Eric Charles Peterson

In situ product removal (ISPR), which emerged in the 1990s, enables real-time extraction of metabolites directly from bioreactors. Since then, ISPR has been applied to diverse compounds, including terpenoids, enhancing yield by mitigating product inhibition and degradation. Among different ISPR approaches, solid-phase extraction offers unique advantages such as mechanical solid-liquid separation to remove culture media and cells, thereby further simplifying downstream processing. However, there are currently no side-by-side comparative ISPR fed-batch studies between solid and liquid extractants. This study investigates the use of solid absorptive polymers as in situ extractants for high-titre production of viridiflorol, a terpenoid flavor. In 10 mL mini bioreactors, ∼5% (w/v) solid extractant (Hytrel 3548) loading achieved comparable titre and yield to its liquid counterpart. Additionally, reducing the polymer size from 4 to 1 mm significantly increased extraction efficiency by up to 52%, highlighting the importance of mass transfer considerations for optimal ISPR efficiency. Upon scale-up to a high-yield fed-batch fermentation, the solid-phase system reached a viridiflorol titre of 10.43 g/L, representing ∼53.5% of the yield with the liquid extractant (19.51 g/L). Crucially, the absence of emulsions allowed for a streamlined downstream workflow for solid extractants, achieving 98% purity via simple recrystallization. The ISPR approach was further validated using another flavor product, α-ionone, demonstrating the broad applicability and transferability of solid-phase ISPR in various fermentation processes.

In view of the growing global need for sustainable protein sources, this study explores the utilization of short-chain fatty acids into single-cell protein using the non-conventional yeast Cyberlindnera jadinii. Short-chain fatty acids can be sustainably produced via anaerobic digestion of organic waste, presenting a promising fermentation substrate for a circular bioeconomy. Cyberlindnera jadinii is demonstrated to be capable of growing on acetate, propionate and butyrate as both a carbon and energy source without strong inhibition. Bioprocess development was conducted in stirred tank bioreactors, where a fed-batch pH-stat bioprocess led to improved efficiency without substrate inhibition. The highest titer of 31.3 ± 1.0 g/L, rate of 0.67 ± 0.02 g/L/h and yield of 0.36 ± 0.01 g/g was achieved with propionate. The resulting biomass contained 41.3% crude protein, and 17.3% crude lipids with 81% unsaturated fatty acids. In contrast to acetate and butyrate, propionate as a substrate led to accumulation of 37% odd-chain fatty acids with titer, rate and yield of 1.74 ± 0.06 g/L, 0.037 ± 0.001 g/L/h and 0.020 ± 0.001 g/g. These findings confirm that short-chain fatty acids are viable fermentation substrates not only for single-cell protein, but also unsaturated and odd-chain fatty acid production with Cyberlindnera jadinii.

Enzymes and biosurfactants, often referred to as “green chemicals,” play pivotal roles in enhancing the washing performance of bio-based detergents—a growing trend driven by environmentally conscious consumers. However, the widespread adoption of such bio-based detergents faces challenges, including high costs, limited efficiency, and the need for ongoing innovations. Bacillus species have long been universally acknowledged and exploited for industrial applications, and Bacillus spp. are largely differentiated from other microorganisms for their enzymatic applications, particularly in detergent production. Recent developments in bio-surfactant production by Bacillus sp. support the adoption of green detergents, and these bacterial biosurfactants are a promising source for detergent manufacturing. This article provides an overview of the current understanding of promising Bacillus species and their potential to advance and accelerate the production of bio-based detergents.

A novel approach for converting non-edible plant biomass into single-cell protein and oil (SCPO) via consolidated bioprocessing has been established, leveraging aerotolerant thermophilic cellulolytic consortia consisting mainly of Thermoanaerobacterium thermosaccharolyticum, Sporolactobacillus spp. and Clostridium sensu stricto to achieve the rapid and complete conversion of crystalline cellulose into a consistent cocktail of lactate, acetate and ethanol. This cocktail is an excellent substrate for cultivating organisms for SCPO production and food and feed applications, including Cyberlindnera jadinii, Yarrowia lipolytica and Corynebacterium glutamicum. Cultivation on this cocktail resulted in yields (Y_X/S) of up to 0.43 ± 0.012 g/g, indicating a yield from cellulose (Y_X/Cellulose) of up to 0.27 ± 0.007 g/g (dwb). The resulting SCPO was rich in protein (42.5% to 57.9%), essential amino acids (27.8% to 43.2%) and lipids (7.9% to 8.4%), with unsaturated fatty acid fractions of up to 89%. Unlike fermentation feedstocks derived from easily digested feedstocks (i.e., food waste), this approach has been applied to cellulosic biomass, and this mixed-culture bioconversion can be carried out without adding expensive enzymes. This two-stage cellulosic bioconversion can unlock non-edible plant biomass as an untapped feedstock for food and feed production, with the potential to strengthen resiliency and circularity in food systems.

Brewer’s spent grain (BSG) and spent coffee ground (SCG) are solid sidestreams from beverage production increasingly being upcycled into food, feed and other value-added products. These solid sidestreams are prone to microbial spoilage, negatively impacting their upcycling potential. Three samples each of BSG and SCG were obtained from generators and recycling facilities in Singapore, and their chemical, elemental, and microbial composition was characterized. The spoilage mechanisms were investigated during storage under anaerobic and aerobic conditions. Bacterial loads of sidestreams were low from craft brewery and café sources (6 log₁₀ CFU/g). The microbiome of BSG from recycling facilities was dominated by Bacilli, and B. coagulans was identified as the most prevalent species. SCG from recycling facilities was dominated by lactic acid bacteria, with L. panis being the most prevalent species. Storage up to 14 days under anaerobic conditions led to further bacterial proliferation mainly by Bacilli, lactic acid bacteria, and acetic acid bacteria, while aerobic storage led to extensive fungal contamination, including potential aflatoxin-producing Aspergillus flavus. The results shed light on the spoilage mechanisms, while highlighting the short shelf-life and food safety risks of BSG and SCG to inform valorization strategies.

Nontraditional yeasts prevalent in tropical agricultural fermentations such as coffee and cocoa are known to contribute to aroma profiles, yet the functional roles and interactions between the associated microbial consortia in a farm fermentation are unclear. Here, boiled green bean extract (GBE) from green coffee beans was developed as a rich screening medium to deconstruct the microbial consortia and their interactions during the fermentation of dried green coffee beans. When cultivated in coculture with S. cerevisiae on GBE, strain-specific groupings with distinct volatile organic profiles were observed for nontraditional yeasts (e.g., Hanseniaspora spp., Pichia kudriavzevii). Further changes are evident when constructed consortia composed of nontraditional yeast, S. cerevisiae, and Lactococcus lactis var. cremoris were cultured in GBE, and a comparison with abiotically acidified GBE suggests that pH plays a major role in the influence of lactic acid bacteria (LAB) on fermentation aromas. This approach represents a tool for the development of starter culture formulations to create different flavor profiles in coffee fermentation.

Raspberry ketone (RK) has high commercial value in the food and healthcare industries. A biological route to this flavour compound is an attractive prospect, considering the need to meet consumer demands and sustainable goals; however, it is yet to become an industrial reality. In this work, fungal production of raspberry ketone (RK) and raspberry compounds (RC) via submerged fermentation of Nidula niveo-tomentosa was characterized in flask, stirred-tank reactor (STR), panel bioreactor (PBR), and fluidized bed reactor (FBR) configurations. The results indicate that the panel bioreactor resulted in larger, floccose pellets accompanied by maximum titres of 20.6 mg/L RK and 50.9 mg/L RC. The stirred-tank bioreactor with impeller mixing yielded compact elliptical pellets, induced the highest volumetric productivity of 2.0 mg L⁻¹ day⁻¹, and showed RK selectivity of 0.45. While differing mixing strategies had clear effects on pellet morphology, RK production presented a more direct positive relationship with cultivation conditions, and showed appropriate mixing and aeration favour RK to raspberry alcohol (RA). Overall, this paper highlights the importance of bioreactor design to fungal fermentation, and gives insight into green and industrial bioproduction of value-added natural compounds.

Production of 2-phenylethanol (2-PE) via Kluyveromyces marxianus is well-established. However, co-culture with other microbes in combination with in situ product recovery (ISPR) yields improved selectivity and volumetric productivity. Fermentation ofK. marxianus (MUCL 53775) with direct inclusion of absorptive polymer Hytrel3548 achieved ISPR, but accumulation of the byproduct phenylethyl acetate (PEA) was strongly favored. Co-culture of K. marxianus (MUCL 53775) with Meyerozyma guilliermondii (MUCL 28072) with ISPR limited PEA production, thereby improving the 2-PE selectivity from 13 to 90%, compared to a pure culture of K. marxianus (MUCL 53775) under similar conditions. This improved the volumetric productivity by 85% compared to 2-PE ISPR with a pure culture of K. marxianus. This is the first report of co-culture in a two-phase fermentation for 2-PE bioproduction and demonstrates that interactions between co-culture and ISPR techniques can modulate bioproduction between 2-PE and byproduct PEA, and this technique will be explored for other strain combinations and for other high-value molecules of interest.

Lignocellulosic biomass is both low cost and abundant, and unlike energy crops, can escape associated ethical dilemmas such as arable land use and food security issues. However, their usage as raw material in a biorefinery implies an inherent upstream pretreatment step to access compounds of interest derived from lignocellulosic biomass. Importantly, the efficiency of this step is determinant for the downstream processes, and while many pretreatment methods have been explored, extrusion is both a very flexible and promising technology. Extrusion is well-known in both the polymer and pharmaceutical industries and has been used since the 18th century. However, as a pretreatment method for lignocellulosic biomass, extrusion is relatively new. The first use for this purpose dates back to the 1990s. Extrusion enjoys a high degree of flexibility due to the many available parameters, but an understanding of extrusion requires a knowledge of these parameters and the different relationships between them. In this paper, we present a concise overview of lignocellulosic biomass extrusion by reviewing key extrusion parameters and their associated extruder design components and operating conditions.