Exopolysaccharide (EPS) Production by Endophytic and Basidiomycete Fungi
by
,
Wai Prathumpai
1,*,
Umpawa Pinruan
2,
Sujinda Sommai
2,
Somjit Komwijit
1 and
Kwanruthai Malairuang
1 1
Biocontrol Technology Research Team, Integrative Crop Biotechnology and Management Research Group, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, 113 Thailand Science Park, Phahonyothin Rd., Khlong Nueng, Khlong Luang, Pathum Thani 12120, Thailand
2
Plant Microbe Interaction Research Team, Integrative Crop Biotechnology and Management Research Group, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, 113 Thailand Science Park, Phahonyothin Rd., Khlong Nueng, Khlong Luang, Pathum Thani 12120, Thailand
*
Author to whom correspondence should be addressed.
Fermentation 2025, 11(4), 183; https://doi.org/10.3390/fermentation11040183
Submission received: 19 February 2025
/
Revised: 13 March 2025
/
Accepted: 14 March 2025
/
Published: 1 April 2025
(This article belongs to the Special Issue Fermentation Processes: Modeling, Optimization and Control: 2nd Edition)
Abstract
:The screening of exopolysaccharides (EPS) produced by 52 isolates of endophytic and basidiomycete fungi was studied on two different media, PDB and PYGM. There were five isolates that could produce dried exopolysaccharide of more than 4 g/L (S. commune LF01962, LF01001, LF01581, Pycnoporus sp. MMCR00271.1, Pestalotiopsis sp. PP0005). The molecular weights of these exopolymers were found to be in the range of 2.5–500 kDa. These five exopolysaccharides, produced by five different fungal isolates, showed non-cytotoxic activity against NCTC clone 929 and HDFn cell lines. The selected fungal isolate of S. commune LF01962 was used for further optimization of different medium compositions affecting exopolysaccharide production using statistical methods. Among four conditions tested in the first step (xylose + peptone, glucose + (NH4)2HPO4, fructose + peptone, and mannose + yeast extract), mannose + yeast extract resulted in the highest exopolysaccharide production of 5.10 ± 2.00 g/L. In the second step using Plackett–Burman design, the optimal medium for S. commune exopolysaccharide production was found to consist of 40 g/L glucose, 5 g/L mannose, 20 g/L (NH4)2HPO4, 5 g/L yeast extract, 3 g/L monosodium glutamate, 0.5 g/L KH2PO4, 0.5 g/L K2HPO4, 0.2 g/L MgSO4, 1 mL/L trace elements, and 3 mL/L vitamin solution, which resulted in 8.16 g/L exopolysaccharide production. Exopolysaccharide production in a 5 L bioreactor using small pellets as seed inoculum was found to produce 18.28 g/L exopolysaccharide.
1. Introduction
Fungal exopolysaccharides (EPSs) are high-molecular-weight polysaccharides composed of sugar monomer subunits that are secreted into the surrounding environments and/or dispersed in their growth media [1]. Some of these possess novel bioactive components, exhibit low toxicity, and have potential applications in various industries including cosmetics, pharmaceuticals, medicine, and food [2,3]. Although EPSs have been produced, isolated, and studied for many different fungi [1,4], novel EPSs have not yet been fully explored due to the high diversity of fungal species in nature [3]. Endophytes, a group of fungi of interest, have the potential to produce EPSs with novel characteristics and properties [5]. The exploration and utilization of microbial polysaccharides for potential industrial applications have significantly increased in recent years [3,4]. Several studies have indicated that endophytes are potent producers of bioactive EPSs with unique properties, structures, and biological activities, making them suitable for applications in cosmetic, pharmaceutical, medical, and food industries [3,5]. Furthermore, many researchers have documented procedures for the production, isolation, and identification of EPS-producing endophytic fungi [1,6,7,8]. Optimization of EPS production by an endophytic fungus, Pestalotiopsis sp. BC55, resulted in the production of 4.320 ± 0.022 g/L of EPS with a molecular weight of approximately 2 × 105 Da. Structural elucidation of the EPS indicated the presence of only (1→3)-linked β-D-glucopyranosyl moieties [1]. Furthermore, the production of EPS by Agrocybe cylindracea reached a maximum of 3.0 g/L within 10 days [6]. A medicinal mushroom, Fomes fomentarius, produced a maximum EPS concentration of 3.64 g/L under optimal culture conditions [6,7]. Schizophyllan, produced by S. commune in an optimized medium in a 5 L fermenter, reached a concentration of 12.80 g/L [8]. Schizophyllan produced by a similar strain of the fungus Schizophyllum commune using cheaply available sago starch as a carbon source was observed to be thermally stable up to 125 °C with a high molecular weight of 14.73 × 103 kDa [9]. Additionally, exopolysaccharides were isolated from the submerged fermentation broth of Morchella conica, and the chemical structure of the isolated polysaccharide was elucidated [10]. In another study, eight endophytes isolated from Piper hispidum Sw., belonging to genera Diaporthe, Marasmius, Phlebia, Phoma, Phyllosticta, and Schizophyllum, were reported to produce EPSs in submerged cultures. These EPSs were rich in glucose (51%) and had a molecular weight of 46.6 kDa [11]. The medicinal mushroom Ganoderma lingzhi yielded EPSs at a concentration of 3.57 ± 0.21 g/L. These EPSs were heteropolysaccharides with high molecular weights (475,000 kDa and 21.6 kDa, 87.97%) and were composed of uronic acid, D-mannose, L-rhamnose, and D-glucose [12]. In submerged culture, Ganoderma lucidum achieved an EPS production of 4.7 g/L when the pH was adjusted from 3.0 to 6.0 after the fourth day [13]. The endophytic fungus Bionectria ochroleuca M21 produced EPS in submerged culture, reaching a production of 2.65 ± 0.16 g/L after 4 days of fermentation in a 5 L bioreactor [14]. Endophytes, therefore, represent a group of fungi of interest that can produce EPSs with the novel characteristics of EPS such as molecular weight distributions, being non-cytotoxic to human cell lines, and having high product yield, etc. This research aimed to study non-cytotoxic EPSs against human cell lines of the threshold of 20% cytotoxicity which serves as a critical point in categorizing exopolysaccharides as toxic to tested cells with different molecular weights and high yields by the fungal potential candidates. These EPSs can then be used for commercial applications in the future.
2. Materials and Methods
2.1. Microorganism and Growth Conditions
Fifty two isolates of mushroom and endophytic fungi (Table 1) were obtained from Biotec Culture Collection (BCC), Pathum Thani, Thailand, and from Plant Microbe Interaction Research Team, Integrative Crop Biotechnology and Management Research Group, National Center for Genetic Engineering and Biotechnology, Pathum Thani, Thailand. All strains were identified by morphological study, phylogeny, and 16 S rRNA gene sequence analysis. Stock cultures were maintained on potato dextrose agar (PDA, DifcoTM and BBLTM, Becton, Dickinson, MD, USA) that was cut into the cryotube containing 10% of glycerol and stored at −80 °C.
2.2. Seed Culture Preparation
Storage stock was activated on PDA medium in a Petri dish for 7 days. The activated culture was transferred and grown in a 250 mL flask containing 50 mL of potato dextrose broth (PDB) at 25 °C on a rotary shaker incubator (200 rpm/min) for 5 days.
2.3. Production of Exopolysaccharide by Selected Fungi
2.3.1. Growth Condition and Exopolysaccharide Extraction
Exopolysaccharide production in submerge cultures was performed in 250 mL flasks containing 50 mL of PDB (standard medium) and Peptone Yeast extract Glucose Medium (PYGM) (10 g/L glucose, 5 g/L bacteriological peptone, 20 g/L yeast extract, 1 g/L KH2PO4, 0.5 g/L MgSO4·7H2O) after inoculation with 10% (v/v) of the seed culture. The experimental cultures were incubated at 25 °C on a rotary incubator (200 rpm/min) for 7 days.
After 7 days, the cultures were vacuum filtered through pre-weight Whatman® No. 1 filter paper. Cold-95% ethanol (4:1) was then added to mycelium-free filtrate for exopolysaccharide precipitation and then the solutions were kept at –20 °C overnight. The precipitants were lyophilized to obtain the exopolysaccharide dried weight. Mycelium on pre-weight filter papers was oven dried at 80 °C for 3 days to obtain the mycelium dried weight [15,16].
2.3.2. In Vitro Cytotoxicity Against Mouse Lung Fibroblasts (NTCT Clone 929) and Human Dermal Fibroblasts, Neonatal (HDFn)
NCTC clone 929 (ATCC, Manassas, VA, USA) and HDFn (Invitrogen, Carlsbad, CA, USA) were grown in Eagle’s Minimum Essential Medium (EMEM) containing 10% FBS and 1 mM pyruvate and were incubated in 5% CO2 chamber at 37 °C. For experiments, NCTC clone 929 and HDFn cells were seeded into 96-well plates at 1 × 103 and 5 × 103 cells/well, respectively. After 48 h of incubation for NCTC clone 929 and 72 h for HDFn, each cell type was challenged with exopolysaccharides at 100, 50, 20, and 10 µg/mL for 48 h. After challenging, cell viability was measured as described by Riss T.L. et al. [17]. Then, the cell viability was measured by MTT assay to obtain the average cell viability number in the tested solution compared with the control. The average of the cell viability was obtained from 8 wells using ellipeticine as a positive control.
2.3.3. Molecular Weight of Exopolysaccharide Measurement
The lyophilized exopolysaccharides were dissolved in 5 mg/mL water and were filtered through a 0.2 µm syringe filter to avoid the insolubilizing agent. The average molecular weight of the exopolysaccharides was determined with High Performance Liquid Chromatography (HPLC), RI detector, using a gel permeation column (PL aquagel-OH MIXED-H; Agilent, Santa Clara, CA, USA) eluted with deionized water at a flow rate of 0.5 mL/min at 80 °C. The standard of 6–450 kDa dextrans was used as references.
2.4. Medium Optimization for Maximized Exopolysaccharide from S. commune BCC 82612
After obtaining the high potential fungal strain for exopolysaccharide production, medium optimization for maximum exopolysaccharide production was performed by Design-Expert®13 (Stat-Ease).
2.4.1. Seed Culture Preparation for S. commune BCC 82612
Storage stock was activated on PDA medium in a Petri dish for 7 days. The activated culture was transferred and grown in a 1000 mL flask containing 200 mL of potato dextrose broth (PDB) at 25 °C on a rotary shaker incubator (200 rpm/min) for 5 days.
After 5 days, a big pellet of seed culture was ready for use. For the mycelial seed culture, the grown culture was homogenized with a sterile blender before use. The small pellet seed culture was prepared by the inoculation of the mycelial seed culture into PDB and incubated on the rotary shaker for 2 days before use.
2.4.2. Selection of Carbon and Nitrogen Sources
Six carbon sources (fructose, glucose, sucrose, maltose, mannose, xylose) and 8 nitrogen sources ((NH4)2HPO4, NH4H2PO4, (NH4)2SO4, bacteriological peptone, casein hydrolysate, KNO3, malt extract, yeast extract) were used in this experiment. A total of 20 g of carbon source and 10 g of nitrogen source were added to the basal medium (0.5 g/L KH2PO4, 0.5 g/L K2HPO4, 0.2 g/L MgSO4). The pH of the experimental medium was adjusted to 5.5. Every combination of each source was performed in triplicate. At the end of the experiment, the mycelium dried weight and exopolysaccharide dried weight were analyzed for the selection of each source.
2.4.3. Screening of the Significant Medium Component by Plackett–Burman Design
A total of 11 variables (Table 2) were used in this experiment to generate a set of 26 experimental designs. All components were added to the basal medium as above with the final pH at 5.5. All of the experiments were carried out in triplicate.
The significant effects of the variables on the production were identified for the isolates based on confidence levels above 95% (p < 0.05).
2.5. Exopolysaccharide Production by S. commune BCC 82612 in Laboratory Scale Bioreactor
Exopolysaccharide production by S. commune BCC 82612 was carried out in a 5 L bioreactor (Satorius) with 4 L of optimized production medium. The bioreactor was equipped with two Rushton type turbines and baffles. The optimized medium (pH 5.5) was sterilized in situ at 121 °C for 15 min. Glucose was sterilized separately and was mixed aseptically with the other components of the medium in the bioreactor. The medium was inoculated with 10% (v/v) inoculum, and fermentation was carried out at 25 °C with uncontrolled pH. The impeller speed was initially adjusted to 100 rpm at the first 2 days of culture and adjusted to 300 after that, and compressed sterile air was sparged into the medium at the rate of 1 vvm. The samples were withdrawn every day and analyzed for dried mycelial weight, dried exopolysaccharide weight, and residual glucose. Three different types of seed cultures (mycelium, small pellet, big pellet) were studied for the production of exopolysaccharides in the laboratory bioreactor.
3. Results
3.1. Screening of the High Potential Fungi for Exopolysaccharide Production
By employing a multi-criteria screening approach involving the ability to produce exopolysaccharides (>4 g/L), assessing toxicity to human cells, and determining molecular weight, this comprehensive selection process ensures that the chosen strains not only exhibit high exopolysaccharide yields but also demonstrate favorable characteristics in terms of safety and structural properties. Ultimately, this systematic screening methodology enables the identification of fungal strains with the most promising attributes for further exploration and potential industrial applications.
3.1.1. Production of Exopolysaccharides in PDB and PYGM Media
Exopolysaccharide and mycelium dried weights were obtained from the experiments that were shown in Table 3. The fungi produced exopolysaccharides with dried weights of 0.01 ± 0.00–4.01 ± 2.32 g/L in PDB medium and 0.11 ± 0.03–6.59 ± 0.98 g/L in PYGM medium. The mycelium dried weights of the fungi of 0.55 ± 0.15–5.01 ± 0.31 and 1.77 ± 0.15–9.76 ± 0.09 g/L were obtained on PDB and PYGM media, respectively. The identification of five strains capable of producing dried exopolysaccharides exceeding 4 g/L showed their remarkable potential for industrial applications and further research. For instance, the high-yield exopolysaccharide-producing strains included S. commune LF01962, LF01001, LF01581, MMCR00333, and Pycnoporus sp. MMCR00271.1. The exopolysaccharides produced by the identified high-yield strains were utilized in subsequent experiments of in vitro human cell toxicity tests and molecular weight determinations.
3.1.2. Cytotoxicity Test of Exopolysaccharides Produced by Fungi Against NTCT Clone 929 and HDFn Cells
Exopolysaccharides produced by the 52 selected fungal strains were challenged with NCTC clone 929 and HDFn cells for 48 h. Cytotoxicity was calculated by normalizing with the non-exopolysaccharides in percentage. At the highest concentration of the challenging reaction, the cytotoxicity of the exopolysaccharides is shown in Table 4. The threshold of 20% cytotoxicity serves as a critical point in categorizing exopolysaccharides as toxic to the tested cells. Exceeding this threshold indicates a significant detrimental effect on cell viability, warranting classification into the toxic category. The identification of non-cytotoxic exopolysaccharides in the experiments signifies their safety for cell viability. On the other hand, the discovery of exopolysaccharides yielding negative results would potentially stimulate cell growth.
3.1.3. Molecular Weight Measurement of Exopolysaccharides
The molecular weight analysis of the exopolysaccharides from S. commune LF01581 and Pycnoporus sp. MMCR00271.1 revealed that their peaks were outside the expected range when compared to the standard dextran with a molecular weight of 6–450 kDa. This observation indicates that these exopolysaccharides may have molecular weights that are significantly higher or lower than the standard range, highlighting potential variations in their structural characteristics. In Table 5, S. commune LF01962, LF01001, and Pestalotiopsis sp. PP0005 exhibit varying molecular weights of the exopolysaccharides. It is interesting to note that Pestalotiopsis sp. PP0005 produced exopolysaccharides with a molecular weight of 494.5 kDa in PYGM medium. However, when cultivated in PBD medium, the molecular weight of the exopolysaccharides exceeded 500 kDa. This shift in molecular weight based on the growth medium suggests the influence of different cultivation conditions on the properties of the exopolysaccharides produced by Pestalotiopsis sp. PP0005. The data on S. commune LF01001 reveal that it produced exopolysaccharides of various sizes in PYGM medium. However, when cultivated in PDB medium, higher molecular weight sizes of the exopolysaccharides were observed, resembling the exopolysaccharides produced by S. commune LF01962, which maintained a consistent molecular weight across both media types. The data in Table 5 indicate that the fungus produced a uniform size of exopolysaccharide on both types of media components. From the results, S. commune LF01962 was chosen as the high potential strain for further study based on the exopolysaccharide production, molecular weight, and cytotoxicity.
3.2. Media Components Optimization for Exopolysaccharides Production by S. commune LF01962
3.2.1. Production of Exopolysaccharides by S. commune LF01962 on Different Carbon and Nitrogen Sources
In the initial step of optimizing the production medium components, S. commune LF01962 underwent screening for suitable carbon and nitrogen sources. The production of exopolysaccharide and mycelium dried weights by S. commune LF01962 is detailed in Table 6. The fungus produced exopolysaccharides ranging from 0.7 g/L to 5.1 g/L using various carbon and nitrogen sources. Notably, there were four conditions that resulted in higher exopolysaccharide production: xylose + peptone, glucose + (NH4)2HPO4, fructose + peptone, and mannose + yeast extract. These successful carbon and nitrogen sources were then utilized as variables along with other factors in the Plackett–Burman design to optimize the significant components of the exopolysaccharide production medium.
3.2.2. Production of Exopolysaccharides by S. commune LF01962 Using Plackettt–Burman Design
In the Plackett–Burman design for the exopolysaccharide and mycelium dried weights of S. commune LF01962 with 26 treatments (Table 7), the fungus produced exopolysaccharides ranging from 0.68 g/L to 8.16 g/L, which was 2.5 times higher than in the enriched medium (PYGM). The ANOVA results (Table 8) from this experiment identified significant factors affecting exopolysaccharide production: glucose, (NH4)2HPO4, yeast extract, monosodium glutamate, and trace elements (p ≤ 0.01). High levels of glucose, diammonium hydrogen phosphate, yeast extract, and monosodium glutamate had a positive impact on exopolysaccharide production, while low levels of trace elements restricted maximum exopolysaccharide production. The optimal medium for S. commune exopolysaccharide production consisted of 40 g/L glucose, 5 g/L mannose, 20 g/L (NH4)2HPO4, 5 g/L yeast extract, 3 g/L monosodium glutamate, 0.5 g/L KH2PO4, 0.5 g/L K2HPO4, 0.2 g/L MgSO4, 1 mL/L trace elements, and 3 mL/L vitamin solution.
3.3. Exopolysaccharide Production by S. commune LF01962 in Laboratory Bioreactor
The validation of the optimal medium for S. commune LF01962 exopolysaccharide production was conducted in a 5 L bioreactor. Three types of inocula were tested: freely dispersed mycelium, small pellets, and big pellets. The freely dispersed mycelium inoculum yielded only 3.7 g/L of exopolysaccharides (Figure 1A), which was lower than the production at the flask scale. Interestingly, the mycelium dried weight decreased during the culture due to the high shear rate of the propeller, which cut the mycelium and reduced the production efficiency.
The study observed that the different pellet forms of the inoculum resulted in higher exopolysaccharide production, with the bigger pellets (pellet size > 5 mm diameter) producing 11.38 g/L (Figure 1C) and the smaller pellets (pellet size > 1 mm diameter) producing 18.28 g/L (Figure 1B). Interestingly, the mycelium dried weight of the pellet form continued to increase during exopolysaccharide production. These findings suggest that the use of small pellet inoculum led to higher exopolysaccharide production compared to other seed inoculum types.
4. Discussions
Endophytic and basidiomycete fungi represent promising groups capable of producing substantial amounts of exobiopolymers with varying chemical structures, cytotoxicities, and molecular weight profiles. In this study involving fifty-two fungal isolates from these groups, screened on PDB and PYGM media (Table 3), five isolates were identified as producing over 4 g/L of non-cytotoxic exopolysaccharides (Table 4) with molecular weights ranging from 2.5 to 5000 kDa (Table 5). The test of non-cytotoxic EPSs against human cell lines of the threshold of 20% cytotoxicity serves as a critical point in categorizing exopolysaccharides as toxic to tested cells with different molecular weights; and most of the selected strains possess non-cytotoxicity after being tested with these human cell lines, proving their potential in human use. The exobiopolymers produced by strains yielding less than 4 g/L were noteworthy for their diverse molecular weight sizes. Different molecular wieghts and structures can lead these exopolysaccharides to be used for different applications [15], and the molucular weight can be reduced by different techniques such as gamma radiation to signify their industrial appication [16]. However, their exclusion from further studies was based on considerations of the growth rates and mycelial yield. The identification of non-cytotoxic exopolysaccharides in the experiments signifies their safety for cell viability; their origins are from natural habitats which are edible mushrooms and which enable them to be used for human food applications, as most of them were non-cytotoxic. The strain S. commune LF01962 was selected for further optimization, focusing on various nutritional factors. It was discovered that the initial step involving mannose/yeast extract had the most significant impact on schizophyllan production, resulting in the highest exobiopolymer yield of 5.10 ± 2.00 g/L (Table 6). Subsequent optimization revealed that an optimal medium comprising 40 g/L glucose, 5 g/L mannose, 20 g/L (NH4)2HPO4, 5 g/L yeast extract, 3 g/L monosodium glutamate, 0.5 g/L KH2PO4, 0.5 g/L K2HPO4, 0.2 g/L MgSO4, 1 mL/L trace element, and 3 mL/L vitamin solution led to a further increase to 8.16 g/L of the exobiopolymer (Table 7). ANOVA analyses of the factorial model for exopolysaccharide production by S. commune LF01962, shown in Table 8, represent a significant model (>99%) with the expression of exopolysaccharides = 38.54A + 9.14B + 0.24C − 41.34D + 60.59E + 11.31F + 0.73G + 2.76H + 21.94J + 20.93K + 11.15L (where A = glucose, B = xylose, C = mannose, D = diammonium hydrogen phosphate, E = yeast extract, F = casein hydrolysate, G = peptone, H = manganese sulfate, J = glutamic acid, K = trace elements, and L = vitamin solutions). Glucose and yeast extract had the highest positive effects but diammonium hydrogen phosphate had the highest negative effects on the exopolysaccharide production of S. commune LF01962. In the evaluation of schizophyllan production in a 5 L bioreactor by S. commune LF01962, three types of seed inoculum, free mycelium (Figure 1A), small pellets (Figure 1B), and big pellets (Figure 1C), were used. Interestingly, the small pellets resulted in the highest schizophyllan production of 18.28 g/L. This highlights that morphological characteristics, such as the size of pellets, can have a significant impact on schizophyllan production in a bioreactor. The study demonstrated that schizophyllan production reached 12.80 g/L in an optimized medium by S. commune in a 5 L fermenter according to Li et al. [18], and schizophyllan was produced by S. commune in a similar study [8,9]. Additionally, another observation by Li et al. [18] reported a slightly higher schizophyllan production of 13.68 g/L. These results show the variations in schizophyllan production levels achieved through different experimental setups and optimization strategies. Schizophyllan’s intriguing biological properties, such as antitumor and immune-stimulating activities, make this exopolysaccharide fascinating due to its unique structure of β-(1-6)-Branched β-(1-3)-glucans, gelation behavior, and natural origin [3]. The study highlighted the highest production levels compared to other reports, indicating its potential for scale-up and future applications in various fields.
5. Conclusions
The study highlighted the potential of endophytic and basidiomycete fungi to produce a variety of exopolysaccharides with non-cytotoxic properties, as they have been part of human food sources for many years. These exopolysaccharides exhibit diverse molecular weight sizes, structures, gelation properties, and compositions, making them versatile for numerous applications. The research, with 52 candidate strains, suggests that these fungi hold promise for further exploration in exopolysaccharide production, ranging from food and feed to cosmetics, pharmaceuticals, medical, and cosmeceutical industries. Notably, the high schizophyllan production of S. commune LF01962 indicates the potential for scalability and future commercial applications.
Author Contributions
Experimental investigation, methodology, data curation, formal analysis, and writing—original draft preparation were performed by S.K., K.M. and S.S.; investigation and methodology were performed by S.K., K.M. and S.S.; supervision was performed by U.P. and W.P.; and conceptualization, funding acquisition, project administration, resources, supervision, and writing—review and editing were performed by W.P. and U.P. All authors have read and agreed to the published version of the manuscript.
Funding
This work was financially supported by TDG-cosmeceuticals platform, National Nanotechnology Center, National Science and Technology Development Agency, Thailand. Funding code P20-50656.
Institutional Review Board Statement
Not applicable.
Informed Consent Statement
Not applicable.
Data Availability Statement
The original contributions presented in this study are included in the article. Further inquiries can be directed to the corresponding author.
Conflicts of Interest
The authors declare no conflicts of interest.
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Figure 1.
Mycelium and exopolysaccharide dried weights and residue glucose during S. commune LF01962 exopolysaccharide production with mycelium (A), small pellets (B), and big pellets (C) inoculum.
Figure 1.
Mycelium and exopolysaccharide dried weights and residue glucose during S. commune LF01962 exopolysaccharide production with mycelium (A), small pellets (B), and big pellets (C) inoculum.
Table 1.
List of 52 strains of endophytic and basidiomycete fungi for exopolysaccharide screening.
| No | Original Code | BCC Code | Microorganisms |
|---|---|---|---|
| 1 | LF00466 | 56724 | Schizophyllum commune |
| 2 | LF00467 | 56725 | Schizophyllum commune |
| 3 | LF00469 | 56727 | Schizophyllum commune |
| 4 | LF00470 | 56728 | Schizophyllum commune |
| 5 | LF00473 | 56731 | Schizophyllum commune |
| 6 | LF00534 | 61999 | Schizophyllum commune |
| 7 | LF00543 | 62007 | Schizophyllum commune |
| 8 | LF01001 | 66090 | Schizophyllum commune |
| 9 | LF01581 | Schizophyllum commune | |
| 10 | LF01962 | 82612 | Schizophyllum commune |
| 11 | MMCR00071 | Schizophyllum commune | |
| 12 | MMCR00176 | Schizophyllum commune | |
| 13 | MMCR00333 | Schizophyllum commune | |
| 14 | MMCR00334 | Schizophyllum commune | |
| 15 | MMCR00336 | Schizophyllum commune | |
| 16 | LF01222 | Auricularia cf. auricula | |
| 17 | LF01580 | Auricularia cf. delicata | |
| 18 | LF01616 | Auricularia cf. polytricha | |
| 19 | MMCR00014 | Auricularia sp. | |
| 20 | MMCR00107 | Auricularia sp. | |
| 21 | MMCR00108 | Auricularia sp. | |
| 22 | MMCR00157.1 | Auricularia sp. | |
| 23 | MMCR00171 | Auricularia sp. | |
| 24 | MMCR00177 | Auricularia sp. | |
| 25 | PHD00142 | Auricularia sp. | |
| 26 | MMCR00214.2 | Calvatia sp. | |
| 27 | MMCR00215.1 | Ganoderma sp. | |
| 28 | MMCR00216.1 | Ganoderma sp. | |
| 29 | MMCR00271.1 | Pycnoporus sp. | |
| 30 | MMCR00309.2 | Coprinus cf. fimetarius | |
| 31 | MMCR00347.1 | Amauroderma sp. | |
| 32 | PP0005 | Pestalotiopsis sp. | |
| 33 | PP0013 | Nigrospora sp. | |
| 34 | PP0049.1 | Mucor-like sp. | |
| 35 | ENFER0001 | Lasiodiplodia-like sp. | |
| 36 | ENFER0002 | Phomopsis sp. | |
| 37 | ENFER0003 | Mucor-like sp. | |
| 38 | ENFER0004 | Unidentified sp. | |
| 39 | ENFER0005 | Unidentified sp. | |
| 40 | ENFER0006 | Unidentified sp. | |
| 41 | ENFER0007 | Unidentified sp. | |
| 42 | ENFER0008 | Unidentified sp. | |
| 43 | ENFER0009 | Unidentified sp. | |
| 44 | MMCR00035 | Panus sp. | |
| 45 | MMCR00041 | Panus sp. | |
| 46 | MMCR00120 | Panus sp. | |
| 47 | MMCR00199 | Lentinus cf. polychrous | |
| 48 | MG0001 | Volvariella volvacea | |
| 49 | MG0002 | Flammulina velutipes | |
| 50 | MCR366.3 | Hericium erinaceus | |
| 51 | MCR369.1 | Pleurotus cf. djamor | |
| 52 | MCR370 | 56724 | Lentinus polychrous |
Table 2.
The selected variables of 26 treatments with different medium composition using Plackett–Burman Design.
Table 2.
The selected variables of 26 treatments with different medium composition using Plackett–Burman Design.
| No. | Factors | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| A | B | C | D | E | F | G | H | J | K | L | |
| g/L | g/L | g/L | g/L | g/L | g/L | g/L | g/L | g/L | ml/L | ml/L | |
| 1 | 40 | 5 | 0 | 20 | 5 | 5 | 0 | 0 | 0 | 3 | 1 |
| 2 | 20 | 5 | 5 | 10 | 5 | 5 | 5 | 0 | 0 | 1 | 3 |
| 3 | 40 | 0 | 5 | 20 | 0 | 5 | 5 | 0.5 | 0 | 1 | 1 |
| 4 | 20 | 5 | 0 | 20 | 5 | 0 | 5 | 0.5 | 3 | 1 | 1 |
| 5 | 20 | 0 | 5 | 10 | 5 | 5 | 0 | 0.5 | 3 | 3 | 1 |
| 6 | 20 | 0 | 0 | 20 | 0 | 5 | 5 | 0 | 3 | 3 | 3 |
| 7 | 40 | 0 | 0 | 10 | 5 | 0 | 5 | 0.5 | 0 | 3 | 3 |
| 8 | 40 | 5 | 0 | 10 | 0 | 5 | 0 | 0.5 | 3 | 1 | 3 |
| 9 | 40 | 5 | 5 | 10 | 0 | 0 | 5 | 0 | 3 | 3 | 1 |
| 10 | 20 | 5 | 5 | 20 | 0 | 0 | 0 | 0.5 | 0 | 3 | 3 |
| 11 | 40 | 0 | 5 | 20 | 5 | 0 | 0 | 0 | 3 | 1 | 3 |
| 12 | 20 | 0 | 0 | 10 | 0 | 0 | 0 | 0 | 0 | 1 | 1 |
| 13 | 30 | 2.5 | 2.5 | 15 | 2.5 | 2.5 | 2.5 | 0.25 | 1.5 | 2 | 2 |
| 14 | 20 | 0 | 5 | 10 | 0 | 0 | 5 | 0.5 | 3 | 1 | 3 |
| 15 | 40 | 0 | 0 | 20 | 0 | 0 | 0 | 0.5 | 3 | 3 | 1 |
| 16 | 20 | 5 | 0 | 10 | 5 | 0 | 0 | 0 | 3 | 3 | 3 |
| 17 | 40 | 0 | 5 | 10 | 0 | 5 | 0 | 0 | 0 | 3 | 3 |
| 18 | 40 | 5 | 0 | 20 | 0 | 0 | 5 | 0 | 0 | 1 | 3 |
| 19 | 40 | 5 | 5 | 10 | 5 | 0 | 0 | 0.5 | 0 | 1 | 1 |
| 20 | 20 | 5 | 5 | 20 | 0 | 5 | 0 | 0 | 3 | 1 | 1 |
| 21 | 20 | 0 | 5 | 20 | 5 | 0 | 5 | 0 | 0 | 3 | 1 |
| 22 | 20 | 0 | 0 | 20 | 5 | 5 | 0 | 0.5 | 0 | 1 | 3 |
| 23 | 40 | 0 | 0 | 10 | 5 | 5 | 5 | 0 | 3 | 1 | 1 |
| 24 | 20 | 5 | 0 | 10 | 0 | 5 | 5 | 0.5 | 0 | 3 | 1 |
| 25 | 40 | 5 | 5 | 20 | 5 | 5 | 5 | 0.5 | 3 | 3 | 3 |
| 26 | 30 | 2.5 | 2.5 | 15 | 2.5 | 2.5 | 2.5 | 0.25 | 1.5 | 2 | 2 |
Remarks: A = glucose, B = Xylose, C = Mannose, D = (NH4)2HPO4, E = yeast extract, F = casein hydrolysate, G = Peptone, H = MnSO4, J = Monosodium glutamate, K = Trace elements, L = Vitamin solution.
Table 3.
Exopolysaccharide and mycelium dried weights produced by 52 fungal strains in different media.
Table 3.
Exopolysaccharide and mycelium dried weights produced by 52 fungal strains in different media.
| No | Code | Genus | Epithet | Mycelium Dried Weight (g/L) | Exopolysaccharide Dried Weight (g/L) | ||
|---|---|---|---|---|---|---|---|
| PDB | PYGM | PDB | PYGM | ||||
| 1 | LF00466 | Schizophyllum | commune | 3.20 ± 0.38 | 8.57 ± 1.34 | 0.10 ± 0.02 | 2.41 ± 0.06 |
| 2 | LF00467 | Schizophyllum | commune | 3.75 ± 0.24 | 10.23 ± 1.64 | 0.37 ± 0.27 | 1.91 ± 0.13 |
| 3 | LF00469 | Schizophyllum | commune | 3.05 ± 0.13 | 7.29 ± 2.11 | 0.02 ± 0.01 | 2.26 ± 0.12 |
| 4 | LF00470 | Schizophyllum | commune | 3.00 ± 0.27 | 11.42 ± 1.59 | 0.17 ± 0.05 | 1.25 ± 0.66 |
| 5 | LF00473 | Schizophyllum | commune | 2.93 ± 0.25 | 10.90 ± 1.33 | 0.25 ± 0.31 | 1.81 ± 0.23 |
| 6 | LF00534 | Schizophyllum | commune | 4.09 ± 0.11 | 13.92 ± 2.53 | 0.64 ± 0.24 | 0.72 ± 0.28 |
| 7 | LF00543 | Schizophyllum | commune | 2.87 ± 2.48 | 12.07 ± 3.78 | 0.01 ± 0.01 | 1.94 ± 0.21 |
| 8 | LF01001 | Schizophyllum | commune | 2.83 ± 0.22 | 13.30 ± 0.34 | 4.01 ± 2.32 | 5.07 ± 0.78 |
| 9 | LF01581 | Schizophyllum | commune | 3.04 ± 0.14 | 8.50 ± 1.00 | 0.32 ± 0.14 | 2.04 ± 0.27 |
| 10 | LF01962 | Schizophyllum | commune | 3.44 ± 0.46 | 11.03 ± 0.63 | 2.72 ± 0.26 | 6.39 ± 0.14 |
| 11 | MMCR00071 | Schizophyllum | commune | 2.58 ± 0.23 | 7.46 ± 0.28 | 0.13 ± 0.04 | 2.07 ± 0.11 |
| 12 | MMCR00176 | Schizophyllum | commune | 3.31 ± 0.39 | 10.64 ± 1.15 | 0.30 ± 0.15 | 1.67 ± 0.61 |
| 13 | MMCR00333 | Schizophyllum | commune | 3.57 ± 0.28 | 12.35 ± 1.54 | 1.70 ± 0.96 | 4.29 ± 0.61 |
| 14 | MMCR00334 | Schizophyllum | commune | 2.95 ± 0.24 | 6.05 ± 1.61 | 0.12 ± 0.04 | 1.56 ± 0.22 |
| 15 | MMCR00336 | Schizophyllum | commune | 2.71 ± 0.09 | 7.66 ± 1.74 | 0.31 ± 0.23 | 1.54 ± 0.88 |
| 16 | LF01222 | Auricularia | cf. auricula | 2.60 ± 0.24 | 5.06 ± 1.51 | 0.31 ± 0.34 | 1.90 ± 0.44 |
| 17 | LF01580 | Auricularia | cf. delicata | 2.61 ± 0.12 | 4.19 ± 0.62 | 0.17 ± 0.07 | 0.93 ± 0.04 |
| 18 | LF01616 | Auricularia | cf. polytricha | 2.39 ± 0.68 | 3.29 ± 0.28 | 0.75 ± 0.19 | 3.01 ± 0.22 |
| 19 | MMCR00014 | Auricularia | sp. | 1.82 ± 0.13 | 5.81 ± 0.94 | 0.05 ± 0.01 | 1.54 ± 0.47 |
| 20 | MMCR00107 | Auricularia | sp. | 3.12 ± 0.42 | 7.69 ± 0.86 | 0.20 ± 0.10 | 1.40 ± 0.46 |
| 21 | MMCR00108 | Auricularia | sp. | 2.99 ± 0.02 | 6.07 ± 0.30 | 1.15 ± 0.10 | 2.62 ± 0.19 |
| 22 | MMCR00157.1 | Auricularia | sp. | 4.39 ± 0.39 | 7.27 ± 2.90 | 0.38 ± 0.19 | 0.11 ± 0.03 |
| 23 | MMCR00171 | Auricularia | sp. | 1.46 ± 0.12 | 3.48 ± 0.78 | 0.08 ± 0.02 | 2.29 ± 0.26 |
| 24 | MMCR00177 | Auricularia | sp. | 1.17 ± 0.14 | 5.65 ± 0.95 | 0.33 ± 0.34 | 2.52 ± 0.18 |
| 25 | PHD00142 | Auricularia | sp. | 0.91 ± 0.26 | 1.84 ± 0.48 | 0.46 ± 0.13 | 2.74 ± 0.29 |
| 26 | MMCR00214.2 | Calvatia | sp. | 0.55 ± 0.15 | 1.32 ± 0.28 | 0.85 ± 0.31 | 3.03 ± 0.23 |
| 27 | MMCR00215.1 | Ganoderma | sp. | 1.64 ± 0.17 | 2.64 ± 0.31 | 0.41 ± 0.18 | 2.89 ± 0.60 |
| 28 | MMCR00216.1 | Ganoderma | sp. | 2.37 ± 0.07 | 7.01 ± 1.68 | 1.80 ± 1.13 | 4.21 ± 0.64 |
| 29 | MMCR00271.1 | Pycnoporus | sp. | 2.63 ± 0.09 | 3.35 ± 0.19 | 0.41 ± 0.13 | 2.82 ± 0.15 |
| 30 | MMCR00309.2 | Coprinus | cf. fimetarius | 2.47 ± 0.40 | 11.43 ± 1.13 | 1.15 ± 0.20 | 1.29 ± 0.16 |
| 31 | MMCR00347.1 | Amauroderma | sp. | 1.29 ± 0.10 | 6.44 ± 2.06 | 0.51 ± 0.06 | 0.38 ± 0.11 |
| 32 | PP0005 | Pestalotiopsis | sp. | 4.73 ± 0.34 | 7.19 ± 0.10 | 0.72 ± 0.27 | 6.59 ± 0.98 |
| 33 | PP0013 | Nigrospora | sp. | 3.50 ± 0.30 | 8.62 ± 0.28 | 0.07 ± 0.04 | 2.05 ± 0.05 |
| 34 | PP0049.1 | Mucor-like | sp. | 5.01 ± 0.31 | 5.79 ± 0.69 | 1.08 ± 0.07 | 2.77 ± 0.25 |
| 35 | ENFER0001 | Lasiodiplodia-like | sp. | 3.70 ± 0.32 | 9.76 ± 0.09 | 0.11 ± 0.04 | 1.55 ± 0.21 |
| 36 | ENFER0002 | Phomopsis | sp. | 3.61 ± 1.05 | 4.52 ± 2.49 | 0.70 ± 0.14 | 1.59 ± 0.59 |
| 37 | ENFER0003 | Mucor-like | sp. | 4.07 ± 0.30 | 5.19 ± 0.23 | 0.42 ± 0.10 | 2.37 ± 0.25 |
| 38 | ENFER0004 | Unidentified | Unidentified | 4.28 ± 0.25 | 8.11 ± 0.25 | 0.01 ± 0.00 | 2.45 ± 0.12 |
| 39 | ENFER0005 | Unidentified | Unidentified | 2.91 ± 0.29 | 7.07 ± 0.11 | 1.74 ± 0.11 | 2.52 ± 0.06 |
| 40 | ENFER0006 | Unidentified | Unidentified | 4.20 ± 0.60 | 6.79 ± 0.06 | 0.50 ± 0.23 | 2.23 ± 0.10 |
| 41 | ENFER0007 | Unidentified | Unidentified | 4.27 ± 0.33 | 6.40 ± 0.17 | 0.18 ± 0.08 | 2.53 ± 0.19 |
| 42 | ENFER0008 | Unidentified | Unidentified | 2.57 ± 0.32 | 5.84 ± 0.08 | 0.90 ± 0.29 | 2.77 ± 0.05 |
| 43 | ENFER0009 | Unidentified | Unidentified | 3.37 ± 0.58 | 6.70 ± 0.46 | 1.21 ± 0.07 | 1.51 ± 0.04 |
| 44 | MMCR00035 | Panus | sp. | 4.55 ± 2.64 | 6.54 ± 2.94 | 0.62 ± 0.32 | 1.72 ± 0.42 |
| 45 | MMCR00041 | Panus | sp. | 1.33 ± 0.24 | 3.20 ± 0.71 | 1.03 ± 0.13 | 2.34 ± 0.20 |
| 46 | MMCR00120 | Panus | sp. | 2.29 ± 0.22 | 6.75 ± 0.69 | 0.73 ± 0.06 | 2.14 ± 0.06 |
| 47 | MMCR00199 | Lentinus | cf. polychrous | 2.10 ± 0.73 | 6.63 ± 0.36 | 0.27 ± 0.05 | 2.21 ± 0.19 |
| 48 | MG0001 | Volvariella | volvacea | 1.60 ± 0.29 | 1.17 ± 0.15 | 0.66 ± 0.20 | 2.67 ± 0.68 |
| 49 | MG0002 | Flammulina | velutipes | 2.52 ± 0.41 | 7.38 ± 1.34 | 0.13 ± 0.06 | 1.95 ± 0.13 |
| 50 | MCR366.3 | Hericium | erinaceus | 1.81 ± 0.13 | 3.22 ± 0.16 | 0.78 ± 0.11 | 1.73 ± 0.21 |
| 51 | MCR369.1 | Pleurotus | cf. djamor | 3.41 ± 0.35 | 12.69 ± 0.96 | 0.11 ± 0.02 | 1.68 ± 0.04 |
| 52 | MCR370 | Lentinus | polychrous | 1.92 ± 0.43 | 7.07 ± 3.98 | 0.20 ± 0.13 | 1.71 ± 0.24 |
Table 4.
Cytotoxicity of 100 mg/mL exopolysaccharides produced by 52 fungal strains against HDFn and NCTC clone 929 cells.
Table 4.
Cytotoxicity of 100 mg/mL exopolysaccharides produced by 52 fungal strains against HDFn and NCTC clone 929 cells.
| No | Code | Genus | Epithet | Cytotoxicity (%) | |||
|---|---|---|---|---|---|---|---|
| HDFn | NCTC Clone 929 | ||||||
| PDB | PYGM | PDB | PYGM | ||||
| 1 | LF00466 | Schizophyllum | commune | −13.4 | −3.13 | 2.11 | 8.72 |
| 2 | LF00467 | Schizophyllum | commune | 14.32 | 15.33 | 4.05 | 3.98 |
| 3 | LF00469 | Schizophyllum | commune | 0 | 5.47 | 0 | 1.6 |
| 4 | LF00470 | Schizophyllum | commune | 10.95 | 6.66 | 5.29 | 4.71 |
| 5 | LF00473 | Schizophyllum | commune | 14.65 | 15.84 | 4.51 | 6.89 |
| 6 | LF00534 | Schizophyllum | commune | 0.07 | −3.73 | 8.03 | 1.7 |
| 7 | LF00543 | Schizophyllum | commune | 0 | −0.64 | 0 | 4.69 |
| 8 | LF01001 | Schizophyllum | commune | 7.98 | 8.19 | 5.18 | 9.52 |
| 9 | LF01581 | Schizophyllum | commune | 6.85 | −0.63 | 4.07 | −1.73 |
| 10 | LF01962 | Schizophyllum | commune | 9.6 | 11.32 | 4.38 | 6.72 |
| 11 | MMCR00071 | Schizophyllum | commune | 12.67 | 1.61 | 11.63 | 15.45 |
| 12 | MMCR00176 | Schizophyllum | commune | 0.38 | 4.65 | 5.78 | 16.06 |
| 13 | MMCR00333 | Schizophyllum | commune | 1.55 | 0.97 | 10.51 | 11.65 |
| 14 | MMCR00334 | Schizophyllum | commune | −6.18 | 7.8 | 7.45 | 12.86 |
| 15 | MMCR00336 | Schizophyllum | commune | 12.34 | 4.9 | 4.54 | 8.52 |
| 16 | LF01222 | Auricularia | cf. auricula | −0.21 | 7.69 | 2.14 | 8.34 |
| 17 | LF01580 | Auricularia | cf. delicata | 9.79 | 6.74 | 9.26 | 4.2 |
| 18 | LF01616 | Auricularia | cf. polytricha | 3.57 | 10.56 | 1.42 | 2.39 |
| 19 | MMCR00014 | Auricularia | sp. | 0 | 8.61 | 0 | −4.78 |
| 20 | MMCR00107 | Auricularia | sp. | 9.45 | 6.51 | −4.76 | −4.73 |
| 21 | MMCR00108 | Auricularia | sp. | 7.79 | 4.53 | −0.02 | −4.7 |
| 22 | MMCR00157.1 | Auricularia | sp. | 13.21 | 4.12 | −3.61 | 4.5 |
| 23 | MMCR00171 | Auricularia | sp. | 0 | 3.53 | 0 | 1.59 |
| 24 | MMCR00177 | Auricularia | sp. | −10.59 | −10.38 | 1.45 | 1.15 |
| 25 | PHD00142 | Auricularia | sp. | −21.4 | −21.88 | 5.93 | 7.62 |
| 26 | MMCR00214.2 | Calvatia | sp. | −11.75 | −14.68 | −2.36 | −2 |
| 27 | MMCR00215.1 | Ganoderma | sp. | −10.31 | −15.9 | 3.8 | −0.26 |
| 28 | MMCR00216.1 | Ganoderma | sp. | −5.1 | −15.56 | 4.45 | 0.06 |
| 29 | MMCR00271.1 | Pycnoporus | sp. | −9.97 | −1.49 | 12.42 | 0.47 |
| 30 | MMCR00309.2 | Coprinus | cf. fimetarius | −15.73 | −9.37 | 4.44 | 6.52 |
| 31 | MMCR00347.1 | Amauroderma | sp. | 5.79 | −4.78 | 2.51 | 9.48 |
| 32 | PP0005 | Pestalotiopsis | sp. | −13.99 | −1.82 | −0.78 | 0.01 |
| 33 | PP0013 | Nigrospora | sp. | −12.39 | −7.84 | 6.02 | 3.71 |
| 34 | PP0049.1 | Mucor-like | sp. | −9.86 | −2.17 | −0.12 | 3.98 |
| 35 | ENFER0001 | Lasiodiplodia-like | sp. | −1.35 | −10.39 | 6.02 | 4.21 |
| 36 | ENFER0002 | Phomopsis | sp. | 3.38 | −1.14 | 7.27 | 4.05 |
| 37 | ENFER0003 | Mucor-like | sp. | 10.11 | 8.43 | 5.01 | 10.01 |
| 38 | ENFER0004 | Unidentified | Unidentified | 0 | 12.06 | 0 | 8.76 |
| 39 | ENFER0005 | Unidentified | Unidentified | 8.14 | 11.7 | 5.63 | 1.95 |
| 40 | ENFER0006 | Unidentified | Unidentified | 1.18 | 4.67 | 2.35 | 4.07 |
| 41 | ENFER0007 | Unidentified | Unidentified | −0.13 | 10.35 | −0.12 | 3.41 |
| 42 | ENFER0008 | Unidentified | Unidentified | 8 | 12.12 | 3.78 | 2.02 |
| 43 | ENFER0009 | Unidentified | Unidentified | 5.81 | 8.97 | 4.35 | −2.37 |
| 44 | MMCR00035 | Panus | sp. | 7.58 | 4.22 | −3.39 | −7.34 |
| 45 | MMCR00041 | Panus | sp. | 1.04 | 10.03 | 4.32 | 1.82 |
| 46 | MMCR00120 | Panus | sp. | 14.99 | 11.76 | −3.67 | −8.52 |
| 47 | MMCR00199 | Lentinus | cf. polychrous | 6.52 | 6.24 | −8.33 | 5.48 |
| 48 | MG0001 | Volvariella | volvacea | 4.97 | 5.69 | −3.22 | 5.29 |
| 49 | MG0002 | Flammulina | velutipes | 5.07 | 11.45 | −1.01 | 2.89 |
| 50 | MCR366.3 | Hericium | erinaceus | 8.2 | 13.94 | 2.77 | 7.92 |
| 51 | MCR369.1 | Pleurotus | cf. djamor | 2.66 | 8.17 | 5.1 | 0.07 |
| 52 | MCR370 | Lentinus | polychrous | 1.65 | 1.51 | 2.11 | 3.41 |
Table 5.
Molecular weights of exopolysaccharides produced by 52 fungal strains in different media.
| No | Code | Genus | Epithet | Molecular Weight (kDa) | |||
|---|---|---|---|---|---|---|---|
| PDB | PYGM | ||||||
| 1 | LF00466 | Schizophyllum | commune | - | - | 20.87 | - |
| 2 | LF00467 | Schizophyllum | commune | - | - | - | - |
| 3 | LF00469 | Schizophyllum | commune | - | - | 1383.87 (9) | 37.64 (91) |
| 4 | LF00470 | Schizophyllum | commune | - | - | - | - |
| 5 | LF00473 | Schizophyllum | commune | - | - | - | - |
| 6 | LF00534 | Schizophyllum | commune | - | - | - | - |
| 7 | LF00543 | Schizophyllum | commune | - | - | - | - |
| 8 | LF01001 | Schizophyllum | commune | 59.39 (35) | 2.45 (65) | 260.82 (25) | 15.83 (75) |
| 9 | LF01581 | Schizophyllum | commune | - | - | - | - |
| 10 | LF01962 | Schizophyllum | commune | 2.56 | - | 18.14 | - |
| 11 | MMCR00071 | Schizophyllum | commune | - | - | - | - |
| 12 | MMCR00176 | Schizophyllum | commune | - | - | - | - |
| 13 | MMCR00333 | Schizophyllum | commune | 0.15 | 2.81 | ||
| 14 | MMCR00334 | Schizophyllum | commune | - | - | - | - |
| 15 | MMCR00336 | Schizophyllum | commune | - | - | - | - |
| 16 | LF01222 | Auricularia | cf. auricula | - | - | - | - |
| 17 | LF01580 | Auricularia | cf. delicata | - | - | - | - |
| 18 | LF01616 | Auricularia | cf. polytricha | - | - | 0.88 | - |
| 19 | MMCR00014 | Auricularia | sp. | - | - | - | - |
| 20 | MMCR00107 | Auricularia | sp. | - | - | - | - |
| 21 | MMCR00108 | Auricularia | sp. | 5.08 | 603.50 (45) | 15.43 (55) | |
| 22 | MMCR00157.1 | Auricularia | sp. | - | - | - | - |
| 23 | MMCR00171 | Auricularia | sp. | - | - | 229.5 | - |
| 24 | MMCR00177 | Auricularia | sp. | - | - | 220.73 (23) | 30.55 (87) |
| 25 | PHD00142 | Auricularia | sp. | - | - | 298.07 | - |
| 26 | MMCR00214.2 | Calvatia | sp. | - | - | 307.76 | 4.73 |
| 27 | MMCR00215.1 | Ganoderma | sp. | - | - | 1234.73 (41) | 19.17 (59) |
| 28 | MMCR00216.1 | Ganoderma | sp. | 2.15 | - | 4.23 (24) | 0.82 (76) |
| 29 | MMCR00271.1 | Pycnoporus | sp. | - | - | 3.39 (64) | 0.72 (46) |
| 30 | MMCR00309.2 | Coprinus | cf. fimetarius | 2.61 | - | - | - |
| 31 | MMCR00347.1 | Amauroderma | sp. | - | - | - | - |
| 32 | PP0005 | Pestalotiopsis | sp. | - | - | 494.5 | - |
| 33 | PP0013 | Nigrospora | sp. | 494.5 | - | 317.76 | - |
| 34 | PP0049.1 | Mucor-like | sp. | - | - | - | - |
| 35 | ENFER0001 | Lasiodiplodia-like | sp. | - | - | - | - |
| 36 | ENFER0002 | Phomopsis | sp. | - | - | - | - |
| 37 | ENFER0003 | Mucor-like | sp. | - | - | 75.44 | - |
| 38 | ENFER0004 | Unidentified | Unidentified | - | - | 216.18 | - |
| 39 | ENFER0005 | Unidentified | Unidentified | 3.98 | - | 458.08 (28) | 273.45 (72) |
| 40 | ENFER0006 | Unidentified | Unidentified | - | - | 175.96 | - |
| 41 | ENFER0007 | Unidentified | Unidentified | - | - | 144.23 | - |
| 42 | ENFER0008 | Unidentified | Unidentified | - | - | 281.55 | - |
| 43 | ENFER0009 | Unidentified | Unidentified | 2.87 | - | - | - |
| 44 | MMCR00035 | Panus | sp. | - | - | - | - |
| 45 | MMCR00041 | Panus | sp. | 4.21 | 503.52 (61) | 335.94 (49) | |
| 46 | MMCR00120 | Panus | sp. | - | - | - | - |
| 47 | MMCR00199 | Lentinus | cf. polychrous | - | - | 1214.29 | - |
| 48 | MG0001 | Volvariella | volvacea | - | - | 234.66 (24) | 2.09 (76) |
| 49 | MG0002 | Flammulina | velutipes | - | - | - | - |
| 50 | MCR366.3 | Hericium | erinaceus | - | - | - | - |
| 51 | MCR369.1 | Pleurotus | cf. djamor | - | - | - | - |
| 52 | MCR370 | Lentinus | polychrous | - | - | - | - |
Remarks: Numbers in brackets are the percentage of molecular weight distribution.
Table 6.
Exopolysaccharide and mycelium dried weight production by S. commune LF01962 on different carbon and nitrogen sources.
Table 6.
Exopolysaccharide and mycelium dried weight production by S. commune LF01962 on different carbon and nitrogen sources.
| Treatments | Exopolysaccharide Weight (g/L) | Mycelium Weight (g/L) | |
|---|---|---|---|
| Carbon Source | Nitrogen Source | ||
| xylose | malt extract | 0.70 ± 0.00 | 5.10 ± 0.10 |
| yeast extract | 3.50 ± 0.70 | 14.20 ± 0.40 | |
| KNO3 | 1.00 ± 0.20 | 4.00 ± 0.50 | |
| casein hydrolysate | 3.00 ± 1.00 | 13.40 ± 1.10 | |
| peptone | 4.80 ± 0.80 | 12.00 ± 0.80 | |
| (NH4)2HPO4 | 1.40 ± 0.60 | 4.8 ± 0.50 | |
| (NH4)2SO4 | 1.20 ± 0.10 | 4.00 ± 0.40 | |
| NH4H2PO4 | 0.60 ± 0.40 | 3.5 ± 0.40 | |
| glucose | malt extract | 0.90 ± 0.00 | 4.40 ± 0.20 |
| yeast extract | 3.50 ± 0.00 | 12.10 ± 1.20 | |
| KNO3 | 1.3 ± 0.10 | 4.90 ± 0.90 | |
| casein hydrolysate | 3.00 ± 0.50 | 11.80 ± 1.30 | |
| peptone | 2.30 ± 0.20 | 9.60 ± 0.50 | |
| (NH4)2HPO4 | 4.00 ± 0.70 | 9.00 ± 1.00 | |
| (NH4)2SO4 | 1.00 ± 0.20 | 6.30 ± 1.00 | |
| NH4H2PO4 | 0.50 ± 0.00 | 4.30 ± 0.30 | |
| fructose | malt extract | 0.80 ± 0.00 | 4.60 ± 0.50 |
| yeast extract | 3.90 ± 0.8 | 14.50 ± 0.40 | |
| KNO3 | 1.30 ± 1.00 | 4.50 ± 0.50 | |
| casein hydrolysate | 3.20 ± 0.50 | 15.20 ± 2.00 | |
| peptone | 4.10 ± 0.60 | 14.20 ± 0.80 | |
| (NH4)2HPO4 | 1.80 ± 0.20 | 8.60 ± 0.20 | |
| (NH4)2SO4 | 1.20 ± 0.10 | 4.20 ± 0.10 | |
| NH4H2PO4 | 0.40 ± 0.10 | 4.00 ± 0.30 | |
| sucrose | malt extract | 1.40 ± 0.20 | 4.10 ± 0.30 |
| yeast extract | 3.10 ± 0.30 | 14.50 ± 1.00 | |
| KNO3 | 2.40 ± 1.2 | 4.70 ± 0.30 | |
| casein hydrolysate | 2.70 ± 0.20 | 15.20 ± 0.50 | |
| peptone | 3.90 ± 0.70 | 14.30 ± 0.40 | |
| (NH4)2HPO4 | 2.60 ± 0.20 | 10.30 ± 0.30 | |
| (NH4)2SO4 | 1.00 ± 0.00 | 5.80 ± 0.10 | |
| NH4H2PO4 | 0.50 ± 0.10 | 3.70 ± 0.30 | |
| maltose | malt extract | 1.00 ± 0.10 | 4.00 ± 0.40 |
| yeast extract | 3.80 ± 0.20 | 14.20 ± 0.60 | |
| KNO3 | 1.20 ± 0.10 | 3.60 ± 0.40 | |
| casein hydrolysate | 3.60 ± 0.30 | 13.40 ± 0.90 | |
| peptone | 3.30 ± 0.50 | 13.30 ± 0.50 | |
| (NH4)2HPO4 | 3.00 ± 0.30 | 9.80 ± 0.50 | |
| (NH4)2SO4 | 1.10 ± 0.00 | 5.10 ± 0.30 | |
| NH4H2PO4 | 0.50 ± 0.50 | 3.90 ± 0.20 | |
| mannose | malt extract | 0.70 ± 0.10 | 4.60 ± 0.50 |
| yeast extract | 5.10 ± 2.00 | 14.30 ± 0.60 | |
| KNO3 | 2.50 ± 0.50 | 4.70 ± 0.40 | |
| casein hydrolysate | 3.10 ± 0.60 | 13.80 ± 0.30 | |
| peptone | 2.70 ± 0.70 | 13.00 ± 0.50 | |
| (NH4)2HPO4 | 2.50 ± 0.20 | 9.70 ± 1.10 | |
| (NH4)2SO4 | 1.30 ± 0.30 | 4.40 ± 0.60 | |
| NH4H2PO4 | 0.60 ± 0.10 | 4.40 ± 0.20 | |
| PDB | 1.40 ± 1.20 | 11.40 ± 1.10 | |
| PYGM | 3.80 ± 0.30 | 8.80 ± 0.00 | |
Table 7.
Exopolysaccharide and mycelium production by S. commune LF01962 using Plackett–Burman design.
Table 7.
Exopolysaccharide and mycelium production by S. commune LF01962 using Plackett–Burman design.
| Treatment | Exopolysaccharide Weight (g/L) | Mycelium Dried Weight (g/L) |
|---|---|---|
| 1 | 7.22 ± 0.18 | 22.00 ± 2.24 |
| 2 | 3.43 ± 0.99 | 22.51 ± 4.33 |
| 3 | 0.96 ± 0.55 | 4.80 ± 2.01 |
| 4 | 1.92 ± 0.8 | 5.97 ± 1.73 |
| 5 | 4.87 ± 0.73 | 18.22 ± 1.69 |
| 6 | 1.93 ± 0.98 | 11.81 ± 2.34 |
| 7 | 5.17 ± 0.4 | 24.58 ± 1.12 |
| 8 | 5.07 ± 0.92 | 21.58 ± 1.54 |
| 9 | 5.26 ± 1.62 | 25.13 ± 0.33 |
| 10 | 0.68 ± 0.29 | 5.11 ± 1.01 |
| 11 | 8.16 ± 1.02 | 22.86 ± 1.01 |
| 12 | 1.52 ± 0.71 | 10.72 ± 2.01 |
| 13 | 5.73 ± 0.43 | 20.48 ± 1.55 |
| 14 | 0.8 ± 0.34 | 10.96 ± 0.80 |
| 15 | 1.26 ± 0.25 | 7.02 ± 3.35 |
| 16 | 5.65 ± 0.27 | 16.24 ± 0.99 |
| 17 | 4.02 ± 0.18 | 19.09 ± 0.71 |
| 18 | 2.51 ± 0.72 | 14.07 ± 0.66 |
| 19 | 4.72 ± 0.59 | 26.39 ± 0.69 |
| 20 | 3.78 ± 1.94 | 12.63 ± 2.5 |
| 21 | 2.81 ± 1.86 | 10.34 ± 3.50 |
| 22 | 0.95 ± 0.37 | 5.87 ± 0.81 |
| 23 | 3.23 ± 1.48 | 22.98 ± 1.50 |
| 24 | 2.84 ± 1.06 | 16.80 ± 1.92 |
| 25 | 1.16 ± 0.55 | 3.77 ± 0.31 |
| 26 | 4.53 ± 2.51 | 18.40 ± 2.51 |
Table 8.
ANOVA of factorial model for exopolysaccharide production by S. commune LF01962.
| Source | Sum of Squares | df | Mean Square | F-Value | p-Value | |
|---|---|---|---|---|---|---|
| Model | 303.15 | 23 | 13.18 | 12.27 | <0.0001 | significant |
| A—Glucose | 38.54 | 1 | 38.54 | 35.87 | <0.0001 | |
| B—Xylose | 9.14 | 1 | 9.14 | 8.51 | 0.0052 | |
| C—Mannose | 0.2367 | 1 | 0.2367 | 0.2203 | 0.6407 | |
| D—Diammonium hydrogen phosphate | 41.34 | 1 | 41.34 | 38.49 | <0.0001 | |
| E—Yeast extract | 60.59 | 1 | 60.59 | 56.40 | <0.0001 | |
| F—Casein hydrolysate | 11.31 | 1 | 11.31 | 10.53 | 0.0020 | |
| G—Peptone | 0.7248 | 1 | 0.7248 | 0.6747 | 0.4151 | |
| H—Manganese sulfate | 2.76 | 1 | 2.76 | 2.57 | 0.1152 | |
| J—Monosodium glutamate | 21.94 | 1 | 21.94 | 20.42 | <0.0001 | |
| K—Trace elements | 20.93 | 1 | 20.93 | 19.49 | <0.0001 | |
| L—Vitamin solution | 11.15 | 1 | 11.15 | 10.38 | 0.0022 | |
| Curvature | 17.95 | 1 | 17.95 | 16.71 | 0.0001 | |
| Pure Error | 56.94 | 53 | 1.07 | |||
| Cor Total | 378.03 | 77 |
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MDPI and ACS Style
Prathumpai, W.; Pinruan, U.; Sommai, S.; Komwijit, S.; Malairuang, K. Exopolysaccharide (EPS) Production by Endophytic and Basidiomycete Fungi. Fermentation 2025, 11, 183. https://doi.org/10.3390/fermentation11040183
AMA Style
Prathumpai W, Pinruan U, Sommai S, Komwijit S, Malairuang K. Exopolysaccharide (EPS) Production by Endophytic and Basidiomycete Fungi. Fermentation. 2025; 11(4):183. https://doi.org/10.3390/fermentation11040183
Chicago/Turabian StylePrathumpai, Wai, Umpawa Pinruan, Sujinda Sommai, Somjit Komwijit, and Kwanruthai Malairuang. 2025. "Exopolysaccharide (EPS) Production by Endophytic and Basidiomycete Fungi" Fermentation 11, no. 4: 183. https://doi.org/10.3390/fermentation11040183
APA StylePrathumpai, W., Pinruan, U., Sommai, S., Komwijit, S., & Malairuang, K. (2025). Exopolysaccharide (EPS) Production by Endophytic and Basidiomycete Fungi. Fermentation, 11(4), 183. https://doi.org/10.3390/fermentation11040183
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