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Purpose Periodontitis is a chronic inflammatory disease associated with microbial accumulation. The purpose of this study was to reuse the agricultural waste to produce cellulose nanofibers (CNF) and further modification of the CNF with κ-carrageenan oligosaccharides (CO) for drug delivery. In addition, this study is focused on the antimicrobial activity of surfactin-loaded CO-CNF towards periodontal pathogens. Materials and methods A chemo-mechanical method was used to extract the CNF and the modification was done by using CO. The studies were further proceeded by adding different quantities of surfactin [50 mg (50 SNPs), 100 mg (100 SNPs), 200 mg (200 SNPs)] into the carrier (CO-CNF). The obtained materials were characterized, and the antimicrobial activity of surfactin-loaded CO-CNF was evaluated. Results The obtained average size of CNF and CO-CNF after ultrasonication was 263 nm and 330 nm, respectively. Microscopic studies suggested that the CNF has a short diameter with long length and CO became cross-linked to form as beads within the CNF network. The addition of CO improved the degradation temperature, crystallinity, and swelling property of CNF. The material has a controlled drug release, and the entrapment efficiency and loading capacity of the drug were 53.15 ± 2.36% and 36.72 ± 1.24%, respectively. It has antioxidant activity and inhibited the growth of periodontal pathogens such as Streptococcus mutans and Porphyromonas gingivalis by preventing the biofilm formation, reducing the metabolic activity, and promoting the oxidative stress. Conclusion The study showed the successful extraction of CNF and modification with CO improved the physical parameters of the CNF. In addition, surfactin-loaded CO-CNF has potential antimicrobial activity against periodontal pathogens. The obtained biomaterial is economically valuable and has great potential for biomedical applications.

Lactic acid has received considerable attention for its various applications. Although lactic acid is usually produced by batch fermentation using free microbes, repeated-batch fermentation using immobilized cells could offer several advantages over this method. In the present study, a novel bacteria immobilization technique using a renewable poly-L-lactic acid (PLLA) microtube array membrane (MTAM) was thoroughly evaluated in terms of its suitability for lactic acid fermentation. A bacteria encapsulation efficiency of 85–90% was obtained using a siphon approach, and bacteria in MTAMs with greater porosity showed greater lactic acid productivity. MTAM-immobilized Lactobacillus acidophilus were evaluated in eight cycles of repeated-batch lactic acid fermentation of commercial medium MRS (4% glucose) and red seaweed Gracilaria hydrolysate. The MTAM exhibited physical stability during lactic acid fermentation and improved glucose consumption and lactic acid production were achieved by immobilized cells. The immobilized bacteria showed a maximum CLA of 37.39 g/L, r PLA of 0.79 g/L·h, and YL/S of 0.94 g/g in MRS fermentation, and a maximum CLA of 27.76 g/L, r PLA of 0.58 g/L·h, and YL/S of 0.92 g/g in seaweed hydrolysate fermentation. Our data indicate that PLLA-MTAM is a novel, promising immobilization technology for lactic acid fermentation.

Drug efflux pumps are one of the major elements used by antibiotic-resistant bacteria. Efflux pump inhibitors (EPIs) are potential therapeutic agents for adjunctive therapy, which can restore the activity of antibiotics that are no longer effective against pathogens. This study evaluated the seaweed compound diphenylmethane (DPM) for its EPI activity. The IC50 and modulation results showed that DPM has no antibacterial activity but can potentiate the activity of antibiotics against drug-resistant E. coli. Time-kill studies reported that a combination of DPM and erythromycin exhibited greater inhibitory activity against drug-resistant Escherichia coli. Dye accumulation and dye efflux studies using Hoechst 33342 and ethidium bromide showed that the addition of DPM significantly increased dye accumulation and reduced dye efflux in drug-resistant E. coli, suggesting its interference with dye translocation by an efflux pump. Using MALDI-TOF, it was observed that the addition of DPM could continuously reduce antibiotic efflux in drug-resistant E. coli. Additionally, DPM did not seem to damage the E. coli membranes, and the cell toxicity test showed that it features mild human-cell toxicity. In conclusion, these findings showed that DPM could serve as a potential EPI for drug-resistant E. coli.

Seaweed oligosaccharides have attracted attention in food, agricultural, and medical applications recently. Compared to red and brown seaweeds, fewer studies have focused on the biological activity of green seaweed’s oligosaccharides. This study aimed to produce bioactive ulvan oligosaccharides via enzymatic hydrolysis from green seaweed Ulva lactuca. Ulvan, a water-soluble polysaccharide, was obtained by hot water extraction. Two isolated marine bacteria, Pseudomonas vesicularis MA103 and Aeromonas salmonicida MAEF108, were used to produce multiple hydrolases, such as ulvanolytic enzymes, amylase, cellulase, and xylanase, to degrade the ulvan extract. An ultrafiltration system was used to separate the enzymatic hydrolysate to acquire the ulvan oligosaccharides (UOS). The characteristics of the ulvan extract and the UOS were determined by yield, reducing sugar, uronic acid, sulfate group, and total phenols. The FT-IR spectrum indicated that the ulvan extract and the UOS presented the bands associated with O-H, C=O, C-O, and S=O stretching. Angiotensin I converting enzyme (ACE) inhibition and antioxidant activities in vitro were evaluated in the ulvan extract and the UOS. These results provide a practical approach to producing bioactive UOS by microbial enzymatic hydrolysis that can benefit the development of seaweed-based products at the industrial scale.

4-Mercapto-4-methyl-2-pentanone (4MMP), a high-impact aroma compound with the box tree and black currant flavors was first identified in wines and could be released by microbial cysteine-S-conjugate β-lyases from its precursors. In this study, various yeasts and bacteria encoding β-lyases were selected to examine their β-lyase activities. A thiol precursor of 4MMP, cysteine-conjugate of 4MMP (cys-4MMP), was synthesized with a purity of >95% in a relatively environmentally friendly approach, and its chemical structure was confirmed by nuclear magnetic resonance spectroscopy. The β-lyase activities of the crude cell extract from the bacteria and yeast strains for different substrates were examined using a colorimetric method. Shewanella putrefaciens cell extract exhibited the highest β-lyase activity for all tested substrates. Additionally, the optimum pH and temperature for their β-lyase activities were determined. To monitor the conversion efficiency of precursor cys-4MMP to 4MMP, liquid chromatography-mass spectrometry was used. Our data indicate that selected bacteria and yeasts could convert cys-4MMP into 4MMP, and S. putrefaciens exhibited the best conversion yield. This study demonstrated the potential use of microbial cell extracts to produce sulfur-containing aroma compounds such as 4MMP.

The aim of this research was to investigate the effects of the addition of 0.5% hydroxymethylfurfural (HMF) and low molecular chitosan on acrylamide and HMF formation in a food model system, which contains 0.5% glucose, asparagine, and HMF within 30 min of heating at 180 °C. At an interval of 10 min, all solutions were evaluated in the following aspects: reducing sugar, asparagine, acrylamide, HMF content, pH, Maillard reaction products, kinematic viscosity, and color. After heating for 10 min, the kinematic viscosity of solutions containing chitosan reduced significantly. The values of the acrylamide, HMF, and absorbance increased at OD294 and OD420 (optical density measured at 294 nm and 420 nm) of solutions. Experimental results showed that low-molecular-weight chitosan might be hydrolyzed into much lower molecular weight, followed by the decrease in kinematic viscosity of the solution at pH lower than 6 and the increase in the formation of acrylamide after heating for 30 min.

This research focuses on the proteolytic capacity of sea bass byproduct (SB) and their hypocholesterolemic activity via the cholesterol micelle formation (CMF) inhibition. SB was fermented with seven mixed lactic acid bacteria for 5 h at 42 °C. The lactic fermented SB was hydrolyzed with Protease N for 6 h under HHP to obtain the SB hydrolysates (HHP-assisted Protease N hydrolysis after fermentation, F-HHP-PN6). The supernatant was separated from the SB hydrolysate and freeze-dried. As the hydrolysis time extended to 6 h, soluble protein content increased from 187.1 to 565.8 mg/g, and peptide content increased from 112.8 to 421.9 mg/g, while inhibition of CMF increased from 75.0% to 88.4%. Decreasing the CMF inhibitory activity from 88.4% to 42.1% by simulated gastrointestinal digestion (FHHP-PN6 was further hydrolyzed by gastrointestinal enzymes, F-HHP-PN6-PP) reduced the CMF inhibitory activity of F-HHP-PN6. Using gel filtration chromatography, the F-HHP-PN6-PP was fractioned into six fractions. The molecular weight of the fifth fraction from F-HHP-PN6-PP was between 340 and 290 Da, and the highest inhibitory efficiency ratio (IER) on CMF was 238.9%/mg/mL. Further purification and identification of new peptides with CMF inhibitory activity presented the peptide sequences in Ser-Ala-Gln, Pro-Trp, and Val-Gly-Gly-Thr, the IERs were 361.7, 3230.0, and 302.9%/mg/mL, respectively.

Periodontitis is an inflammatory disease that can lead to the periodontal pocket formation and tooth loss. This study was aimed to develop antimicrobials loaded hydrogels composed of cellulose nanofibers (CNF) and κ-carrageenan oligosaccharides (CO) nanoparticles for the treatment of periodontitis. Two antimicrobial agents such as surfactin and Herbmedotcin were selected as the therapeutic agents and the hydrogels were formulated based on the increasing concentration of surfactin. The proposed material has high thermal stability, controlled release, and water absorption capacity. This study was proceeded by investigating the in vitro antibacterial and anti-inflammatory properties of the hydrogels. This material has strong antibacterial activity against periodontal pathogens such as Streptococcus mutans, Porphyromonas gingivalis, Fusobacterium nucleatum, and Pseudomonas aeruginosa. Moreover, a significant increase in malondialdehyde (MDA) production and a decrease in biofilm formation and metabolic activity of the bacteria was observed in the presence of hydrogel. Besides, it reduced the reactive oxygen species (ROS) generation, transcription factor, and cytokines production in human gingival fibroblast cells (HGF) under inflammatory conditions. In conclusion, the hydrogels were successfully developed and proven to have antibacterial and anti-inflammatory properties for the treatment of periodontitis. Thus, it can be used as an excellent candidate for periodontitis treatment.

Biodegradable polylactic acid material is manufactured from lactic acid, mainly produced by microbial fermentation. The high production cost of lactic acid still remains the major limitation for its application, indicating that the cost of carbon sources for the production of lactic acid has to be minimized. In addition, a lack of source availability of food crop and lignocellulosic biomass has encouraged researchers and industries to explore new feedstocks for microbial lactic acid fermentation. Seaweeds have attracted considerable attention as a carbon source for microbial fermentation owing to their non-terrestrial origin, fast growth, and photoautotrophic nature. The proximate compositions study of red, brown, and green seaweeds indicated that Gracilaria sp. has the highest carbohydrate content. The conditions were optimized for the saccharification of the seaweeds, and the results indicated that Gracilaria sp. yielded the highest reducing sugar content. Optimal lactic acid fermentation parameters, such as cell inoculum, agitation, and temperature, were determined to be 6% (v/v), 0 rpm, and 30 °C, respectively. Gracilaria sp. hydrolysates fermented by lactic acid bacteria at optimal conditions yielded a final lactic acid concentration of 19.32 g/L.

Hollow, poly(L-lactic acid) microtube array membranes (MTAM) were used in preparing membranes that contained immobilized yeast cells. To evaluate the performance of the developed system for continuous and fed-batch fermentation, a gas chromatography/milli-whistle device was used to on-line monitor the production of ethanol. The milli-whistle was connected to the outlet of a GC capillary, and when the fermentation gases and the GC carrier gas passed through it, a sound with a fundamental frequency was produced. The online data obtained for frequency-change vs. retention time can be recorded after a fast Fourier transform. In typical bioethanol fermentation, the yeast cells cannot be recycled, whereas the artificial yeast-MTAMs can be. The hollow-MTAM containing immobilized yeast cells significantly enhanced to bioethanol productivity, and represent a novel, promising technology for bioethanol fermentation. Our data indicate that the gas chromatography/milli-whistle device, which is economical and stable, is a very useful detector for long-term monitoring.

In this study, the low-molecular-weight (LMW) fucoidan, rich in fucose and sulfate, was extracted and purified from the edible brown seaweed, Laminaria japonica. In this study, we orally administered LMW fucoidan to mice for 6 weeks. We then examined fucoidan&rsquo, s effects on innate immunity, adaptive immunity, and Mycoplasma pneumoniae (MP)-antigen-stimulated immune responses. Our data showed that LMW fucoidan stimulated the innate immune system by increasing splenocyte proliferation, natural killer (NK) cell activity, and phagocytic activity. LMW fucoidan also increased interleukin (IL)-2, IL-4, and interferon (IFN)-&gamma, secretion by splenocytes and immunoglobulin (Ig)-G and IgA content in serum, which help regulate adaptive immune cell functions, and decreased allergen-specific IgE. In MP-antigen-stimulated immune responses, the IgM and IgG content in the serum were significantly higher in the LMW fucoidan group after MP-antigen stimulation. Our study provides further information about the immunomodulatory effects of LMW fucoidan and highlights a potential role in preventing M. pneumoniae infection.

Multidrug-resistant pathogens are a significant clinical problem. Efflux pump inhibitors (EPIs) can restore the activities of existing antibiotics by interfering with drug efflux pumps located in bacterial cell membranes. Seaweeds are important sources of biologically active metabolites of natural origin; however, their potential as EPIs remains uninvestigated. Here, functional extracts from the brown seaweeds Laminaria japonica and Sargassum horneri and the red seaweeds Gracilaria sp. and Porphyra dentata were evaluated as potential EPIs against drug-resistant Escherichia coli. All these extracts were found to potentiate the activities of drugs in modulation tests, although not to the same extent. Synergistic effects of the extracts and the drug clarithromycin were observed from the onset of Time-kill assays, with no evidence of bacterial regrowth. Ethidium bromide accumulation studies revealed that the efflux decreased in the presence of each extract, as indicated by the presence of EPIs. Most identified EPIs that have been discovered to date have aromatic structures, and the seaweed extracts were found to contain various terpenes, terpenoids, phenolic compounds, indoles, pyrrole derivatives, alkaloids, and halogenated aromatic compounds. Our study highlights the potential of these compounds of the seaweeds as drug EPIs.

Cell immobilization is a way to isolate or localize intact cells in a certain space and maintain their catalytic activity. Immobilized cells can effectively reduce the negative effects of inhibitors and the processing cost of inoculum preparation for continuous or fed-batch fermentation of microorganisms. In this study, a novel yeast immobilization technique using renewable poly- l -lactic acid (PLLA) microtube array membrane (MTAM) was thoroughly evaluated for bioethanol fermentation. PLLA-MTAM was shown to be stable in 15% (v/v) ethanol solution during shaking cultivation. A yeast encapsulation efficiency of 67–70% was obtained, and the yeasts in MTAMs with greater porosity showed greater bioethanol productivity. The MTAM-immobilized Kluyveromyces marxianus , prepared using in situ and siphon methods, were evaluated using 5% (w/v) glucose fermentation. Improved glucose consumption and bioethanol production were observed in batch bioethanol fermentation. In 7 cycles during repeated-batch fermentation, the immobilized yeasts prepared using the in situ method showed a maximum C EtOH of 24.23 g/L, maximum Y P/S of 0.48 g/g, and r P EtOH of 2.69 g/L h. Our data indicated that the PLLA-MTAM immobilized yeasts significantly enhanced bioethanol productivity and was a novel, promising technology for bioethanol fermentation.

The gene agaA, of the isolated marine bacterium Pseudomonas vesicularis MA103, comprised 2958-bp nucleotides encoding a putative agarase AgaA of 985 amino acids, which was predicted to contain a signal peptide of 29 amino acids in the N-terminus, a catalytic domain of glycoside hydrolase 16 (GH16) family, a bacterial immunoglobulin group 2 (Big 2), and three carbohydrate binding modules 6 (CBM 6). The gene agaA was cloned and overexpressed in Escherichia coli, and the optimum temperatures for AgaA overexpression were 16, 20 and 24 °C. The agaA was cloned without its signal peptide for cytosolic production overexpression, whereas it was cloned with the heterologous signal peptide PelB and its endogenous signal peptide for periplasmic and extracellular productions, respectively. Extracellular and periplasmic rAgaA showed greater activity than that of cytosolic rAgaA, indicating that membrane translocation of AgaA may encourage proper protein folding. Time-course hydrolysis of agarose by rAgaA was accomplished and the products were analyzed using thin layer chromatography and matrix-assisted laser desorption inoization-time of flight mass spectrometry, indicating that AgaA from P. vesicularis was an endo-type β-1,4 agarase that cleaved agarose into neoagarotetraose and neoagarohexaose as the final products.

One-step sulfuric acid saccharification of the red alga Pterocladiella capillacea was optimized, and various detoxification methods (neutralization, overliming, and electrodialysis) of the acid hydrolysate were evaluated for fermentation with the thermotolerant yeast Kluyveromyces marxianus. A proximate composition analysis indicated that P. capillacea was rich in carbohydrates. A significant galactose recovery of 81.1 ± 5% was also achieved under the conditions of a 12% (w/v) biomass load, 5% (v/v) sulfuric acid, 121°C, and hydrolysis for 30 min. Among the various detoxification methods, electrodialysis was identified as the most suitable for fermentable sugar recovery and organic acid removal (100% reduction of formic and levulinic acids), even though it failed to reduce the amount of the inhibitor 5-HMF. As a result, K. marxianus fermentation with the electrodialyzed acid hydrolysate of P. capillacea resulted in the best ethanol levels and fermentation efficiency.

Despite the prevalence and severe effects of depression, the efficacy of currently available antidepressants is often inconsistent, and many exert undesirable side effects. Administration of food containing gamma-aminobutyric acid (GABA) has been proposed as an alternative treatment for depression. Therefore, this study ferments soy-based milk enriched in GABA and tests its anti-depressant activity in Sprague-Dawley (SD) rats. First, Lactobacillus brevis FPA 3709 isolated from fish intestine was selected for its ability to produce GABA during cultivation in MRS broth with monosodium glutamate (MSG). Second, the conditions (substrate composition and treatment, initial pH, and time and temperature of fermentation) for obtaining the highest synthesis of GABA by the selected microorganism were established. Finally, cooked black soybean milk supplemented with 1% MSG, 1% brown sugar, and 0.1% peptone (without adjusting the initial pH level) was fermented (37 °C, 48 h) by L. brevis FPA 3709. SD rats subjected to a forced swimming test (FST) and fed with fermented black soybean milk showed immobility times that were not significantly different from that of rats administered with fluoxetine, a common antidepressant drug, without exhibiting the side effects of lost appetites and reduced body weight typical of fluoxetine use. Fermented black soybean milk enriched with GABA is a good candidate as an alternative treatment of depression.

For a long time, fucoidan has been well known for its pharmacological activities, and recently low molecular weight fucoidan (LMF) has been used in food supplements and pharmaceutical products. In the present study, LMF was extracted from Laminaria japonica by enzyme hydrolysis. The toxicity of LMF in mouse and rat models was determined by many methods, such as total arsenic content, bacterial reverse mutation assay, chromosome aberration assay, and in vivo micronucleus assay. The present findings showed that LMF at 5000 μg/mL exhibited no mutagenicity. It also produced no formatting disruption of red blood cells in vivo. At 2000 mg/kg BW/day there were no toxicological indications. LMF is expected to be used as a safe food supplement.

Porphyra dentata, an edible red macroalgae, is used as a folk medicine in Asia. This study evaluatedin vitroandin vivothe protective effect of a sterol fraction fromP. dentataagainst breast cancer linked to tumor-induced myeloid derived-suppressor cells (MDSCs). A sterol fraction containing cholesterol,β-sitosterol, and campesterol was prepared by solvent fractionation of methanol extract ofP. dentata in silicagel column chromatography. This sterol fractionin vitrosignificantly inhibited cell growth and induced apoptosis in 4T1 cancer cells. Intraperitoneal injection of this sterol fraction at 10 and 25 mg/kg body weight into 4T1 cell-implanted tumor BALB/c mice significantly inhibited the growth of tumor nodules and increased the survival rate of mice. This sterol fraction significantly decreased the reactive oxygen species (ROS) and arginase activity of MDSCs in tumor-bearing mice. Therefore, the sterol fraction fromP. dentatashowed potential for protecting an organism from 4T1 cell-based tumor genesis.