Sustainable biosurfactant production from secondary feedstock-recent advances, process optimization and perspectives.

Biosurfactants have garnered increased attention lately due to their superiority of their properties over fossil-derived counterparts. While the cost of production remains a significant hurdle to surpass synthetic surfactants, biosurfactants have been anticipated to gain a larger market share in the coming decades. Among these, glycolipids, a type of low-molecular-weight biosurfactant, stand out for their efficacy in reducing surface and interfacial tension, which made them highly sought-after for various surfactant-related applications. Glycolipids are composed of hydrophilic carbohydrate moieties linked to hydrophobic fatty acid chains through ester bonds that mainly include rhamnolipids, trehalose lipids, sophorolipids, and mannosylerythritol lipids. This review highlights the current landscape of glycolipids and covers specific glycolipid productivity and the diverse range of products found in the global market. Applications such as bioremediation, food processing, petroleum refining, biomedical uses, and increasing agriculture output have been discussed. Additionally, the latest advancements in production cost reduction for glycolipid and the challenges of utilizing second-generation feedstocks for sustainable production are also thoroughly examined. Overall, this review proposes a balance between environmental advantages, economic viability, and societal benefits through the optimized integration of secondary feedstocks in biosurfactant production.

Optimal preparation of food waste to increase its utility for sophorolipid production by Starmerella bombicola.

Secondary feedstocks, such as food waste (FW), have been used for yeasts (e.g. Starmerella bombicola) to produce sophorolipids (SLs), which are commercially available biosurfactants. However, the quality of FW varies by location and season and may contains chemicals that inhibit SLs production. Therefore, it is crucial to identify such inhibitors and, if possible, remove them, to ensure efficient utilization. In this study, large scale FW was first analysed to determine the concentration of potential inhibitors. Lacticacid, acetic acid and ethanol were identified and found to be inhibitors of the growth of S. bombicola and its SLs production. Various methods were then evaluated for their ability to remove these inhibitors. Finally, a simple and effective strategy for removing inhibitors from FW was developed that complied with the 12 principles of green chemistry and could be adopted by industry for high SLs production.

Biorefinery potential of chemically enhanced primary treatment sewage sludge to representative value-added chemicals - A de novo angle for wastewater treatment.

Chemically enhanced primary treatment (CEPT) is an emerging sewage treatment strategy due to its high efficiency and small land requirement. CEPT sludge can be easily dewatered and used for energy recovery through incineration. However, with large amount of reusable nutrients (40% organic carbon, 23% lipids, and 17% protein), the value of CEPT sludge may have been underestimated. In this study, the biorefinery potential of CEPT sludge has been proven via production of 28.9 g/L ethanol or 50.3 g/L lactic acid (LA) or 1.43 filter paper unit (FPU)/mL cellulase from 10 g of CEPT sludge experiment. Inhibition on cell growth and potential inhibitors from plasticizers, pharmaceuticals, and surfactants were determined. Nevertheless, production titer was not affected or performed even better than the non-inhibitors controls. CEPT sludge showed significant potential in biochemical conversion, and the related products may offer an opportunity to support wastewater treatment toward sustainability and carbon neutrality.

Guiding environmental sustainability of emerging bioconversion technology for waste-derived sophorolipid production by adopting a dynamic life cycle assessment (dLCA) approach.

Microbial biosurfactants are surface-active molecules that are naturally produced by a range of microorganisms. They have certain advantages over chemical surfactants, such as lower toxicity, higher biodegradability, anti-tumor, and anti-microbial properties. Sophorolipids (SLs) in particular are one of the most promising biosurfactants, as they hold the largest share of the biosurfactant market. Currently, researchers are developing novel approaches for SL production that utilize renewable feedstocks and advanced separation technologies. However, challenges still exist regarding consumption of materials, enzymes, and electricity, that are primarily fossil based. Researchers lack a clear understanding of the associated environmental impacts. It is imperative to quantify and optimize the environmental impacts associated with this emerging technology very early in its design phase to guide a sustainable scale-up. It is necessary to take a collaborative perspective, wherein life cycle assessment (LCA) experts work with experimentalists, to quantify environmental impacts and provide recommendations for improvements in the novel waste-derived SL production pathways. Studies that have analyzed the environmental sustainability of microbial biosurfactant production are very scarce in literature. Hence, in this work, we explore the possibility of applying LCA to evaluate the environmental sustainability of SL production. A dynamic LCA (dLCA) framework that quantifies the environmental impacts of a process in an iterative manner, is proposed and applied to evaluate SL production. The first traversal of the dLCA was associated with the selection of an optimal feedstock, and results identified food waste as a promising feedstock. The second traversal compared fermentation coupled with alternative separation techniques, and highlighted that the fed-batch fermentation of food waste integrated with the in-situ separation technique resulted in less environmental impacts. These results will guide experimentalists to further optimize those processes, and improve the environmental sustainability of SL production. Resultant datasets can be iteratively used in subsequent traversals to account for technological changes and mitigate the corresponding impacts before scaling up.

Techno-economic evaluation of a biorefinery applying food waste for sophorolipid production - A case study for Hong Kong.

This study evaluates the techno-economic feasibility of sophorolipid (SL) production process that co-utilizes food waste, glucose and oleic acid as substrates. Two variables are considered in terms of (a) Plant construction: Purchasing equipment either from the US or Mainland China and (b) Production: to produce SL crystals (about 97% active) or a concentrated SL liquid/syrup (about 78% active). Hence, four scenarios are generated: Scenario I: equipment made in the USA + SL crystals; Scenario II: equipment made in the USA + SL syrup; Scenario III: equipment made in China + SL crystals; Scenario IV: equipment made in China + SL syrup. It is found that all scenarios are economically feasible and Scenario I has the highest net profit. Scenario III has the highest internal rate of return, net present value and the shortest payback period at a 7% discount rate. Finally, comparison of food waste-related techno-economic studies was conducted.

Aqueous mercury adsorption by activated carbons.

Due to serious public health threats resulting from mercury pollution and its rapid distribution in our food chain through the contamination of water bodies, stringent regulations have been enacted on mercury-laden wastewater discharge. Activated carbons have been widely used in the removal of mercuric ions from aqueous effluents. The surface and textural characteristics of activated carbons are the two decisive factors in their efficiency in mercury removal from wastewater. Herein, the structural properties and binding affinity of mercuric ions from effluents have been presented. Also, specific attention has been directed to the effect of sulfur-containing functional moieties on enhancing the mercury adsorption. It has been demonstrated that surface area, pore size, pore size distribution and surface functional groups should collectively be taken into consideration in designing the optimal mercury removal process. Moreover, the mercury adsorption mechanism has been addressed using equilibrium adsorption isotherm, thermodynamic and kinetic studies. Further recommendations have been proposed with the aim of increasing the mercury removal efficiency using carbon activation processes with lower energy input, while achieving similar or even higher efficiencies.

Enumeration and functional investigation of endothelial progenitor cells in neovascularization of diabetic foot ulcer rats with a Chinese 2-herb formula.

BACKBROUND: We investigated the effect of a Chinese 2-herb formula (NF3) on the enumeration and angiogenic differentiation of endothelial progenitor cells (EPCs) in diabetic foot ulcer rats. METHODS: EPCs and stromal cell-derived factor-1α (SDF-1α) were quantified by flow cytometry and ELISA, respectively. In vitro angiogenesis assays included proliferation, adhesion, migration and tube formation. RESULTS: Our result demonstrated that NF3 (0.98 g/kg) could significantly enhance the circulating CD34(+) /VEGFR2(+) /CD45(-) EPCs levels in diabetic foot ulcer rats by 60% (P < 0.05) through the partial elevation of SDF-1α, restoring the mobilization ability of EPCs for wound neovascularization. We successfully isolated the BM-derived EPCs to study their angiogenic potential after NF3 treatment. BM-derived EPCs significantly expressed cell surface markers of CD34, CD146 and VEGFR2 (P < 0.05 - 0.01). NF3 could significantly stimulate the proliferation and attachment ability of EPCs dose-dependently (P < 0.01-0.001). Besides, NF3 could significantly augment EPCs migration (P < 0.001) and tube formation (P < 0.01-0.001). CONCLUSIONS: NF3 modulated diabetic wound healing through regulation of systemic EPCs level and increase in local vascular formation.

Transport in Caco-2 cell monolayers of antidiabetic cucurbitane triterpenoids from Momordica charantia fruits.

Bitter melon, the fruit of Momordica charantia L. (Cucurbitaceae), is a widely-used treatment for diabetes in traditional medicine systems throughout the world. Various compounds have been shown to be responsible for this reputed activity, and, in particular, cucurbitane triterpenoids are thought to play a significant role. The objective of this study was to investigate the gastrointestinal transport of a triterpenoid-enriched n-butanol extract of M. charantia using a two-compartment transwell human intestinal epithelial cell Caco-2 monolayer system, simulating the intestinal barrier. Eleven triterpenoids in this extract were transported from the apical to basolateral direction across Caco-2 cell monolayers, and were identified or tentatively identified by HPLC-TOF-MS. Cucurbitane triterpenoids permeated to the basolateral side with apparent permeability coefficient (P app) values for 3-ß-7-ß,25-trihydroxycucurbita-5,23(E)-dien-19-al and momordicines I and II at 9.02 × 10(-6), 8.12 × 10(-6), and 1.68 × 10(-6)cm/s, respectively. Also, small amounts of these triterpenoids were absorbed inside the Caco-2 cells. This is the first report of the transport of the reputed antidiabetic cucurbitane triterpenoids in human intestinal epithelial cell monolayers. Our findings, therefore, further support the hypothesis that cucurbitane triterpenoids from bitter melon may explain, at least in part, the antidiabetic activity of this plant in vivo.

Anti-dermatophytic activity of bakuchiol: in vitro mechanistic studies and in vivo tinea pedis-inhibiting activity in a guinea pig model.

Bakuchiol was an active antifungal compound isolated from Psoraleae Fructus by means of bioassay-guided fractionation in our previous study. The present work aimed to investigate the underlying mechanisms and the therapeutic effect of bakuchiol in Trichophyton mentagrophytes-induced tinea pedis. After exposure to bakuchiol at 0.25-fold, 0.5-fold and 1-fold of minimum inhibitory concentration (MIC) (3.91 µg/ml) for 24h, the fungal conidia of T. mentagrophytes demonstrated a significant dose-dependent increase in membrane permeability. Moreover, bakuchiol at 1-fold MIC elicited a 187% elevation in reactive oxygen species (ROS) level in fungal cells after a 3-h incubation. However, bakuchiol did not induce DNA fragmentation. In a guinea pig model of tinea pedis, bakuchiol at 1%, 5% or 10% (w/w) concentration in aqueous cream could significantly reduce the fungal burden of infected feet (p<0.01-0.05). In conclusion, this is the first report to demonstrate that bakuchiol is effective in relieving tinea pedis and in inhibiting the growth of the dermatophyte T. mentagrophytes by increasing fungal membrane permeability and ROS generation, but not via induction of DNA fragmentation.

Potent airway smooth muscle relaxant effect of cynatratoside B, a steroidal glycoside isolated from Cynanchum stauntonii.

The dried roots of Cynanchum stauntonii in having cough-relieving efficacy are commonly included in traditional antitussive formulas. The active components in a C. stauntonii root extract responsible for airway relaxation were isolated using an ex vivo bioassay-guided fractionation method, in which subfractions were evaluated for their inhibitory effects on the contraction of isolated rat tracheal rings by isometric tension measurements. A steroidal glycoside, cynatratoside B (1), identified by LC-MS and NMR spectroscopic analysis, was shown to have potent inhibition on acetylcholine- and carbachol-induced tracheal contractions. The present data provide scientific evidence to support the traditional use of C. stauntonii as an antitussive herbal medicine.

A Chinese 2-herb formula (NF3) promotes hindlimb ischemia-induced neovascularization and wound healing of diabetic rats.

Diabetic foot ulcer is closely associated with peripheral vascular disease. Enhancement of tissue oxidative stress, reduction of nitric oxide (NO) and angiogenic growth factors, and abnormal matrix metalloproteinase (MMP) activity are pathophysiological factors in post-ischemic neovascularization and diabetic wound healing. Our previous study demonstrated that the Chinese 2-herb formula, NF3, showed significant wound healing effects on diabetic foot ulcer rats. A novel rat diabetic foot ulcer with hindlimb ischemia model was established in order to strengthen our claims on the diabetic wound healing and post-ischemic neovascularization effects of NF3. Our results demonstrate that NF3 can significantly reduce the wound area of the diabetic foot ulcer rat with hindlimb ischemia by 21.6% (p<0.05) compared with the control group. In addition, flow cytometric analysis revealed that NF3 could boost circulating EPC levels for local wound vessel incorporation. Immunohistochemical analysis showed that NF3 could significantly augment blood vessel density, VEGF and eNOS expression, and attenuate tissue oxidative stress of ischemic muscles (p<0.001). NF3 significantly stimulated MMP activity involved in angiogenesis. Our study shows, for the first time, the beneficial effects of NF3 in wound healing and post-ischemic neovascularization in diabetes.

Synergistic interaction between Astragali Radix and Rehmanniae Radix in a Chinese herbal formula to promote diabetic wound healing.

ETHNOPHARMACOLOGICAL RELEVANCE: Astragali Radix (AR) and Rehmanniae Radix (RR) are two traditional Chinese medicines widely used in China for treating diabetes mellitus and its complications, such as diabetic foot ulcer. AIM OF STUDY: In our previous study, a herbal formula NF3 comprising AR and RR in the ratio of 2:1 was found effective in enhancing diabetic wound healing in rats through the actions of tissue regeneration, angiogenesis promotion and inflammation inhibition. The aims of the present study were to investigate the herb-herb interaction (or the possible synergistic effect) between AR and RR in NF3 to promote diabetic wound healing and to identify the principal herb in the formula by evaluating the potencies of individual AR and RR in different mechanistic studies. MATERIALS AND METHODS: A chemically induced diabetic foot ulcer rat model was used to examine the wound healing effect of NF3 and its individual herbs AR and RR. For mechanistic studies, murine macrophage cell (RAW 264.7) inflammation, human fibroblast (Hs27) proliferation and human endothelial cell (HMEC-1) migration assays were adopted to investigate the anti-inflammatory, granulation formation and angiogenesis-promoting activities of the herbal extracts, respectively. RESULTS: In the foot ulcer animal model, neither AR nor RR at clinical relevant dose (0.98g/kg) promoted diabetic wound healing. However, when they were used in combination as NF3, synergistic interaction was demonstrated, of which NF3 could significantly reduce the wound area of rats when compared to water group (p<0.01). For anti-inflammation and granulation formation, AR was more effective than RR in inhibiting lipopolysaccharide (LPS)-induced nitric oxide production from RAW 264.7 cells and promoting Hs27 fibroblast proliferation. In the aspect of angiogenesis promotion, only NF3 promoted cell migration of HMEC-1 cells. CONCLUSIONS: AR plays a preeminent role in the anti-inflammatory and fibroblast-proliferating activities of NF3. The inclusion of RR, however, is crucial for NF3 to exert its overall wound-healing as well as the underlying angiogenesis-promoting effects. The results of present study justified the combined usage of AR and RR in the ratio of 2:1 as NF3 to treat diabetic foot ulcer and illustrated that AR is the principal herb in this herbal formula.

Screening for anti-inflammatory and bronchorelaxant activities of 12 commonly used Chinese herbal medicines.

The use of health supplements derived from medicinal herbs as self-medication for the relief of respiratory tract pathology symptoms is increasing in Chinese communities as air pollution is worsening. Twelve herbs from two formulae of our previous studies were evaluated for their anti-inflammatory, immunomodulatory and bronchorelaxant activities in this study. Among the extracts tested, those of Herba Schizonepetae and Radix Glycyrrhizae showed significant inhibitory effects on LPS-induced nitric oxide production (p < 0.05) in mouse macrophage RAW264.7 cells, suggesting their anti-inflammatory activities. Radix Scutellariae and Radix Glycyrrhizae extracts showed significant inhibitory effects on phytohaemagglutinin-induced proliferation in human peripheral blood mononuclear cells (p < 0.05). These extracts also showed inhibition of TNF-α, IFN-γ and IL-10 production. For the bronchorelaxant assay, Rhizoma Cynanchi Stauntonii and Radix Glycyrrhizae extracts showed potent attenuation of the acetylcholine- and carbachol-induced contractions in rat trachea (p < 0.05), implying their relaxant activities. In conclusion, Herba Schizonepetae, Radix Glycyrrhizae, Radix Scutellariae and Rhizoma Cynanchi Stauntonii extracts were demonstrated to exert anti-inflammatory, immunomodulatory and bronchorelaxant activities, which may help to ameliorate the symptoms of respiratory tract pathologies. The findings have thus provided some scientific evidence on the efficacy and mechanisms of action of these herbs, which are useful for the further development of clinical applications.

Stachyose: One of the Active Fibroblast-proliferating Components in the Root of Rehmanniae Radix ( dì huáng).

This study aimed to investigate and compare the fibroblast-proliferating activities of different Rehmanniae Radix (RR) samples and its chemical components using human normal fibroblast cells Hs27. Those active components were quantified in differently treated RR samples using UPLC so as to correlate activity with component content. Our results showed that dried RR aqueous extract exhibited the most potent fibroblast-proliferating activity. Stronger effect was observed when ethanol with heating was applied in the extraction process. Stachyose and verbascoside were demonstrated for their first time to exhibit significant stimulatory effects on fibroblast proliferation. However, the proliferating effect of dried RR extract did not correlate with the stachyose content, and verbascoside was not responsible for the fibroblast proliferative effect of RR since it was undetectable in all samples. In conclusion, stachyose only contributed in part to the activity of RR, suggesting that other active components might be present and yet to be found.

The in vivo and in vitro diabetic wound healing effects of a 2-herb formula and its mechanisms of action.

ETHNOPHARMACOLOGICAL RELEVANCE: The herbs Radix Astragali (RA) and Radix Rehmanniae (RR) have long been used in traditional Chinese Medicine and serve as the principal herbs in treating diabetic foot ulcer. AIM OF STUDY: Diabetic complications, such as foot ulcer, impose major public health burdens worldwide. In our previous clinical studies, two Chinese medicine formulae F1 and F2 have achieved over 80% limb salvage. A simplified 2-herb formula (NF3) comprising of RA and RR in the ratio of 2:1 was used for further study. NF3 was examined for the ulcer healing effect in diabetic rats, and its potential mechanisms of action in fibroblast proliferation, angiogenesis and anti-inflammation in vitro. MATERIALS AND METHODS: A chemically induced diabetic foot ulcer rat model was used for studying the wound healing effect. In the in vitro mechanistic studies, human fibroblast cells (Hs27), human umbilical vein endothelial cells (HUVEC) and mouse macrophage cells (RAW264.7) were assessed for tissue regeneration, angiogenesis and anti-inflammatory activities, respectively. RESULTS: Our in vivo results demonstrated a significant reduction of wound area at day 8 in NF3 (0.98g/kg) group as compared to control (p<0.01). NF3 could significantly stimulate Hs27 proliferation in a dose dependent manner (p<0.05). Besides, NF3 could significantly increase the cell migration and tube formation (p<0.05-0.001) of HUVEC in the angiogenesis study. Furthermore, significant inhibition of nitric oxide production (p<0.01) was found in NF3-treated macrophage cells, suggesting its anti-inflammatory activity. CONCLUSIONS: Our study presents for the first time scientific evidence towards the efficacy of the two-herb formula NF3 in enhancing diabetic wound healing through the actions of tissue regeneration, angiogenesis and anti-inflammation.