Pomegranate (Punica granatum L.) belongs to the Punicaceae plant family. It is an important fruit due to its nutritional and medicinal properties. Pomegranates are widely distributed around the world and, therefore, have a broad genetic diversity, resulting in differences in their phytochemical composition. The scientific community has focused on the positive health effects of pomegranate as a whole, but the different varieties have rarely been compared according to their bioactive compounds and bioactivity. This review aims to provide a holistic overview of the current knowledge on the bioactivity of pomegranate trees, with an emphasis on differentiating both the varieties and the different plant parts. This review intends to provide a general and organized overview of the accumulated knowledge on pomegranates, the identification of the most bioactive varieties, their potential consumption pathways and seeks to provide knowledge on the present gaps to guide future research.

The synthesis of silver nanoparticles (SNP) from plants is a simple, fast and environmentally safe route. In the present study, the aqueous extract of fresh leaves from Leea coccinea L. was evaluated as a possible source of reducing and stabilizing agents to obtain SNP. The synthesized SNP were characterized by spectroscopic techniques such as UV–visible spectrophotometry and Fourier transform infrared spectroscopy (FTIR), scanning electron and confocal microscopies and the antimicrobial activity against Xanthomonas phaseoli pv. phaseoli was evaluated using agar diffusion methods. The results showed that the evaluated extract was promising for the green synthesis of the SNP, which was visually identified by the formation of a dark-brown complex and the presence of a peak of maximum absorption at 470 nm in a UV–VIS spectrum. FTIR spectrum of SNP showed main characteristic signals of aromatic compounds, carboxylic group among others confirmed by phytochemical screening that made evident the presence of flavonoids, phenols, leucoanthocyanidins, terpenes and steroids groups. Fluorescent SNP with high degree of agglomeration were observed by the microscopical technics used. A promising antibacterial activity of SNP was shown by a zone of microbial growth inhibition. These results suggested the need for going deeper in the physico-chemical characterization and kinetic studies, as well as the biological evaluations to make possible the use of this plant source in the future development of antibacterial formulations for bean seed protection.

Hepatocellular carcinoma (HCC) is one of the most prevalent and deadliest cancers. In this study, the anti-tumor effect of singular degree of polymerization (DP) chitooligosaccharides (COS) (DP 2–5) and the underlay molecular mechanisms were investigated on HCC cell line HepG2. MTT assay showed that (GlcN)₅ have the best anti-proliferation effect among the different DP of COS (DP2-5). Furthermore, the administration of (GlcN)₅ could decrease mitochondrial membrane potential, release cytochrome c into cytoplasm, activate the cleavage of Caspases9/3, thus inducing mitochondrial-mediated apoptosis in HepG2 cells (accounting for 24.57 ± 2.25%). In addition, (GlcN)₅ treatment could increase the accumulation of autophagosomes. Further investigation showed that (GlcN)₅ suppressed protective autophagy at the fusion of autophagosomes and lysosomes. Moreover, the inhibition of protective autophagy flux by (GlcN)₅ could further decrease cell viability and increase the apoptosis rate. Our findings suggested that (GlcN)₅ suppressed HepG2 proliferation through inducing apoptosis via the intrinsic pathway and impairing cell-protective autophagy. COS might have the potential to be an agent for lowering the risk of HCC.

The effects of wheat gluten hydrolysates (WGH) and their ethanol elution fractions obtained on XAD-16 resin on physiological activity and fermentation performance of brewer’s yeast during very-high-gravity (VHG) worts fermentation were investigated. The results showed that the addition of WGH and their elution fractions in VHG worts significantly enhanced yeast biomass and viability, and further increased the fermentability, ethanol yield and productivity of yeast. Supplementation with 40% ethanol fraction exhibited the highest biomass (6.9 g/L dry cell), cell viability, fermentability (82.05%), ethanol titer (12.19%, v/v) and ethanol productivity during VHG worts fermentation. In addition, 40% ethanol fraction supplementation also caused the most consumption of amino acid and the highest accumulation of intracellular glycerol and trehalose, 15.39% of increase in cell-membrane integrity, 39.61% of enhancement in mitochondrial membrane potential (MMP), and 18.94% of reduction in intracellular reactive oxygen species (ROS) level in yeast under VHG conditions. Therefore, WGH supplementation was an efficient method to improve fermentation performance of brewer’s yeast during VHG worts.

Knowledge of the interactions between soil systems, management practices, and climatic extremes are critical for prescription-based sustainable practices that reduce environmental pollution/footprints, disruption of food supply chains, food contamination, and thus improve socio-economic wellbeing. Soil quality status and dynamics under climate change present both a hazard which may not be remedied by simply adding chemicals or improved by crop varieties, and an opportunity (e.g., by indicating impact of a shift in land use) although the specifics remain debatable. This entry not only revisits the science of soil quality determination but also explicates on intricacies of monitoring using big data generated continuously and integrated using the “internet of things.” Indeed, relaying credible soil quality information especially for heterogeneous soils at field scale is constrained by challenges ranging from data artifacts and acquisition timing differences, vague baselines, validation challenges, scarcity of robust standard algorithms, and decision support tools. With the advent of digital technology, modern communication networks, and advancement in variable rate technologies (VRT), a new era has dawned for developing automated scalable and synthesized soil quality metrics. However, before digital technology becomes the routine tool for soil quality sensing and monitoring, there is need to understand the issues and concerns. This contribution not only exemplifies a unique application of digital technology to detect residue cover but also deliberates on the following questions: (1) is digital agriculture the missing link for integrating, understanding the interconnectivity, and ascertaining the provenance between soil quality, agronomic production, environmental health, and climate dynamics? and (2) what are the technological gaps?

The efficacy of alcohol/sugar aqueous biphasic (ABS) system on protein extraction was investigated. A model protein, bovine serum albumin (BSA), was adopted to evaluate the effects of types and concentration of phase-forming components, protein concentration, and system pH on the protein partition efficiency. The 1-propanol/maltose ABS exhibited an overall better partition efficiency of BSA to the alcohol-rich top phase. A maximum partition coefficient (K) of 20.01 ± 0.05 and recovery yield (Y) of 95.42% ± 0.01% of BSA were achieved with 35% (w/w) 1-propanol/22% (w/w) maltose ABS at pH 5.0 for 10% (w/w) BSA load. The K and Y of BSA in 1-propanol/maltose ABS was slightly improved with the addition of 3% (w/w) of ionic liquid, 1-butyl-3-methylimidazolium bromide ([Bmim]Br) as the adjuvant that could provide protein stabilizing effect. The Fourier Transform Infrared Spectrum (FTIR) analysis revealed that the protein structure remained unaltered upon the separation process.

The anaerobic process is considered to be a sustainable technology for the treatment of wastewaters rich in organic matter mainly due to its lower energy consumption and production of value-added products such as biogas and organic fertilizer. However, it cannot be seen as providing ‘complete’ environmental solution as its treated effluents would typically not meet the desired discharge limits in terms of residual carbon, nutrients and other pollutants. This has given impetus to subsequent post treatment in order to meet the environmental standards and protect the receiving water bodies and environment. The aim of this study was to evaluate the post-treatment potential of a pilot scale two-stage horizontal subsurface flow constructed wetland (HSSFCW) system planted with Cyperus alternifolius and Typha latifolia, respectively, for enhanced removal of residual carbon and nutrient from an up-flow anaerobic sludge blanket (UASB) reactor treated brewery effluent. A pilot scale two-stage HSSFCW was integrated with the UASB reactor, and its performance efficiency was assessed for the removal of total suspended solids (TSS), chemical oxygen demand (COD), total nitrogen (TN), ammonium–nitrogen (NH₄–N), total phosphorous (TP), and orthophosphate (PO₄ ³⁻). Macrophytes aboveground biomass and nutrient accumulation potential were also determined following standard methods. The results from this study showed that Cyperus alternifolius planted CW cell removed 68.5% TSS, 74.2% COD, 55.7% TN, 68.6% NH₄–N, 41.1% TP and 48.1% PO₄ ³⁻. Moreover, further polishing with Typha latifolia planted CW cell enhanced the removal efficiencies to 89% TSS, 92% COD, 83.6% TN, 92.9% NH₄ ^–N, 74.4% TP, and 79.5% PO₄ ³⁻. Strong linearity and Pearson correlation was found between macrophyte biomass and nutrient accumulation in each CW cell (Cyperus alternifolius: R ² = 0.91, r = 0.97 for TN; R ² = 0.92, r = 0.96 for TP; and Typha latifolia: R ² = 0.96, r = 0.98 for TN and TP), and showed substantial nutrient reduction with cumulative nutrient accumulation of 1290 gTNm⁻² and 708.7 gTPm⁻² in the complete system. The performance of the pilot CW system as a tertiary treatment for brewery wastewater showed that the effluent meets the permissible discharge standards throughout the year excluding phosphorous.

Conversion of the abundant agricultural residual cotton stalk (CS) into useful chemicals or functional materials could alleviate the fossil fuels caused energy shortages and environmental crises. Although some advances have been achieved, less attention has been paid to the plant tissues effect. In this study, the plant tissue of CS was changed by part degradation of some components (hemicelluloses and lignin, for example) with the aid of acid/base (or both). The pretreated CS was transformed into hydrochar by hydrothermal carbonization (HTC) method. Morphological and chemical compositions of CS hydrochar were analyzed by various techniques, including elemental analysis, Fourier transform infrared (FTIR), BET analysis, X-ray photoelectron spectroscopy (XPS) and X-ray diffraction (XRD). Methylene blue (MB) removal of prepared CS hydrochar was used to evaluate CS hydrochar pollutions adsorption capacity. Results reveal acid/base (or both) pretreatment is beneficial for CS raw material to prepare high-quality CS hydrochar. The effects of some parameters, such as initial MB concentration, temperature, pH value and recyclability on the adsorption of MB onto both acid and base-pretreated CS hydrochar (CS-H₂SO₄ + NaOH-HTC) were studied. The present work exhibits the importance of agricultural waste biomass material plant tissues on its derived materials, which will have a positive effect on the direct utilization of waste biomass.

The accumulation of petrochemical plastic waste is detrimental to the environment. Polyhydroxyalkanoates (PHAs) are bacterial-derived polymers utilized for the production of bioplastics. PHA-plastics exhibit mechanical and thermal properties similar to conventional plastics. However, high production cost and obtaining high PHA yield and productivity impedes the widespread use of bioplastics. This study demonstrates the concept of cyclic fed-batch fermentation (CFBF) for enhanced PHA productivity by Bacillus thuringiensis using a glucose-rich hydrolyzate as the sole carbon source. The statistically optimized fermentation conditions used to obtain high cell density biomass (OD₆₀₀ of 2.4175) were: 8.77 g L⁻¹ yeast extract; 66.63% hydrolyzate (v/v); a fermentation pH of 7.18; and an incubation time of 27.22 h. The CFBF comprised three cycles of 29 h, 52 h, and 65 h, respectively. After the third cyclic event, cell biomass of 20.99 g L⁻¹, PHA concentration of 14.28 g L⁻¹, PHA yield of 68.03%, and PHA productivity of 0.219 g L⁻¹ h⁻¹ was achieved. This cyclic strategy yielded an almost threefold increase in biomass concentration and a fourfold increase in PHA concentration compared with batch fermentation. FTIR spectra of the extracted PHAs display prominent peaks at the wavelengths unique to PHAs. A copolymer was elucidated after the first cyclic event, whereas, after cycles CFBF 2–4, a terpolymer was noted. The PHAs obtained after CFBF cycle 3 have a slightly higher thermal stability compared with commercial PHB. The cyclic events decreased the melting temperature and degree of crystallinity of the PHAs. The approach used in this study demonstrates the possibility of coupling fermentation strategies with hydrolyzate derived from lignocellulosic waste as an alternative feedstock to obtain high cell density biomass and enhanced PHA productivity.

Nicotinamide adenine dinucleotide phosphate (NADPH), as a well-known cofactor, is widely used in the most of enzymatic redox reactions, playing an important role in industrial catalysis. However, the absence of a comparable method for efficient NADP⁺ to NADPH cofactor regeneration radically impairs efficient green chemical synthesis. Alcohol dehydrogenase (ADH) enzymes, allowing the in situ regeneration of the redox cofactor NADPH with high specific activity and easy by-product separation process, are provided with great industrial application potential and research attention. Accordingly, herein a NADP⁺-specific ADH from Clostridium beijerinckii was selected to be engineered for cofactor recycle, using an automated algorithm named Protein Repair One-stop Shop (PROSS). The mutant CbADH-6M (S24P/G182A/G196A/H222D/S250E/S254R) exhibited a favorable soluble and highly active expression with an activity of 46.3 U/mL, which was 16 times higher than the wild type (2.9 U/mL), and a more stable protein conformation with an enhanced thermal stability: Δ ${T}_{1/2}^{60\mathrm{min}}$=  + 3.6 °C (temperature of 50% inactivation after incubation for 60 min). Furthermore, the activity of CbADH-6M was up-graded to 2401.8 U/mL by high cell density fermentation strategy using recombinant Escherichia coli, demonstrating its industrial potential. Finally, the superb efficiency for NADPH regeneration of the mutant enzyme was testified in the synthesis of some fine chiral aromatic alcohols coupling with another ADH from Lactobacillus kefir (LkADH).

Programming non-canonical organisms is more attractive due to the prospect of high-value chemical production. Among all, Shewanella oneidensis MR-1 possesses outstanding heme synthesis ability and is well-known for electron transfer, thus has high potential in microbial fuel cell and bioremediation. However, heme, as the final product of C4 and C5 pathways, is regulated by heme cluster for the high-value 5-aminolevulinic acid (ALA) for cancer photodynamic therapy, which has never been explored in MR-1. Herein, the heme metabolism in MR-1 was firstly optimized for ALA production. We applied CRISPR interference (CRISPRi) targeted on the genes to fine-tune carbon flux in TCA cycle and redirected the carbon out-flux from heme, leading to a significant change in the amino acid profiles, while downregulation of the essential hemB showed a 2-fold increasing ALA production via the C5 pathway. In contrast, the modular design including of glucokinase, GroELS chaperone, and ALA synthase from Rhodobacter capsulatus enhanced ALA production markedly in the C4 pathway. By integrating gene cluster under dual T7 promoters, we obtained a new strain M::TRG, which significantly improved ALA production by 145-fold. We rewired the metabolic flux of MR-1 through this modular design and successfully produced the high-value ALA compound at the first time.

Biomass is one of the most abundant renewable energy resources on the earth, which is also considered as one of the most promising alternatives to traditional fuel energy. In recent years, microbial fuel cell (MFC) which can directly convert the chemical energy from organic compounds into electric energy has been developed. By using MFC, biomass energy could be directly harvested with the form of electricity, the most convenient, wide-spread, and clean energy. Therefore, MFC was considered as another promising way to harness the sustainable energies in biomass and added new dimension to the biomass energy industry. In this review, the pretreatment methods for biomass towards electricity harvesting with MFC, and the microorganisms utilized in biomass-fueled MFC were summarized. Further, strategies for improving the performance of biomass-fueled MFC as well as future perspectives were highlighted.

Effective nutrient transport and appropriate mechanical stimulation play important roles in production of tissue-engineered bone grafts. In this study, an experimental set-up for magnetic-driven dynamic culture of cells was designed to mimic the microenvironment of the bone tissue. Here, its ability to contribute to osteogenic differentiation was investigated by inoculating human umbilical cord mesenchymal stem cells (HUMSCs) on magnetic scaffolds. The cytocompatibility of the developed magnetic scaffolds was verified for HUMSCs. Magnetic scaffolds seeded with HUMSCs were exposed to magnetic fields. The results showed that magnetic fields did not affect cell activity and promoted HUMSCs osteogenic differentiation. The magnetic scaffolds were magnetically driven for dynamic culture in the experimental set-up to evaluate the influence of HUMSCs osteoblast differentiation. The results indicated that magnetic-driven dynamic culture increased cell alkaline phosphatase (ALP) activity (p < 0.05) and calcium release (p < 0.05) compared with static culture. The effect was demonstrated in the expression of bone-associated genes. Overall, this study showed that magnetic-driven dynamic culture is a promising tool for regenerative bone engineering.

The capacity of different Bacillus species to produce large amounts of extracellular enzymes and ability to ferment various substrates at a wide range of pH and temperature has placed them among the most promising hosts for the industrial production of many improved and novel products. The global interest in prebiotics, for example, xylooligosaccharides (XOs) is ever increasing, rousing the quest for various forms with expanded productivity. This article provides an overview of xylanase producing bacilli, with more emphasis on their capacity to be used in the production of the XOs, followed by the purification strategies, characteristics and application of XOs from bacilli. The large-scale production of XOs is carried out from a number of xylan-rich lignocellulosic materials by chemical or enzymatic hydrolysis followed by purification through chromatography, vacuum evaporation, solvent extraction or membrane separation methods. Utilization of XOs in the production of functional products as food ingredients brings well-being to individuals by improving defense system and eliminating pathogens. In addition to the effects related to health, a variety of other biological impacts have also been discussed.

In plants, viral diseases are second only to fungal diseases in terms of occurrence, and cause substantial damage to agricultural crops. The aqueous extracts of shell ginger, Alpinia zerumbet exhibit inhibitory effects against virus infections in belonging to the Solanaceae family. In this study, we isolated an anti-plant-virus molecule from the extracts using a conventional method involving a combination of reversed phase column chromatography, dialysis, and lyophilization. The anti-plant-virus molecule was identified as proanthocyanidin, which mostly consisted of epicatechin and exhibited more than 40 degrees of polymerization.

Esters are widely used in plastics, textile fibers, and general petrochemicals. Usually, esters are produced via chemical synthesis or enzymatic processes from the corresponding alcohols and acids. However, the fermentative production of esters from alcohols and/or acids has recently also become feasible. Here we report a cognate microbial consortium capable of producing butyl butyrate. This microbial consortium consists of two engineered butyrate- and butanol-producing E. coli strains with nearly identical genetic background. The pathways for the synthesis of butyrate and butanol from butyryl-CoA in the respective E. coli strains, together with a lipase-catalyzed esterification reaction, created a “diamond-shaped” consortium. The concentration of butyrate and butanol in the fermentation vessel could be altered by adjusting the inoculation ratios of each E. coli strain in the consortium. After optimization, the consortium produced 7.2 g/L butyl butyrate with a yield of 0.12 g/g glucose without the exogenous addition of butanol or butyrate. To our best knowledge, this is the highest titer and yield of butyl butyrate produced by E. coli reported to date. This study thus provides a new way for the biotechnological production of esters.

Considering global developments like climate change and the depletion of fossil resources, the use of new and sustainable feedstocks such as lignocellulosic biomass becomes inevitable. Green waste comprises heterogeneous lignocellulosic biomass with low lignin content, which does not stem from agricultural processes or purposeful cultivation and therefore mainly arises in urban areas. So far, the majority of green waste is being composted or serves as feedstock for energy production. Here, the hitherto untapped potential of green waste for material utilization instead of conventional recycling is reviewed. Green waste is a promising starting material for the direct extraction of valuable compounds, the chemical and fermentative conversion into basic chemicals as well as the manufacturing of functional materials like electrodes for electro-biotechnological applications through carbonization. This review serves as a solid foundation for further work on the valorization of green waste.

The development of yeast that converts raw corn or cassava starch to ethanol without adding the exogenous α-amylase and/or glucoamylase would reduce the overall ethanol production cost. In this study, two copies of codon-optimized Saccharomycopsis fibuligera glucoamylase genes were integrated into the genome of the industrial Saccharomyces cerevisiae strain CCTCC M94055, and the resulting strain CIBTS1522 showed comparable basic growth characters with the parental strain. We systemically evaluated the fermentation performance of the CIBTS1522 strain using the raw corn or cassava starch at small and commercial-scale, and observed that a reduction of at least 40% of the dose of glucoamylase was possible when using the CIBTS1522 yeast under real ethanol production condition. Next, we measured the effect of the nitrogen source, the phosphorous source, metal ions, and industrial microbial enzymes on the strain’s cell wet weight and ethanol content, the nitrogen source and acid protease showed a positive effect on these parameters. Finally, orthogonal tests for some other factors including urea, acid protease, inoculum size, and glucoamylase addition were conducted to further optimize the ethanol production. Taken together, the CIBTS1522 strain was identified as an ideal candidate for the bioethanol industry and a better fermentation performance could be achieved by modifying the industrial culture media and condition.

Biomass may ignite due to biological oxidation and chemical oxidation. If this phenomenon (spontaneous ignition) is controlled, it would be possible to produce biochar at a lower cost without the need for an external heat resource. We investigated if self-heating could be controlled by using sawdust and bark chips. When sawdust and bark chips were used under controlled conditions, the bark chips temperature increased to the torrefaction temperature. The ash content of bark chips was ~ 2%d.b. higher than that of sawdust; consequently, the inorganic substances contained in the bark chips might affect the self-heating. Self-heating was suppressed when inorganic substances were removed by washing with water. Therefore, the inorganic substances in the biomass might have affected self-heating. The inorganic element contents of the bark chips were measured by inductively coupled plasma optical emission spectrometry before and after washing. The potassium content of the bark chips was reduced remarkably by washing, and there was a possible influence of potassium on self-heating. Finally, the effect of moisture content on self-heating was investigated to obtain stable reactivity. Thus, at a moisture content of 40%w.b., a steady self-heating behavior may be realized.

(R)-mandelic acid is an industrially important chemical, especially used for producing antibiotics. Its chemical synthesis often uses highly toxic cyanide to produce its racemic form, followed by kinetic resolution with 50% maximum yield. Here we report a green and sustainable biocatalytic method for producing (R)-mandelic acid from easily available styrene, biobased L-phenylalanine, and renewable feedstocks such as glycerol and glucose, respectively. An epoxidation-hydrolysis-double oxidation artificial enzyme cascade was developed to produce (R)-mandelic acid at 1.52 g/L from styrene with > 99% ee. Incorporation of deamination and decarboxylation into the above cascade enables direct conversion of L-phenylalanine to (R)-mandelic acid at 913 mg/L and > 99% ee. Expressing the five-enzyme cascade in an L-phenylalanine-overproducing E. coli NST74 strain led to the direct synthesis of (R)-mandelic acid from glycerol or glucose, affording 228 or 152 mg/L product via fermentation. Moreover, coupling of E. coli cells expressing L-phenylalanine biosynthesis pathway with E. coli cells expressing the artificial enzyme cascade enabled the production of 760 or 455 mg/L (R)-mandelic acid from glycerol or glucose. These simple, safe, and green methods show great potential in producing (R)-mandelic acid from renewable feedstocks.

The development of biosimilar products or follow-on biologics has been flourishing in recent years because of their lower price than the originators. In this study, a multivariate data analysis method based on JMP software was proposed to assess the glycosylation pattern similarity of antibody candidates from different conditions in optimization experiments with a reference. A specific distance was generated by this method and indicated the glycoform similarity between the biosimilar and the reference. This method can be applied to analyze the similarity of other physicochemical and functional characteristics between follow-on biologics and originators. Then, the design of experimental methods can be realized to optimize the conditions of cell culture to attain similar antibody candidates. A higher concentration of GlcNAc added to the basal media made the glycan of the antibody more similar to the glycan of the reference in this study.

Considering the expected increasing demand for cellulose fibers in the near future and that its major source is wood pulp, alternative sources such as vegetable wastes from agricultural activities and agro-food industries are currently being sought to prevent deforestation. In the present study, cellulose was successfully isolated from six agroindustrial residues: corncob, corn husk, grape stalk, pomegranate peel, marc of strawberry-tree fruit and fava pod. Cellulose fibers were characterized by Fourier-transform infrared spectroscopy, thermogravimetric analysis, stereomicroscopy and scanning electron microscopy (SEM). Despite the evident morphological differences among the extracted celluloses, results revealed similar compositional and thermal properties with the wood-derived commercial microcrystalline cellulose used as a control. Trace amounts of lignin or hemicellulose were detected in all cellulose samples, with the exception of corncob cellulose, that exhibited the greatest extraction yield (26%) and morphological similarities to wood-derived microcrystalline cellulose, visible through SEM. Furthermore, corncob cellulose was found to have thermal properties (T_Onset of 307.17 °C, TD of 330.31 °C, and ΔH of 306.04 kJ/kg) suitable for biomedical applications.

An active site is normally located inside enzymes, hence substrates should go through a tunnel to access the active site. Tunnel engineering is a powerful strategy for refining the catalytic properties of enzymes. Here, P450_BsβHI (Q85H/V170I) derived from hydroxylase P450_Bsβ from Bacillus subtilis was chosen as the study model, which is reported as a potential decarboxylase. However, this enzyme showed low decarboxylase activity towards long-chain fatty acids. Here, a tunnel engineering campaign was performed for modulating the substrate preference and improving the decarboxylation activity of P450_BsβHI. The finally obtained BsβHI-F79A variant had a 15.2-fold improved conversion for palmitic acid; BsβHI-F173V variant had a 3.9-fold improved conversion for pentadecanoic acid. The study demonstrates how the substrate preference can be modulated by tunnel engineering strategy.

Under the optimal conditions of immobilization and fermentation, the highest LA yield of 0.966 ± 0.006 g/g fructose and production rate of 2.426 ± 0.018 g/(L × h) with an error of -0.5% and -0.2% to the predicted results were obtained from batch fermentation by the CS film-coated SA-PVA immobilized L. pentosus cells. The LA yield and production rate of these immobilized cells were 2.7% and 10.1% higher than that of normal SA-PVA immobilized cells respectively, and they were 5.7% and 48.4% higher than that of free cells, respectively. The effect of temperature on different types of immobilized cells and free cells was significantly different, but the effect of pH on different types of cells was not much different. The kinetic models could effectively describe the different fermentation performances of three types of cells. The immobilized cells have excellent reusability to conduct 9 runs of repeated batch fermentation.

Extensive decoration of backbones is a major factor resulting in resistance of enzymatic conversion in hemicellulose and other branched polysaccharides. Employing debranching enzymes is the main strategy to overcome this kind of recalcitrance at present. A carbohydrate-binding module (CBM) is a contiguous amino acid sequence that can promote the binding of enzymes to various carbohydrates, thereby facilitating enzymatic hydrolysis. According to previous studies, CBMs can be classified into four types based on their preference in ligand type, where Type III and IV CBMs prefer to branched polysaccharides than the linear and thus are able to specifically enhance the hydrolysis of substrates containing side chains. With a role in dominating the hydrolysis of branched substrates, Type III and IV CBMs could represent a non-catalytic approach in overcoming side-chain recalcitrance.

In integrated bioprocessing applications, expanded bed adsorption (EBA) chromatography presents an opportunity to harvest biomolecules directly from the crude feedstock. However, unfavorable biomass interactions with adsorbent usually leads to fouling, which reduces its protein binding capacity as it alters column hydrodynamics and binding site availability. In this work, a detailed study on biomass adhesion behavior of four different industrially relevant microorganisms on 26 different, most commonly occurring adsorbent surfaces with varying degrees of surface energy and surface charge has been conducted. The results showed the derivation of a relative “stickiness” factor for every microorganism, which further classifies each organism based on their general degree of adhesion to surfaces with respect to one another. The obtained results can help to better understand the effect of biomass homogenization on biomass–adsorbent interactions in EBA. The data of surface energy and charge for the surfaces investigated in this work can be used to calculate the stickiness factor of other microorganisms of interest and may assist in the development of novel adsorbent materials for EBA chromatography.

Biomethanation is of great interest as it can transform CO₂ to methane under ambient conditions. In particular, genetically engineered bacterium of Rhodopseudomonas palustris showed great promise for one-step biomethanation powered by solar energy, which is attractive for CO₂ fixation as well as solar energy storage. However, biomethanation with R. palustris under visible light is inefficient due to its poor visible light response. In this study, CdS quantum dots with excellent visible light response were prepared and R. palustris/CdS hybrid cells were constructed. Interestingly, this bio-nano-hybrid cells showed high cell viability without significant cell damage, and the biomethanation performance of was enhanced about ~ 79% compared to that of the bare R. palustris cells. Moreover, the effects of different parameters on the methane production of this bio-nano-hybrid cells were determined, and the methane production rate was further improved by parameter optimization. This work demonstrated an efficient approach to reinforce the biomethanation of bacteria under unfavorable light wavelength, which would be helpful to extend the light spectra for photo-driven biomethanation.

Lactic acid has become one of the most important chemical substances used in various sectors. Its global market demand has significantly increased in recent years, with a CAGR of 18.7% from 2019 to 2025. Fermentation has been considered the preferred method for producing high-purity lactic acid in the industry over chemical synthesis. However, the recovery and separation of lactic acid from microbial fermentation media are relatively complicated and expensive, especially in the process relating to second-generation (2G) lactic acid recovery. This article reviews the development and progress related to lactic acid separation and recovery from fermentation broth. Various aspects are discussed thoroughly, such as the mechanism of lactic acid production through fermentation, the crucial factors that influence the fermentation process, and the separation and recovery process of conventional and advanced lactic acid separation methods. This review's highlight is the recovery of lactic acid by adsorption technique using ion-exchange resins with a brief focus on the potential of in-site separation strategies alongside the important factors that influenced the lactic acid recovery process by ion exchange. Apart from that, other lactic acid separation techniques, such as chemical neutralization, liquid–liquid extraction, membrane separation, and distillation, are also thoroughly reviewed.

ε-Caprolactone is a monomer of poly(ε-caprolactone) which has been widely used in tissue engineering due to its biodegradability and biocompatibility. To meet the massive demand for this monomer, an efficient whole-cell biocatalytic approach was constructed to boost the ε-caprolactone production using cyclohexanol as substrate. Combining an alcohol dehydrogenase (ADH) with a cyclohexanone monooxygenase (CHMO) in Escherichia coli, a self-sufficient NADPH-cofactor regeneration system was obtained. Furthermore, some improved variants with the better substrate tolerance and higher catalytic ability to ε-caprolactone production were designed by regulating the ribosome binding sites. The best mutant strain exhibited an ε-caprolactone yield of 0.80 mol/mol using 60 mM cyclohexanol as substrate, while the starting strain only got a conversion of 0.38 mol/mol when 20 mM cyclohexanol was supplemented. The engineered whole-cell biocatalyst was used in four sequential batches to achieve a production of 126 mM ε-caprolactone with a high molar yield of 0.78 mol/mol.

Microalgae biomass exploitation as a carbon–neutral energy source is currently limited by several factors, productivity being one of the most relevant. Due to the high absorption properties of light-harvesting antenna, photosynthetic cells tend to capture an excessive amount of energy that cannot be entirely channeled through the electron transfer chain that ends up dissipated as heat and fluorescence, reducing the overall light use efficiency. Aiming to minimize this hurdle, in this work we studied the effect of decreasing concentrations of Magnesium (Mg²⁺) on the chlorophyll a content, photosynthetic performance, biomass and lipid production of autotrophic cultures of Botryococcus braunii LB 572. We also performed, for the first time, a comparative lipidomic analysis to identify the influence of limited Mg²⁺ supply on the lipid profile of this algae. The results indicated that a level of 0.0037 g L⁻¹ MgSO₄ caused a significant decline on chlorophyll a content with a concomitant 2.3-fold reduction in the biomass absorption coefficient. In addition, the Mg²⁺ limitation caused a decrease in the total carbohydrate content and triggered lipid accumulation, achieving levels of up to 53% DCW, whereas the biomass productivity remained similar for all tested conditions. The lipidome analysis revealed that the lowest Mg²⁺ concentrations also caused a differential lipid profile distribution, with an enrichment of neutral lipids and an increase of structural lipids. In that sense, we showed that Mg²⁺ limitation represents an alternative optimization approach that not only enhances accumulation of neutral lipids in B. braunii cells but also may potentially lead to a better areal biomass productivity due to the reduction in the cellular light absorption properties of the cells.

Sawdust, cotton stalk and groundnut shell were used for removal of methylene blue from aqueous solution using batch sorption. Effect of initial dye concentration, temperature, and particle size of sorbents on methylene blue removal was investigated. Sorption capacity increases with rise in initial dye concentration and temperature. Impact of particle size on sorption of methylene blue was investigated and indicated that removal of dye increases with decrease in particle size of sorbents. Maximum sorption for sawdust, cotton stalks and groundnut shell were 9.22 mg g⁻¹, 8.37 mg g⁻¹ and 8.20 mg g⁻¹ respectively; at 60 °C and 100 ppm initial dye concentration. Sorption isotherms were analyzed using fundamental Freundlich isotherm. Subsequently, sips isotherm model was employed for better fitting. Kinetic study shows that, biosorption process is pseudo-second-order in nature. During the course of this study, adsorption dynamics revealed that film diffusion was key step for biosorption. In addition, thermodynamics of sorption was studied; and it was found that Gibbs free energy (∆G°) decreases with increase in temperature. Sawdust was found to be best among all the sorbents. Therefore, column studies and breakthrough curve modelling were performed using sawdust. Furthermore, it was estimated that a scaled-up column using sawdust can treat 6672 L of wastewater in 24 h with 80% efficiency.

A wide variety of biomass is available all around the world. Most of the biomass exists as a by-product from manufacturing industries. Pulp and paper mills contribute to a higher amount of these biomasses mostly discarded in the landfills creating an environmental burden. Biomasses from other sources have been used to produce different kinds and grades of biomaterials such as those used in industrial and medical applications. The present review aims to investigate the availability of biomass from pulp and paper mills and show sustainable routes for the production of high value-added biomaterials. The study reveals that using conventional and integrated biorefinery technology the ample variety and quantity of waste generated from pulp and paper mills can be converted into wealth. As per the findings of the current review, it is shown that high-performance carbon fiber and bioplastic can be manufactured from black liquor of pulping waste; the cellulosic waste from sawdust and sludge can be utilized for the synthesis of CNC and regenerated fibers such as viscose rayon and acetate; the mineral-based pulping wastes and fly ash can be used for manufacturing of different kinds of biocomposites. The different biomaterials obtained from the pulp and paper mill biomass can be used for versatile applications including conventional, high performance, and smart materials. Through customization and optimization of the conversion techniques and product manufacturing schemes, a variety of engineering materials can be obtained from pulp and paper mill wastes realizing the current global waste to wealth developmental approach.

Spirulina platensis protein hydrolysates were prepared by digesting protein extracts with papain, and the hydrolysates were separated into 30, 10, and 3 kDa weights using membrane ultrafiltration. The 0–3 kDa low-molecular-weight Spirulina peptides (LMWSPs) proved the highest chemical antioxidant activity by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability, hydroxyl radical (·OH) scavenging activities and total antioxidant capacity. Cellular antioxidant ability of LMWPs fractions against 2000 μg/mL H₂O₂ induced oxidative damage of L02 cells were investigated. The MTT assay results displayed that LMWSPs at different concentrations (0–1000 μg/mL) had proliferation effect on the L02 cells and that treatment of the L02 cells with the 1000 μg/mL LMWSPs (0–3 kDa) significantly prevented H₂O₂-induced oxidative damage compared with control cells. Moreover, the 2′,7′-dichlorofluorescein diacetate (DCFH-DA) fluorescent probe assay showed that the levels of ROS and NO were significantly lower in the experimental group that was treated with the peptides for 24 h than in the control group. Furthermore, using the corresponding kits, the treatment inhibited the reduction of SOD activity and the increase of MDA contents in the L02 cells. Therefore, LMWSPs (0–3 kDa) may have potential applications in antioxidant and liver health products.

In this study, introduction of a viable cell sensor and electronic nose into ethanol fermentation was investigated, which could be used in real-time and on-line monitoring of the amount of living cells and product content, respectively. Compared to the conventional off-line biomass determination, the capacitance value exhibited a completely consistent trend with colony forming units, indicating that the capacitance value could reflect the living cells in the fermentation broth. On the other hand, in comparison to the results of off-line determination by high-performance liquid chromatography, the ethanol concentration measured by electronic nose presented an excellent consistency, so as to realize the on-line monitoring during the whole process. On this basis, a dynamic feeding strategy of glucose guided by the changes of living cells and ethanol content was developed. And consequently, the ethanol concentration, productivity and yield were enhanced by 15.4%, 15.9% and 9.0%, respectively. The advanced sensors adopted herein to monitor the key parameters of ethanol fermentation process could be readily extended to an industrial scale and other similar fermentation processes.

Red alga dulse contains xylan with β(1→3)/β(1→4) linkages. We previously prepared xylooligosaccharides (XOSs) from dulse xylan; however, the product contained many d-xylose residues and fewer XOSs with β(1→3) linkages. To improve the efficiency of XOS production, we prepared two recombinant endoxylanases from Streptomyces thermogriseus (StXyl10 and StXyl11). Comparing the k_cat/K_m values for dulse xylan, this value from StXyl10 was approximately two times higher than that from StXyl11. We then determined the suitable conditions for XOS production. As a result, dulse XOS was prepared by the successive hydrolysis of 10 mg/mL dulse xylan by 0.5 μg/mL StXyl10 for 4 h at 50 °C and then 2.0 μg/mL StXyl11 for 36 h at 60 °C. Xylan was converted into 95.8% XOS, including 59.7% XOS with a β(1→3) linkage and 0.97% d-xylose. Our study provides useful information for the production of XOSs with β(1→3) linkages.

The management of municipal solid waste is a major logistic and environmental problem worldwide. Nonetheless, the organic fraction of municipal solid waste (OFMSW) is a valuable source of nutrients which can be used for a variety of purposes, according to the Circular Economy paradigm. Among the possible applications, the bioproduction of a biodegradable polyester, poly(3-hydroxybutyrate) [P(3HB)], using OFMSW as carbon platform is a promising strategy. Here, an economic and environmental assessment of bacterial P(3HB) production from OFMSW is presented based on previously published results. The SuperPro Designer® software was used to simulate P(3HB) production under our experimental parameters. Two scenarios were proposed depending on the fermentation medium: (1) enzymatic hydrolysate of OFMSW supplemented with glucose and plum waste juice; and (2) basal medium supplemented with glucose and plum waste juice. According to our results, both scenarios are not economically feasible under our experimental parameters. In Scenario 1, the low fermentation yield, the cost of the enzymes, the labour cost and the energy consumption are the factors that most contribute to that result. In Scenario 2, the cost of the extraction solvent and the low fermentation yield are the most limiting factors. The possibility of using process waste as raw material for the generation of other products must be investigated to enhance economic feasibility. From an environmental viewpoint, the photochemical oxidation potential (derived from the use of anisole as extraction solvent) and the generation of acid rain and global warming effect (caused by the burning of fuels for power generation) are the most relevant impacts associated to P(3HB) production under our experimental parameters.

In this study, a four-enzyme cascade pathway was developed and reconstructed in vivo for the production of d-p-hydroxyphenylglycine (D-HPG), a valuable intermediate used to produce β-lactam antibiotics and in fine-chemical synthesis, from l-tyrosine. In this pathway, catalytic conversion of the intermediate 4-hydroxyphenylglyoxalate by meso-diaminopimelate dehydrogenase from Corynebacterium glutamicum (CgDAPDH) was identified as the rate-limiting step, followed by application of a mechanism-guided “conformation rotation” strategy to decrease the hydride-transfer distance d_{(C6HDAP−C4NNADP)} and increase CgDAPDH activity. Introduction of the best variant generated by protein engineering (CgDAPDH^{BC621/D120S/W144S/I169P} with 5.32 ± 0.85 U·mg⁻¹ specific activity) into the designed pathway resulted in a D-HPG titer of 42.69 g/L from 50-g/L l-tyrosine in 24 h, with 92.5% conversion, 71.5% isolated yield, and > 99% enantiomeric excess in a 3-L fermenter. This four-enzyme cascade provides an efficient enzymatic approach for the industrial production of D-HPG from cheap amino acids.

Photorespiration consumes fixed carbon and energy generated from photosynthesis to recycle glycolate and dissipate excess energy. The aim of this study was to investigate whether we can use the energy that is otherwise consumed by photorespiration to improve the production of chemicals which requires energy input. To this end, we designed and introduced an isoprene synthetic pathway, which requires ATP and NADPH input, into the cyanobacterium Synechocystis sp. 6803. We then deleted the glcD1 and glcD2 genes which encode glycolate dehydrogenase to impair photorespiration in isoprene-producing strain of Synechocystis. Production of isoprene in glcD1/glcD2 disrupted strain doubled, and stoichiometric analysis indicated that the energy saved from the impaired photorespiration was redirected to increase production of isoprene. Thus, we demonstrate we can use the energy consumed by photorespiration of cyanobacteria to increase the energy-dependent production of chemicals from CO₂.

A novel biosynthesis of dual reduced graphene oxide/silver nanocomposites (rGO/AgNC) using the crude metabolite of Escherichia coli D8 (MF06257) strain and sunlight is introduced in this work. Physicochemical analysis of these rGO/AgNC revealed that they are sheet-like structures having spherically shaped silver nanoparticles (AgNPs) with an average particle size of 8 to 17 nm, and their absorption peak ranged from 350 to 450 nm. The biosynthesized rGO/AgNC were characterized by UV–vis and FT-IR spectra, X-ray diffraction, Zeta potential and transmission electron microscopy. After the injection of these nanocomposites to mice, their uptake by the kidney and liver has been proven by the ultrastructural observation and estimation of the hepatic and renal silver content. These nanocomposites caused a moderate toxicity for both organs. Changes in the liver and kidney functions and histopathological effects had been observed. The rGO/AgNC revealed a remarkable antitumor effect. They showed a dose-dependent cytotoxic effect on Ehrlich ascites carcinoma (EAC) cells in vitro. Treatment of mice bearing EAC tumors intraperitoneally with 10 mg/kg rGO/AgNC showed an antiproliferative effect on EAC cells, reduced ascites volume, and maintained mice survival. The results indicate that this green synergy of silver nanoparticles with reduced graphene oxide may have a promising potential in cancer therapy.

Rubber seeds are a by-product of rubber production and are rich in oil and protein. Upgrading of rubber seeds to produce proteins, oils and feedstock can generate additional revenue for rubber production and reduce waste. The present study investigates the effects of different pre-treatments and extraction methods to determine the optimal methods to produce oil and protein from rubber seed kernels. Mechanical expulsion using a screw press and solvent extraction using n-hexane were employed for oil separation. The highest oil recovery efficiency of 95.12% was obtained using rubber seed meal that was pre-dried at 105 ℃. The sequential water–alkaline treatment was ideal for achieving high protein recovery while reducing the protein denaturation that can result from high operating temperatures and organic solvent contact. Over 90% of the total protein from rubber seed kernels could be recovered. Separating oil from kernels using hexane followed by protein extraction from the meals by enzymatic treatment provides a suitable method for comprehensive utilization of rubber seeds.

Argonaute proteins (Agos) from thermophiles function as endonucleases via guide-target base-pairing cleavage for host defense. Since guides play a key role in regulating the catalytic specificity of Agos, elucidating its underlying molecular mechanisms would promote the application of Agos in the medical sciences. Here, we reveal that an Ago from Pyrococcus furiosus (PfAgo) showed a stepwise endonuclease activity, which was demonstrated through a double-stranded DNA cleavage directed by a single guide DNA (gDNA) rather than a canonical pair of gDNAs. We validated that the cleavage products with 5'-phosphorylated ends can be used as a new guide to induce a new round of cleavage. Based on the reprogrammable capacity of Ago’s stepwise activity, we established a rapid and specific platform for unambiguous multiplex gene detection, termed Renewed-gDNA Assisted DNA cleavage by Argonaute (RADAR). Combined with a pre-amplification step, RADAR achieved sensitivity at the femtomolar level and specificity with at least a di-nucleotide resolution. Furthermore, RADAR simultaneously discriminated among multiple target sequences simply by corresponding multiple guides. We successfully distinguished four human papillomavirus serotypes from patient samples in a single reaction. Our technique, based on the unique properties of Ago, provides a versatile and sensitive method for molecular diagnosis.

Carotenoids are a large family of health-beneficial compounds that have been widely used in the food and nutraceutical industries. There have been extensive studies to engineer Saccharomyces cerevisiae for the production of carotenoids, which already gained high level. However, it was difficult to discover new targets that were relevant to the accumulation of carotenoids. Herein, a new, ethanol-induced adaptive laboratory evolution was applied to boost carotenoid accumulation in a carotenoid producer BL03-D-4, subsequently, an evolved strain M3 was obtained with a 5.1-fold increase in carotenoid yield. Through whole-genome resequencing and reverse engineering, loss-of-function mutation of phosphofructokinase 1 (PFK1) was revealed as the major cause of increased carotenoid yield. Transcriptome analysis was conducted to reveal the potential mechanisms for improved yield, and strengthening of gluconeogenesis and downregulation of cell wall-related genes were observed in M3. This study provided a classic case where the appropriate selective pressure could be employed to improve carotenoid yield using adaptive evolution and elucidated the causal mutation of evolved strain.

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.

Flavone C-arabinosides/xylosides are plant-originated glycoconjugates with various bioactivities. However, the potential utility of these molecules is hindered by their low abundance in nature. Engineering biosynthesis pathway in heterologous bacterial chassis provides a sustainable source of these C-glycosides. We previously reported bifunctional C-glucosyl/C-arabinosyltransferases in Oryza sativa japonica and O. sativa indica, which influence the C-glycoside spectrum in different rice varieties. In this study, we proved the C-arabinosyl-transferring activity of rice C-glycosyltransferases (CGTs) on the mono-C-glucoside substrate nothofagin, followed by taking advantage of specific CGTs and introducing heterologous UDP-pentose supply, to realize the production of eight different C-arabinosides/xylosides in recombinant E. coli. Fed-batch fermentation and precursor supplement maximized the titer of rice-originated C-arabinosides to 20–110 mg/L in an E. coli chassis. The optimized final titer of schaftoside and apigenin di-C-arabinoside reached 19.87 and 113.16 mg/L, respectively. We demonstrate here the success of de novo bio-production of C-arabinosylated and C-xylosylated flavones by heterologous pathway reconstitution. These results lay a foundation for further optimal manufacture of complex flavonoid compounds in microbial cell factories.

Butyl butyrate (BB) is an important chemical with versatile applications in beverage, food and cosmetics industries. Since chemical synthesis of BB may cause adverse impacts on the environment, biotechnology is an emerging alternative approach for microbial esters biosynthesis. BB can be synthesized by using a single Clostridium strain natively producing butanol or butyrate, with exogenously supplemented butyrate or butanol, in the presence of lipase. Recently, E. coli strains have been engineered to produce BB, but the titer and yield remained very low. This review highlighted a new trend of developing cognate microbial consortium for BB production and associated challenges, and end up with new prospects for further improvement for microbial BB biosynthesis.