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With the global population continuously rising, efficient bioconversion of inedible agricultural by-products is crucial for human food and energy sustainability. We here propose solid-state fermentation approaches to efficiently convert biopolymers into oligomers/monomers by accelerating the natural degradation process of the versatile Streptomyces sp. strain SCUT-3. Using fish skin as a representative by-product, 54.3 g amino acids and 14.7 g peptides (91 % < 2500 Da) were recovered from 89.0 g protein in 100 g tilapia skin sample by collagenase-overexpressed SCUT-3 for seven days at a 1:4 substrate:liquid ratio. Fish skin collagen hydrolysates exhibited excellent anti-oxidation, anti-hypertension, scratch-repairing, anti-aging, anti-ultraviolet radiation, and anti-inflammation effects on human skin fibroblasts In vitro and zebrafish larvae in vivo, indicating their potential applications in healthcare/skincare and anti-atopic dermatitis. As Laozi said, the divine law follows nature. This study underscores the efficacy of genetically engineered SCUT-3 according to its natural biomass utilization laws in large-scale biopolymer conversion.

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The management of rural waste, particularly agri-food waste, poses a major challenge to the ecosystem health. This study investigated the efficacy of black soldier fly larvae (Hermetia illucens L., BSFL) bioconversion for agri-food waste under independent treatment or co-treatment strategies using chicken manure and food waste as a model system. The results showed a synergistic effect of co-treating agri-food waste from different sources. The co-treatment strategy enhanced bioconversion efficiency, resulting in a 1.31-fold waste reduction rate and a 1.93-fold bioconversion rate. Additionally, larval growth performance and biomass quality of BSFL were improved, while lauric acid and oleic acid were enriched in the larval fat from the co-treatment strategy. 16S rRNA amplicon sequencing revealed that the co-treatment strategy reshaped both the residue and larval gut microbiota, with distinct enrichment of taxonomical biomarkers. Furthermore, under this strategy, metabolic functions of the residue microbiota were significantly activated, especially carbohydrate, amino acid, and lipid metabolism were enhanced by 16.3%, 23.5%, and 20.2%, respectively. The early colonization of lactic acid bacteria (Weisella and Aerococcus) in the residue, coupled with a symbiotic relationship between Enterococcus in the larval gut and the host, likely promoted organic matter degradation and larval growth performance. Scaling up the findings to a national level in China suggests that the co-treatment strategy can increase waste reduction quantity by 86,329 tonnes annually and produce more larval protein and fat with a market value of approximately US$237 million. Therefore, co-treatment of agri-food waste streams using BSFL presents a sustainable solution for rural waste management that potentially contributes to the achievement of SDG2 (Zero Hunger), SDG3 (Good Health and Well-Being), and SDG12 (Responsible Consumption and Production).

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Organic waste including food garbage (FG) forms a major part of man-made problems that are highly associated with global pollution. This includes emission of greenhouse gases (GHGs) and foul odor which negatively affect human health. Interestingly, bioconversion of FG by black soldier fly larvae (BSFL) has been reported to reduce foul odors released from decaying FG. This paper will give overview on the potential of BSFL in lowering putrid odors from FGs. Thus, various bioconversion treatment methods of managing FG including were compared and discussed. The life cycle and role of BSF in reducing putrid odors from biowastes were also discussed in detail. Lastly, the potential utilization of BSFL in controlling odors and GHGs as well as the economic value of products derived from BSFL bioconversion were also discussed. BSFL inoculation slightly reduces odor compounds by modifying odor-producing compounds and microbes in FG. However, BSFL effectiveness is highly influenced by FG decomposition rate.

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A key aspect of sustainable bioeconomy is the recirculation of renewable, agricultural waste streams as substrates for microbial production of high-value compounds. One approach is the bioconversion of corn stover, an abundant maize crop byproduct, using the fungal maize pathogen Ustilago maydis. U. maydis is already used as a unicellular biocatalyst in the production of several industrially-relevant compounds using plant biomass hydrolysates. In this study, we demonstrate that U. maydis can grow using untreated corn stover as its sole carbon source. We developed a small-scale bioreactor platform to investigate U. maydis processing of corn stover, combining online monitoring of fungal growth and metabolic activity profiles with biochemical analyses of the pre- and post-fermentation residues. Our results reveal that U. maydis primarily utilizes soluble sugars i.e., glucose, sucrose and fructose present in corn stover, with only limited exploitation of the abundant lignocellulosic carbohydrates. Thus, we further explored the biotechnological potential of enhancing U. maydis´ lignocellulosic utilization. Additive performance improvements of up to 120 % were achieved when using a maize mutant with increased biomass digestibility, co-fermentation with a commercial cellulolytic enzyme cocktail, and exploiting engineered fungal strains expressing diverse lignocellulose-degrading enzymes. This work represents a key step towards scaling up the production of sustainable compounds from corn stover using U. maydis and provides a tool for the detailed monitoring of the fungal processing of plant biomass substrates.

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Curcumin is a plant-derived secondary metabolite exhibiting antitumor, neuroprotective, antidiabetic activities, and so on. We previously isolated Escherichia coli as an enterobacterium exhibiting curcumin-converting activity from human feces, and discovered an enzyme showing this activity (CurA) and named it NADPH-dependent curcumin/dihydrocurcumin reductase. From soil, here, we isolated a curcumin-degrading microorganism (No. 34) using the screening medium containing curcumin as the sole carbon source and identified as Rhodococcus sp. A curcumin-degrading enzyme designated as CurH was purified from this strain and characterized, and compared with CurA. CurH catalyzed hydrolytic cleavage of a carbon-carbon bond in the ß-diketone moiety of curcumin and its analogs, yielding two products bearing a methyl ketone terminus and a carboxylic acid terminus, respectively. These findings demonstrated that a curcumin degradation reaction catalyzed by CurH in the soil environment was completely different from the one catalyzed by CurA in the human microbiome. Of all the curcumin analogs tested, suitable substrates for the enzyme were curcuminoids (i.e., curcumin and bisdemethoxycurcumin) and tetrahydrocurcuminoids. Thus, we named this enzyme curcuminoid hydrolase. The deduced amino acid sequence of curH exhibited similarity to those of members of acetyl-CoA C-acetyltransferase family. Considering results of oxygen isotope analyses and a series of site-directed mutagenesis experiments on our enzyme, we propose a possible catalytic mechanism of CurH, which is unique and distinct from those of enzymes degrading ß-diketone moieties such as ß-diketone hydrolases known so far.

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Heavy metals emanate from diverse anthropogenic activities and the top soil in the vicinity of these activities acts as an immediate sink and facilitates diffusion of heavy metals into the food chain. In the semi-arid plains of India, Prosopis juliflora is the most common and dominant weed along the motorways and barren lands including industrial environs. This investigation hypothesizes the adaptive nature of Prosopis juliflora in the metal enriched soils and attempts to understand its hyper-accumulating potential of metals besides bioconversion/detoxification capability. Prosopis juliflora samples (root, stem, leaves, and pods) from 100 sites in the environs of anthropogenic activities (vehicular emissions and industrial operations) were analyzed for heavy metal concentrations (Cu, Fe, Cr, Cd, Ni, Pb). Prosopis juliflora accumulate metals at the rate of 0.138 mg/kg/day DW for Copper (Cu), Fe: 0.142 mg/kg/day DW, Cr: 0.114 mg/kg/day DW, Ni: 0.048 mg/kg/day DW, Pb: 0.052 mg/kg/day DW, Cd: 0.009 mg/kg/day DW. Furthermore, X-ray Photoelectron Spectroscopy (XPS) metal oxidation state analysis revealed that in the pods of Prosopis juliflora heavy metals (Fe, Cr, Pb) largely existed in non-toxic form (toxic:non-toxic - 3:6), while in the under canopy soil, metals predominantly existed in toxic form (toxic:non-toxic - 7:2); conclusively XPS results ascertains the heavy metal bioconversion/detoxification potential of the plant. These findings suggest that presence of Prosopis juliflora coppice in the barren landscapes across the transportation corridors and metal based industrial zones may ideally favor phyto-remediation of heavy metals.

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Industrial wastewater, often characterized by its proximity to neutral pH, presents a promising opportunity for fungal utilization despite the prevalent preference of fungi for acidic conditions. This review addresses this discrepancy, highlighting the potential of certain industrial wastewaters, particularly those with low pH levels, for fungal biorefinery. Additionally, the economic implications of biomass recovery and compound separation, factors that require explicit were emphasized. Through an in-depth analysis of various industrial sectors, including food processing, textiles, pharmaceuticals, and paper-pulp, this study explores how filamentous fungi can effectively harness the nutrient-rich content of wastewaters to produce valuable resources. The pivotal role of ligninolytic enzymes synthesized by fungi in wastewater purification is examined, as well as their ability to absorb metal contaminants. Furthermore, the diverse benefits of fungal biorefinery are underscored, including the production of protein-rich single-cell protein, biolipids, enzymes, and organic acids, which not only enhance environmental sustainability but also foster economic growth. Finally, the challenges associated with scaling up fungal biorefinery processes for wastewater treatment are critically evaluated, providing valuable insights for future research and industrial implementation. This comprehensive analysis aims to elucidate the potential of fungal biorefinery in addressing industrial wastewater challenges while promoting sustainable resource utilization.

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The biological conversion of methane under ambient conditions can be performed by methanotrophs that utilize methane as both a sole source of energy and a carbon source. However, compared to the established microbial chassis used for general fermentation with sugar as a feedstock, the productivity of methanotrophs is low. The fundamental knowledge of their metabolic or cellular bottlenecks is limited. In this review, the industrial-scale potential of methane bioconversion was evaluated. In particular, the enzyme kinetics associated with the oxidation and assimilation of methane were investigated to evaluate the potential of methane fermentation. The kinetics of enzymes involved in methane metabolism were compared with those used in the metabolic processes of traditional fermentation (glycolysis). Through this analysis, the current limitations of methane metabolism were identified. Methods for increasing the efficiency of methane bioconversion and directions for the industrial application of methane-based fermentation were discussed.

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One-pot enzymatic synthesis is flourishing in synthetic chemistry, heralding a sustainable and green era. Recent advancements enable the creation of complex enzymatic prosthetic groups and regeneration of enzymatic cofactors such as S-adenosylmethionine. The next frontier is to develop the effective and innovative cofactors for essential micronutrients, metabolic modulators, and biomedicines.

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Methanol is a promising feedstock for biomanufacturing, but the efficiency of methanol-based bioprocesses is limited by the low rate of methanol utilization pathways and methanol toxicity. Yeast diversity is an attractive biological resource to develop efficient bioprocesses since any effort with strain improvement is more deserving if applied to innate robust strains with relevant catabolic and biosynthetic potential. The present study is in line with such rational and describes the isolation and molecular identification of seven isolates of the methylotrophic species Candida boidinii from waters derived from the traditional curation of olives, in different years, and from contaminated superficial soil near fuel stations. The yeast microbiota from those habitats was also characterized. The four C. boidinii isolates obtained from the curation of olives' water exhibited significantly higher maximum specific growth rates (range 0.15-0.19 h-1), compared with the three isolates obtained from the fuel contaminated soils (range 0.05-0.06 h-1) when grown on methanol as the sole C-source (1% (v/v), in shake flasks, at 30°C). The isolates exhibit significant robustness towards methanol toxicity that increases as the cultivation temperature decreases from 30°C to 25°C. The better methanol-based growth performance exhibited by C. boidinii isolates from olives´ soaking waters could not be essentially attributed to higher methanol tolerance. These methanol-efficient catabolizing isolates are proposed as a promising platform to develop methanol-based bioprocesses.

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Rearing scale may influence black soldier fly (BSF) larvae traits when they are fed on a single diet, but different feeding substrates have not been tested yet. This study evaluated the effects of wheat starch processing by-products-based diets on growth performance, bioconversion efficiency, and nutritional profile of BSF larvae reared in different scales. Four diets (D1 and D2 [isonitrogenous, isolipidic and isoenergetic]; D3 and D4 [displaying 1:1 and 1:2 as protein to carbohydrate ratios, respectively]) were tested at 3 rearing scales (4 replicate boxes/diet, with a constant volume [0.84 cm3]/larva and feed [0.7 g]/larva): 1) small (S; 12 × 12 cm, substrate height: 4 cm, 686 6-day-old larvae (6-DOL)/box), 2) medium (M, 32 × 21 cm, substrate height: 7 cm, 5 600 6-DOL/box), and 3) large (L, 60 × 40 cm, substrate height: 7 cm, 20 000 6-DOL/box). Larval weight was recorded at the beginning of trial and every 4 days, and growth rate (GR), specific growth rate (SGR), feed conversion ratio (FCR), survival, bioconversion efficiency corrected for residue (BER), reduction rate (RR), and waste reduction index (WRI) calculated at the end of larval growth (frass DM ≥ 55%). Substrate pH, T and height were measured at the beginning, every 4 days, and end of trial. Larval proximate composition was analysed at the end of trial. Data were analysed by generalised linear mixed model (SPSS software, P < 0.05). The D1 larvae showed higher weight, GR, SGR and WRI (along with higher substrate T) than D2 at M scale, while increased SGR and FCR - as well as decreased survival, RR and WRI - were observed in D2 larvae at S scale (P < 0.05). Larval CP and ether extract (EE) contents were influenced by M and L scales only, being higher in D2 group than in D1 (P < 0.001). Differently, decreased ash was recorded in D2 larvae when reared at S and M scales, while L scale revealed higher ash in D2 group than D1 (P < 0.001). The D3 larvae displayed greater weight, SGR, survival, RR and WRI (along with greater substrate T) than D4 at M scale, with increased survival and substrate T being also highlighted in L scale (P < 0.05). The D3 larvae also showed lower DM and EE - as well as higher CP - than D4 at all the rearing scales (P < 0.001). In conclusion, D1 and D3 led to better BSF larval growth performance, bioconversion efficiency and nutritional profile mainly at M and L scales, as a consequence of their ability to facilitate larval aggregation and, in turn, allow achieving a higher substrate T.

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Seaweed, a valuable marine resource widely cultivated worldwide, can be vulnerable to stress and microbiome alterations, resulting in the decay of seaweeds and substantial economic losses. To investigate the seaweed-microbiome interaction, our study aimed to isolate marine bacteria and fungi that can cause Ice-Ice disease and evaluate their enzymatic characteristics for potential application in bioethanol production from seaweed biomass. Three red seaweed species (Gracilaria edulis, Kappaphycus alvarezii, and Eucheuma cottonii) were obtained for our study and placed in separate culture tanks. Among the 18 isolated marine microbial species, 12 tested positive for agar and carrageenan activity: six exhibited both activities, three displayed only agar activity, and three only carrageenan activity. DNA sequencing of the positive microbes identified ten bacteria and two yeast species. The 3,5-Dinitrosalicylic acid (DNSA) assay results revealed that the identified bacterial Caldibacillus kokeshiiformis strain FJAT-47861 exhibited the highest carrageenase activity (0.76 units/ml), while the yeast Pichia fermentans strain PM79 demonstrated the highest agarase activity (0.52 units/ml). Notably, Pichia fermentans strain PM79 exhibited the highest overall agarase and carrageenase activity, averaging 0.63 units/ml. The average carrageenase activity of all six positive microbes was 1.5 times higher than their agarase activity. These findings suggest that the 12 isolated microbes hold potential for bioethanol production from macroalgae, as their agarase and carrageenase activity indicates their ability to break down seaweed cell wall carbohydrates, causing ice-ice disease. Moreover, these results provide exciting prospects for harnessing the bioconversion capabilities of these microbes, paving the way for sustainable and efficient bioethanol production from seaweed resources. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-024-01205-w.

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Astaxanthin is a red xanthophyll with high economic and industrial value in the pharmaceutical, nutraceutical, cosmetic and food industries. In recent years, the biotechnological production of astaxanthin has attracted much attention as a sustainable alternative to the predominating petrochemical-dependent chemical synthesis. In this regard, Xanthophyllomyces dendrorhous is regarded as a promising microorganism for industrial production of astaxanthin. Unfortunately, biotechnological production of the carotenoid is currently expensive. The present study investigated soy molasses (SM) and residual brewers' yeast as cheap fermentation feedstocks for the cultivation of X. dendrorhous and astaxanthin production. Yeast extract was obtained from residual brewers' yeast using various techniques and then combined with SM to formulate a two-component growth medium which was subsequently used to cultivate X. dendrorhous. Generally, the yeast extract produced from residual brewers' yeast supported X. dendrorhous growth and astaxanthin production at levels comparable to those seen with commercial yeast extract. Overall, cultivating X. dendrorhous in an SM-based medium containing 5% SM and 0.2% yeast extract obtained from residual brewers' yeast resulted in significantly higher (> 20% more) biomass accumulation compared to the control media (YPD). A similar slightly higher astaxanthin output (up to 14% more) was recorded in the SM-based medium compared to YPD. The formulated cultivation medium in this study provides an opportunity to reduce the production cost of astaxanthin from X. dendrorhous while simultaneously reducing the environmental impact related to the disposal of the industrial waste used as feedstock. KEY POINTS: • Cheap culture media were formulated from soy molasses and brewers' spent yeast • The formulated medium resulted in at least 20% more biomass than the control • Up to 14% more astaxanthin was produced in molasses-based medium.

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Methane, a potent greenhouse gas, requires sustainable mitigation strategies. Here, the microbial upcycling of methane to phytoene, a valuable colorless carotenoid with applications in the cosmeceutical industry was demonstrated. To achieve this goal, a stepwise metabolic engineering approach was employed in Methylocystis sp. MJC1, a methane-oxidizing bacterium. The incorporation of crtE and crtB genes from Deinococcus radiodurans R1 established the phytoene biosynthetic pathway. This pathway was fine-tuned through promoter optimization, resulting in a phytoene production of 450 µg/L from 37 mmol/L methane. Disrupting the ackA gene reduced a by-product, acetate, by 50 % and increased phytoene production by 56 %. Furthermore, overexpressing the dxs gene boosted phytoene titer 3-fold. The optimized strain produced 15 mg/L phytoene from 2 mol/L methane in fed-batch fermentation, a 4-fold increase in phytoene titer and 4-fold in yield. This demonstrates Methylocystis sp. MJC1's potential for efficient phytoene production and presents a novel approach for greenhouse gas reduction.

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Mesophilic enzymes, which are active at moderate temperatures, may dominate enzymatic reactions even in the presence of thermophilic crude enzymes. This study was conducted to investigate this hypothesis with mesophilic inositol dehydrogenases (IolG and IolX) produced in Geobacillus kaustophilus HTA426. To ensure the efficient production of mesophilic enzymes, we first screened for promoters induced at moderate temperatures using transcriptome analysis and identified four genes highly expressed at 30°C in the thermophile. We further characterized these promoters using fluorescent reporter assays to determine that the mti3 promoter could direct efficient gene expression at 40°C. We cloned the promoter into an Escherichia coli-Geobacillus shuttle plasmid and confirmed that the resulting vector functioned in G. kaustophilus and other thermophiles. We then used this vector for the cooperative expression of the iolG and iolX genes from Bacillus subtilis 168. G. kaustophilus cells carrying the expression vector were incubated at 60°C for cellular propagation and then at 40°C for the production of IolG and IolX. When the cells were permeabilized, IolG and IolX acted as catalysts to convert exogenous myo-inositol into scyllo-inositol at 30°C. In a scaled-up reaction, 10 g of myo-inositol was converted to 1.8 g of scyllo-inositol, which was further purified to yield 970 mg of pure powder. Notably, myo-inositol was degraded by intrinsic enzymes of G. kaustophilus at 60°C but not at 30°C, supporting our initial hypothesis. We indicate that this approach is useful for preparing enzyme cocktails without the need for purification. IMPORTANCE: Enzyme cocktails are commonly employed for cell-free chemical synthesis; however, their preparation involves cumbersome processes. This study affirms that mesophilic enzymes in thermophilic crude extracts can function as specific catalysts at moderate temperatures, akin to enzyme cocktails. The catalyst was prepared by permeabilizing cells without the need for concentration, extraction, or purification processes; hence, its preparation was considerably simpler compared with conventional methods for enzyme cocktails. This approach was employed to produce pure scyllo-inositol from an economical substrate. Notably, this marks the first large-scale preparation of pure scyllo-inositol, holding potential pharmaceutical significance as scyllo-inositol serves as a promising agent for certain diseases but is currently expensive. Moreover, this approach holds promise for application in pathway engineering within living cells. The envisioned pathway is designed without chromosomal modification and is simply regulated by switching culture temperatures. Consequently, this study introduces a novel platform for both whole-cell and cell-free synthetic systems.

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Enzymatic saccharification is used to convert polysaccharides in lignocellulosic biomass to sugars which are then converted to ethanol or other bio-based fermentation products. The efficacy of commercial cellulase preparations can potentially increase if lytic polysaccharide monooxygenase (LPMO) is included. However, as LPMO requires both a reductant and an oxidant, such as molecular oxygen, a reevaluation of process configurations and conditions is warranted. Saccharification and fermentation of pretreated softwood was investigated in demonstration-scale experiments with 10 m3 bioreactors using an LPMO-containing cellulase preparation, a xylose-utilizing yeast, and either simultaneous saccharification and fermentation (SSF) or hybrid hydrolysis and fermentation (HHF) with a 24-hour or 48-hour initial phase and with 0.15 vvm aeration before addition of the yeast. The conditions used for HHF, especially with 48 h initial phase, resulted in better glucan conversion, but in poorer ethanol productivity and in poorer initial ethanol yield on consumed sugars than the SSF. In the SSF, hexose sugars such as glucose and mannose were consumed faster than xylose, but, in the end of the fermentation >90% of the xylose had been consumed. Chemical analysis of inhibitory pretreatment by-products indicated that the concentrations of heteroaromatic aldehydes (such as furfural), aromatic aldehydes, and an aromatic ketone decreased as a consequence of the aeration. This was attributed mainly to evaporation caused by the gas flow. The results indicate that further research is needed to fully exploit the advantages of LPMO without compromising fermentation conditions.

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Utilizing carbon dioxide (CO2) for valuable chemical production is key to a circular economy. Current processes are costly due to limited microorganism use, low-value products, and the need for affordable energy. This study addresses these challenges by using industrial contaminants like thiosulfate (S2O32-) for CO2 conversion into ectoines. Ectoines, are important ingredients as pharmaceuticals and cosmetics. Here, six microbial genomes were identified as potential candidates to valorize CO2 and S2O32- into ectoine. After laboratory validation at 3 % NaCl, the fastest-growing strain, Guyparkeria halophila, was optimized at 6 %, 9 %, and 15 % NaCl, showing the highest specific ectoine contents (mgEct gbiomass-1) at 15 %. Batch bioreactors, combining optimal conditions, achieved maximum specific ectoine contents of 47 %. These results not only constitute the highest ectoine content so far reported by autotrophs and most of heterotrophs, but also the first proof of a novel valorization platform for CO2 and S2O32-, focused on pharmaceuticals production.

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In a world with a population exceeding 8 billion people and continuing to grow, pollution from food and plastic waste is causing long-term issues in ecosystems. Potential solutions may be found by exploiting insect-based bioconversion. In this context, we investigated the impact of polyvinyl chloride microparticles (PVC-MPs) on the development of Hermetia illucens (black soldier fly; BSF) and its midgut bacterial and fungal microbiota. The impact of PVC-MPs was evaluated feeding BSF larvae with a PVC-MPs-supplemented diet. The larvae exposed to different PVC-MPs concentrations (2.5%, 5%, 10% and 20% w/w) developed into adults with no significant increase in pupal mortality. Faster development and smaller pupae were observed when 20% PVC-MPs was provided. The BSF larvae ingest PVC-MPs, resulting in a reduction in MPs size. Larvae exposed to PVC-MPs did not exhibit differences in gut morphology. Regarding the impact of PVC-MPs on the structure of both bacterial and fungal communities, the overall alpha- and beta-diversity did not exhibit significant changes. However, the presence of PVC-MPs significantly affected the relative abundances of Enterobacteriaceae and Paenibacillaceae among the bacteria and of Dipodascaceae and Plectospharellaceae among the fungi (including yeast and filamentous life forms), suggesting that PVC-MP contamination has a taxa-dependent impact. These results indicate that BSF larvae can tolerate PVC-MPs in their diet, supporting the potential use of these insects in organic waste management, even in the presence of high levels of PVC-MP contamination.

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