Explore the vast range of titles available.

Biopharmaceuticals have revolutionized the field of medicine in the types of active ingredient molecules and treatable indications. Adoption of Quality by Design and Process Analytical Technology (PAT) frameworks has helped the biopharmaceutical field to realize consistent product quality, process intensification, and real-time control. As part of the PAT strategy, Raman spectroscopy offers many benefits and is used successfully in bioprocessing from single-cell analysis to cGMP process control. Since first introduced in 2011 for industrial bioprocessing applications, Raman has become a first-choice PAT for monitoring and controlling upstream bioprocesses because it facilitates advanced process control and enables consistent process quality. This paper will discuss new frontiers in extending these successes in upstream from scale-down to commercial manufacturing. New reports concerning the use of Raman spectroscopy in the basic science of single cells and downstream process monitoring illustrate industrial recognition of Raman’s value throughout a biopharmaceutical product’s lifecycle. Finally, we draw upon a nearly 90-year history in biological Raman spectroscopy to provide the basis for laboratory and in-line measurements of protein quality, including higher-order structure and composition modifications, to support formulation development.

Scaling up industrial microbial processes for commercial production is a high-stakes endeavor, requiring time and investment often exceeding that for laboratory microbe and process development. Omissions, oversights and errors can be costly, even fatal to the program. Approached properly, scale-up can be executed successfully. Three guiding principles are provided as a basis: begin with the end in mind; be diligent in the details; prepare for the unexpected. A detailed roadmap builds on these principles. There is a special emphasis on the fermentation step, which is usually the costliest and also impacts downstream processing. Examples of common scale-up mistakes and the recommended approaches are given. It is advised that engineering resources skilled in integrated process development and scale-up be engaged from the very beginning of microbe and process development to guide ongoing R&D, thus ensuring a smooth and profitable path to the large-scale commercial end.

Synergistic microbial communities are ubiquitous in nature and exhibit appealing features, such as sophisticated metabolic capabilities and robustness. This has inspired fast-growing interest in engineering synthetic microbial consortia for biotechnology development. However, there are relatively few reports of their use in real-world applications, and achieving population stability and regulation has proven to be challenging. In this work, we bridge ecology theory with engineering principles to develop robust synthetic fungal-bacterial consortia for efficient biosynthesis of valuable products from lignocellulosic feedstocks. The required biological functions are divided between two specialists: the fungus Trichoderma reesei , which secretes cellulase enzymes to hydrolyze lignocellulosic biomass into soluble saccharides, and the bacterium Escherichia coli , which metabolizes soluble saccharides into desired products. We developed and experimentally validated a comprehensive mathematical model for T. reesei / E. coli consortia, providing insights on key determinants of the system’s performance. To illustrate the bioprocessing potential of this consortium, we demonstrate direct conversion of microcrystalline cellulose and pretreated corn stover to isobutanol. Without costly nutrient supplementation, we achieved titers up to 1.88 g/L and yields up to 62% of theoretical maximum. In addition, we show that cooperator–cheater dynamics within T. reesei / E. coli consortia lead to stable population equilibria and provide a mechanism for tuning composition. Although we offer isobutanol production as a proof-of-concept application, our modular system could be readily adapted for production of many other valuable biochemicals.

Biobutanol production directly from lignocellulose, known as consolidated bioprocessing (CBP), is expected to be much less expensive than a process where hydrolytic enzyme production, cellulose saccharification, and microbial fermentation are accomplished separately. However, few microbes possess both cellulolytic and solventogenic properties in nature. Current research aims to endow cellulolytic microorganisms with butanol-producing ability or to set up microbial consortia for CBP. This review comprehensively details current achievements attempting to confer butanol-generating ability, not only to cellulolytic Clostridium strains but also to microbial consortia, to address and overcome major challenges in butanol production from cellulose. Recent advances in improving cellulosome activities within cellulolytic Clostridium strains are also emphasized. Isobutanol and butanol can be produced from cellulose in mesophilic Clostridium cellulolyticum and Clostridium cellulovorans by introducing butanol synthesis modules. Thermophilic Clostridium thermocellum shows increased cellulose degradation efficiency. Microbial consortia including cellulolytic and solventogenic strains represent a simple and efficient method to achieve butanol production from cellulose in CBP. Multivariate modular metabolic engineering (MMME) provides a versatile tool to optimize the pathway and strain interactions. Cohesin fusion of β-glucosidase into the Clostridium cellulosome can eliminate the cellobiose inhibition effect, improving the cellulose degradation efficiency of C. thermocellum. Lytic polysaccharide monooxygenases (LPMOs) have been described as ‘cellulase boosters’. Integrating bacterial LPMOs into cellulosomes increases the levels of soluble sugars released from cellulose.

Despite the lack of implementation of consolidated bioprocessing (CBP) at an industrial scale, this bioconversion strategy still holds significant potential as an economically viable solution for converting lignocellulosic biomass (LCB) into biofuels and green chemicals, provided an appropriate organism can be isolated or engineered. The use of Saccharomyces cerevisiae for this purpose requires, among other things, the development of a cellulase expression system within the yeast. Over the past three decades, numerous studies have reported the expression of cellulase-encoding genes, both individually and in combination, in S. cerevisiae . Various strategies have emerged to produce a core set of cellulases, with differing degrees of success. While one-step conversion of cellulosic substrates to ethanol has been reported, the resulting titers and productivities fall well below industrial requirements. In this review, we examine the strategies employed for cellulase expression in yeast, highlighting the successes in developing basic cellulolytic CBP-enabled yeasts. We also summarize recent advancements in rational strain design and engineering, exploring how these approaches can be further enhanced through modern synthetic biology tools to optimize CBP-enabled yeast strains for potential industrial applications. Key points • S. cerevisiae’s lack of cellulolytic ability warrants its engineering for industry. • Advancements in the expression of core sets of cellulases have been reported. • Rational engineering is needed to enhance cellulase secretion and strain robustness. • Insights gained from omics strategies will direct the future development of CBP strains. Graphical Abstract

The impact of climate change presents an opportunity for orphan crops like millet to contribute to sustainable food systems. A prime example of an orphan crop with special traits and the potential to develop climate-smart agriculture is Barnyard millet (BYM). Alkaloids, steroids, polysaccharides, glycosides, tannins, phenols, dietary lignans and flavonoids are just a few of the antioxidants that are abundant in BYM. BYM’s antioxidant potential, prebiotic status, anti-inflammatory, hypoglycemic, antibacterial and anticancerous properties help it to combat a myriad of diseases. The antinutritional compounds present in BYM such phytic acid, tannins, polyphenols, and amylase inhibitors limit the absorption of minerals because they form complexes with dietary minerals like calcium, zinc, magnesium, and iron and make them inaccessible for absorption. Various processing methods like dehulling, soaking, heating, gamma irradiation, cold plasma processing, fermentation might enhance the nutritional and technological functional qualities of BYM. The bioactives in BYM can improve their bioavailability, in vitro digestibility, efficiency, structural modification and stability by biological processing techniques such as germination and fermentation employing microbial strains. BYM straw’s high cellulose and hemicellulose percentage makes C5 and C6 sugars accessible for bioconversion into bioethanol.

Background Consolidated bioprocessing, which combines saccharolytic and fermentative abilities in a single microorganism, is receiving increased attention to decrease environmental and economic costs in lignocellulosic biorefineries. Nevertheless, the economic viability of lignocellulosic ethanol is also dependent of an efficient utilization of the hemicellulosic fraction, which contains xylose as a major component in concentrations that can reach up to 40% of the total biomass in hardwoods and agricultural residues. This major bottleneck is mainly due to the necessity of chemical/enzymatic treatments to hydrolyze hemicellulose into fermentable sugars and to the fact that xylose is not readily consumed by Saccharomyces cerevisiae—the most used organism for large-scale ethanol production. In this work, industrial S. cerevisiae strains, presenting robust traits such as thermotolerance and improved resistance to inhibitors, were evaluated as hosts for the cell-surface display of hemicellulolytic enzymes and optimized xylose assimilation, aiming at the development of whole-cell biocatalysts for consolidated bioprocessing of corn cob-derived hemicellulose. Results These modifications allowed the direct production of ethanol from non-detoxified hemicellulosic liquor obtained by hydrothermal pretreatment of corn cob, reaching an ethanol titer of 11.1 g/L corresponding to a yield of 0.328 g/g of potential xylose and glucose, without the need for external hydrolytic catalysts. Also, consolidated bioprocessing of pretreated corn cob was found to be more efficient for hemicellulosic ethanol production than simultaneous saccharification and fermentation with addition of commercial hemicellulases. Conclusions These results show the potential of industrial S. cerevisiae strains for the design of whole-cell biocatalysts and paves the way for the development of more efficient consolidated bioprocesses for lignocellulosic biomass valorization, further decreasing environmental and economic costs.

Increased energy demands in today’s world have led to the exploitation of fossil resources as fuel. Fossil resources are not only on the verge of extinction but also causing environmental and economic issues. Due to these reasons, scientists have started focusing their interest on other eco-friendly processes to biofuel and recently, second-generation biorefinery is gaining much more attention. In second-generation biorefinery, the main objective is the valorization of lignocellulosic biomass cost-effectively. Therefore, many scientists started different bioprocessing techniques like Consolidated Bioprocessing (CBP) to produce ethanol by using a single or plethora of microorganisms to produce ethanol in a single process. In this review, in-depth study on CBP is assessed as well as biofuel’s socio-economic value and a brief study of biorefineries. The study not only involves innovative approaches used in CBP but their effect on society and economic aspects.

One approach to reducing the costs of advanced biofuel production from cellulosic biomass is to engineer a single microorganism to both digest plant biomass and produce hydrocarbons that have the properties of petrochemical fuels. Such an organism would require pathways for hydrocarbon production and the capacity to secrete sufficient enzymes to efficiently hydrolyze cellulose and hemicellulose. To demonstrate how one might engineer and coordinate all of the necessary components for a biomass-degrading, hydrocarbon-producing microorganism, we engineered a microorganism naïve to both processes, Escherichia coli, to grow using both the cellulose and hemicellulose fractions of several types of plant biomass pretreated with ionic liquids. Our engineered strains express cellulase, xylanase, beta-glucosidase, and xylobiosidase enzymes under control of native E. coli promoters selected to optimize growth on model cellulosic and hemicellulosic substrates. Furthermore, our strains grow using either the cellulose or hemicellulose components of ionic liquid-pretreated biomass or on both components when combined as a coculture. Both cellulolytic and hemicellulolytic strains were further engineered with three biofuel synthesis pathways to demonstrate the production of fuel substitutes or precursors suitable for gasoline, diesel, and jet engines directly from ionic liquid-treated switchgrass without externally supplied hydrolase enzymes. This demonstration represents a major advance toward realizing a consolidated bioprocess. With improvements in both biofuel synthesis pathways and biomass digestion capabilities, our approach could provide an economical route to production of advanced biofuels.

There is a renewed interest in acetone-butanol-ethanol (ABE) fermentation from renewable substrates for the sustainable and environment-friendly production of biofuel and platform chemicals. However, the ABE fermentation is associated with several challenges due to the presence of heterogeneous components in the renewable substrates and the intrinsic characteristics of ABE fermentation process. Hence, there is a need to select optimal substrates and modify their characteristics suitable for the ABE fermentation process or microbial strain. This “designed biomass” can be used to establish the consolidated bioprocessing systems. As there are very few reports on designed biomass, the main objectives of this review are to summarize the main challenges associated with ABE fermentation from renewable substrates and to introduce feasible strategies for designing the substrates through pretreatment and hydrolysis technologies as well as through the establishment of consolidated bioprocessing systems. This review offers new insights on improving the efficiency of ABE fermentation from designed renewable substrates.