioplastics, brewer’s spent grain, circular economy, solid-state fermentation, waste to product

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Authors: Zelikha Labbani

Abstract:

Many biotechnology methods have been used in plant breeding programs. The in vitro isolated microspore culture is the one of these methods. For durum wheat, the use of this technology has been limited for a long time due to the low number of embryos produced and also most regeneration plants are albina. The objective of this paper is to show that using isolated microspores culture on durum wheat is possible due to the development of the new methods using the new pretreatment of the microspores before their isolation and cultivation.

Keywords: isolated microspore culture, pretreatments, in vitro embryogenesis, plant breeding program

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Authors: Godfrey Muiya Mukala

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This research looks at an array of fields in Sub-Saharan Africa comprising agriculture, food enterprises, medicine, organisms genetically modified, microbiology, and nanotechnology that can be gained from biotechnological research and development. Findings into dangerous organisms, mainly bacterial germs, rickettsia, fungi, parasites, or organisms that are genetically engineered, have immensely posed questions attributed to the biological danger they bring forth to human beings and the environment because of their uncertainties. In addition, the recurrence of previously managed diseases or the inception of new diseases are connected to biosafety challenges, especially in rural set-ups in low and middle-income countries. Notably, biotechnology laboratories are required to adopt biosafety measures to protect their workforce, community, environment, and ecosystem from unforeseen materials and organisms. Sensitization and inclusion of educational frameworks for laboratory workers are essential to acquiring a solid knowledge of harmful biological agents. This is in addition to human pathogenicity, susceptibility, and epidemiology to the biological data used in research and development. This article reviews and analyzes research intending to identify the proper implementation of universally accepted practices in laboratory safety and biological hazards. This research identifies ideal microbiological methods, adequate containment equipment, sufficient resources, safety barriers, specific training, and education of the laboratory workforce to decrease and contain biological hazards. Subsequently, knowledge of standardized microbiological techniques and processes, in addition to the employment of containment facilities, protective barriers, and equipment, is far-reaching in preventing occupational infections. Similarly, reduction of risks and prevention may be attained by training, education, and research on biohazards, pathogenicity, and epidemiology of the relevant microorganisms. In this technique, medical professionals in rural setups may adopt the knowledge acquired from the past to project possible concerns in the future.

Keywords: sub-saharan africa, biotechnology, laboratory, infections, health

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Authors: Jožef Ritonja

Abstract:

The biochemical industry is one of the most important modern industries. Biochemical reactors are crucial devices of the biochemical industry. The essential bioprocess carried out in bioreactors is the fermentation process. A thorough insight into the fermentation process and the knowledge how to control it are essential for effective use of bioreactors to produce high quality and quantitatively enough products. The development of the control system starts with the determination of a mathematical model that describes the steady state and dynamic properties of the controlled plant satisfactorily, and is suitable for the development of the control system. The paper analyses the fermentation process in bioreactors thoroughly, using existing mathematical models. Most existing mathematical models do not allow the design of a control system for controlling the fermentation process in batch bioreactors. Due to this, a mathematical model was developed and presented that allows the development of a control system for batch bioreactors. Based on the developed mathematical model, a control system was designed to ensure optimal response of the biochemical quantities in the fermentation process. Due to the time-varying and non-linear nature of the controlled plant, the conventional control system with a proportional-integral-differential controller with constant parameters does not provide the desired transient response. The improved adaptive control system was proposed to improve the dynamics of the fermentation. The use of the adaptive control is suggested because the parameters’ variations of the fermentation process are very slow. The developed control system was tested to produce dairy products in the laboratory bioreactor. A carbon dioxide concentration was chosen as the controlled variable. The carbon dioxide concentration correlates well with the other, for the quality of the fermentation process in significant quantities. The level of the carbon dioxide concentration gives important information about the fermentation process. The obtained results showed that the designed control system provides minimum error between reference and actual values of carbon dioxide concentration during a transient response and in a steady state. The recommended control system makes reference signal tracking much more efficient than the currently used conventional control systems which are based on linear control theory. The proposed control system represents a very effective solution for the improvement of the milk fermentation process.

Keywords: biochemical reactor, fermentation process, modelling, adaptive control

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