The ability of gellan gum-immobilised cells of the heavy metal-tolerant bacterium Alcaligenes sp. AQ05-001 to utilise both heavy metal-free and heavy metal-polluted feathers (HMPFs) as substrates to produce keratinase enzyme was studied. Optimisation of the media pH, incubation temperature and immobilisation parameters (bead size, bead number, gellan gum concentration) was determined for the best possible production of keratinase using the one-factor-at-a-time technique. The results showed that the immobilised cells could tolerate a broader range of heavy metal concentrations and produced higher keratinase activity at a gellan gum concentration of 0.8% (w/v), a bead size of 3 mm, bead number of 250, pH of 8 and temperature of 30 °C. The entrapped bacterium was used repeatedly for ten cycles to produce keratinase using feathers polluted with 25 ppm of Co, Cu and Ag as substrates without the need for desorption. However, its inability to tolerate/utilise feathers polluted with Hg, Pb, and Zn above 5 ppm, and Ag and Cd above 10 ppm resulted in a considerable decrease in keratinase production. Furthermore, the immobilised cells could retain approximately 95% of their keratinase production capacity when 5 ppm of Co, Cu, and Ag, and 10 ppm of As and Cd were used to pollute feathers. When the feathers containing a mixture of Ag, Co, and Cu at 25 ppm each and Hg, Ni, Pb, and Zn at 5 ppm each were used as substrates, the immobilised cells maintained their operational stability and biological activity (keratinase production) at the end of 3rd and 4th cycles, respectively. The study indicates that HMPF can be effectively utilised as a substrate by the immobilised-cell system of Alcaligenes sp. AQ05-001 for the semi-continuous production of keratinase enzyme.
Nephrotic syndrome affects both children and adults. Idiopathic nephrotic syndrome is reported to be one of the most frequent renal pathologies in childhood. Nephrotic children are at high risk for severe pneumococcal infections as one of the life-threatening complications of nephrotic syndrome due to involvement of the immunosuppressive regimen and the acquired immune deficiency induced by nephrotic syndrome including decreased plasma IgG and low complement system components. Aiming to prevent pneumococcal infection is of paramount importance especially in this era of ever-increasing pneumococcal resistance to penicillins and cephalosporins. The pneumococcal vaccines currently available are inactivated vaccines-the two main forms in use are polysaccharide vaccines and conjugated vaccines. However, the data supporting the use of these vaccines and to guide the timing and dosage recommendations is still limited for nephrotic children. Thus, this review discusses the evidences of immunogenicity and safety profile of both vaccinations on nephrotic patients as well as the effect of nephrotic syndrome treatment on vaccine seroresponses.
The demand for fresh, safe, and healthy fruits by consumers has increased, which concurrently occurs with an increase in initiatives on reducing food wastage. Starfruit consists of good nutrition and valuable sensory attributes, but its shelf life is short and can only be preseved for a few days at ambient storage. This research was conducted to determine the effectiveness of synergistic edible coatings (pectin [Pe] and maltodextrin [M] and 100, 200, and 300 ppm of sodium chloride [SC]) on the quality and safety criterion of starfruits throughout a shelf life analysis of 14 days at ambient temperature. Consumer acceptability of the edible-coated starfruit was also evaluated. The coating process was performed using a dipping method. The uncoated (control) and coated samples were evaluated for the characteristics of weight, pH, total soluble solids (TSS), water activity, color, texture, microbial growth, FTIR, and sensory evaluation. From the results, the starfruit coated with Pe + M + 100 ppm SC had a significantly lower weight-loss trend compared to the other samples. On day 14, pH of the coated starfruits were 3.02, 3.14, and 3.31 for 100, 200, and 300 ppm of SC, respectively, were found to be significantly different (p < 0.05) from the control (pH 3.49). The control had a significantly higher value of total soluble solids (6.00 ⁰Brix) compared to the coated starfruits (6.00, 5.47, and 5.33 ⁰Brix, respectively). The coated samples have significantly higher values of firmness than control especially in initial days of storage. It was observed that Pe + M + 100 ppm SC could minimize the spoilage of fruits by reducing the growth of yeast and mold, as well as bacteria, up to 0.86 and 2.02 log CFU/ml, respectively. FTIR results confirmed the presence of the coating on the starfruit. In the sensory evaluation, no significance different (p > 0.05) were obtained for all the sensory attributes and overall acceptability for day 0 and 3. In conclusion, starfruit coated with synergistic Pe + M + 100 ppm SC appeared to be the best sample in extending its shelf life and maintaining the physicochemical characteristics of starfruits up to more than 14 days.
Fungi of the Trichoderma species are valued industrial enzymes in support of the 'zero-waste' technology to convert agro-industrial biomass into valuable products, i.e. nanocellulose (NC). In this study, an in silico approach using substrate docking and molecular dynamic (MD) simulation was used to predict the order of which the multilayers of cellulosic polymers, i.e. lignin, hemicellulose and cellulose in oil palm leaves (OPL) are degraded by fungal enzymes, endocellulase and exocellulase. The study aimed to establish the catalytic tendencies of the enzymes to optimally degrade the cellulosic components of OPL for high yield production of NC. Energy minimized endocellulase and exocellulase models revealed satisfactory scores of PROCHECK (90.0% and 91.2%), Verify3D (97.23% and 98.85%) and ERRAT (95.24% and 91.00%) assessments. Active site prediction by blind docking, COACH meta-server and multiple sequence alignment indicated the catalytic triads for endocellulase and exocellulase were Ser116-His205-Glu249 and Ser382-Arg124-Asp385, respectively. Binding energy of endocellulase docked with hemicellulose (-6.0 kcal mol-1) was the most favourable followed by lignin (-5.6 kcal mol-1) and cellulose (-4.4 kcal mol-1). Exocellulase, contrarily, bonded favorably with lignin (-8.7 kcal mol-1), closely followed by cellulose (-8.5 kcal mol-1) and hemicellulose (-8.4 kcal mol-1). MDs simulations showed that interactions of complexes, endocellulase-hemicellulose and the exocellulase-cellulose being the most stable. Thus, the findings of the study successfully identified the specific actions of sugar-acting enzymes for NC production. Communicated by Ramaswamy H. Sarma.
Nowadays, many of the tropical fruits have been commercialized worldwide due to increasing demand. In 2018, global tropical fruit has reached an unprecedented peak of 7.1 million tonnes. As such, a lot of large scale farming has been initiated to cultivate the fruit for commercialization. The nature of tropical fruit is perishable make the fruit easily undergo post-harvest losses especially when the fruit travels in a long distance for distribution. Losses of tropical fruit is estimated around 18-28% after harvesting. Then, the losses will continually develop during the trading process. Applying fruit coating on the fruit can minimize substantial privation. This article compendiously reviews the needs of coating and discuss different types of coating materials. The efficiency of different coating materials; polysaccharide, protein, lipid and composite based coating on tropical fruit is highlighted. There are various types of coating available for major fruit such as banana, mango, pineapple and avocado that can effectively extend the post-harvest life, minimize water loss, reduce chilling injuries and fight against post-harvest disease. Coating from minor fruit such as durian, rambutan, passion-fruit and mangosteen are still limited especially made from lipid and protein coating. In choosing the most appropriate coating for tropical, the nature of fruit needs to be understood. In addition, the chemistry of coating components and techniques of application is important in modulating the fruit quality.
The aim was to explore the utilization of tea leaf waste fibers (TLWF) as a source for the production of cellulose nanocrystals (CNC). TLWF was first treated with alkaline, followed by bleaching before being hydrolyzed with concentrated sulfuric acid. The materials attained after each step of chemical treatments were characterized and their chemical compositions were studied. The structure analysis was examined by Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction (XRD). From FTIR analysis, two peaks at 1716 and 1207 cm-1-which represent C=O stretching and C⁻O stretching, respectively-disappeared in the spectra after the alkaline and bleaching treatments indicated that hemicellulose and lignin were almost entirely discarded from the fiber. The surface morphology of TLWF before and after chemical treatments was investigated by scanning electron microscopy (SEM) while the dimension of CNC was determined by transmission electron microscopy (TEM). The extraction of CNC increased the surface roughness and the crystallinity index of fiber from 41.5% to 83.1%. Morphological characterization from TEM revealed the appearance of needle-like shaped CNCs with average diameter of 7.97 nm. The promising results from all the analyses justify TLWF as a principal source of natural materials which can produce CNC.
The aim was to explore the utilization of oil palm mesocarp fiber (OPMF) as a source for the production of cellulose nanocrystals (CNC). OPMF was first treated with alkali and then bleached before the production of CNC by acid hydrolysis (H₂SO₄). The produced materials were characterized using Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), thermogravimetric analysis (TGA), a scanning electron microscope (SEM) and a transmission electron microscope (TEM). It was proven that acid hydrolysis can increase the crystallinity of bleached OPMF and reduce the dimension of cellulose to nano scale. Changes in the peaks of the FTIR spectrum at 2852 (C-H stretching), 1732 (C=O stretching) and 1234 cm-1 (C-O stretching) indicated that the alkali treatment completely removed hemicelluloses and lignin from the fiber surface. This can be seen from the thermogram obtained from the TGA characterization. Morphological characterization clearly showed the formation of rod-shaped CNCs. The promising results prove that OPMF is a valuable source for the production of CNC.
Bacteria are considered as the major cell factories, which can effectively convert nitrogen and carbon sources to a wide variety of extracellular and intracellular biopolymers like polyamides, polysaccharides, polyphosphates, polyesters, proteinaceous compounds, and extracellular DNA. Bacterial biopolymers find applications in pathogenicity, and their diverse materialistic and chemical properties make them suitable to be used in medicinal industries. When these biopolymer compounds are obtained from pathogenic bacteria, they serve as important virulence factors, but when they are produced by non-pathogenic bacteria, they act as food components or biomaterials. There have been interdisciplinary studies going on to focus on the molecular mechanism of synthesis of bacterial biopolymers and identification of new targets for antimicrobial drugs, utilizing synthetic biology for designing and production of innovative biomaterials. This review sheds light on the mechanism of synthesis of bacterial biopolymers and its necessary modifications to be used as cell based micro-factories for the production of tailor-made biomaterials for high-end applications and their role in pathogenesis.
Fucoidan is a sulfated polysaccharide that consists mainly of fucose and is found in brown seaweeds. In this study, fucoidan was extracted from Sargassum binderi (Fsar) from Malaysia and subsequently characterized in terms of composition, structure and toxicology. It was found that the molecular weight, polydispersity index, monosaccharide profile and degree of sulfation of Fsar differed from those of commercial food-grade fucoidan (Fysk). NMR analysis suggested that the main structure of Fsar was →3)fuc-2-OSO3-(1→3)fuc-2-OSO3-(1→. A cytotoxicity study employing up to 200 mg/mL Sargassum binderi extract showed that cell inhibition was less than 50% (IC50), while acute toxicity results classified S. binderi as category 5 (unclassified) according to the OECD Guideline 423, as no mortality was observed at the highest dosage (2,000 mg/kg). Both toxicity results showed that this material is safe to be consumed. The chemical characteristics and non-toxicity of Fsar demonstrate its potential in biological and food product applications.
Protein-based films are thin and flexible films derived from protein sources. They are
completely biodegradable and used in food engineering, packaging, drug recovery, and other
applications. In food packaging, gelatin is widely used due to properties such as low cost,
availability, functional attributes, mechanical (flexibility and tension) and optical (brightness
and opacity) strength, barrier against gas flow, and structural resistance to water and
microorganisms. Therefore, this paper reviews the characterisation of biodegradable
protein-based films from gelatin alternatives, mainly from fish and chicken skin, as food
packaging materials. The properties of film packaging derived from gelatin alternatives were
compared with films derived from mammalian gelatin. The findings showed that the blended
gelatin alternatives with polysaccharide improved physical properties such as water vapour
permeability, gas permeability, light transmission and transparency, thermal properties,
microstructure, colour, and heat sealability. Moreover, improvements in mechanical
properties such as tensile strength and elongation at break were also investigated. This review
also comes out with suggestions for future research on the compatibility between gelatin films
and food ingredients. This paper provides a comprehensive overview that promotes the
development of biodegradable blended films from gelatin alternatives for packaging
applications in the food industry and related fields.
This article presents experimental data on oil palm biomass (oil palm leaves, oil palm trunk and empty fruit bunch) handsheet production characterization by biodelignification treatment using Bacillus cereus extracted from termite gut (Coptotermus curvignathus). It associates the lignocellulose chemical composition obtained via technical association pulp and paper industry TAPPI T 222 om-02 testing on lignin content reduction determination, holocellulose and hemicellulose content determination (Kurscher-Hoffner method). Several data obtained for handsheet characterization presents brightness, opacity, contrast ratio, din transparency, thickness, bursting and tearing indexes are collected. Handsheet surface morphology was also observed on ratio of gaps differences between fiber bonding conducted using scanning electron microscope (SEM) and ImageJ software. The raw data findings supplement chemical composition analysis for both untreated and treated substrates on handsheet quality performance check as presented in the research article "Bio-Mechanical Pulping of Bacteria Pre-Treatment on Oil Palm Biomass for Handsheet Production" [1]. For understanding correlations into the difference among lignocellulose content composition which affect the handsheet formation and mechanical strength refer to article from this research [1]. This dataset is made publicly available for optimizing alternative waste material reuse in the pulp and paper industrial section.
Unfavorable side effects of available antipsychotics limit the use of conventional delivery systems, where limited exposure of the drugs to the systemic circulation could reduce the associated risks. The potential of intranasal delivery is gaining interest to treat brain disorders by delivering the drugs directly to the brain circumventing the tight junctions of the blood-brain barrier with limited systemic exposure of the entrapped therapeutic. Therefore, the present research was aimed to fabricate, optimize and investigate the therapeutic efficacy of amisulpride (AMS)-loaded intranasal in situ nanoemulgel (AMS-NG) in the treatment of schizophrenia. In this context, AMS nanoemulsion (AMS-NE) was prepared by employing aqueous-titration method and optimized using Box-Behnken statistical design. The optimized nanoemulsion was subjected to evaluation of globule size, transmittance, zeta potential, and mucoadhesive strength, which were found to be 92.15 nm, 99.57%, -18.22 mV, and 8.90 g, respectively. The AMS-NE was converted to AMS-NG using poloxamer 407 and gellan gum. Following pharmacokinetic evaluation in Wistar rats, the brain Cmax for intranasal AMS-NG was found to be 1.48-folds and 3.39-folds higher when compared to intranasal AMS-NE and intravenous AMS-NE, respectively. Moreover, behavioral investigations of developed formulations were devoid of any extrapyramidal side effects in the experimental model. Finally, outcomes of the in vivo hematological study confirmed that intranasal administration of formulation for 28 days did not alter leukocytes and agranulocytes count. In conclusion, the promising results of the developed and optimized intranasal AMS-NG could provide a novel platform for the effective and safe delivery of AMS in schizophrenic patients.
In this study, we evaluated and characterized microbial cellulose produced from Kombucha after eighth day of fermentation by employing SEM, FTIR, X-ray diffractometry, adsorption isotherm, and by measuring the swelling properties. Results on SEM revealed microbial cellulose layer to be composed of a compact cellulose ultrafine network like structure. FTIR spectra showed the presence of a characteristic region of anomeric carbons (960 – 730 cm-1), wherein a band at 891.59 cm-1 confirmed the presence of β, 1-4 linkages. Results of FTIR spectra also showed microbial cellulose to be free from contaminants such as lignin or hemicellulose, which are often present in plant cellulose. X-ray diffraction studies exhibited the overall degree of crystallinity index for MCC to be slightly lower than that of microbial cellulose. Results on swelling properties indicated microbial cellulose to possess higher fiber liquid retention values (10-160%) compared to commercial MCC (5-70%). The adsorption isotherm curves showed similarities between microbial cellulose with that of pure crystalline substance. Overall, results obtained in this study were comparable with the commercial microcrystalline cellulose, indicating that the process developed by us can be explored industrially on a pilot scale.
The aim of this work is to study the effect of hydrocolloids; guar gum (GG), xanthan gum (XG) and carboxymethyl cellulose (CMC) on the physicochemical properties, microbiological quality and sensory properties in order to investigate the potential of applying fermented cassava (tapai ubi) in ice cream. Fermented cassava ice cream (FCI) incorporated with the three types of hydrocolloid was prepared and the protein content, pH value, overrun, colour, hardness, microstructure, FTIR spectrum and sensory acceptance of all samples were determined. Fermented cassava ice cream incorporated with XG showed the highest protein content (14.88%), pH value (pH 6.07), and overrun value (4.27%) as compared to the fermented cassava ice cream incorporated with GG and CMC. Meanwhile, ice cream incorporated with GG possessed the highest L* (94.43) and hardness (3693.15 g) value as compared to XG and CMC. The microstructure study showed that the difference in uniformity at the interface obtained with different types of the hydrocolloids added demonstrated the effect of fat absorption at the air interfaces. The FTIR spectrum investigated indicated that the addition of the fermented cassava to FCI had increased the OH group in the ice cream as compared to the control. All samples were microbial safe as the total plate counts in all samples were below the standard as prescribed in Food Act 1983 with no presence of E. coli . In conclusion, fermented cassava ice cream with XG showed the good quality in terms of its pH value, overrun, total plate count and overall acceptability.
The use of component from Ganoderma lucidum as prebiotic source is interesting as the G. lucidum itself was known for more than a decade in the traditional Chinese medicine. In this work, Ganoderma lucidum crude polysaccharides (GLCP) and Polysaccharide-fraction number 2 (PF-2) were used as carbon sources in the fermentation with Bifidobacterium sp. The results showed the potential of prebiotic effect of the G. lucidum extract in batch-culture fermentation based on increment in the growth of bacteria used (0.4 – 1.5 log10 CFU/mL) after 18h fermentation. Fermentation was further done using faecal materials as bacterial inocula and bacterial growth changes were examined using real-time PCR. The results showed the ability of GLCP and PF-2 to support the growth of Bifidobacterium genus with 0.3 and 0.7 log10 cells/ml increased, respectively. Interestingly, Lactobacillus which is known as beneficial bacterial genus also showed growth increment with 0.7 and 1 log10 cells/ml increased. The competition for carbon sources thus inhibits the growth of potentially harmful genus, Salmonella (0.3 and 0.5 log10 cells/ml) in comparison to the control.
To understand the variations in the decomposability of tropical peat soil following deforestation for an oil palm plantation, a field incubation experiment was conducted in Sarawak, Malaysia. Peat soils collected from three types of primary forest, namely Mixed Peat Swamp (MPS; Gonystylus-Dactylocladus-Neoscrotechinia association), Alan Batu (ABt; Shorea albida-Gonstylus-Strenonurus association), and Alan Bunga (ABg; Shorea albida association), were packed in polyvinyl chloride pipes and installed in an oil palm plantation. Carbon dioxide (CO2) and methane (CH4) fluxes from soil were monthly measured for 3years. Environmental variables including soil temperature, soil moisture content, and groundwater table were also monitored. The pH, loss on ignition, and total carbon (C) content were similar among the three soils, while total N content was larger in the MPS than in the ABg soils. Based on13C nuclear magnetic resonance (NMR) spectroscopy, C composition of the MPS and ABg soils was characterized by the largest proportion of C present as alkyl C and O-alkyl C, respectively. The C composition of the ABt soil was intermediate between the MPS and ABg soils. The CO2fluxes from the three soils ranged from 78 to 625mgCm-2h-1with a negative correlation to groundwater level. The CH4fluxes ranged from -67 to 653μgCm-2h-1. Both total CO2and CH4fluxes were larger in the order ABg>ABt>MPS (P<0.05). Annual rate of peat decomposition as was estimated from cumulative C loss differed up to 2 times, and the rate constant in exponential decay model was 0.033y-1for the MPS soil and 0.066y-1for the ABg soil. The field incubation results of the three forest peat soils seem to reflect the difference in the labile organic matter content, represented by polysaccharides.
This paper reports on the extraction, partial characterization and the potential application of crude polysaccharides from defatted coconut residue as a prebiotic. The coconut residue was defatted and extracted to obtain the crude polysaccharides and its physicochemical properties were determined. The crude polysaccharides were assessed for monosaccharide composition, total carbohydrate content, reducing sugar concentration and protein content determination. The functional group and structural elucidation of crude polysaccharides was also done using Fourier transform infrared spectra analysis. The product was then subjected to artificial human gastric juice treatment to determine digestibility. Finally, an in vitro proliferation and acid production by two probiotic bacteria namely Lactobacillus casei Shirota and Lactobacillus bulgaricus were included in this study. It was found that the defatted coconut residue contained ash (0.54%), moisture (55.42%), protein (1.69%), crude fat (17.26%) and carbohydrate (25.73%). The percentage of crude polysaccharides extracted was 0.73 ± 0.04. The two fractions of monosaccharides obtained were glucose and fructose. Total carbohydrate content of DCR was 13.35% (w/v). The quantitative value of the reducing sugars obtained was 20.71%. Protein content in the crude polysaccharides was 0.009% and the peaks which indicated the presence of protein were observed at around 1640 cm(-1) (amide I) and 1530 cm(-1) (amide II). DCR crude polysaccharides were highly resistant (88%) to hydrolysis when subjected to artificial human gastric juice. The product was found to markedly stimulate two tested probiotics to proliferate and produce organic acids. All the above findings are supportive of the fact that polysaccharides extracted from DCR, an industrial waste, have a vast potential to be exploited as novel prebiotics.
Lignocellulosic biomasses such as wheat straw are widely used as a feedstock for biogas production. However, these biomasses are mainly composed of a compact fibre structure and therefore, it is recommended to treat them prior to its usage for biogas production in order to improve their bioavailability. The aim of this work is to evaluate, in terms of performance stability, methane yield and economic feasibility, two different scenarios: a mesophilic codigestion of wheat straw and animal manure with or without a low-energy demand alkaline pre-treatment (0.08gKOHgTS-1of wheat straw, for 24h and at 25°C). Besides this, said pre-treatment was also analysed based on the improvement of the bioavailable carbohydrate content in the untreated versus the pre-treated wheat straw. The results pointed out that pre-treated wheat straw prompted a more stable performance (in terms of pH and alkalinity) and an improved methane yield (128% increment) of the mesophilic codigestion process, in comparison to the "untreated" scenario. The pre-treatment increased the content of cellulose, hemicellulose and other compounds (waxes, pectin, oil, etc.) in the liquid fraction, from 5% to 60%, from 11.5% to 39.1% TS and from 57% to 79% of the TS in the liquid fraction for the untreated and pre-treated wheat straws, respectively. Finally, the pre-treated scenario gained an energy surplus of a factor 13.5 and achieved a positive net benefit of 90.4€tVS-WS-1d-1, being a favourable case for an eventual scale-up of the combined process.
The β-1,6-glucanases are ubiquitous enzymes which appear to be implicated in the morphogenesis and have the ability to become virulence factor in plant-fungal symbiotic interaction. To our knowledge, no report on ß-1,6-glucanases purification from Trichoderma longibrachiatum has been made, although it has been proven to have a significant effect as a biocontrol agent for several diseases. Therefore, the aim of this study was to purify β-1,6- glucanase from T. longibrachiatum T28, with an assessment on the physicochemical properties and substrate specificity. β-1,3-glucanase enzyme, from the culture filtrate of T. longibrachiatum T28, was successively purified through precipitation with 80% acetone, followed by anionexchange chromatography on Neobar AQ and chromatofocusing on a Mono P HR 5/20 column. (One β-1,6-glucanase) band at 42kDa in size was purified, as shown by the SDS-PAGE. The physicochemical evaluation showed an optimum pH of 5 and optimum temperature of 50°C for enzyme activity with an ability to maintain 100% enzyme stability. Enzyme activity was slightly reduced by 10-20% in the presence of 20 mM of Zn2+, Ca2+, Co2+, Mg2+, Cu2+, Mn2+ and Fe2+. The highest β-1,6-glucanase hydrolysis activity was obtained on pustulan due to the similarity of β-glucosidic bonds followed by laminarin, glucan and cellulose. Therefore, it can be concluded that the characterization of ß-1,6-glucanase secreted by T. longibrachiatum in term of molecular weight, responsed to selected physicochemical factors and the substrate specificity are approximately identical to other Trichoderma sp.
Lignosus rhinocerotis has a long history of use by the indigenous community within East Asia to treat a range of health conditions including asthma and chronic cough. To date, there is limited scientific evidence to support its therapeutic effects in relieving these airways conditions. In this study, we examined the effects of the different molecular weight fractions [high-molecular-weight (HMW), medium-molecular-weight (MMW), and low-molecular-weight (LMW)] obtained from the cold water sclerotial extract (CWE) of L. rhinocerotis on airways patency using airway segments isolated from Sprague Dawley rat in an organ bath set-up. It is demonstrated that the HMW and MMW fractions exhibited higher efficacy in relaxing the pre-contracted airways when compared to the CWE and LMW fraction. In addition, the HMW fraction markedly supressed carbachol-, 5-hydroxytrptamine-, and calcium-induced airway contractions. CWE demonstrated a lower efficacy than the HMW fraction but it also significantly attenuated carbachol- and calcium-induced airway contractions. Results showed that the bronchorelaxation effect of CWE and fractions is mediated via blockade of extracellular Ca2+ influx. The composition analysis revealed the following parts of carbohydrate and proteins, respectively: HMW fraction: 71 and 4%; MMW fraction: 35 and 1%; and LMW fraction: 22 and 0.3%. Our results strongly suggest that the polysaccharide-protein complex or proteins found in the HMW and MMW fractions is likely to contribute to the bronchorelaxation effect of CWE.