BACKGROUND: An important step in replacing petrochemical products with sustainable, cost-effective alternatives is the use of feedstocks other than, e.g., pure glucose in the fermentative production of platform chemicals. Ustilaginaceae offer the advantages of a wide substrate spectrum and naturally produce a versatile range of value-added compounds under nitrogen limitation. A promising candidate is the dicarboxylic acid malic acid, which may be applied as an acidulant in the food industry, a chelating agent in pharmaceuticals, or in biobased polymer production. However, fermentable residue streams from the food and agricultural industry with high nitrogen content, e.g., sugar beet molasses, are unsuited for processes with Ustilaginaceae, as they result in low product yields due to high biomass and low product formation.
RESULTS: This study uncovers challenges in evaluating complex feedstock applicability for microbial production processes, highlighting the role of secondary substrate limitations, internal storage molecules, and incomplete assimilation of these substrates. A microliter-scale screening method with online monitoring of microbial respiration was developed using malic acid production with Ustilago trichophora on molasses as an application example. Investigation into nitrogen, phosphate, sulphate, and magnesium limitations on a defined minimal medium demonstrated successful malic acid production under nitrogen and phosphate limitation. Furthermore, a reduction of nitrogen and phosphate in the elemental composition of U. trichophora was revealed under the respective secondary substrate limitation. These adaptive changes in combination with the intricate metabolic response hinder mathematical prediction of product formation and make the presented screening methodology for complex feedstocks imperative. In the next step, the screening was transferred to a molasses-based complex medium. It was determined that the organism assimilated only 25% and 50% of the elemental nitrogen and phosphorus present in molasses, respectively. Due to the overall low content of bioavailable phosphorus in molasses, the replacement of the state-of-the-art nitrogen limitation was shown to increase malic acid production by 65%.
CONCLUSIONS: The identification of phosphate as a superior secondary substrate limitation for enhanced malic acid production opens up new opportunities for the effective utilization of molasses as a more sustainable and cost-effective substrate than, e.g., pure glucose for biobased platform chemical production.

In the present study, the nutritional and sensory properties of spelt cookies without wild garlic, cookies with fresh wild garlic, cookies with osmodehydrated wild garlic in sugar beet molasses, and cookies with osmodehydrated wild garlic in an aqueous solution of sucrose and salt were evaluated and compared. The tested cookie samples were characterized in terms of total antioxidative activity, the total content of phenols, flavonoids, and thiosulfates, the presence of dominant phenols, the content of betaine and dietary fiber, antioxidant activity after in vitro digestion, and sensory attributes for appearance, taste, smell, and texture. The results proved that the addition of wild garlic leaves osmodehydrated in molasses provided the cookies with the best nutritional and bioactive properties: 1.75 times higher total phenols content, 2.4 times higher total flavonoids content, 1.52 times higher total thiosulfates content, and 1.56 times higher betaine content, and a total quality increase of 54% compared to the control cookies. The cookies enriched with osmodehydrated wild garlic in molasses were rated as pleasant and acceptable, but also more complex compared to other cookies. The production of this nutritionally and sensory-improved cookie would contribute to expanding the assortment of flour confectionery products, especially for consumers who care about health and nutrition.

The widespread adoption of Poly(3-hydroxybutyrate) (PHB) encounters challenges due to its higher production costs compared to conventional plastics. To overcome this obstacle, this study investigates the use of low-cost raw materials and optimized production methods. Specifically, food processing byproducts such as corn germ and corn bran were utilized as solid substrates through solid-state fermentation, enriched with molasses and cheese whey. Employing the One Factor at a Time technique, we examined the effects of substrate composition, temperature, initial substrate moisture, molasses, and cheese whey on PHB production at the flask scale. Subsequently, experiments were conducted at the bioreactor scale to evaluate the influence of aeration. In flask-scale experiments, the highest PHB yield, reaching 4.1 (g/kg Initial Dry Weight Substrate) (IDWS) after 72 hours, was achieved using a substrate comprising a 1:1 mass ratio of corn germ to corn bran supplemented with 20 % (v/w) cheese whey. Furthermore, PHB production in a 0.5-L packed-bed bioreactor yielded a maximum of 8.4 (g/kg IDWS), indicating a more than 100 % increase in yield after 72 hours, with optimal results achieved at an aeration rate of 0.5 l/(kg IDWS. h).

Pomegranate molasses is made from concentrated pomegranate juice with nothing added. Due to its nutritional value, limitation in production, and high production cost, this product may be adulterated by date syrup. This study was done to differentiate various types of pomegranate molasses and investigate the possibility of nonauthenticity detection in pomegranate molasses samples using the hyperspectral imaging (HSI) technique compared with physicochemical measurement analysis. The physicochemical properties (brix index, sucrose, acidity, ash content, pH, and formalin index) of 24 samples were measured as the reference analysis method, and it was found that the formalin index was a good factor for pomegranate molasses authenticity evaluation. Additionally, an HSI system (400-1000 nm) was used as a nondestructive and rapid screening method to capture spectral data of the samples. The evolutionary wavelength selection algorithm was applied to select effective wavelengths in sample clustering based on the obtained Davies-Bouldin index. Next, principal component analysis was used to visually interpret the spectral data of the sample when using the selected wavelengths and the whole spectra of the samples. Finally, an automatic clustering algorithm by the artificial bee colony as an unsupervised method was developed for the clustering of the authentic and nonauthentic samples. The method did not need descriptively labeled samples and obtained agreed satisfactorily with the degree of nonauthenticity in the samples. This study showed that the developed HSI technique coupled with an automatic clustering algorithm could detect date syrup nonauthenticity in pomegranate samples from the level of 5% adulteration.

BACKGROUND: Saccharomyces cerevisiae is an important microorganism in ethanol synthesis, and with sugarcane molasses as the feedstock, ethanol is being synthesized sustainably to meet growing demands. However, high-concentration ethanol fermentation based on high-concentration sugarcane molasses-which is needed for reduced energy consumption of ethanol distillation at industrial scale-is yet to be achieved.
RESULTS: In the present study, to identify the main limiting factors of this process, adaptive laboratory evolution and high-throughput screening (Py-Fe3+) based on ARTP (atmospheric and room-temperature plasma) mutagenesis were applied. We identified high osmotic pressure, high temperature, high alcohol levels, and high concentrations of K+, Ca2+, K+ and Ca2+ (K+&Ca2+), and sugarcane molasses as the main limiting factors. The robust S. cerevisiae strains of NGT-F1, NGW-F1, NGC-F1, NGK+, NGCa2+ NGK+&Ca2+-F1, and NGTM-F1 exhibited high tolerance to the respective limiting factor and exhibited increased yield. Subsequently, ethanol synthesis, cell morphology, comparative genomics, and gene ontology (GO) enrichment analysis were performed in a molasses broth containing 250 g/L total fermentable sugars (TFS). Additionally, S. cerevisiae NGTM-F1 was used with 250 g/L (TFS) sugarcane molasses to synthesize ethanol in a 5-L fermenter, giving a yield of 111.65 g/L, the conversion of sugar to alcohol reached 95.53%. It is the highest level of physical mutagenesis yield at present.
CONCLUSIONS: Our results showed that K+ and Ca2+ ions primarily limited the efficient production of ethanol. Then, subsequent comparative transcriptomic GO and pathway analyses showed that the co-presence of K+ and Ca2+ exerted the most prominent limitation on efficient ethanol production. The results of this study might prove useful by promoting the development and utilization of green fuel bio-manufactured from molasses.

This study aimed to evaluate the effect of including soybean molasses (SM) on performance, blood parameters, carcass traits, meat quality, fatty acid, and muscle (longissimus thoracis) transcriptomic profiles of castrated lambs. Twenty Dorper × Santa Inês lambs (20.06 ± 0.76 kg body weight [BW]) were assigned to a randomized block design, stratified by BW, with the following treatments: CON: 0 g/kg of SM and SM20: 200 g/kg of SM on dry matter basis, allocated in individual pens. The diet consisted of 840 g/kg concentrate and 160 g/kg corn silage for 76 d, with the first 12 d as an adaptation period and the remaining 64 d on the finishing diet. The SM20 diet increased blood urea concentration (P = 0.03) while reduced glucose concentration (P = 0.04). Lambs fed SM showed higher subcutaneous fat deposition (P = 0.04) and higher subcutaneous adipocyte diameter (P < 0.01), in addition to reduced meat lipid oxidation (P < 0.01). SM reduced the quantity of branched-chain fatty acids in longissimus thoracis (P = 0.05) and increased the quantity of saturated fatty acids (P = 0.01). In the transcriptomic analysis, 294 genes were identified as differentially expressed, which belong to pathways such as oxidative phosphorylation, citric acid cycle, and monosaccharide metabolic process. In conclusion, diet with SM increased carcass fat deposition, reduced lipid oxidation, and changed the energy metabolism, supporting its use in ruminant nutrition.
This study investigated the effects of incorporating soybean molasses (SM) into the diet of castrated lambs on various aspects of their performance and meat quality. Twenty lambs were divided into two groups: one was fed a control diet without SM whereas the other was fed a similar diet but containing 20% of SM. The feeding trial lasted for 76 d. Results showed that the SM inclusion in the diet led to increased blood urea levels and decreased glucose concentrations. SM inclusion also resulted in lambs with higher levels of subcutaneous fat and larger adipocytes, while reducing meat lipid oxidation. Moreover, SM altered fatty acid composition in the meat, decreasing branched-chain fatty acids and increasing saturated fatty acids. In agreement with these findings, transcriptomic analysis revealed a significant change in the expression of genes related to energy metabolism in the muscle of lambs fed SM. In conclusion, incorporating SM in lamb’s diet increased fat deposition, improved meat quality, and induced a transcriptomic change in the muscle energy metabolism, supporting its potential use in ruminant nutrition.

The objective of this experiment was to investigate the effects of Lactobacillus plantarum and molasses on the nutrient composition, fermentation quality, bacterial count, aerobic stability, and microflora of alfalfa silage in sandy grasslands. The experimental treatments included control (CK), 106 CFU/g Lactobacillus plantarum (L), 5% molasses (M), and 106 CFU/g Lactobacillus plantarum + 5% molasses (LM). The nutrient composition, fermentation quality, bacterial count, aerobic stability, and microflora were determined after 14 days and 56 days of ensiling, respectively. The results showed that the addition of L, M, and LM reduced dry matter loss (DM), neutral detergent fiber (NDF), and acid detergent fiber (ADF) content, and increased water-soluble carbohydrates (WSC) and ether extract (EE) content, compared to the CK group. Meanwhile, more lactic acid (LA) and accelerated fermentation were observed, causing the pH value to drop below 4.5 in the L, M, and LM groups after 56 days of ensiling. The addition of L, M, and LM promoted lactic acid bacteria (LAB), and inhibited yeast. The addition of L significantly increased the content of acetic acid (AA). In terms of microflora, the addition of L, M, and LM made Firmicutes become the dominant bacterial phylum earlier, while Lactobacillus, Weissella, and Pediococcus had a higher abundance. According to the result of Pearson\'s correlation, there is a very significant negative correlation between pH value and Lactobacillus (P < 0.01) and a very significant positive correlation between pH value and Lactococcus, Enterobacter, Enterococcus, and Leuconostoc (P < 0.01), which may be inhibited by Lactobacillus under the decreased pH value. The results of the prediction of microbial genes indicated that the addition of M could enhance the carbohydrate metabolism and membrane transport metabolism, which may contribute to LA production by LAB metabolism. In general, L, M and LM all improved the fermentation quality and reduced the loss of nutrients to varying degrees, but considering the fermentation quality, the overall effects of M and LM were better than L. M and LM are recommended to be used as silage additives in the process of alfalfa silage in sandy grasslands to improve the quality.

Production of carotenoids by yeast fermentation is an advantaged technology due to its easy scaling and safety. Nevertheless, carotenoid production needs an economic culture medium and other efficient yeast stains. The study aims to isolate and identify a yeast strain capable of producing carotenoids using a cost-effective substrate. A new strain was identified as Rhodotorula toruloides L/24-26-1, which can produce carotenoids at different pretreated and unpretreated sugarcane molasses concentrations (40 and 80 g/L). The highest biomass concentration (18.6 ± 0.6 g/L) was reached in the culture using 80 g/L of hydrolyzed molasses. On the other hand, the carotenoid accumulation reached the maximum value using pretreated molasses at 40 g/L (715.4 ± 15.1 µg/g d.w). In this case, the β-carotene was 1.5 times higher than that on the control medium. The yeast growth in molasses was not correlated with carotenoid production. The most outstanding production of The DPPH, ABTS, and FRAP tests demonstrated the antioxidant activity of the obtained carotenogenic extracts. This research demonstrated the R. toruloides L/24-26-1 strain biotechnological potential for carotenoid compounds. The yeast produces carotenoids with antioxidant activity in an inexpensive medium, such as sulfuric acid pretreated and unpretreated molasses.

Fructooligosaccharides (FOS) are a type of important prebiotics and produced by transfructosylating enzymes. In this study, sugarcane molasses was used as the substrate for production of transfructosylating enzymes by Aureobasidium pullulans FRR 5284. NaNO3 was a superior nitrogen source to yeast extract for production of transfructosylating enzymes by A. pullulans FRR 5284 and decreasing the ratio of NaNO3 to yeast extract nitrogen from 1:0 to 1:1 resulted in the reduction of the total transfructosylating activity from 109.8 U/mL to 82.5 U/mL. The addition of only 4.4 g/L NaNO3 into molasses-based medium containing 100 g/L mono- and di-saccharides resulted in total transfructosylating activity of 123.8 U/mL. Scale-up of the A. pullulans FRR 5284 transfructosylating enzyme production process from shake flasks to 1 L bioreactors improved the enzyme activity and productivity to 171.7 U/mL and 3.58 U/mL/h, 39% and 108% higher than those achieved from shake flasks, respectively. Sucrose (500 g/L) was used as a substrate for extracellular, intracellular, and total A. pullulans FRR 5284 transfructosylating enzymes, with a maximum yield of 61%. Intracellular, extracellular, and total A. pullulans FRR 5284 transfructosylating enzymes from different production systems resulted in different FOS profiles, indicating that FOS profiles can be controlled by adjusting intracellular and extracellular enzyme ratios and, hence prebiotic activity.

This study endeavoured to capitalize on prickly pear by-products for the optimization of molasses formulation, targeting elevated antioxidant capacities and superior sugar contents. Through robust statistical modelling, the optimal cooking parameters-temperature (70-80 °C) and duration (60-90 min)-were determined, guided by responses of antioxidant activity and Brix value. A D-Optimal mixture design further delineated the ideal proportions of molasses components (pulp, peel, and seeds). Characterization revealed that peel harboured the highest concentrations of total polyphenols (396.41 mg GAE/100g FW) and flavonoids (234.26 mg CE/100g FW), emphasizing its antioxidant potential (DPPH inhibition IC50: 12.72 µg/ml). The optimal cooking conditions were established at 78.35 °C for 79.70 min, with predictive equations guiding ingredient proportions (0.265 g pulp, 0.710 g peel, 0.025 g seed). Intriguingly, while peel inclusion enhanced total sugar content and antioxidant activity, seed incorporation exerted a contrasting effect by reducing total sugar content and limiting antioxidant activity.