Maneeruttanarungroj, Cherdsak

Preferred name

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School of Science

cherdsak.ma@kmitl.ac.th

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Ethanol-enhanced biohydrogen production and metabolomic response in the green microalga Micractinium sp. KLSc62
(2025-02-17)
Klinsalee, Rachaneekorn
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Laokua, Natwikar
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Rittiyan, Nutnicha
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Kornrawudaphikasama, Yosita
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Tonawut, Yothawut
Hydrogen (H<inf>2</inf>) is recognized as a viable clean energy option due to its high energy density and minimal environmental impact. Green microalgae have garnered attention as a potential source of hydrogen production because of their ability to produce photosynthetic hydrogen and their environmental benefits. In a recent study, thirteen species of microalgae were screened for their growth potential and hydrogen production when supplemented with ethanol. Among the species evaluated, Micractinium sp. KLSc62 (initially named as G4) yielded the highest hydrogen production rate of 6090.5 ± 28.6 μmol mg⁻1 Chl with the supplementation of 30 mM ethanol over a 7-day period. Further analysis revealed that ethanol was utilized during H<inf>2</inf> production process, leading to the formation of acetaldehyde as an intermediate before acetate was produced. Metabolic profiling identified 44 metabolites that were up- or down-regulated, categorized into four distinct groups. These findings highlight the potential of Micractinium sp. KLSc62 as an alternative energy producer and a promising candidate for future metabolite production in various chemical applications, emphasizing its utility as a cellular factory.
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Freshwater green alga Chlorella sp. KLSc59 produced all forms of omega-3 oil: ALA, EPA, and DHA
(2024-06-01)
Preechaphonkul, Nathanan
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Sirikwanpong, Sukrit
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Maneeruttanarungroj, Cherdsak
Omega-3 fats are known to be the essential nutrient for human health, obtainable only through food sources. Algae and phytoplankton are primary sources that can synthesize various useful metabolites, such as carotenoids, antioxidants, and omega-3 oil. As the trend toward healthy alternative food options grows, this study aimed to screen eight strains of freshwater microalgae that might be promising sources of omega-3 oil. The results showed that three algal strains-BNG2, Chlorella sp. KLSc59, and Chlorella sp. KLSc61-had the ability to produce all forms of omega-3 oil, including alpha-linolenic acid (ALA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA). Among these strains, Chlorella sp. KLSc59 exhibited the most consistent omega-3 fatty acids-production when detected with gas chromatography equipped with flame ionization detector. Under optimal conditions (30 % flask volume, 10 mM CaCl<inf>2</inf>, pH of 6.0, and light incubation at 162 μmol photons m−2 s−1) for 5 days, Chlorella sp. KLSc59 produced the highest amount of omega-3 fat, yielding 21.74 ± 0.63 % of the total peak area (equivalent to 18.43 ± 0.6 μg/mg DCW), including 21.53 ± 0.66 % (equivalent to 18.26 ± 0.7 μg/mg DCW) ALA, 0.03 ± 0.2 % (equivalent to 0.02 ± 0.0 μg/mg DCW) EPA, and 0.18 ± 0.01 % (equivalent to 0.15 ± 0.0 μg/mg DCW) DHA, with an omega-6 to omega-3 ratio of 2.28. This study highlights the potential use of Chlorella sp. KLSc59 for omega-3 oil production and its role as an alternative source of omega-3 oil for the next generation.
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Bioprospecting of cyanobacteria from Thai karstic caves as potential producers of phenolic compounds with antioxidant capacity
(2026-04-01)
Suphan, Sutthawan
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Kula, Kasinee
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Lomthong, Thanasak
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Sujarit, Kanaporn
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Maneeruttanarungroj, Cherdsak
Cyanobacteria are highly adaptable microorganisms that can survive in extreme environments. Survival of cyanobacteria in assorted habitats has positively impelled them to produce a range of bioactive compounds, which show a variety of beneficial biological activities. The karstic caves are harsh environments with low light and nutrient input and serve as a unique habitat for exploring cyanobacterial biodiversity as producers of natural products of economic and ecological importance. A total of 86 cyanobacterial strains isolated from 23 karstic caves in four regions of Thailand showed photosynthetic pigments correlated with growth performance and exhibited potential for producing phenolic compounds with significant antioxidant capacity. Based on DPPH scavenging efficiency per unit concentration of crude extract, Leptolyngbya sp. LKK14, Nostoc sp. SW02, and Leptolyngbya sp. LP01 exhibited the superior antioxidant potency. The highest contents of total phenolics (42.10 mgGAE/gCE) and flavonoids (395.90 mgQE/gCE) were found in Leptolyngbya sp. LP01 extract. The presence of phenolic compounds correlated with antioxidant activity; Leptolyngbya sp. LP01 showed the highest antioxidant activity, as measured by DPPH, ABTS, and FRAP assays, with respective values of 15.79, 20.87, and 42.69 mgAAE/gCE. Nine phenolic compounds were tentatively identified based on MS/MS fragmentation patterns compared with library data by LC-ESI-QTOF-MS/MS approach, suggesting that the extract of Leptolyngbya sp. LP01 contains valuable biological antioxidants. This study contributes to our current understanding of cyanobacterial biodiversity in poorly studied habitats, particularly in terms of their biotechnological potential as producers of secondary metabolites suitable for application in the food, pharmaceutical, and cosmetic industries.
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Discovery and characterization of bioactive compounds from Limnophila aromatica: nevadensin and related flavonoids as potent antimicrobial agents
(2025-08-01)
Maswanna, Thanaporn
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Maneeruttanarungroj, Cherdsak
Limnophila aromatica, a traditional medicinal plant, has been previously reported to possess notable antimicrobial properties. However, the specific bioactive constituents responsible for this activity remain largely unidentified. This study aimed to isolate, identify, and evaluate the antibacterial potential of compounds from a 100% ethanolic extract of L. aromatica. The crude ethanolic extract exhibited the highest antibacterial activity against Methicillin-resistant Staphylococcus aureus (MRSA) and was subsequently subjected to Diaion HP-20 column chromatography, followed by preparative HPLC. Seven major peaks were identified using UV-Vis spectra, LC-QTOF-MS, and NMR analyses. The antibacterial efficacy of these isolated compounds was assessed using disc diffusion and broth microdilution assays against Bacillus subtilis, S. aureus, MRSA, Escherichia coli, and Pseudomonas aeruginosa. The isolated compounds were identified as norethindrone acetate, isothymusin, nevadensin, gardenin B, 5,3’-dihydroxy-7,8,2’-trimethoxyisoflavone, jasmolin II, and oleanolic acid. Nevadensin, the predominant compound (79.55%), demonstrated potent bactericidal activity against B. subtilis, S. aureus, and MRSA. Jasmolin II and gardenin B also exhibited promising antibacterial effects. Although the disc diffusion assay was limited by compound diffusion, the broth microdilution method confirmed significant MIC values, ranging from 0.59 to 2.86 mg/ml while the MBC values ranged from 0.59 to 12.72 mg/ml. This study highlights nevadensin and other flavonoids from L. aromatica as potential candidates for the development of alternative antibacterial therapies, particularly against drug-resistant Gram-positive bacteria.
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In silico encrypted peptide from green alga Tetraspora sp. CU2551 showed high antimicrobial activities
(2024-03-01)
Tonawut, Yothawut
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Rittiyan, Nutnicha
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Kornrawudaphikasama, Yosita
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Klinsalee, Rachaneekorn
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Laokua, Natwikar
The green alga Tetraspora sp. CU2551 has been previously identified as a biohydrogen producer for an alternative option for sustainable energy production development. Its biomass still contains a source of valuable biomolecules, especially proteins. Crude algal protein showed significant antimicrobial activity against Gram-positive bacteria (Bacillus subtilis TISTR 1248 and Staphylococcus aureus TISTR 746) and Gram-negative bacteria (Escherichia coli TISTR 074 and Pseudomonas aeruginosa TISTR 2370), representing a clear zone of 19.0 ± 0.1, 18.3 ± 0.5, 17.7 ± 0.5, and 19.0 ± 0.5, respectively. Further investigation revealed that encrypted peptides from pepsin-hydrolyzed crude proteins increased the inhibition activity against these bacteria by 3.4–10.5 %. The hydrolyzed peptides were fractionated using a Sephadex G-25 column, and peptides in fraction 17 showed the highest antimicrobial activity. Through peptide identification using LC-MS and prediction of obtained peptide primary sequences with four bioinformatic tools (DBAASP, AMPDiscover, iAMPpred, and AMPscanner), five peptides were selected and synthesized for activity evaluation. The PT11 peptide (RYGAAMAVGIACA) demonstrated the highest antimicrobial activity among the five peptides tested and compared favorably to previously reported algal encrypted peptides. The findings of this research demonstrate that the encrypted PT11 peptide derived from Tetraspora sp. CU2551 has promising potential as an antimicrobial agent. Additionally, we suggest that PT11 might be used for other things besides this, like animal feed.
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Lactobacillus plantarum JCM 1149 Growth Enhancement by using Chlorella sp. KLSc61-pretreated Cells
(2025-06-01)
Khanrin, Lalita
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Boonyakorn, Phonwimon
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Wongsariya, Karn
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Kittiwongwattana, Chokchai
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Maneeruttanarungroj, Cherdsak
Dry microalgal biomass was previously tested as prebiotic to enhance the growth of probiotic bacteria. However, the drying process could be ineffective for scaling up probiotic production. This study aimed to investigate the use of fresh, pretreated microalgal biomass to promote the growth of Lactobacillus plantarum JCM 1149. Chlorella sp. KLSc61 cells were pretreated by three different methods: physical treatment with microwave radiation at power levels of 300, 500, and 700 W; chemical treatment with 0.1 M citric acid and 0.5 M sodium hydroxide; and biological treatment with cellulase enzyme. The 2.5% pretreated Chlorella cells were then added to L. plantarum JCM 1149 culture, and the growth was observed at 37 °C for 24 h of incubation. The results showed that, during the log phase (6-10 h), Chlorella cells pretreated with microwave radiation at 700 W were the most effective in promoting L. plantarum JCM 1149 growth, which was 1.4- and 1.5-fold of L. plantarum JCM 1149 without adding Chlorella and with untreated cells, respectively. Extension of the pretreatment time by microwave radiation at 700 W from up to 2 min increased the growth of L. plantarum JCM 1149 up to 1.8-fold of pretreatment time by microwave radiation at 700 W 1 min, compared to the control groups. Additionally, increasing the amount of Chlorella biomass up to 5% (w/v) extended the log phase of L. plantarum JCM 1149 and increased cell accumulation during the stationary phase. Unlike dry microalgal biomass, the simplicity of fresh, pretreated Chlorella biomass shown in this study may facilitate large-scale, commercial production of L. plantarum strains as probiotics.
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Purification and Identification of Antimicrobial Protein from the Green Alga Tetraspora Sp. CU2551
(2026-02-01)
Klinsanit, Tanaporn
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Tonawut, Yothawut
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Laokua, Natwikar
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Pansomsuay, Rawirat
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Khetkorn, Wanthanee
Antimicrobial resistance (AMR) is an escalating global health threat, necessitating the discovery of novel antimicrobial agents. Algae-derived proteins and peptides have gained increasing attention for their bioactive potential; however, standardized protocols for investigating antimicrobial peptides (AMPs) in green algae, particularly Tetraspora sp. CU2551, remain limited. This protocol describes a step-by-step workflow for the isolation, purification, and characterization of antimicrobial intact proteins from the green alga Tetraspora sp. CU2551. The procedure integrates optimized protein extraction, chromatographic fractionation, electrophoretic separation, antimicrobial activity screening, and protein identification. Protein extraction conditions are first optimized to reduce background inhibitory effects originating from buffer components. When the identity of the active peptide is unknown, a pull-down assay is applied to assess protein binding across different ion-exchange resins and to guide the selection of an appropriate purification matrix. DEAE-Sepharose ion-exchange chromatography is a suitable method for enriching antimicrobial protein fractions. All fractions are systematically evaluated for antimicrobial activity and analyzed by SDS-PAGE. The protein band corresponding to the highest antimicrobial activity, along with a distinct electrophoretic profile, is excised and subjected to LC-MS/MALDI-TOF analysis for protein identification. The workflow is further complemented by in silico analyses to predict antimicrobial peptide-related properties using publicly available bioinformatic tools. This protocol provides a versatile framework for antimicrobial protein discovery and can be readily adapted to other algal species and related biotechnological applications.
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Hydrochloric acid producing higher purity of glucosamine than sulfuric acid: A comparison study with different detection approaches
(2024-06-01)
Kornrawudaphikasama, Yosita
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Laokua, Natwikar
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Rittiyan, Nutnicha
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Klinsalee, Rachaneekorn
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Tonawut, Yothawut
In this study, HCl and H<inf>2</inf>SO<inf>4</inf> were used to compare the best conditions for producing glucosamine from the dry chitin of shrimp shells. The results showed that for HCl hydrolysis, the most favored conditions were 12 M, 80 °C, 2 hours, and a chitin-to-acid volume ratio of 1:20, whereas, for H<inf>2</inf>SO<inf>4</inf> hydrolysis, the most favored conditions were 6 M, 90 °C, 5 hours, and a chitin to acid volume ratio of 1:20. HCl produced pured glucosamine with a higher yield than H<inf>2</inf>SO<inf>4</inf>. Additionally, our findings indicated that glucosamine could be detected using a UV detector with a weak signal, whereas we recommended using an RI detector for a comparably stronger signal. Our production maximum yield of 283.9 ± 13.8 mg Gln g-1 chitin from HCl hydrolysis was comparable to that of other studies, with a comparison between using HCl and H<inf>2</inf>SO<inf>4</inf> being highlighted.
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Purification, characterization, and structural insights of Exo-1,4-β-D-glucosaminidase from Amycolatopsis sp. KLSc63: predictive modeling with glucosamine and N-acetyl glucosamine dimers
(2026-12-01)
Kornrawudaphikasama, Yosita
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Maneeruttanarungroj, Cherdsak
Chitin, a long-chain polysaccharide, is a major component in the exoskeletons of arthropods and the cell walls of fungi. Its derivative, chitosan, is widely used in various fields due to its solubility and versatility. This study focuses on the purification and biochemical characterization of exo-1,4-β-D-glucosaminidase from Amycolatopsis sp. KLSc63, an enzyme crucial for the degradation of chitin and chitosan. The enzyme was purified using anion exchange chromatography and characterized for its activity on colloidal chitin and chitosan solutions. Optimal activity was observed at pH 5.0 and temperatures of 40–45 °C for colloidal chitin and pH 4.0–6.0 at 55 °C for chitosan solution. The enzyme’s molecular weight was approximately 94 kDa. Various metal ions and surfactants significantly influenced enzyme activity, with Mn²⁺ at 1 mM concentration notably enhancing both activities. Structural modeling and docking studies confirmed the enzyme’s substrate specificity and binding interactions. These findings highlight the potential applications of exo-1,4-β-D-glucosaminidase in industrial processes, waste management, and the production of bioactive compounds.
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Proteomic insights into high biohydrogen production by Chlorella sp. KLSc61 under potassium deprivation: upregulation of carbohydrate synthesis proteins
(2025-12-01)
Laokua, Natwikar
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Dejtisakdi, Wipawee
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Maneeruttanarungroj, Cherdsak
Microalgae produce hydrogen by utilizing light energy to split water molecules and this produced hydrogen is considered as a promising alternative energy resource. This study investigated the proteomic response of the microalga, Chlorella sp. KLSc61 under an optimal condition for increasing hydrogen production. Chlorella KLSc61 cells were cultured in potassium deprivation medium (TAP-K medium) with an initial pH of 9.0 and added 25 mM ethanol, they were cultivated and exposed to the light intensity of 54 μmol photons m−2 s−1 at 35 °C. By day 7, under this growth condition, Chlorella cells produced a maximum H<inf>2</inf> yield of 19,600 ± 0.3 mmol H<inf>2</inf> mg−1 chlorophyll, which was 2.5 times greater than that under normal TAP condition. Under an increase of hydrogen production, Chlorella KLSc61 cells were both round- and oval-shaped with an average cell diameter of 10.0 ± 0.5 μm, along with variations of chloroplast distribution within the cytoplasm, whereas Chlorella cells in normal TAP medium showed only round shape with an average cell diameter of 5.0 ± 0.5 μm. Proteins from cells grown under normal and high hydrogen conditions were subjected to proteomic analysis, with the results presented as a cluster heat map of proteomics profiles from five different conditions. A total of 736 protein expression patterns were classified into six different expressed protein groups, which included both up- and down-regulated proteins. Carbohydrate synthesis proteins, including starch synthase, glucose-6-phosphate isomerase, glycerol-3-phosphate dehydrogenase, phosphoglucomutase, and UDP-glucuronate decarboxylase, were abundant during the high H<inf>2</inf> production. These findings allow us to take the next step of those protein overexpression into Chlorella KLSc61 cells to enhance in vivo hydrogen production and this strain could be used as a hydrogen production platform.

Maneeruttanarungroj, Cherdsak

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