Pilot-scale Production Research Articles

Valorization of waste Fe-rich sludge as erdite/KFeS2 rods under atmosphere condition and evaluation of their mutual transformation.

Pilot scale production of one-dimensional (FeS2)nn- rods was performed by using an automatic 20L vessel at 80°C under atmosphere condition with the resource utilization of Fe-rich sludge. The sludge was simulated at lab-scale with chemical pure of ferric trichloride. After the sludge treatment, the corresponding rods were not formed at room temperature. But by heating at 80°C, erdite rod was well-crystallized after 0.5h by only adding Na halite, and KFeS2 rod was crystallized weakly after 2h and highly at 10h with the addition of K halite. After 48h heating, the rods grow radially to 300nm for erdite, but to 5μm for KFeS2. However, at room temperature, erdite rod was converted to high crystallized KFeS2 in KOH water or ethanol solution, whilst the conversion of KFeS2 rod to erdite also occurred in NaOH water solution, but terminated in NaOH ethanol solution, without any morphology change. It is also noted that with the presence of both Na and K halite, the rod was an intermediate of erdite to KFeS2 with 1μm length after heating at 100°C but converted to 10-μm-length KFeS2 crystal at the temperature of > 120°C. The thermodynamic results confirmed that during the rod polymerization, the Fe(OH)3HS- formation was the sole rate-limiting step and showed a positive Gibbs value of 6.45kJ/mol at room temperature and negative values at the temperature of > 48°C. In summary, this method not only enabled the vaporization of waste Fe-rich sludge as value-added rods without generating any secondary waste but also showed a new route for the in situ conversion of erdite/KFeS2 rods at room temperature.

Pilot-Scale Production of the Natural Colorant Laetiporic Acid, Its Stability and Potential Applications

Laetiporus sulphureus, a wood-decaying basidiomycete, produces yellow-orange pigments in fruiting bodies and, as was recently shown, in submerged cultivated mycelia. Out of four strains, the most potent laetiporic acid producer was identified and its yield compared in different media. The complex Moser b medium was replaced by potato dextrose broth, achieving higher yields at a lower cost. Cultivation was then scaled up from shake flask to a 7 L stirred tank bioreactor. Optimization of parameters led to increased product concentrations up to 1 g L−1, the highest yield reported so far. An in situ product recovery strategy with a biphasic system was established, increasing the yield by 19% on the shake flask scale. A crude ethanolic extract of the biomass was examined for color stability and application trials. In contrast to what has been suggested in the past, the pigment showed limited long-term stability to oxygen and light, but was stable under storage in the dark at 4 °C under nitrogen. The orange extract was successfully incorporated into different matrices like foods, cosmetics and textiles. Laetiporic acid can potentially replace petrochemical based synthetic dyes, and can thus support the development of a circular bioeconomy.

Numerical optimization approach for the development of Giloy stem (Tinospora cordifolia) extract based functional yoghurt

AbstractFunctional yoghurt is a prominent fermented milk product. It is popular among all age groups and has considerable significance due to the presence of active nutrients to fulfill the human dietary needs. Tinospora cordifolia (giloy) is extensively discussed during the recent pandemic owing to its high antioxidant, anti‐inflammatory, and strong immunity boosting properties. This investigation has been aimed to optimize functional yoghurt impregnated with ultrasound assisted giloy stem extract. Central composite rotatable design (CCRD) was used for experimental design of process parameters (giloy stem extract, total soluble solid, and incubation temperature). Functional yoghurt formulation was optimized based on different (whiteness index, overall acceptability, pH, syneresis, firmness, DPPH scavenging activity, and complex viscosity) responses. The results revealed that giloy stem extract and total soluble solid exhibited maximum effects on all the responses. Numerical optimization revealed the optimum combination of giloy stem extract, total soluble solid, and incubation temperature as 0.08%, 9.1%, and 45°C, respectively. At this optimum condition, the whiteness index, overall acceptability (OA), pH, syneresis, firmness, DPPH scavenging activity, and complex viscosity values of optimized functional yoghurt were 84.513, 8.567, 4.193, 47.268%, 281.19 N, 56.78%, and 76.89 cP, respectively. This study established the effect of giloy extract addition, total soluble solids, and incubation temperature on yoghurt preparation and subsequent changes in its properties.Novelty impact statement Giloy stem extract (GSE) enriched functional yoghurt was developed having high antioxidant activities. Ultrasound assisted GSE improved the texture and overall desirability of functional yoghurt. Derived the optimum conditions which might be replicated and scaled up for pilot scale production.

Production and Characterization of Indole Acetic Acid of Endophytic Bacteria isolated from Kalanchoe pinnata (Lam.) in Optimized Media

Objectives: To optimize, quantify and characterize indole acetic acid (IAA) production from endophtic bacteria isolated from the leaves of Kalanchoe pinnata (Lam.). Methods: The selected endophytic bacteria are identified by biochemical, morphological and 16SrRNA sequencing analysis. Endophytes are screened for IAA using Salkowski reagent and media optimization is carried out for maximum IAA yield and quantification was done with UV Spectrophotometer at 30 and 120 minutes. The mean  standard error of each sample for three triplicates is calculated from mean value of the samples followed by normal distribution. The characterization of IAA was studied with ethyl acetate extract of endophytes using TLC and HPLC. Findings: The selected endophytes are identified as Bacillus thuringiensis, Bacillus paranthracis, Staphylococcus xylosus and Bacillus cereus. Optimized yeast malt dextrose (YMD) broth containing mannitol, ammonium nitrate, and 0.2% of L-tryptophan at pH 8 and 30◦ C temperature for 72 h was used to obtain maximum IAA production from these endophytes. B. paranthracis produced 82.47330.0728mg/ml and 83.56670.457mg/ml, S. xylosus produced 93.120.261mg/ml and 94.76670.285mg/ml, B. cereus produced 24.4 0.113mg/ml and 25.0667 0.581mg/ml and B. thrungiensis produced 4.13330.131mg/ml and 50.226mg/ml of IAA at 30 and 120 minutes, respectively. B. paranthracis and S. xylosus were producing potential IAA with Rf 0.91 and 0.95 respectively compared with standard Rf 0.95. The HPLC analysis of Bacillus paranthracis and Staphylococcus xylosus were confirmed for the presence of IAA with the retention time of 5.053 and 5.58 minutes respectively compared to the retention time of the standard. Novelty: Endophytic bacteria notable in the leaves of Kalanchoe pinnata (Lam.) produce indole acetic acid in measurable quantity by UV Spectrometry and HPLC analysis, that can be replace for unstable and expensive synthetic IAA using costly chemicals. This is best pilot scale production of IAA using endophytes in the field of white biotechnology. Keywords: Endophytes; Kalanchoe pinnata; Indole Acetic Acid; TLC; HPLC

Pilot scale production and characterization of next generation high molecular weight and tense quaternary state polymerized human hemoglobin.

Polymerized human hemoglobin (PolyhHb) is being studied as a possible red blood cell (RBC) substitute for use in scenarios where blood is not available. While the oxygen (O2 ) carrying capacity of PolyhHb makes it appealing as an O2 therapeutic, the commercial PolyhHb PolyHeme® (Northfield Laboratories Inc.) was never approved for clinical use due to the presence of large quantities of low molecular weight (LMW) polymerichemoglobin (Hb) species (<500 kDa), which have been shown to elicit vasoconstriction, systemic hypertension, and oxidative tissue injury in vivo. Previous bench-top scale studies in our lab demonstrated the ability to synthesize and purify PolyhHb using a two-stage tangential flow filtrationpurification process to remove almost all undesirable Hb species (>0.2 µm and <500 kDa) in the material, to create a product that should be safer for transfusion. Therefore, to enable future large animal studies and eventual human clinical trials, PolyhHb synthesis and purification processes need to be scaled up to the pilot scale. Hence in this study, we describe the pilot scale synthesis and purification of PolyhHb. Characterization of pilot scale PolyhHb showed that PolyhHb could be successfully produced to yield biophysical properties conducive for its use as an RBC substitute. Size exclusion high performance liquid chromatography showed that pilot scale PolyhHb yielded a high molecular weight Hb polymer containing a small percentage of LMW Hb species (<500 kDa). Additionally, the auto-oxidation rate of pilot scale PolyhHb was even lower than that of previous generations of PolyhHb. Taken together, these results demonstrate that PolyhHb has the ability to be seamlessly manufactured at the pilot scale to enable future large animal studies and clinical trials.

Pilot scale production of Hermetia illucens (L.) larvae and frass using former foodstuffs

The food and feed sector requires new sustainable sources of protein and innovative solutions for upcycling of food waste (former foodstuffs), which today is downcycled into energy or even wasted. This study aimed at evaluating the use of former foodstuff waste streams as feed substrate for Hermetia illucens (L.) larvae (black soldier fly larvae, BSFL) under long-term and semi-industrial conditions. Different foodstuff-based mixtures and different stocking BSFL densities were used during 20 batches, and quality and safety assessments were performed on the main outputs, namely BSFL production performance, frass impurities, larval and frass nutrient profiles and heavy metal content. About 1400 kg of former foodstuffs (fresh weight) were used to produce 239 kg BSFL and 230 kg frass. The production of BSFL reared on former foodstuffs was highly efficient, with feed conversion rates (FCR) ranging between 2.3 and 5.5 (dry matter basis). The optimization experiment revealed that former foodstuffs-based mixture and high larval density (10 larvae/cm2) lead to highly efficient (FCR: 2.6) and heavy metal-free production of BSFL and frass. The quality of the derived BSFL meal was high in terms of protein and amino acids. Furthermore, the quality of the technical frass was high in terms of N, P, and K levels and minimal packaging material residuals (<2.65%). This investigation suggests that nutrients in former foodstuffs can be successfully and safely recycled in production of BSFL.

Model-assisted process design for better evaluation and scaling up of continuous downstream bioprocessing

Continuous process is a promising alternative for tradition batch process in biomanufacturing, which has higher productivity and lower material consumption. However, despite the maturation of necessary technologies for continuous process, there are few discussion about optimization of full continuous process. One possible reason is that the continuous process is such a complex and interacted process that the traditional experiment-based optimization approach is not sufficient anymore. To address that problem, the process simulation tool SuperPro Designer and continuous capture chromatography model were integrated into a model-assisted design platform for better development of continuous process. The influences of different continuous capture operation modes and sub-lot number under various upstream conditions were analyzed for pilot-scale production. The best combination of operation mode and sub-lot number were determined for the highest equipment utilization without any conflict. After the process optimization, the equipment utilization of continuous process was significantly improved to 27.3%. Then, a pilot-scale case study was carried out to validate the proposed approach. The results showed that the scaling up and process design of continuous process were successful. No time conflict and process failure happened and the final product met the release specification. Finally, the cost of goods was evaluated with SuperPro Designer, and the results showed a 17.4% cost reduction for optimized continuous downstream process compared to batch process, which is promoting for the future industrial applications.

Degradation of sulfamethazine using sludge-derived photocatalysts from dyeing industry and livestock farm: preparation and mechanism

Excess sludge disposal and management has become a global issue of scientific and public concern owing to its high yield and toxicity. In this study, for the first time, we prepared industrial and livestock sludge-derived photocatalysts using dyeing sludge and swine sludge as raw materials, respectively, using a facile pyrolysis coupled ball-milling process without additives. The photocatalytic performances of the two types of photocatalysts were evaluated using sulfamethazine as the target contaminant. The surface morphology and functional groups of the catalysts were analyzed using field-emission scanning electron microscopy combined with energy-dispersive X-ray spectroscopy and Fourier transform infrared spectroscopy. The sulfamethazine removal rate was 91.5 % under optimal conditions. Degradation intermediates were detected, and a possible degradation pathway was proposed. Major reactive oxygen species in the photocatalytic systems were investigated using electron spin resonance spectroscopy and radical trapping experiments. The results revealed that·O2– and h+ played main roles in the degradation of SMZ. Furthermore, a pilot-scale experimental setup was designed and manufactured to realize pilot-scale production. Economic analysis of the pilot-scale experiment suggested that the developed photocatalysts are comparatively suitable for practical applications.

Modeling the Succinic Acid Bioprocess: A Review

Succinic acid has attracted much interest as a key platform chemical that can be obtained in high titers from biomass through sustainable fermentation processes, thus boosting the bioeconomy as a critical production strategy for the future. After several years of development of the production of succinic acid, many studies on lab or pilot scale production have been reported. The relevant experimental data reveal underlying physical and chemical dynamic phenomena. To take advantage of this vast, but disperse, kinetic information, a number of mathematical kinetic models of the unstructured non-segregated type have been proposed in the first place. These relatively simple models feature critical aspects of interest for the design, control, optimization and operation of this key bioprocess. This review includes a detailed description of the phenomena involved in the bioprocesses and how they reflect on the most important and recent models based on macroscopic and metabolic chemical kinetics, and in some cases even coupling mass transport.

Vaccination with Omicron Inactivated Vaccine in Pre-vaccinated Mice Protects against SARS-CoV-2 Prototype and Omicron Variants.

In response to the fast-waning immune response and the great threat of the Omicron variant of concern (VOC) to the public, we report the pilot-scale production of an inactivated Omicron vaccine candidate that induces high levels of neutralizing antibody titers to protect against the Omicron virus. Here, we demonstrate that the inactivated Omicron vaccine is safe and effective in recalling immune responses to the HB02, Omicron, and Delta viruses after one or two doses of BBIBP-CorV. In addition, the efficient productivity and good genetic stability of the manufactured inactivated vaccine is proved. These results support the further evaluation of the Omicron vaccine in a clinical trial.

Evaluation of Conditions to Improve Biomass Production by Submerged Culture of Ganoderma sp.

In the present investigation, the conditions for in vitro submerged culture of a native strain of Ganoderma sp. were evaluated. Different culture medium ingredients, inoculum concentrations, inoculation methods, configuration, and airflows were evaluated to improve biomass production. The addition of thiamine and olive oil to the culture medium increased biomass production, as well as inoculating 6.6 g/L since there are no significant differences in biomass growth according to inoculum origin (pre-inoculum, discs or with spores). The best configuration of the 3 L stirred tank bioreactor was using three impellers and a porous air diffuser of 0.25 volume per volume per minute (vvm), the dry biomass concentration was 22.6 g/L after 12 days of cultivation at 30 °C, much higher than other investigations. This study provides relevant information for pilot-scale production of this fungus for future secondary metabolites. The culture medium was optimized, and it was defined that the concentration and origin of the inoculum did not influence the growth of Biomass, but the aeration and the configuration of the system allowed the establishment of protocols for the cultivation of Ganoderma sp.

Resistance of DISCOVR algae strains to deleterious species

The goal of the DISCOVR consortium is to improve the annualized areal production of open algal production systems through a process of algal strain evaluation and selection combined with testing in outdoor pilot scale production systems. The role of Sandia National Laboratories, in this evaluation process, has been to screen the selected subset of algal strains for resistance to grazing. Sandia tested 9 marine and 8 freshwater strains of algae against a panel of 14 grazer species, (5 marine, 5 freshwater and 4 euryhaline). A total of 153 algal/grazer pairs at two grazer concentrations for a total of 306 assays in triplicate. The relative resistance of each algal strain to each grazer strain was expressed as the decimal fraction of its specific growth rate 29in the presence of the grazer divided by the growth rate of the algae alone. Of the strains tested, those with the greatest resistance to the grazer panel were the freshwater strains Acutodesmus obliquus UTEX 393 and Scenedesmus obliquus. DOE 152z, and the marine strains Micractinium sp 14-F2 and Scenedesmus sp. 46B-D3. Additionally, these assays revealed the unexpected levels of variation in grazer resistance between taxonomically related organisms.

Natural pigment from Monascus: The production and therapeutic significance.

The present review highlights the advantages of using natural colorant over the synthetic one. We have discussed the fermentation parameters that can enhance the productivity of Monascus pigment on agricultural wastes. Food industry is looking for natural colours because these can enhance the esthetic value, attractiveness, and acceptability of food while remaining nontoxic. Many synthetic food colours (Azorubine Carmoisine, quinoline) have been prohibited due to their toxicity and carcinogenicity. Increasing consumer awareness towards the food safety has forced the manufacturing industries to look for suitable alternatives. In addition to safety, natural colorants have been found to have nutritional and therapeutic significance. Among the natural colorants, microbial pigments can be considered as a viable option because of scalability, easier production, no seasonal dependence, cheaper raw materials and easier extraction. Fungi such as Monascus have a long history of safety and therefore can be used for production of biopigments. The present review summarizes the predicted biosynthetic pathways and pigment gene clusters in Monascus purpureus. The challenges faced during the pilot-scale production of Monascus biopigment and taming it by us of low-cost agro-industrial substrates for solid state fermentation has been suggested. Keeping in mind, therapeutic properties of Monascus pigments and their derivatives, they have huge potential for industrial and pharmaceutical application. Though the natural pigments have wide scope in the food industry. However, stabilization of pigment is the greatest challenge and attempts are being made to overcome this by complexion with hydrocolloids or metals and by microencapsulation.

Off-spec fly ash-based lightweight aggregate properties and their influence on the fresh, mechanical, and hydration properties of lightweight concrete: A comparative study

An off-spec fly ash-based spherical lightweight aggregate (LWA), designated as Spherical Porous Reactive Aggregate (SPoRA), was manufactured through a lab pilot-scale production and its engineering properties, including specific gravity, dry rodded unit weight, water absorption, mechanical performance, and pore structure, were evaluated. Using SPoRA, lightweight concrete (LWC) samples were made and their fresh, mechanical, and hydration properties were assessed and compared with LWC samples made using two commercial LWA available in the US market. The results indicated that fine and coarse SPoRA had 72 h water absorption capacities of 16.4 % and 20.9 %, respectively, which were higher than that of the two commercial LWAs. Higher saturated surface dry specific gravity of SPoRA compared to commercial LWAs led to a higher fresh density for the corresponding LWC. Using X-ray computed tomography, large spherical type pores were observed in SPoRA similar to those in the commercial slate-based LWA. The pore size distribution of SPoRA, characterized by a dynamic vapor sorption analyzer, indicated that more than 97 % of the pores had diameters greater than 50 nm. SPoRA’s average bulk crushing strength was 6.8 MPa which was smaller than commercial LWA and was potentially attributed to the differences in manufacturing processes. Nonetheless, SPoRA LWC with 28 days compressive strength of 29 MPa passed the ASTM C330 requirement and had a comparable strength with LWC prepared with the commercial LWAs indicating lesser importance for the LWA strength. SPoRA LWC had a flowability of 5 % compared to LWC prepared with commercial LWAs having flowability of ≥ 18 %. This observation was potentially related to the fluxing agent used in the SPoRA production. SPoRA had a 70 % degree of hydration at 12 days which was comparable with that of LWC prepared with commercial LWA.

Comparing environmental impacts of direct compaction versus wet granulation tableting methods for drugs with poor flowability by life cycle assessment

Pharmaceutical tablet productions by direct compaction (DC) are more environmentally sustainable than wet granulation (WG) owed to DC’s lower energy consumption. For drug particles with poor flowability, however, the environmental benefits of DC become questionable because DC of such drugs requires either pre-compaction treatment, increased excipients’ proportion in the tablets, or using excipients with unfavorable sustainability profiles. Using ibuprofen (IBU) as the model drug with poor flowability, we performed cradle-to-gate life cycle assessment (LCA) using ReCiPe 2016 method to characterize the environmental impacts of DC and WG tablet productions. Material and energy flow data from laboratory-scale (1 and 2.2 kg IBU) and simulated pilot-scale (25 kg IBU) productions were utilized in the LCA. Despite the increased proportion of excipients with less-than-ideal sustainability profile in DC tablets, the environmental impacts of DC tablet production remained smaller than WG tablet production across different production scales, as the impacts were governed by process-level energy consumption. The impacts of DC and WG tablet productions, nevertheless, became closer in magnitude with increasing production scale attributed to superior improvements in the energy requirement and yield of WG tablets. Thus, the environmental beneftis of DC tablets over WG tablets was diminished for drugs with poor flowability. • Environmental impact of tablet production is centered on human/ecosystem toxicity. • Environmental impact of tablet production is ruled by process-level energy usage. • DC tablet has less environmental impact than WG tablet for poorly flowable drugs. • Environmental impacts of DC and WG tablets become more comparable at larger scales.

Enrichment of the protein content of the macroalgae Saccharina latissima and Palmaria palmata

The large brown seaweeds (kelps) are potential sources of protein for animal feed. They have lower protein contents than most red and green algae, but due to potential for large-scale production, they may represent a significant future protein source. The impact of pH, temperature and polysaccharide-degrading enzymes on the solubility and extraction yields of protein from wet Saccharina latissima biomass was investigated. The protein solubility increased with increasing pH and reached maximum of 23% at pH 11, determined as total amino acids (TAA). The enzyme treatments increased the release of soluble compounds by 30–35%. The highest protein yield obtained was 19%, using a ratio of water to wet seaweed of 1:1 for extraction. Even if the yields can be increased by increasing the water amounts used for extraction, the majority of the protein would remain in the insoluble residue after separation. The strategy for production of a larger quantity of protein-enriched biomass was therefore to maintain the insoluble fraction as the product. A pilot scale production was carried out, also including the red algae Palmaria palmata. In total 750 kg S. latissima and 195 kg P. palmata were processed. The protein content in the product increased from 10 to 20% of dry weight (dw) for S. latissima and from 12 to 28% for P. palmata, with yields of 79 and 69%, respectively. The ash content was reduced from 44 to 26% and from 12 to 5% of dw, respectively, for the two species. The main protein loss was free amino acids, which constituted approximately 10% of TAA in the feedstocks. Less essential than non-essential amino acids were lost, thus, the essential amino acids were enriched in the product.

Pilot-Scale Production of A. platensis: Protein Isolation Following an Ultrasound-Assisted Strategy and Assessment of Techno-functional Properties

Pilot-Scale Production of A. platensis: Protein Isolation Following an Ultrasound-Assisted Strategy and Assessment of Techno-functional Properties

A pilot plant scale testing of the application of seaweed‐based natural coating and modified atmosphere packaging for shelf‐life extension of fresh‐cut apple

Codium tomentosum hydroethanolic extract was obtained using a pilot solid–liquid extractor to validate the anti-browning functionality of the extract under industrial conditions. Fresh-cut apple slices were coated by immersion in: (1) a seaweed extract solution (0.5% w/v) and (2) a commercial coating, and the two sets of samples were compared with a control (immersion in water). Packaged samples were stored, under ambient and modified atmosphere conditions at 4°C. After 30 days of storage, the samples that were coated with the seaweed extract and packaged under modified atmosphere, demonstrated lower peroxidase activity and polyphenol oxidation when compared with the samples treated with the commercial additive. These results confirm, at pilot scale and under industrial production conditions, the efficacy of the seaweed extract as a bio-based substitute for the synthetic coatings, which are currently used to prevent browning in fresh-cut apples. Novelty impact statement Fresh-cut fruits are subjected to processing operations leading to a decrease in nutritional and organoleptic properties. It is therefore necessary to adopt strategies to delay the degradative processes. In this study, the efficacy of a pilot-scale production and industrial application of a coating formulated with Codium tomentosum seaweed extract has been established for the first time. This seaweed extract possesses the potential to prevent browning development in fresh-cut apples under industrial operating conditions.

Why Catechin and Epicatechin from Early Hopping Impact the Color of Aged Dry-Hopped Beers while Flavan-3-ol Oligomers from Late and Dry Hopping Increase Colloidal Instability

Dry hopping imparts distinct aromas but also a series of non-volatile compounds suspected of causing flavor and physical instability during beer storage. In this work, color, chill haze, total polyphenols, total flavanoids, and flavan-3-ol monomers (catechin and epicatechin) and oligomers (procyanidin dimers and trimers) were monitored in five commercial pale-colored Belgian dry-hopped beers over 24 months of storage at 20 °C in the dark. Fresh dry-hopped beers contained unusually high levels of flavan-3-ol monomers (up to 6.6 mg/L) and oligomers (up to 14.1 and 10.2 mg/L dimers and trimers, respectively). The increase in color intensity during storage (up to 6.4°EBC) correlated with fresh beer monomer levels, while the oligomer content correlated with chill haze formation (up to 25.7°EBC). The evolution of these two physical attributes also correlated with the level of total polyphenols in the fresh beers. In a pilot-scale production, kettle hopping was shown to impart either monomers (early) or oligomers (late), while dry hopping promoted efficient extraction of both monomers and dimers (extraction yields of 62 and 74%, respectively). Dry hopping thus plays an important role in color and chill haze increase.

Development of a High-Performance Proton Exchange Membrane: From Structural Optimization to Quantity Production

Quantity production of low-cost and high-performance proton exchange membranes (PEMs) used in hydrogen fuel cells is the centerpiece step toward the hydrogen future. Herein, we developed a facile synthesis approach for this task. The prepared PEM breaks through the unfavorable trade-off between thermal-dimensional stability and proton conductivity due to the adequately designed hierarchical polymer structure composed of flexible ionic side-chains anchored onto twisted rigid backbone. Microscale topology structure analyses and molecular dynamics simulations indicate that the hydrated ionic groups self-assemble to form well-connected proton nanochannels. More importantly, the quantity production of the PEM is allowed with a pilot-scale production line. The PEM reaches a peak power density of 1.2 W cm–2 under the realistic low fuel gas flow in a single-cell of H2/O2 fuel cell. Additionally, the PEM maintains profitable fuel cell performance under lower relative humidity (1.1 and 0.9 W cm–2 peak power density at 80 and 60% relative humidity, respectively). In short, we report that a canonical form from the structural optimization of a PEM to the pilot-scale production in which the micromorphology-proton conduction relationship is fully explored.