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β-glucan derived from the black yeast Aureobasidium pullulans (A. pullulans) is one of the natural products attracting attention due to its high potential for application as a functional food and pharmaceutical. Our team of researchers obtained a highly soluble, low-molecular-weight β-glucan from the fermentation culture medium of A. pullulans via mechanochemical ball milling method, that is, the low-molecular-weight A. pullulans-fermented β-D-glucan (LMW-AP-FBG). We investigated the anti-inflammatory effect of LMW-AP-FBG using lipopolysaccharide (LPS)-stimulated murine macrophages (RAW264.7 cells) in the current study. LMW-AP-FBG altered LPS-stimulated inflammatory responses by reducing the release of inflammatory mediators such as nitric oxide (NO), interleukin (IL)-1β, IL-6 and tumor necrosis factor-α. As well, the mitogen-activated protein kinases (MAPKs) and nuclear factor-κB (NF-κB) signaling pathways were downregulated by LMW-AP-FBG. Furthermore, LMW-AP-FBG markedly reduced LPS-induced expression of cell surface molecules, CD14, CD86, and MHC class II, which mediate macrophage activation. These findings suggest that LMW-AP-FBG can be used as an effective immune modulator to attenuate the progression of inflammatory disease.

The beneficial immunomodulation effects of a biological response modifier glucan (BRMG) produced by two strains of Aureobasidium pullulans, AFO-202 and N-163, have already been reported. Herein, we compared their efficacy on immune-inflammatory parameters in Sprague Dawley (SD) rats. This study was performed on four groups of healthy SD rats, n=6 in each group: Group 1, euthanised on Day 0 for baseline values; Group 2, control (drinking water); Group 3, AFO-202 beta glucan, 200 mg/kg/day; and Group 4, N-163 beta glucan, 300 mg/kg/day. The neutrophil to lymphocyte ratio (NLR) decreased and leukocyte-to C-reactive protein ratio (LeCR) increased in Group 3 (AFO-202) at 15 and 29 days whereas the lymphocyte to C-reactive protein ratio (LCR) increased in group 4 (N-163), within the normal physiological range. These promising results warrant further investigations in larger numbers of healthy and diseased models to develop appropriate strategies for balancing immune system dysregulation.

Xylooligosaccharides (XOS) are non-digestible and fermentable oligomers that stand out for their efficient production by enzymatic hydrolysis and beneficial effects on human health. This study aimed to investigate the influence of the main reaction parameters of the beechwood xylan hydrolysis using crude xylanase from Aureobasidium pullulans CCT 1261, thus achieving the maximum XOS production. The effects of temperature (40 to 50 °C), reaction time (12 to 48 h), type of agitation, substrate concentration (1 to 6%, w/v), xylanase loading (100 to 300 U/g xylan), and pH (4.0 to 6.0) on the XOS production were fully evaluated. The most suitable conditions for XOS production included orbital shaking of 180 rpm, 40 °C, and 24 h of reaction. High contents of total XOS (10.1 mg/mL) and XOS with degree of polymerization (DP) of 2-3 (9.7 mg/mL), besides to a high percentage of XOS (99.1%), were obtained at 6% (w/v) of beechwood xylan, xylanase loading of 260 U/g xylan, and pH 6.0. The establishment of the best hydrolysis conditions allowed increasing both the content of total XOS 1.5-fold and the percentage of XOS by 9.4%, when compared to the initial production (6.7 mg/mL and 89.7%, respectively). Thus, this study established an efficient enzymatic hydrolysis process that results in a hydrolysate containing XOS with potential prebiotic character (i.e., rich in XOS with DP 2-3) and low xylose amounts.

In this study, cost-effective substrates such as cassava starch, corn steep liquor (CSL) and soybean meal hydrolysate (SMH) were used for pullulan production by Aureobasidium pullulans CCTCC M 2012259. The medium was optimized using response surface methodology (RSM) and artificial neural network (ANN) approaches, and analysis of variance indicated that the ANN model achieved higher prediction accuracy. The optimal medium predicted by ANN was used to produce high molecular weight pullulan in high yield. SMH substrates increased both biomass and pullulan titer, while CSL substrates maintained higher pullulan molecular weight. Results of kinetic parameters, key enzyme activities and intracellular uridine diphosphate glucose contents revealed the physiological mechanism of changes in pullulan titer and molecular weight using different substrates. Economic analysis of batch pullulan production using different substrates was performed, and the cost of nutrimental materials for CSL and SMH substrates was decreased by 46.1% and 49.9%, respectively, compared to the control using glucose and yeast extract as substrates, which could improve the competitiveness of pullulan against other polysaccharides in industrial applications.

To improve β-1,3-1,6-D-glucan (β-glucan) production by Aureobasidium pullulans, an Agrobacterium tumefaciens-mediated transformation method was developed to screen a mutant A. pullulans CGMCC 19650. Based on thermal asymmetric-interlaced PCR detection, DNA sequencing, BLAST analysis, and quantitative real-time PCR assay, the T-DNA was identified to be inserted in the coding region of mal31 gene, which encodes a sugar transporter involved in pullulan biosynthesis in the mutant. The maximal biomass and β-glucan production under batch fermentation were significantly increased by 47.6% and 78.6%, respectively, while pullulan production was decreased by 41.7% in the mutant, as compared to the parental strain A. pullulans CCTCC M 2012259. Analysis of the physiological mechanism of these changes revealed that mal31 gene disruption increased the transcriptional levels of pgm2, ugp, fks1, and kre6 genes; increased the amounts of key enzymes associated with UDPG and β-glucan biosynthesis; and improved intracellular UDPG contents and energy supply, all of which favored β-glucan production. However, the T-DNA insertion decreased the transcriptional levels of ags2 genes, and reduced the biosynthetic capability to form pullulan, resulting in the decrease in pullulan production. This study not only provides an effective approach for improved β-glucan production by A. pullulans, but also presents an accurate and useful gene for metabolic engineering of the producer for efficient polysaccharide production. KEY POINTS: • A mutant A. pullulans CGMCC 19650 was screened by using the ATMT method. • The mal31 gene encoding a sugar transporter was disrupted in the mutant. • β-Glucan produced by the mutant was significantly improved.

In this study, crude and partially purified xylanase from Aureobasidium pullulans CCT 1261 were characterized and applied in beechwood xylan hydrolysis to obtain xylooligosaccharides (XOS). Fractional precipitation (0–30 %/30–60 %) with ammonium sulfate allowed partial purification of xylanases in 6.8-fold and recovery of 69.4 %. Both extracts exhibited optimum activity at 50 °C and pH 4.5, and greater stability at 40 °C within the studied range. The thermostability parameters (kd, t1/2, ΔH*, ΔG*, ΔS*) of the extracts indicated the enzyme denaturation by the increment in temperature from 40 °C to 60 °C; its process was endothermic and led to protein unfolding in a more random structure. Crude xylanase showed a lower value of the Michaelis-Menten constant (25 mg/mL) than the partially purified enzyme (50 mg/mL). In hydrolysis reactions, total XOS contents (7.7 mg/mL and 7.9 mg/mL) and XOS yields (25.7 % and 26.5 %) were equal statistically using the crude or partially purified extracts, differing only in the amount of xylose released. The hydrolysates were mainly composed of potential prebiotics, i.e., xylobiose (50.9 % and 53.8 %) and xylotriose (30.9 % and 32.8 %). Based on these results, the use of crude and partially purified xylanase from A. pullulans showed a promising perspective for xylan hydrolysis to produce XOS.

Xylanases are hydrolase enzymes mainly produced by microorganisms and applied in several industrial segments. In this paper, the effects of different pretreatments of oat hulls were evaluated on the production of xylanase by Aureobasidium pullulans CCT 1261 under submerged cultivation. Oat hulls were subjected to several pretreatments: alkaline (0.5, 1, 2, 3, and 4% of NaOH, w/v), alkaline hydrogen peroxide (7.5% of H2O2, v/v), ultrasound-assisted alkaline (1% of NaOH, w/v, 90 W, 20 kHz), and sequential alkaline and ultrasound procedures. Both untreated and pretreated oat hulls were used as substrates for xylanase production by the strain CCT 1261. Changes caused by the pretreatments were observed through the characterization of oat hulls (i.e. lignocellulosic composition and scanning electron microscopy) and then related to the enzymatic production. Sequential alkaline and ultrasound pretreatments on oat hulls promoted the highest xylanase production (30.2 U/mL) and productivity (0.42 U/mL.h) in comparison to the other strategies evaluated. The sequential pretreatment of oat hulls also increased xylanase production more than 11-fold when compared to the untreated substrate. The effectiveness of the sequential procedure, and consequent increase in xylanase production, were attributed to the physical modification of the biomass surface by the ultrasound waves, as well as the lignin removal and hemicellulose maintenance by the alkaline pretreatment with 1% NaOH. Sequential pretreatment of oat hulls allowed the production of microbial xylanases in a more sustainable way using an agricultural by-product as substrate.

The microbiota of winter wheat seeds from the North-East of Ukraine was studied by a biological method. Its considerable variability is established over three years (2017–2019). The effect of the treatment agents on most microorganisms of wheat seed microbiota in Ukraine, rather than on its genera and species, is shown. It has been proven that fungicides deleted some species and did not affect the development of others. Chemicals replaced some species or genera of fungi with others or even other microorganisms. Biological seed treatment (Phytosporin-M) has caused less microbiota change than chemical treatment (Maxim 0.25 FS, Rostock, Kinto Duo). Fungicides have replaced the dominance of Alternaria spp. (2017 – 57.8%, 2018 – 63.5%) for the dominance of yeast (Rostock – 54%) and Aureobasidium pullulans (Maxim 0.25 FS – 84.2%) in 2017, bacteria (Maxim 0.25 FS – 72.3%, Rostock – 53.8%) – in 2018. A. pullulans dominated in the microbiota of winter wheat seeds in 2019. The highest amount of A. pullulans was noted for the treatment of seeds by Phytosporin-M (85.9%). The biological seed treatment reduced the amount of Nigrospora spp. and Alternaria spp. Several times (3 and 5, respectively), chemical agents did not give Nigrospora spp. germination reduced the amount of A. pullulans, Alternaria spp. in 2019. Maxim 0.25 FS, Rostock 50%, and Kinto Duo delayed seed germination and seedling development on agar medium and soil, whereas Phytosporin-M – on the contrary, promoted the growth of seedlings and significantly exceeded control. Keywords: seed microbiota, winter wheat, seed treatment, seed-borne fungi, fungicides References Adhikari, P., Khatri-Chhetri, G., Shrestha, S., & Marahatta, S. (2016). Study on prevalence of mycoflora in wheat seeds. Turkish Journal of Agriculture - Food Science and Technology, 4 (1), 31–35. https://doi.org/10.24925/turjaf.v4i1.31-35.509 Adhikari, P., Khatri-Chhetri, G., Shrestha, S., & Marahatta, S. (2018). In-vitro study on prevalence of mycoflora in wheat seeds. Journal of the Institute of Agriculture and Animal Science, 33, 27–34. https://doi.org/10.3126/jiaas.v33i0.20679 Andersen, B. Nielsen, K.F., Fernandez, P.V, & Patriarca, A. (2015).Characterization of Alternaria strains from Argentinean blueberry, tomato, walnut and wheat. International Journal of Food Microbiology, 196, 1–10. https://doi.org/10.1016/j.ijfoodmicro.2014.11.029 Bankina, B., Bimsteine, G., Neusa-Luca, I., Roga, A., & Fridmanis, D. (2017). What influences the composition of fungi in wheat grains? ActaAgrobotanica, 70(4), 1726. https://doi.org/10.5586/aa.1726 Baybakova, E.V., Nefed'eva, E.E., & Belopukhov, S.L. (2016). Assessment of the influence of modern protectants on the germination of seeds and growth of seedlings of grain crops. Proceedings of Universities. Applied Chemistry and Biotechnology, 6(3), 57–64 (in Russian).

The combined biochemical impact of carbon, nitrogen and temperature on the biosynthesis of the antifungal volatile organic compounds (VOCs): ethanol, 2-methyl-1-propanol, 3-methyl-1-butanol and 2-phenylethanol produced by Aureobasidium pullulans A1 and A3 was investigated using a Box-Behnken experimental design and response surface methodology (RSM). Normalized peak areas derived from solid phase micro extraction-gas chromatography-mass spectrometry (SPME-GC-MS) analysis, indicated that initial carbon content had a significant influence on the biosynthesis of ethanol and alcohols with greater than three carbon atoms. This result suggests a dominant activity of the A. pullulans anabolic pathway to biosynthesize three higher alcohols via de novo biosynthesis of amino acids from sugar metabolism. Low concentrations of carbon (3-13 g l-1 ) with nitrogen as both ammonium and amino acids in the growth medium resulted in a higher number of significant linear and quadratic relationships. Nitrogen availability and growth temperature had significant negative linear and quadratic correlations with VOCs biosynthesis in most instances. Isolate-dependant metabolic response was evident for all abiotic parameters tested on alcohol production. The findings of this study offer new perspectives to improve the production of key antifungal compounds by antagonists in biological control systems.

Pullulan is an imperative microbial exo-polymer commercially produced by yeast like fungus Aureobasidium pullulans. Its structure contains maltosyl repeating units which comprises two α-(1 → 4) linked glucopyranose rings attached to one glucopyranose ring through α-(1 → 6) glycosidic bond. The co-existence of α-(1 → 6) and α-(1 → 4) glycosidic linkages endows distinctive physico-chemical properties to pullulan. It is highly biocompatible, non-toxic and non-carcinogenic in nature. It is extremely resistant to any mutagenicity or immunogenicity. The unique properties of pullulan make it a potent candidate for biomedical applications viz. drug delivery, gene delivery, tissue engineering, molecular chaperon, plasma expander, vaccination, etc. This review highlights the potential of pullulan in biomedical research and development.

Pepaya adalah buah klimakterik yang memiliki umur simpan pendek dan potensi gangguan penyakit antraknosa pada saat tahap pascapanen yang disebabkan oleh patogen Colletotrichum gloeosporioides. Tujuan dari penelitian ini adalah mengkaji keefektifan beberapa spesies khamir antagonis untuk memperpanjang umur simpan dan mengendalikan penyakit antraknosa pada pascapanen buah pepaya Callina. Penelitian ini dilaksanakan di kebun pepaya Desa Kanaga, Kabupaten Lebak, Banten pada November 2018 sampai Juli 2019 dan Laboratorium Pascapanen AGH, IPB pada bulan Maret sampai dengan Juli 2019. Penelitian ini menggunakan rancangan kelompok lengkap teracak dengan perlakuan lima jenis khamir yaitu Cryptococcus albidus Yp, Aureobasidium pullulans Dmg 30 DEP, Rhodotorula minuta Dmg 16 BEP, Candida tropicalis Lm 13 BE, dan Pseudozyma hubeiensis Dmg 18 BEP, dan sebagai pembanding perlakuan fungisida berbahan aktif azoksistrobin serta tanpa perlakuan sebagai kontrol. Hasil penelitian menunjukkan penggunaan khamir Candida tropicalis, Aureobasidium pullulans, dan Cryptococcus albidus dapat memperpanjang umur simpan buah pepaya Callina masing-masing 12.6, 12.4, dan 12.2 hari, lebih lama dibandingkan kontrol yang hanya 7 hari, dan efektif mengendalikan penyakit antraknosa dengan tingkat penekanan secara berturut-turut 58.33%, 54.17%, dan 50.00%, selama penyimpanan. Penggunaan khamir antagonis tidak mempengaruhi mutu fisik dan kimia buah pepaya Callina. Kata kunci: buah klimakterik, Colletotrichum gloeosporioides, pascapanen

Extracellular rhodanese obtained from Aureobasidium pullulans was employed in both free and immobilized forms for the biodegradation of cyanide present in cassava processing mill effluent (CPME). Crosslinking with glutaraldehyde (at an optimum concentration of 5% v/v) before entrapment in alginate beads resulted in the highest immobilization yield of 94.5% and reduced enzyme leakage of 1.8%. Rhodanese immobilized by cross-linking before entrapment (cbe) retained about 46% of its initial activity after eight cycles of catalysis compared to the entrapment in alginate alone (eaa) which lost more than 79% after the fifth catalytic cycle. A cross-examination of thermodynamic ( Δ G d * , Δ S d * , Δ H d * ) kinetic ( k d , t 1 / 2 , D and z - values ) parameters at 30-70 °C showed that cbe displayed a higher resistance to thermal inactivation when compared to the free enzyme (fe) and (eaa). The efficiency of cyanide biodegradation from the CPME by the fe, eaa and cbe were 55, 62, and 74% respectively after 6 h. Biodegradation of cyanide using the cbe was monitored using FTIR spectroscopy. Rhodanese immobilized via cbe had a higher resistance to thermal denaturation over other enzyme forms. Hence, this makes cbe adaptable for large-scale detoxification of cyanide from CPME.

With the COVID-19 pandemic causing huge threat to public health and definite treatment modalities and preventive vaccines yet to be arrived at, some of the key indicators of relevance to its prognosis have started emerging. One such independent predictor of outcome has been fasting plasma glucose (FPG) at the time of admission. Earlier, co-morbidities such as diabetes also have been reported to have a risk of relatively increased mortality due to COVID-19. In this background, we herein report on the beneficial effects of Biological response modifier glucan (BRMG) secreted by the black yeast Aureobasidium pullulans AFO-202 which has been proven to bring under control blood sugar levels in human subjects and also has potential in enhancing & regulating the immune parameters in relevance to COVID-19. We further recommend that this BRMG be tried in clinical studies of COVID-19 to provide a prophylactic effect for validation.

Background: The alleviating effects of diabetic nephropathy and hepatopathy of β-glucan were evaluated in this study. Objective: The anti-diabetic effects of β -glucan from Aureobasidium pullulans were assessed in a streptozotocin (STZ)-induced rat diabetes model at 62.5 and 125 mg/kg doses. In addition, the possibility of changes in the effects of β-glucan according to the severity of diabetes was also assessed at one dosage (62.5 mg/kg): severe, >360 mg/dL; slight, 130-200 mg/dL. Methods: Test articles were administered orally to STZ-induced diabetic rats from 21 days after STZ dosing for 4 weeks. Each of five or six female rats per group was selected using blood glucose levels at 21 days after STZ dosing. Changes in body weight were recorded during the study, along with blood glucose, blood urea nitrogen, creatinine, and serum aspartate aminotransferase and alanine aminotransferase levels. On the day of sacrifice, livers and kidneys were weighed and observed microscopically for changes in the percentage of degenerative regions and numbers of degenerative tubules in the kidney. Results: β-glucan showed no hypoglycemic effects in the STZ-induced diabetic rat model. However, it had favorable effects on decreasing diabetic complications related to diabetic nephropathy and hepatopathy. Conclusion: Based on the results of this study, it was concluded that β-glucan showed favorable effects in decreasing diabetic complications in STZ-induced rat diabetes model.

β-poly(L-malic acid) (PMLA) has attracted industrial interest for its potential applications in medicine and other industries. For a sustainable PMLA production, it requires replacing/reducing the CaCO3 usage, since the residual CaCO3 impeded the cells’ utilization, and a large amount of commercially useless gypsum was accumulated. In this study, it was found that more glucose was converted into CO2 using soluble alkalis compared with CaCO3 usage. Moreover, since the high ion strength and respiration effect of soluble alkalis also inhibited PMLA production, they could not effectively replace CaCO3. Furthermore, comparing the fermentations with different neutralizers (soluble alkali vs. CaCO3), it was found that the differential genes are mainly involved in the pathway of starch and sucrose metabolism, pentose and glucuronate interconversions, histidine metabolism, ascorbate and aldarate metabolism, and phagosome. In detail, in the case with CaCO3, 562 genes were downregulated and 262 genes were upregulated, and especially, those genes involved in energy production and conversion were downregulated by 26.7%. Therefore, the irreplaceability of CaCO3 was caused by its effect on the PMLA metabolic pathway rather than its usage as neutralizer. Finally, a combined pH shift control strategy with CaCO3 addition was developed. After the fermentation, 64.8 g/L PMLA and 38.9 g/L biomass were obtained with undetectable CaCO3 and less CO2 emission. • The effect of CaCO3 on PMLA metabolic pathway resulted in its irreplaceability. • A pH shift control strategy with CaCO 3 addition was developed. • Undetectable CaCO 3 and less CO 2 emission were detected with the new strategy.

Background: Poly-L-malic acid (PLMA) comprises aliphatic polyester polymers with broad applications in pharmaceutical industries. The fungal microorganisms are among the best natural sources recruited to supply L-malic acid (MA) as a precursor of PLMA. In this study, we investigated MA production ability of 7 clinical isolated of the fungus Aureobasidium pullulans. Materials and Methods: Seven clinical isolates of A. pullulans acquired from Westerdijk Fungal Biodiversity Institute were studied, and the isolate with the highest total MA production was selected for the optimization process. We tried to optimize the output by applying different concentrations of CaCO3 in fungus medium (1.5%, 3%, and 6%) and various incubation temperatures (27°C, 32°C, and 37°C) during 3, 7, and 14 days. Results: Intra-strains variation was significantly strong (P0.05). Conclusion: Overall, we obtained the highest MA production in Sabouraud dextrose agar (SDA)medium with 3% CaCO3 at 27°C after 7 days of incubation. Our study indicated that the fermentation period and CaCO3 concentration significantly alter MA production in A. pullulans var. melanigenum.

Aureobasidium pullulans LB83 was evaluated for cellulase production under submerged fermentation conditions. Different process variables such as carbon sources (corn cob, sugarcane bagasse, and sug...

Cellulose is the most abundant renewable carbon source found in nature and is of great interest to various industries, including the food industry. Generally, microorganisms can be used to convert ...

Low-temperature red winemaking is mainly used for enhancing the wine aroma; however, colour and phenolic extraction is reduced under this condition. This work evaluated the effects of the pectinoly...

The crude β-xylosidase from Aureobasidium pullulans CBS 135684 was immobilized on the surface of the modified spent expanded perlite via (3-aminopropyl) triethoxy-silane and glutaraldehyde cross-linking reaction. Over 80% of the initial free enzyme activity was detected on the bound enzyme. The immobilized β-xylosidase (204.7 ± 6.5 U g-1 of perlite) exhibited transxylosylation activity when sugars from black liquor xylan hydrolysate and a variety of alcohols (C1-6) were used as donor and acceptor molecules, respectively, yielding a number of alkyl xylosides. The synthesized hexyl xyloside exhibited some interesting properties comparable with those of a commercial hexyl glucoside, including an antioxidant activity (IC50 = 8.9 ± 2.0 mg mL-1), wetting time (43 s), and critical micelle concentration value (252 mmol L-1). Optimization for hexyl xyloside synthesis was performed via statistical analysis using a Box and Behnken design. The maximum predicted yield of hexyl xyloside (499.0 ± 1.1 mg g-1 xylan hydrolysate) was obtained with 10% (w/v) xylan hydrolysate, 10 U g-1 immobilized β-xylosidase, and 13% (v/v) hexanol at 70 °C, pH 6.0 for 4 h, and it was validated by the following experimental yield (498.9 ± 4.8 mg g-1 xylan hydrolysate). The maximum production yield remained relatively stable after the immobilized enzyme was reused for 12 cycles with a 471.5 ± 15.1 mg g-1 hexyl xyloside yield (94.4% of the first cycle). Thus, the immobilized β-xylosidase has a high potential as a robust catalyst for application in alkyl xyloside production.

Chitin-glucan complex (CGC) copolymer is a cell wall component in fungi and yeasts, used in medicine and cosmetics applications. This study investigated the production of CGC from Aureobasidium pullulans KSY-0516, which is used in the industrial production of β-1,3/1,6-glucan. The amino acid composition of the A. pullulans fermentation by-products was determined using an amino acid analyzer. The total amount of amino acid was 6.67 g/100 g of by-product. Additionally, the recovery of the β-glucan complex from the A. pullulans fermentation by-products was 8.44 g/100 g of by-product using ethanol precipitation. The CGC yield was determined from the by-products after yeast fermentation. The recovery of CGC was 14.83% for the yeast cell wall obtained by alkaline extraction. The CGC and β-glucan were characterized using Fourier transform infrared analysis, and compared with crab shell chitin and laminarin. This study aimed to examine the potential, as a useful substrate for biotechnology, by producing CGC from yeast cultivation by-products.

This study involved the design and fabrication of airlift fermenters for the mass propagation of a consortium of three microorganisms (Pseudomonas aeruginosa, Pseudomonas fluorescence and Aureobasidium pullulans). The efficacy of the blend in the bioremediation of simulated crude oil polluted water systems environments was also tested with a view to generating more data on crude oil degrading microorganisms for bioremediation purposes. The organisms were separately grown on a gyratory shaker at 120 rpm and at 30 °C until notable growths were observed. Equal volume of the grown organisms (500 ml) of mixed culture was used as inoculum for the 4.5 L of medium in the 7 L airlift fermenter. Thereafter, this was transferred into the 45 litres medium in the 70 L fermenter until significant growth was observed at room temperature. Culture broths were withdrawn at intervals for the determination of biomass and residual hydrocarbon concentrations. The product from the 70 L fermenter was introduced into simulated polluted water systems from which samples were withdrawn weekly. Analyses for residual hydrocarbon contents were carried out using appropriate analytical techniques. The results showed that 78.5% hydrocarbon removal was observed in the 7 L fermenter after 72 h, while that of the 70 L fermenter was 97.8% after 180 h of fermentation. Polluted water sample treated with the microbes experienced 96.4% removal after 10 weeks of treatment. It was observed further that the application of NPK fertilizer biostimulant aided microbial activities in the removal of petroleum hydrocarbons than urea, cow dung and poultry droppings. The study has demonstrated that effective bioremediation of crude oil polluted water systems could be achieved through the application of biostimulants with mass propagated crude oil degrading organisms using airlift fermenter systems. Keywords: Pseudomonas aeruginosa; Pseudomonas fluorescence; Aureobasidium pullulans; Airlift fermenter; Bioremediation; Biostimulation; Bioaugmentation.

The interest in pullulan produced from industrial wastes has substantially increased due to its industrial applications. The major constraint in pullulan production is expensive and low yield. In this study, attempts were made on the evaluation of media components for maximum pullulan production from Aureobasidium pullulans MTCC 1991 using cassava waste in solid-state fermentation. A statistical approach was scrutinized to evaluate the effects of yeast extract, K2HPO4, (NH4)2SO4, MgSO4⋅7H2O, and NaCl on pullulan production. The optimal conditions eliciting the maximum pullulan yield (6.45 g pullulan/L of culture broth) were found to be 0.6 g/L yeast extract, 3 g/L K2HPO4, 0.2 g/L (NH4)2SO4, 0.3 g/L MgSO4⋅7H2O, and 1 g/L NaCl. The produced pullulan was extracted using the solvent precipitation method and characterized by Fourier transform infrared spectroscopy and proton nuclear magnetic resonance spectroscopy spectroscopic analysis that confirmed pullulan with linear α (1–6) linkage. The results revealed that cassava waste could be used as an effective carbon source for the production of pullulan.

Determination of creatinine concentration in biological fluids is becoming increasingly relevant as a clinical test. Various chemical and physico-chemical methods have been proposed to determine creatinine concentration in biological samples, including various modifications of the well-known Jaffe color reaction, although it has several drawbacks, namely, a low specificity and sensitivity to positive and negative interference from related substances. In addition, the available methods are time consuming, require a complicated sample preparation procedure, expensive reagents, highly qualified personnel and, most importantly, do not allow real-time analysis. Therefore, increasing attention has been paid to enzymatic methods for the determination of creatinine, particularly, using microbial creatinine deiminase. In this regard, the search for new microorganisms – effective producers of specific creatinine deiminases – is an actual task for microbial enzymology and analytical biotechnology. We have analyzed creatinine deiminase activity for 10 strains of different molds: Botrytis aclada , Botrytis allii 100(5), Monilinia fructicola , Sporobolomyces salmonicolor , Totula sp., Stachybotris chartarum , Pichiapini , Aspergillus oryzea , Trichoderma lignorum , Aspergillus glaucus , Trichothecium roseum , Aureobasidium pullulans . Among the strains tested, the enzyme activity was detected in the following species: M. fructicola, A. pullulans, B. allii, T. roseum, and C. g lobosum , although only two fungi, B. allii and C. globosum , were able to use creatinine as the sole nitrogen source. Creatinine deiminase activity in cell free extracts of B. allii and C. globosum , grown in glucose medium supplemented with creatinine (as the sole carbon and nitrogen source, respectively) was 2.5 and 1.5 times higher, related to that for the strains grown in glucose-sodium nitrate medium. Therefore, the fungi B. allii and C. globosum can be used for isolation of creatinine deiminase to be used in construction of biosensors selective to creatinine.

Bioorganic asymmetric reduction of carbonyl compounds is one of the most important fundamental and practical reactions for producing chiral alcohols. The stereoselective bioreduction of prochiral ketones of benzofuran derivatives in the presence of yeast-like fungus Aureobasidium pullulans contained in the antifungal Boni Protect agent was studied. Biotransformations were carried out under moderate conditions in an aqueous and two-phase system and without multiplication of the bioreagent. Despite similar chemical structure, each of the used ketone has been reduced with varying efficiency and selectivity. One of the reasons for these results is the presence of a whole set of oxidoreductases in A. pullulans cells that are sensitive to the smallest changes in the structure of prochiral substrate. The unsymmetrical methyl ketones were biotransformed with the highest selectivity. Aureobasidium pullulans microorganism is less effective in the reduction of unsymmetrical halomethyl ketones. The presence of a heteroatom in the alkyl group significantly decreases the selectivity of the process. Finally, as a result of the preferred hydride ion transfer from the dihydropyridine ring of the cofactor to the carbonyl double bond on the re side, secondary alcohols of the S and R configuration were obtained with moderate to high enantioselectivity (55-99%).

Aureobasidium pullulans YTP6-14 was demonstrated to be an excellent multiple biosurfactant producer utilizing cheap carbon sources available in Thailand, including glycerol and cassava flour hydrolysate. A. pullulans YTP6-14 maximally produced 1.81 g/l biosurfactant in an aqueous layer (BS-AQ) in a medium containing glycerol, and 7.37 or 6.37 g/l biosurfactant in a heavy oil layer (BS-HO) in cassava flour hydrolysate or a glucose containing medium, respectively. Each BS-AQ and BS-HO had critical micelle concentration values of 41.32 mg/l and 13.51 mg/l, and both biosurfactants formed a stable food oil emulsion and reduced the amount of biofilms formed by Streptococcus sobrinus and Streptococcus mutans. BS-AQ and BS-HO were mainly composed of liamocins or exophilins and massoia lactone, respectively.

Pullulan has many useful characteristics but, its high cost limits its potential applications. In the present work, kitchen waste (KW), which otherwise has zero commercial value, was evaluated for the economical production of pullulan. Before fermentation, the KW was hydrolyzed into free sugars using an in-house produced cocktail of enzymes. During hydrolysis, 46 ± 3.5 g/l and 31 ± 2.2 g/l of total reducing sugars and glucose were released, respectively. Hydrolyzed kitchen waste was then used as substrate for fermentation by Aureobasidium pullulans MTCC 2013 yielding 20.46 ± 2.01 g/l pullulan. Further, effect of different nitrogen sources was evaluated and yeast extract (3%) was found to be the best, yielding (24.77 ± 1.06 g/l) exopolysaccharide (EPS). The pullulan produced from KW was characterized in terms of organoleptic properties, physical strength, Fourier-transform infrared spectroscopy (FTIR), and H nuclear magnetic resonance (H NMR) analysis. The results corroborated well with commercial pullulan. The biodegradable nature and water solubility of the film developed from pullulan was also confirmed. To the best of our knowledge, this is the first report on the validation of the biodegradability of in-house produced pullulan. Thus, kitchen waste appears to be a promising option for economical pullulan production. Additionally, the method may also prove to be helpful for managing the increasing load of municipal solid waste in an eco-friendly and scientific way.

A colour-variant strain of Aureobasidium pullulans FB-1 was used for the production of pullulan in a stirred tank reactor in a medium composed of sucrose, ammonium sulphate, yeast extract, K2HPO4, NaCl and MgSO4. Maximum pullulan production (4.8%, w/v) was obtained at an agitation rate of 300 rpm and aeration of 0.75 vvm in a stirred tank reactor (1 L) with a fermentation time of 6 days. Fourier transformed infrared and NMR spectrum of crude pullulan from A. pullulans FB-1 revealed solely α-(1→6) linked maltosyl units, in accord with the generally accepted structure of pullulan. Hydrolysis of crude pullulan, i.e. digestion with pullulanase also confirmed the identity as well as homogeneity of the produced pullulan. XRD analysis of crude pullulan verified its amorphous nature. Thermal analysis showed melting range of crude pullulan from 245.99–295.71 °C.

Bacterial cellulose (BC11 Uncommon abbreviation used: BC – Bacterial Cellulose) is a biomaterial produced by various strains of microorganisms. BC has improved strength and unique structural properties as compared to plant cellulose, thus has many usages in the food and pharmaceutical industries. In our previous study, a novel co-culture agitated fermentation of Komagataeibacter hansenii, a BC producer, with Aureobasidium pullulans, a producer of pullulan polysaccharide, had been demonstrated where the BC produced exhibited improved mechanical properties. Therefore, this study is undertaken to analyze BC production under different medium composition using response surface methodology (RSM) in shake-flasks and benchtop bioreactors. A verified local high point provided 22.4% higher BC production and 4.5- to 6- folds higher elastic moduli in shake-flasks and bioreactors compared to the baseline media. Overall, the study had revealed the potential of the co-culturing method to enhance BC production while maintaining the desired mechanical properties of BC produced in shake-flasks and larger scale bioreactors.

The ability of Aureobasidium pullulans NCYC 4012 to produce gluconic acid (GA) was firstly studied in flask batch cultures and it was found that GA productivity was not affected by initial glucose concentration, but highest substrate uptake rate was observed at glucose concentration of 80gL-1. The effect of oxygen mass transfer, varied by changing agitation and aeration rates or total air pressure, was evaluated in batch cultures of A. pullulans in stirred tank bioreactors. Higher productivities were attained in conditions of high oxygen mass transfer and it was proven that the use of increased total pressure of 4bar is an effective means of oxygen supply to A. pullulans cultures. At increased pressure, cells induced the antioxidant intracellular enzyme superoxide dismutase to cope with oxidative stress due to high oxygenation conditions. On the other hand, extracellular glucose oxidase activity was reduced by highly oxygenated conditions, suggesting opposite effects of oxygen on this enzyme secretion and GA production., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit and the Doctoral grant (SFRH/BD/129475/2017)., info:eu-repo/semantics/publishedVersion

Fructooligosaccharides (FOS) can be used as feed prebiotics, but are limited by high production costs. In this study, low-cost sugarcane molasses was used to produce whole-cell biocatalysts containing transfructosylating enzymes by Aureobasidium pullulans FRR 5284, followed by FOS production from molasses using the whole-cells of A. pullulans. A. pullulans in molasses-based medium produced cells and broth with a total transfructosylating activity of 123.6 U/mL compared to 61.0 and 85.8 U/mL in synthetic molasses-based and sucrose-based media, respectively. It was found that inclusion of glucose in sucrose medium reduced both transfructosylating and hydrolytic activities of the produced cells and broth. With the use of pure glucose medium, cells and broth had very low levels of transfructosylating activities and hydrolytic activities were not detected. These results indicated that A. pullulans FRR 5284 produced both constitutive and inducible enzymes in sucrose-rich media, such as molasses while it only produced constitutive enzymes in the glucose media. Furthermore, treatment of FOS solutions generated from sucrose-rich solutions using an invertase-deficient Saccharomyces yeast converted glucose to ethanol and acetic acid and improved FOS content in total sugars by 20–30%. Treated FOS derived from molasses improved the in vitro growth of nine probiotic strains by 9–63% compared to a commercial FOS in 12 h incubation. This study demonstrated the potential of using molasses to produce FOS for feed application.

Objective In this pilot clinical study, we report the beneficial effects of beta glucans derived from two strains AFO-202 and N-163 of a black yeast Aureobasidium pullulans on the biomarkers for cytokine storm and coagulopathy in COVID-19 patients. Methods A total of 24 RT-PCR positive COVID-19 patients were recruited and randomly divided into three groups (Gr): Gr. 1 control (n=8) – Standard treatment; Gr. 2: Standard treatment + AFO-202 beta glucan (n=8); and Gr. 3, Standard treatment + combination of AFO-202 and N-163 beta glucans (n=8) for 30 days. Results There was no mortality or requirement of ventilation of the subjects in any of the groups. There was a decrease in D-Dimer values (751 ng/ml to 143.89 ng/ml) and IL-6 values (7.395 pg/ml to 3.16 pg/ml) in Gr. 1 in 15 days but the levels increased to abnormal levels on day 30 (D-Dimer: 202.5 ng/ml; IL-6 55.37 pg/ml); which steadily decreased up to day 30 in groups 2 (D-dimer: 560.99 ng/dl to 79.615; IL-6: 26.18 pg/ml to 3.41 pg/ml) and 3 (D-dimer: 1614 ng/dl to 164.25 ng/dl; IL-6: 6.25 pg/ml to 0.5 pg/ml). The same trend was observed with ESR. LCR and LeCR increased while NLR decreased significantly in Gr. 3. CD4+ and CD8+ T cell count showed relatively higher increase in Gr.3. There was no difference in CRP within the groups. Conclusion As these beta glucans are well known food supplements with a track record for safety, larger multi-centric clinical studies are recommended to validate their use as an adjunct in the management of COVID-19 and the ensuing long COVID-19 syndrome., Graphical Abstract

A black yeast, Aureobasidium pullulans, extracellularly produces β-(1,3), (1,6)-D-glucan (β-glucan) under certain conditions. The β-glucan is known to be an immunomodulatory agent, and β-glucan enriched A. pullulans cultured fluid (AP-CF) is used in supplements to maintain human health. Concanavalin A (ConA) is a lectin, and when injected it is known to cause T cell mediated autoimmune hepatitis in mice. The present study investigated the effects of oral administration of AP-CF on ConA injection in mice. The results demonstrated that increases in serum alanine transaminase (ALT) levels after ConA injection were significantly suppressed in an AP-CF administered group of mice. To understand the mechanism of the ALT lowering effects of AP-CF, we used Foxp3 (forkhead box P3) knock-in mice which express the green fluorescent protein (GFP) in Foxp3 induced cells, and the effects of AP-CF on the regulatory T cell (Treg) populations were investigated. The results show that the basal level of Foxp3+ Treg populations in peripheral blood lymphocytes, liver infiltrating lymphocytes, and splenocytes was decreased after 7 days of administration of AP-CF. These findings suggest that oral administration of AP-CF suppresses the basal level of inflammation, and that it may be postulated to be involved in the ALT lowering effects of AP-CF.

Polymalic acid (PMA) is a novel biopolymer produced by the polyextremotolerant fungus Aureobasidium pullulans. In this study, a GATA-family transcriptional factor, Gat1, which regulates nitrogen uptake and PMA biosynthesis, was investigated. PMA production increased to 11.2% in the mutant overexpressing gat1 but decreased to 49.1% of the PMA titre when gat1 was knocked out from the genome of A. pullulans. Comparative transcriptome analysis of wild-type and mutant strains (∆gat1 and OE::gat1) revealed that 23 common differentially expressed genes were related to oxidative phosphorylation, ribosome biogenesis, and nitrogen metabolism. Under nitrogen-limited stress, regardless of the preferred nitrogen (glutamine, Gln) or non-preferred nitrogen (proline, Pro), 70% of Gat1 in the cells was located in the nucleus-cytoplasm, which resulted in an increase in nitrogen uptake and PMA biosynthesis regulation. Quantitative RT-PCR revealed that glucosekinase (GLK) in the glycolytic pathway and malate synthase (MLS) in the glyoxylate shunt pathway may be cross-regulated by Gat1 and nitrogen concentration (Gln or Pro), Therefore, glk was overexpressed in mutant strain (OE::gat1), which resulted in an increased PMA titre and yield of 12.6% and 13.0% respectively. These findings indicate that Gat1 may play an important role in the dual regulation of the nitrogen and carbon metabolisms in PMA biosynthesis.

A low molecular weight intracellular and extracellular fructosyltransferase was purified from the wild (Wt) and improved mutant-type (Mt) Aureobasidium pullulans and its characteristics and thermod...

Any life-threatening alteration in environmental circumstances from the optimal imposes stress on an organism. Microorganisms possess various strategies, including pigment production, to resist stress conditions in their surrounding environments. In the present study, yeast form of fungus Aureobasidium pullulans either melanized or albino was exposed to different stresses, such as salinity and antifungals (fluconazole, itraconazole, and terbinafine) as well as gamma irradiation. Results revealed that at 800 μg/ml or less of antifungal terbinafine, a positive impact was achieved on the viability of A. pullulans yeast cells, in the cultivation medium wasn't supplemented with melanin precursor (L-3,4 dihydroxyphenylalanine; L-DOPA). This enhancement in the growth yield was accompanied by increases in both internal and external melanin yields by eight and six folds, respectively. In addition, melanin is synthesized during terbinafine stress in a pathway independent from L-DOPA, but based on the excess of acetyl-CoA to tolerate squalene accumulation by feedback inhibition. Moreover, the melanized yeast cells show greater viable stability against gamma irradiation as well as pigment productivity than albino cells. The main objective of this study is to create new composites to use them as shielding materials. Theses composites based on fungal melanin's chelating activity with nanoparticles of bismuth, lead, and silver. The mass attenuation coefficient of Melanin-Bismuth composite is nearly double the mass attenuation of lead at energy = 0.662 MeV; a higher increase in mass attenuation is observed with higher gamma intensities; 1.17 and 1.33 MeV. Coupling synthesis of composites containing natural microbial products with nanotechnology is a promising field to manufacture new radiation shielding materials.

The production of cell-wall degrading enzymes (CWDE) such as cellulase and pectinase by Aureobasidium pullulans NAC8 through induction using orange peels was investigated for the potential ...

Pullulan is a microbial exopolysaccharide produced from Aureobasidium pullulans by submerged fermentation of a medium supplemented with carbon, nitrogen and other essential nutrients. These nutrients are expensive which increase the cost of pullulan production. The requirement of alternative cost-effective substrates for pullulan production is a prerequisite. Agro-based industries generate a large volume of solid/liquid waste and its accumulation generates a severe environmental impact. These wastes are composed of carbohydrates, proteins and other constituents, and can be used as substrates for the development of low-cost processes for the production of various microbial products. This could be a good environmental friendly waste management system. Pullulan production from agro-industrial wastes can be carried out by both submerged and solid-state fermentation by A. pullulans. Owing to its unique properties, pullulan has wide applications in many food-based industries. This review highlights pullulan production from agro-industrial wastes and potential applications of pullulan in various food industries.

Trilepisium madagascariense fruits are carbohydrate-rich and this study directly fermented the fruit wastes into bioethanol without the need for nutrient supplementation. The total reducing sugar (TRS) present in the mesocarp and seed of T. madagascariense fruit wastes (Tmfw) was fermented to bioethanol using Aureobasidium pullulans. Bioethanol production by A. pullulans was also optimized using Box-Behnken response surface methodology (RSM). The TRS in the mesocarp and seed of Tmfw were 11.2 ± 0.8 and 17.1 ± 1.2 g/L, respectively and further hydrolysis with cellulase resulted in increased TRS indicating the presence of cellulose. Pre-optimization, the bioethanol yield (Yps) and volumetric productivity (Qp) obtained from the fermentation of the seed by A. pullulans were 0.57 ± 0.03 g/g and 0.21 ± 0.02 g/L-1h-1, respectively. The optimum conditions for maximum bioethanol production were pH (5.95), time (24 h) and substrate concentration (5 g/L) resulting in Yps, Qp of 0.66 ± 0.06 g/g and 0.27 + 0.01 g/L-1h-1, respectively after model validation. Tmfw served as a suitable, cheap, non-toxic and readily available substrate especially in Nigeria to produce bioethanol while A. pullulans is a fungus that might be utilized for large-scale industrial bioethanol production.

Aureobasidium pullulans strains L1 and L8 were used as an alternative strategy against Neofusicoccum parvum, pathogen of apple wood and fruit. Both strains displayed a great ability to reduce pathogen conidia germination of almost 50% and totally stop its sporification in wood of ‘Gala’ and ‘Fuji’ apple cultivars. In addition, by in vitro tests the antagonistic activity displayed by non-volatile metabolites produced by both yeasts was highlighted by 70% of reduction of the pathogen mycelial growth. On apple fruit, the pathogen severity was reduced by 66% and 50% in ‘Gala’, and 70% and 60% in ‘Fuji’ by L1 and L8 respectively. In wood, no statistical differences were observed between L1, L8 and the control, except for ‘Fuji’ wood, where L8 strain displayed a wood lesion reduction of 23% with respect to the untreated control. Infrared analysis showed how the presence of both strains on woody tissues helped preserving cellulose content, preventing in this way the faster wood degradation by the pathogen. In addition, L1 and L8 simply colonized the wood surface, probably displaying an endophytic attitude, an essential requirement for controlling N. parvum in field.

The fungi Aureobasidium pullulans, Mortierella humilis, Trichoderma harzianum and Phoma glomerata were used to investigate the formation of selenium- and tellurium-containing nanoparticles during growth on selenium- and tellurium-containing media. Most organisms were able to grow on both selenium- and tellurium-containing media at concentrations of 1 mM resulting in extensive precipitation of elemental selenium and tellurium on fungal surfaces as observed by the red and black colour changes. Red or black deposits were confirmed as elemental selenium and tellurium, respectively. Selenium oxide and tellurium oxide were also found after growth of Trichoderma harzianum with 1 mM selenite and tellurite as well as the formation of elemental selenium and tellurium. The hyphal matrix provided nucleation sites for metalloid deposition with extracellular protein and extracellular polymeric substances localizing the resultant Se or Te nanoparticles. These findings are relevant to remedial treatments for selenium and tellurium and to novel approaches for selenium and tellurium biorecovery.

Since cold atmospheric pressure plasma (CAP) has not only bactericidal activity but also fungicidal activity without toxic residues and thermal damage, it is considered as an alternative method for sterilization of fungi on the surfaces of perishable foodstuffs and human bodies. Aureobasidium pullulans is a ubiquitous yeast-like fungus and called black yeast because it produces melanin, a dark biological pigment. It is well known that various melanized fungi show hyper-resistance to extreme stress conditions including high levels of radioactivity. Curiously, however, there is very little information about the fungicidal effects of CAP on melanized fungi. Therefore, we herein investigated the effects of CAP on A. pullulans, using cold atmospheric argon plasma (Ar plasma). We found that ammonium sulfate repressed the synthesis of melanin in A. pullulans as well as Aureobasidium melanogenum. Although the non-melanized A. pullulans cells were efficiently killed by the exposure of Ar plasma, the melanized cells showed the significant resistance to Ar plasma as well as to hydrogen peroxide and thermal stress. In order to improve the fungicidal efficacy of Ar plasma, we examined the combination of Ar plasma and Fenton reaction. We realized that FeCl2 and FeSO4 significantly improved the sterilization efficacy of Ar plasma on the melanized A. pullulans.

Liamocins are biosurfactants produced by the fungus Aureobasidium pullulans. A. pullulans belongs to the black yeasts and is known for its ability to produce pullulan and melanin. However, the production of liamocins has not been investigated intensively. Initially, HPLC methods for the quantification of liamocin and the identification of liamocin congeners were established. Eleven congeners could be detected, differing in the polyol head groups arabitol or mannitol. In addition, headless molecules, so-called exophilins, were also identified. The HPLC method reported here allows quick and reliable quantification of all identified congeners, an often-overlooked prerequisite for the investigation of valuable product formation. Liamocin synthesis was optimized during cultivation in lab-scale fermenters. While the pH can be kept constant, the best strategy for liamocin synthesis consists of a growth phase at neutral pH and a subsequent production phase induced by a manual pH shift to pH 3.5. Finally, combining increased nitrogen availability with a pulsed fed-batch fermentation, cell growth, and liamocin titers could be enhanced. Here, the maximal titers of above 10 g/L that were reached are the highest reported to date for liamocin synthesis using A. pullulans in lab-scale fermenters.

The Kururu no β-glu® (KBG) is a commercial hydrothermal-treated Aureobasidium pullulans β-glucan produced by a unique hydrothermal process that results in high solubility of the β-glucan. In this study, we examined the biological activities of this reagent. RAW264.7 cells do not express Dictin-1 on the cell surface, but cells still respond to various pathogen molecular patterns. Lipopolysaccharide (LPS) induced nitrogen oxide (NO) synthesis and TNF−α production in RAW264.7 cells, and those were suppressed by KBG in a dose-dependent manner. The major signaling cell surface receptor respond to LPS is the TLR4/MD-2 complex. The UT12 antibody against to the TLR4/MD-2 complex mimics LPS function and induces cell responses. NO generation and TNF-α production were similarly induced in cells by stimulation with the antibody, but those were not suppressed by KBG. Cell responses induced by other TLR ligands, such as CPG (TLR9 ligand) and Pam3CSK4 (TLR1/TLR2 ligand), were also suppressed by KBG. Therefore, the target molecule for KBG is different from TLR receptors and Dictin-1. Although we also examined the suppressive activities of several other β-glucan products, comparable activities were not detected with other reagents. A unique hydrothermal process may produce the active reagent. Reprocessing KBG increased low molecular weight fractions, and suppressive activities were markedly enhanced. Therefore, low molecular weight fractions obtained by hydrothermal processing of KBG may result in potential reagents that control inflammation induced by various pathogens.