Bioprocess Research Articles

Evaluation of environmental impact on cocoa production and processing under life cycle assessment method: From beans to liquor

This study aimed to identify the environmental impact on cocoa production and processing based on the life cycle assessment method (LCA). The major contribution of this study is to evaluate the environmental performance of a chocolate liquor in order to reveal the significant environmental hotspots along the entire supply chain. This is achieved through two main steps. First, the LCA method is applied to identify the environmental implications of managing the production and processing cocoa to become chocolate liquor. The LCA is performed based on the international ISO 14040–14043 series. Secondly, a sensitivity analysis was carried out to evaluate the impact of five proposed improvement options to discover whether any occurring potential environmental impacts will be reduced. The kind of scenarios are: reducing pesticides by 50 % (option 1); using compost instead of fertilizer (option 2); avoid burning stubble (option 3), conserving soil water (option 4) and applying alternative energy sources (option 5). As a result, the most significant environmental impact for the upstream and core processing was human toxicity potential (HTP) and cumulative energy demand (CED), 89 % and 59 %, respectively. In addition, the impact of the proposed improvement has significantly reduced environmental impacts (HTP, 68 %) and achieved savings (IDR 18800, 17.7 %). Survey data includes the entire supply chain, with a detailed focus on production and processing stages for an Indonesian chocolate company. This study only dealt with cocoa beans that generated 1 kg of chocolate liquor.

The effects of dextran in residual impurity on trehalose crystallization and formula in food preservation

The residual dextran impurities in the upstream process significantly impact the crystallization of starch-based functional sugar and the related food properties. This study intends to reveal the mechanism of dextran’s influence on trehalose crystallization, and build a relationship among the dextran in syrup and the physicochemical and functional properties of trehalose. Instead of incorporating into the crystal lattice, dextran changes the assembly rate of trehalose molecules on crystal surface. The different sensitivity and adsorption capacity of the crystal surface to the chain length of dextran determines the growth rate of crystal surfaces, resulting in different crystal morphology. The bulk trehalose crystals, which were obtained from syrups with short chain dextran, have excellent powder properties, including best flowability (35◦), highest crystal strength (2.7 N), lowest caking rate (62.22 %), and the most uniform mixing with other sweeteners (sucrose/xylitol) in food formulations, achieving more stable starch preservation.

Upstream and downstream implementation arrangements in two‐level games. A focus on administrative simplification in the Italian National Recovery and Resilience Plan

AbstractThe onset of the European NGEU program represented for European member states a formidable opportunity for post‐pandemic recovery and yet a significant challenge at the same time: to receive and retain EU funds, each state had to promptly draw up a National Recovery and Resilience Plan—NRRP) and commit to a pressing timetable for its implementation. Regarding this challenge, the Italian government's response is a case in point: first, Italy is by far the largest beneficiary of NGEU funds; second, it has long had a reputation for being laggard in both the implementation of European directives and the spending of cohesion policy structural funds; and the formulation of the NRRP, the design of the governance in charge of its implementation, and implementation itself, occurred at a particular moment in the country's political life. Based on these premises, the article examines the upstream process by which the Italian government designed the implementation arrangements for the adoption of simplification policies under the NRRP and their downstream recalibration in the first two years, taking the implementation arrangements as the dependent variable. Analytically, the focus is on the interplay between the pressures of EU timetables and internal political dynamics, be they the legacy or strategic political considerations on the part of national policymakers, in determining the design and eventual re‐design of implementation arrangements as the plan unfolds.

Transboundary hazard and downstream impact of glacial lakes in Hindu-Kush Karakoram Himalayas

Glacial Lake Outburst Floods (GLOFs) can generate catastrophic flash floods when the damming structure is breached or overtopped. Some of these glacial lakes are located in transboundary regions where floods originating from the lake in one country could inundate a neighboring country, devastating the population and infrastructure of both nations and influencing socio-political relationships. Therefore, assessing the lakes' hazard is crucial. This study investigates transboundary glacial lakes, considering their GLOF hazard, including potential mass movement intrusion, moraine's stability, upstream and downstream process cascades, downstream flood extents, and the exposure and vulnerability of the downstream infrastructure and affected population. GLOF exposure assessments were carried out to identify exposed buildings, bridges, and hydropower systems in transboundary regions. China currently has the highest number of transboundary lakes, with most of them potentially impacting India and Nepal. Most of the transboundary lakes in China, and many in India and Nepal, are susceptible to mass movements. Among the 230 transboundary glacial lakes in the Hindu Kush Karakoram Himalaya, 55 lakes can potentially impact other glacial lakes along their flow path, creating a cascade of events. Five transboundary lakes could potentially impact over 1000 buildings, and 16 lakes could impact over 500 buildings. A total of 35 lakes can impact at least one hydropower station along their flow path, and 4 lakes can impact two hydropower stations. This research emphasizes the critical importance of conducting comprehensive risk analyses of GLOFs in transboundary regions to inform policy-makers. It calls for investing in broad-scale assessments and data-driven decision-making for mitigating and adapting to GLOF risks effectively. Finally, by raising awareness among policy-makers, the study aims to drive actions that safeguard communities and infrastructure vulnerable to GLOF.

Coordinate regulation mechanism of primary recrystallization texture and microstructure of grain-oriented silicon steel

Texture control, which is affected by composition, hot rolling, cold rolling, and annealing, is the core element to improve the magnetic properties of grain-oriented silicon steel and is one of the key scientific problems in developing electrical steel technology. The research and control of first factors have been very mature, and the integrated regulation of upstream and downstream processes has become a breakthrough point for improving texture. For the common grain-oriented silicon steel based on the “two-stage cold rolling method”, primary recrystallization is the stage achievement of smelting, hot rolling, first cold rolling, and decarburization annealing. The grain size and texture characteristics of the primary recrystallization are the only standards to measure the pre-process technology and is the pointer of whether the secondary recrystallization can occur in the subsequent process or not, which are extremely important to improve the magnetic properties. This paper studied the hot rolling, the first cold rolling and decarburization annealing of grain-oriented silicon steel by using XRD, EBSD technologies. The results showed that the hot rolled plate is not as thinner as better, only the optimum ratio of the thickness of hot rolled plate and the first cold reduction rate can obtain optimal primary recrystallization. The minimum iron loss value and maximum magnetic induction intensity are obtained under the primary cold rolling reduction rate controlled at 77.27%, the average grain size of the primary recrystallization is 14 µm, the iron loss P1.7/50 is reduced to 1.02 W/Kg, and the magnetic induction intensity B8 can reach 1.910 T.

On the fate of deep eutectic solvents after their use as reaction media: the CO2 production during downstream and ultimate disposal

What can we do with Deep Eutectic Solvents (DES) once they have been used? What can be their disposal and final fate? This perspective adds some considerations to stimulate the debate.

Proceedings of the 2023 Viral Clearance Symposium, Session 7: Up- and Downstream Virus Retentive Filtration.

Session 7 of the 2023 Viral Clearance Symposium reviewed progresses in virus retentive filtrations applied to both upstream and downstream processing. Upstream topics included investigations and applications of media viral filtration for upstream cell culture viral risk mitigation. Downstream topics included evaluation of viral breakthrough in continuous processing using surrogate particles and demonstration of extensive viral filtration cycling with flow interruptions and long duration in connected process. Reuse of viral filters with proposed procedures was successfully demonstrated amid the supply chain challenge encountered during the pandemic. Discussions and additional considerations for the topics were also provided.

Designing profitable supply chains for lithium-ion battery recycling in the United States

<abstract> <p>Recycling spent lithium-ion batteries (LIBs) has attracted lots of attention recently, due to the increasing demand for critical materials contained in LIBs, putting high pressure on their geological reserves. We evaluated the potential of bioleaching technology as a sustainable solution for recycling spent LIBs to help inform decision-making processes for stakeholders involved in LIB recycling supply chains. A supply chain model was developed to include required upstream processes with the objective of maximizing economic feasibility of LIB recycling through the technology. The model has been applied to the U.S. and an optimal supply chain configuration was identified, considering the major factors affecting the economic viability of the technology. The net present value of the supply chain was estimated to be ＄18.4 billion for operating over 10 years, achieving the maximum processing capacity of 900,000 tons of black mass per year. The economic viability of the technology was identified to be highly sensitive to the cost associated with purchasing black mass, which accounted for more than 60% of the total supply chain cost. The breakeven price of black mass was identified as ＄8.7/kg over which the supply chain was not economically sustainable. Additionally, we examined the non-cooperative scenarios where each tier tries to maximize its own profit to demonstrate how the overall profitability of the supply chain changes with different pricing strategies of sortation facilities and acid producers. We estimated that the maximum prices of non-recyclable paper and acid that the supply chain could tolerate were ＄0.89/kg and ＄8.5/kg, respectively, beyond which the supply chain was no longer sustainable.</p> </abstract>

Disposable Bioreactors Used in Process Development and Production Processes with Plant Cell and Tissue Cultures.

The bioreactor is the centerpiece of the upstream processing in any biotechnological production process. Its design, the cultivation parameters, the production cell line, and the culture medium all have a major influence on the efficiency of the process and the result of the cultivation. Disposable bioreactors have been used for the past 20 years, playing a major role in process development and commercial production of high-value substances at medium scales.Our review deals with scalable, disposable bioreactors that have proven to be useful for the cultivation of plant cell and tissue cultures. Based on the definitions of terms and a categorization approach, the most commonly used, commercially available, disposable bioreactor types are presented below. The focus is on wave-mixed, stirred, and orbitally shaken bioreactors. In addition to their instrumentation and bioengineering characteristics, cultivation results are discussed, and emerging trends for the development of disposable bioreactors for plant cell and tissue cultures are also addressed.

IDENTIFIKASI MISKONSEPSI MATERI SEL PADA PESERTA DIDIK DI KELAS XI IPA MAN 2 KOTA PADANG

Cell material is one of the biological materials that contains many abstract concepts and objects, this causes students to have difficulty understanding the concept and has an impact on the low learning outcomes of students. Misconceptions arise when learners develop their own understanding of a concept, but the concept is wrong and different from the actual concept. Therefore, it is necessary to identify the level of understanding of the concept of students. Identification is done using a Multiple Choice test with CRI which aims to determine the percentage level of understanding of students on cell material. Students' understanding is categorized into understanding the concept, misconceptions and not understanding the concept. The results showed that the percentage of students' misconceptions on cell organelles was 34.03%, the concept of differences between animal cells and plant cells was 31.48%, the concept of cell types was 28.47%, the concept of cell components was 35.42% and the concept of cell bioprocessing was 36.11%. The average misconception of students on cell material is 33.10%. The average percentage of students' misconceptions is classified as low.

Upstream and Downstream Bioprocessing in Enzyme Technology.

The development of biotransformation must integrate upstream and downstream processes. Upstream bioprocessing will influence downstream bioprocessing. It is essential to consider this because downstream processes can constitute the highest cost in bioprocessing. This review comprehensively overviews the most critical aspects of upstream and downstream bioprocessing in enzymatic biocatalysis. The main upstream processes discussed are enzyme production, enzyme immobilization methodologies, solvent selection, and statistical optimization methodologies. The main downstream processes reviewed in this work are biocatalyst recovery and product separation and purification. The correct selection and combination of upstream and downstream methodologies will allow the development of a sustainable and highly productive system.

Techno-economic analysis as a valuable tool for the selection of high-potential extractive fermentation approaches for large-scale prodigiosin production

Prodigiosin is a promising microbial red pigment with several therapeutic properties. Unfortunately, production through fermentation is hampered by product inhibition, resulting in low titers, high production costs, and an elevated price of the final product. It is of the upmost importance to develop cost-effective strategies for the intensification of this metabolite. Previously, we developed a strategy for prodigiosin production using a low-cost medium successfully coupled to a mineral oil extractive phase. In the present work, we analyze through economical modelling the developed strategies, while considering the upstream and downstream processing, aiming to evaluate the economic feasibility of each and determine the best approach for large scale production. As a production benchmark, a reported method for prodigiosin extractive fermentation using micellar ATPS (81 % recovery) was included. From an economic perspective, the best approach resulted in processing only the mineral oil fraction at continuous operation, obtaining a 4.73-fold and 16.77-fold cost per mg decrease when compared to batch and the micellar approach, respectively. The optimum operational parameters to generate 1 kg of prodigiosin at continuous scale were 2,431 L reactor volume with a 91.02 L/h flow. Based on the obtained results, it is possible to propose the mineral oil approach as a cost-effective strategy for simultaneous prodigiosin production and direct extraction. This approach yields higher upstream yields, a relatively high product purity level (∼90 %), and results in a competitive production cost of USD $25 x 10-4 per mg.

Mulberry silkworm biotechnology as the basis of a bioindustrial platform. Improvements on the upstream processing stage (USP)

Molecular engineering is an upstream engineering method for creating functional materials and devices using molecules, and atoms as building blocks. In 2000, Japan became the first country to genetically modify silkworms (Bombyx mori, hereinafter referred to as B. mori). The subsequent research in the development of new materials expanded the possibilities of using sericulture products, characterizing this situation as a "sericulture revolution". In the Russian Federation, molecular engineering in life sciences is aimed at solving the tasks of developing world-class technological platforms to create tools to produce new molecules (biopolymers, proteins, enzymes), bioproducts, cells and organisms. The main "bottom-up" approaches applied in the upstream process (USP) stage of silk production to improve the production and economic performance and quality characteristics of raw materials have been studied. The variety of improvement methods includes: the possibility of using artificial nutrient medium, molecular engineering based on transient expression techniques or stable germline transformation, genetic selection methods, breeding management, etc. The advantages of the mulberry silkworm (B. mori) such as low breeding costs, significantly higher production yield compared to other protein expression systems, favor its use as an effective producer of recombinant proteins, antimicrobial peptides and biologically active substances. USP stage biotechnologies enable the production of new raw materials for downstream processing (DSP) to produce a wide range of products that contribute to improving the quality of human life. A set of biotechnological solutions forms the modern basis of the mulberry silkworm bioindustrial platform.

Continuous biomanufacturing in upstream and downstream processing

Abstract Continuous bioprocesses have become a significant technological change in regulated industries, with process analytical technology (PAT) and quality-by-design (QbD) being essential for enabling continuous biomanufacturing. PAT and QbD are associated with process automation and control, providing real-time key process information. Continuous manufacturing eliminates hold times and reduces processing times, providing benefits such as improved product quality, reduced waste, lower costs, and increased manufacturing flexibility and agility. Over the past decade, advancements in science and engineering, along with the adoption of QbD and the advancement of PAT, have progressed the scientific and regulatory readiness for continuous manufacturing. Regulatory authorities support the implementation of continuous manufacturing using science- and risk-based approaches, providing a great deal of potential to address issues of agility, flexibility, cost, and robustness in the development of pharmaceutical manufacturing processes.

Astaxanthin from microalgae: A review on structure, biosynthesis, production strategies and application

Astaxanthin is a red-colored secondary metabolite with excellent antioxidant properties, typically finds application as foods, feed, cosmetics, nutraceuticals, and medications. Astaxanthin is usually produced synthetically using chemicals and costs less as compared to the natural astaxanthin obtained from fish, shrimps, and microorganisms. Over the decades, astaxanthin has been naturally synthesized from Haematococcus pluvialis in commercial scales and remains exceptional, attributed to its higher bioactive properties as compared to synthetic astaxanthin. However, the production cost of algal astaxanthin is still high due to several bottlenecks prevailing in the upstream and downstream processes. To that end, the present study intends to review the recent trends and advancements in astaxanthin production from microalgae. The structure of astaxanthin, sources, production strategies of microalgal astaxanthin, and factors influencing the synthesis of microalgal astaxanthin were discussed while detailing the pathway involved in astaxanthin biosynthesis. The study also discusses the relevant downstream process used in commercial scales and details the applications of astaxanthin in various health related issues.

Challenges and Opportunities in the Process Development of Chimeric Vaccines.

Vaccines are integral to human life to protect them from life-threatening diseases. However, conventional vaccines often suffer limitations like inefficiency, safety concerns, unavailability for non-culturable microbes, and genetic variability among pathogens. Chimeric vaccines combine multiple antigen-encoding genes of similar or different microbial strains to protect against hyper-evolving drug-resistant pathogens. The outbreaks of dreadful diseases have led researchers to develop economical chimeric vaccines that can cater to a large population in a shorter time. The process development begins with computationally aided omics-based approaches to design chimeric vaccines. Furthermore, developing these vaccines requires optimizing upstream and downstream processes for mass production at an industrial scale. Owing to the complex structures and complicated bioprocessing of evolving pathogens, various high-throughput process technologies have come up with added advantages. Recent advancements in high-throughput tools, process analytical technology (PAT), quality-by-design (QbD), design of experiments (DoE), modeling and simulations, single-use technology, and integrated continuous bioprocessing have made scalable production more convenient and economical. The paradigm shift to innovative strategies requires significant attention to deal with major health threats at the global scale. This review outlines the challenges and emerging avenues in the bioprocess development of chimeric vaccines.

Producing protein-based products from microalgae cultivated on anaerobically digested abattoir effluent: Process integration and techno-economic analysis

The integration of microalgae production with anaerobically digested effluents might result in development of sustainable processes not only to remove nitrogen but also to produce value-added products rich in protein. This work analyses the feasibility of producing protein-based products derived from microalgae grown on anaerobically digested wastewater by performing techno-economic evaluation. The results show that it would be possible to propose an upstream process flow diagram producing 2.7 kg microalgae from 1 m−3 wastewater with possible production cost of 1.8–8.1 USD kg−1 depending on the scale of anaerobic digested effluent. It would be possible to produce protein isolate, concentrate, and animal feed from anaerobically digested effluents with possible production cost of 33 USD kg−1, 28 USD kg−1, and 6 USD kg−1 respectively at the scale of 1600 m3 d−1 effluent. Although currently the production of protein-based products might not be feasible, increasing the scale of production might result in some feasible outcomes. However, the associated costs regarding collecting more AD wastewater from different sources should be evaluated to increase the scale of production.

Online multimethod platform for comprehensive characterization of monoclonal antibodies in cell culture fluid from a single sample injection - Intact protein workflow

BackgroundTherapeutic monoclonal antibodies (mAbs) comprise a large structural variability with respect to charge, size and post-translational modifications. These critical quality attributes (CQAs) need to be assessed during and after the production of mAbs. This normally requires off-line purification and sample preparation as well as several chromatographic selectivities, which makes the whole process time-consuming and error-prone. To improve on this, we developed an integrated and automated multi-dimensional analytical platform for the simultaneous assessment of multiple CQAs of mAbs in cell culture fluid (CCF) from upstream processes. ResultsThe on-line system allows mAb characterization at the intact level, combining protein A affinity chromatography (ProtA) with size-exclusion, ion-exchange, and reversed-phase liquid chromatographic modes with UV and mass spectrometric detection. Multiple heart cuts of a single mAb elution band from ProtA are stored in 20-μL loops and successively sent to the multimethod options in the second dimension. ProtA loading and elution conditions and their compatibility with second-dimension LC modes were studied and optimized. Subsequently, heart-cutting and valve-switching schemes were investigated to achieve effective and reproducible analyses. The applicability of the developed workflow was demonstrated by the direct analysis (i.e. not requiring off-line sample preparation) of a therapeutic mAb in CCF, obtaining useful information on accurate molecular mass, glycosylation, and charge and size variants of the mAb product at the same time and in just over 1 h. SignificanceThe developed multidimensional platform is the first system that allows for multiple fractions from a single ProtA band to be characterized using different chromatographic selectivities in a single run allowing direct correlation between CQAs. The performance of the system is comparable to established off-line methods, fully compatible with upstream process samples, and provides a significant time-reduction of the characterization procedure.

Simplified Synthesis of Renieramycin T Derivatives to Target Cancer Stem Cells via β-Catenin Proteasomal Degradation in Human Lung Cancer.

Cancer stem cells (CSCs) found within cancer tissue play a pivotal role in its resistance to therapy and its potential to metastasize, contributing to elevated mortality rates among patients. Significant strides in understanding the molecular foundations of CSCs have led to preclinical investigations and clinical trials focused on CSC regulator β-catenin signaling targeted interventions in malignancies. As part of the ongoing advancements in marine-organism-derived compound development, it was observed that among the six analogs of Renieramycin T (RT), a potential lead alkaloid from the blue sponge Xestospongia sp., the compound DH_32, displayed the most robust anti-cancer activity in lung cancer A549, H23, and H292 cells. In various lung cancer cell lines, DH_32 exhibited the highest efficacy, with IC50 values of 4.06 ± 0.24 μM, 2.07 ± 0.11 μM, and 1.46 ± 0.06 μM in A549, H23, and H292 cells, respectively. In contrast, parental RT compounds had IC50 values of 5.76 ± 0.23 μM, 2.93 ± 0.07 μM, and 1.52 ± 0.05 μM in the same order. Furthermore, at a dosage of 25 nM, DH_32 showed a stronger ability to inhibit colony formation compared to the lead compound, RT. DH_32 was capable of inducing apoptosis in lung cancer cells, as demonstrated by increased PARP cleavage and reduced levels of the proapoptotic protein Bcl2. Our discovery confirms that DH_32 treatment of lung cancer cells led to a reduced level of CD133, which is associated with the suppression of stem-cell-related transcription factors like OCT4. Moreover, DH_32 significantly suppressed the ability of tumor spheroids to form compared to the original RT compound. Additionally, DH_32 inhibited CSCs by promoting the degradation of β-catenin through ubiquitin-proteasomal pathways. In computational molecular docking, a high-affinity interaction was observed between DH_32 (grid score = -35.559 kcal/mol) and β-catenin, indicating a stronger binding interaction compared to the reference compound R9Q (grid score = -29.044 kcal/mol). In summary, DH_32, a newly developed derivative of the right-half analog of RT, effectively inhibited the initiation of lung cancer spheroids and the self-renewal of lung cancer cells through the upstream process of β-catenin ubiquitin-proteasomal degradation.

Optimum formulation design and properties of drilling fluids incorporated with green uncoated and polymer-coated magnetite nanoparticles

Nanomaterials are materials that possess unique properties due to their high specific surface area and quantum effects. Nanomaterials have diverse applications in different fields including the petroleum and gas industry as additives. One of the classes of nanomaterials that currently have potential usage in the downstream, midstream, and upstream processes of the petroleum industry is nanoparticles (NPs). Among the upstream processes in the petroleum and gas industry is the drilling operations. It is popular that the most critical features that ensure the success of a drilling operation are the rheological and filtration loss characteristics of the drilling fluid. The current work deals with the synthesis of green uncoated and polymer-coated green magnetite nanoparticles (MNPs). The MNPs were characterized and assessed as rheology and filtration loss modifiers for water-based drilling fluids. The optimum formulation design of drilling fluids incorporated the MNPs for high-performance drilling fluids in terms of density, rheological, filtration loss, and sagging properties was identified. The effect of temperature (ambient − 80 °C), and aging time (6–248 h) on the investigated properties were evaluated. The results confirmed that optimum values for plastic viscosity, apparent viscosity, yield Point, gel strength (10sec), gel strength (10 min), mud thickness, and sag index were 13.77 cP, 69.69 cP, 89.87 lb/100ft2, 86.75 lb/100ft2, 128.38 lb/100ft2, ≤ 1 mm, and 0.511, respectively. Most of those values could be reached using an optimum formulation involving 0.92 % MNPs at ambient temperature. Increasing the temperature displays a decrease in the values while increasing the aging time displays an increase in the values. Drilling fluids with MNPs showed insignificant changes in the investigated properties with increasing temperature in particular those incorporated with polymer-coated MNPs compared to the water-based drilling fluids.