Pellet diameter of <i>Ganoderma lucidum</i> in a repeated-batch fermentation for the trio total production of biomass-exopolysaccharide-endopolysaccharide and its anti-oral cancer beta-glucan response

Nur Raihan Abdullah; Faez Sharif; Nur Hafizah Azizan; Ismail Fitri Mohd Hafidz; Sugenendran Supramani; Siti Rokhiyah Ahmad Usuldin; Rahayu Ahmad; Wan Abd Al Qadr Imad Wan-Mohtar; Nur Raihan Abdullah; Faez Sharif; Nur Hafizah Azizan; Ismail Fitri Mohd Hafidz; Sugenendran Supramani; Siti Rokhiyah Ahmad Usuldin; Rahayu Ahmad; Wan Abd Al Qadr Imad Wan-Mohtar

doi:10.3934/microbiol.2020023

AIMS Microbiology

2020, Volume 6, Issue 4: 379-400. doi: 10.3934/microbiol.2020023

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Research article

Pellet diameter of Ganoderma lucidum in a repeated-batch fermentation for the trio total production of biomass-exopolysaccharide-endopolysaccharide and its anti-oral cancer beta-glucan response

Nur Raihan Abdullah ^1,2,
Faez Sharif ²,
Nur Hafizah Azizan ²,
Ismail Fitri Mohd Hafidz ²,
Sugenendran Supramani ¹,
Siti Rokhiyah Ahmad Usuldin ³,
Rahayu Ahmad ⁴,
Wan Abd Al Qadr Imad Wan-Mohtar ^{1,5
,
,}

1.
Functional Omics and Bioprocess Development Laboratory, Institute of Biological Sciences, Faculty of Science, Universiti of Malaya, 50603, Kuala Lumpur, Malaysia
2.
Department of Biotechnology, Kulliyyah of Science, International Islamic University Malaysia, 25200 Kuantan, Pahang, Malaysia
3.
Agro-Biotechnology Institute, Malaysia (ABI), National Institutes Biotechnology Malaysia (NIBM), C/O HQ MARDI, 43400, Serdang, Selangor, Malaysia
4.
Halal Action Laboratory, Kolej Genius Insan, University Sains Islam Malaysia, Bandar Baru Nilai, 71800, Nilai, Negeri Sembilan, Malaysia
5.
Bioscience Research Institute, Athlone Institute of Technology, Ireland

Received: 12 June 2020 Accepted: 19 October 2020 Published: 22 October 2020

The pellet morphology and diameter range (DR) of Ganoderma lucidum were observed in a repeated-batch fermentation (RBF) for the trio total production of biomass, exopolysaccharide (EPS) and endopolysaccharide (ENS). Two factors were involved in RBF; broth replacement ratio (BRR: 60%, 75% and 90%) and broth replacement time point (BRTP: log, transition and stationary phase) in days. In RBF, 34.31 g/L of biomass favoured small-compact pellets with DR of 20.67 µm< d < 24.00 µm (75% BRR, day 11 of BRTP). EPS production of 4.34 g/L was prone to ovoid-starburst pellets with DR of 34.33 µm< d <35.67 µm (75% BRR, day 13 of BRTP). Meanwhile, the highest 2.43 g/L of ENS production favoured large-hollow pellets with DR of 34.00 µm< d < 38.67 µm (90% BRR, day 13 of BRTP). In addition, RBF successfully shortened the biomass-EPS–ENS fermentation period (31, 33 and 35 days) from batch to 5 days, in seven consecutive cycles of RBF. In a FTIR detection, β-glucan (BG) from EPS and ENS extracts were associated with β-glycosidic linkages (2925 cm⁻¹, 1635 cm⁻¹, 1077 cm⁻¹, 920 cm⁻¹ and 800 cm⁻¹ wavelengths) with similar ¹H NMR spectral behaviour (4.58, 3.87 and 3.81 ppm). Meanwhile, 4 mg/L of BG gave negative cytotoxic effects on normal gingival cell line (hGF) but induced antiproliferation (IC₅₀ = 0.23 mg/mL) against cancerous oral Asian cellosaurus cell line (ORL-48). Together, this study proved that G. lucidum mycelial pellets could withstand seven cycles of long fermentation condition and possessed anti-oral cancer beta-glucan, which suits large-scale natural drug fermentation.
- Ganoderma lucidum,
- pellet morphology,
- repeated-batch fermentation,
- exopolysaccharide,
- endopolysaccharide,
- anti-oral cancer
Citation: Nur Raihan Abdullah, Faez Sharif, Nur Hafizah Azizan, Ismail Fitri Mohd Hafidz, Sugenendran Supramani, Siti Rokhiyah Ahmad Usuldin, Rahayu Ahmad, Wan Abd Al Qadr Imad Wan-Mohtar. Pellet diameter of Ganoderma lucidum in a repeated-batch fermentation for the trio total production of biomass-exopolysaccharide-endopolysaccharide and its anti-oral cancer beta-glucan response[J]. AIMS Microbiology, 2020, 6(4): 379-400. doi: 10.3934/microbiol.2020023

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Abstract

The pellet morphology and diameter range (DR) of Ganoderma lucidum were observed in a repeated-batch fermentation (RBF) for the trio total production of biomass, exopolysaccharide (EPS) and endopolysaccharide (ENS). Two factors were involved in RBF; broth replacement ratio (BRR: 60%, 75% and 90%) and broth replacement time point (BRTP: log, transition and stationary phase) in days. In RBF, 34.31 g/L of biomass favoured small-compact pellets with DR of 20.67 µm< d < 24.00 µm (75% BRR, day 11 of BRTP). EPS production of 4.34 g/L was prone to ovoid-starburst pellets with DR of 34.33 µm< d <35.67 µm (75% BRR, day 13 of BRTP). Meanwhile, the highest 2.43 g/L of ENS production favoured large-hollow pellets with DR of 34.00 µm< d < 38.67 µm (90% BRR, day 13 of BRTP). In addition, RBF successfully shortened the biomass-EPS–ENS fermentation period (31, 33 and 35 days) from batch to 5 days, in seven consecutive cycles of RBF. In a FTIR detection, β-glucan (BG) from EPS and ENS extracts were associated with β-glycosidic linkages (2925 cm⁻¹, 1635 cm⁻¹, 1077 cm⁻¹, 920 cm⁻¹ and 800 cm⁻¹ wavelengths) with similar ¹H NMR spectral behaviour (4.58, 3.87 and 3.81 ppm). Meanwhile, 4 mg/L of BG gave negative cytotoxic effects on normal gingival cell line (hGF) but induced antiproliferation (IC₅₀ = 0.23 mg/mL) against cancerous oral Asian cellosaurus cell line (ORL-48). Together, this study proved that G. lucidum mycelial pellets could withstand seven cycles of long fermentation condition and possessed anti-oral cancer beta-glucan, which suits large-scale natural drug fermentation.

Abbreviation EPS: Exopolysaccharide; ENS: Endopolysaccharide; GL: ; g: Grams; L: Litre; mL: Millilitre; v/v: Volume per volume; PDA: Potato dextrose agar; RBF: Repeated batch fermentation; RPM: Revolutions per minute; µm: Micrometres; SSF: Solid state fermentation; SLF: Submerged liquid fermentation; BRTP: Broth replacement time point; BRR: Broth replacement ratio;
Acknowledgments

This work was supported by the Ministry of Higher Education Malaysia under the Southeast and South Asia and Taiwan Universities [SATU Joint Research Scheme] [RU Geran ST002-2020: Dr Wan-Mohtar] and Fundamental Research Grant Scheme [FRGS 19-140-0749: Dr Mohd Faez Sharif].

Compliance with ethical standards

The written article complies with ethical standards.

Conflict of interest

There is no conflict of interest for this journal article.

References

[1]	Vunduk J, Wan-Mohtar WAAQI, Mohamad SA, et al. (2019) Polysaccharides of Pleurotus flabellatus strain Mynuk produced by submerged fermentation as a promising novel tool against adhesion and biofilm formation of foodborne pathogens. LWT Food Sci Technol 112. doi: 10.1016/j.lwt.2019.05.119
[2]	Ahmad Usuldin SR, Mahmud N, Ilham Z, et al. (2020) In-depth spectral characterization of antioxidative (1,3)-β-D-glucan from the mycelium of an identified tiger milk mushroom Lignosus rhinocerus strain ABI in a stirred-tank bioreactor. Biocatal Agric Biotechnol 23. doi: 10.1016/j.bcab.2019.101455
[3]	Hassan NA, Supramani S, Azzimi Sohedein MN, et al. (2019) Efficient biomass-exopolysaccharide production from an identified wild-Serbian Ganoderma lucidum strain BGF4A1 mycelium in a controlled submerged fermentation. Biocatal Agric Biotechnol 21. doi: 10.1016/j.bcab.2019.101305
[4]	Supramani S, Jailani N, Ramarao K, et al. (2019a) Pellet diameter and morphology of European Ganoderma pfeifferi in a repeated-batch fermentation for exopolysaccharide production. Biocatal Agric Biotechnol 19. doi: 10.1016/j.bcab.2019.101118
[5]	Wan-Mohtar WAAQI, Ab Kadir S, Saari N (2016) The morphology of Ganoderma lucidum mycelium in a repeated-batch fermentation for exopolysaccharide production. Biotechnol Reports 11: 2-11. doi: 10.1016/j.btre.2016.05.005
[6]	Kozarski M, Klaus A, Jakovljević D, et al. (2019) Ganoderma lucidum as a cosmeceutical: Antiradical potential and inhibitory effect on hyperpigmentation and skin extracellular matrix degradation enzymes. Arch Biol Sci 71: 253-264. doi: 10.2298/ABS181217007K
[7]	Ziegenbein FC, Hanssen HP, König WA (2006) Secondary metabolites from Ganoderma lucidum and Spongiporus leucomallellus. Phytochemistry 67: 202-211. doi: 10.1016/j.phytochem.2005.10.025
[8]	Zhong JJ, Xiao JH (2009) Secondary metabolites from higher fungi: discovery, bioactivity, and bioproduction. Adv Biochem Eng Biotechnol 113: 79-150.
[9]	Liu J, Yang F, Ye LB, et al. (2004) Possible mode of action of antiherpetic activities of a proteoglycan isolated from the mycelia of Ganoderma lucidum in vitro. J Ethnopharmacol 95: 265-272. doi: 10.1016/j.jep.2004.07.010
[10]	Wan-Mohtar WAAQI, Young L, Abbott GM, et al. (2016) Antimicrobial properties and cytotoxicity of sulfated (1,3)-β-D-glucan from the mycelium of the mushroom Ganoderma lucidum. J Microbiol Biotechnol 26: 999-1010. doi: 10.4014/jmb.1510.10018
[11]	Wachtel-Galor S, Tomlinson B, Benzie IFF (2004) Ganoderma lucidum (‘Lingzhi’), a Chinese medicinal mushroom: biomarker responses in a controlled human supplementation study. Br J Nutr 91: 263-269. doi: 10.1079/BJN20041039
[12]	Chen TW, Wong YK, Lee SS (1991) In vitro cytotoxicity of Ganoderma lucidum on oral cancer cells. Zhonghua Yi Xue Za Zhi (Taipei) 48: 54-58.
[13]	Wan Mohtar WAAQI, Latif NA, Harvey LM, et al. (2016) Production of exopolysaccharide by Ganoderma lucidum in a repeated-batch fermentation. Biocatal Agric Biotechnol 6: 91-101. doi: 10.1016/j.bcab.2016.02.011
[14]	Supramani S, Ahmad R, Ilham Z, et al. (2019b) Optimisation of biomass, exopolysaccharide and intracellular polysaccharide production from the mycelium of an identified Ganoderma lucidum strain QRS 5120 using response surface methodology. AIMS Microbiol 5: 19-38. doi: 10.3934/microbiol.2019.1.19
[15]	Ubaidillah NHN, Abdullah N, Sabaratnama V (2015) Isolation of the intracellular and extracellular polysaccharides of Ganoderma neojaponicum (Imazeki) and characterization of their immunomodulatory properties. Electron J Biotechnol 18: 188-195. doi: 10.1016/j.ejbt.2015.03.006
[16]	Stanbury PF, Whitaker A, Hall SJ (2017) Principles of Fermentation Technology, Elsevier.
[17]	Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31: 426-428. doi: 10.1021/ac60147a030
[18]	Diamantopoulou P, Papanikolaou S, Kapoti M, et al. (2012) Mushroom polysaccharides and lipids synthesized in liquid agitated and static cultures. Part I: Screening various mushroom species. Appl Biochem Biotechnol 167: 536-551. doi: 10.1007/s12010-012-9713-9
[19]	Diamantopoulou P, Papanikolaou S, Komaitis M, et al. (2014) Patterns of major metabolites biosynthesis by different mushroom fungi grown on glucose-based submerged cultures. Bioprocess Biosyst Eng 37: 1385-1400. doi: 10.1007/s00449-013-1112-2
[20]	Athenaki M, Gardeli C, Diamantopoulou P, et al. (2018) Lipids from yeasts and fungi: physiology, production and analytical considerations. J Appl Microbiol 124: 336-367. doi: 10.1111/jam.13633
[21]	Fazenda ML, Harvey LM, McNeil B (2010) Effects of dissolved oxygen on fungal morphology and process rheology during fed-batch processing of Ganoderma lucidum. J Microbiol Biotechnol 20: 844-851. doi: 10.4014/jmb.0911.11020
[22]	Ding Zhongyang, Wang Qiong, Peng Lin, et al. (2012) Relationship between mycelium morphology and extracellular polysaccharide production of medicinal mushroom Ganoderma lucidum in submerged culture. J Med Plants Res 6: 2868-2874.
[23]	Berovič M, Popovic M (2018) Submerged cultivation of Ganoderma lucidum biomass in stirred tank reactor. Chem Biochem Eng Q 32: 465-472. doi: 10.15255/CABEQ.2018.1371
[24]	Fang QH, Zhong JJ (2002) Effect of initial pH on production of ganoderic acid and polysaccharide by submerged fermentation of Ganoderma lucidum. Process Biochem 37: 769-774. doi: 10.1016/S0032-9592(01)00278-3
[25]	Synytsya A, Novak M (2014) Structural analysis of glucans. Ann Transl Med 2: 1-14.
[26]	Wang J, Zhang L, Yu Y, et al. (2009) Enhancement of antitumor activities in sulfated and carboxymethylated polysaccharides of Ganoderma lucidum. J Agric Food Chem 57: 10565-10572. doi: 10.1021/jf902597w
[27]	Paulo EM, Boffo EF, Branco A, et al. (2012) Production, extraction and characterization of exopolysaccharides produced by the native Leuconostoc pseudomesenteroides R2 strain. An Acad Bras Cienc 84: 495-507. doi: 10.1590/S0001-37652012000200018
[28]	Miao M, Li R, Jiang B, et al. (2014) Structure and digestibility of endosperm water-soluble a-glucans from different sugary maize mutants. Food Chem 143: 156-162. doi: 10.1016/j.foodchem.2013.07.109
[29]	Osińska-Jaroszuk M, Jaszek M, Mizerska-Dudka M, et al. (2014) Exopolysaccharide from Ganoderma applanatum as a promising bioactive compound with cytostatic and antibacterial properties. Biomed Res Int 2014: 743812-743812. doi: 10.1155/2014/743812
[30]	Gurst JE (1991) NMR and the structure of d-glucose. J Chem Educ 68: 1003-1004. doi: 10.1021/ed068p1003
[31]	Zeng Z, Xiao K (2020) Ganoderma lucidum Polysaccharide (GLP) Inhibited the progression of oral squamous cell carcinoma via the miR-188/BCL9/ β-catenin pathway. Adv Polym Technol 2020: 1-6.
[32]	Taufek NM, Harith HH, Hafiz M, et al. (2020) Performance of mycelial biomass and exopolysaccharide from Malaysian Ganoderma lucidum for the fungivore red hybrid Tilapia ( Oreochromis sp.) in Zebrafish embryo. Aquac Reports 17: 100322. doi: 10.1016/j.aqrep.2020.100322

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