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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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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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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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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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Characterization and exploration of biological properties of phycobiliproteins purified from Thai karstic cave cyanobacterium Nostoc sp. SW02
(2023-09-01)
Suphan, Sutthawan
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Limrujiwat, Kittakorn
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Kula, Kasinee
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Maneeruttanarungroj, Cherdsak
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Raksajit, Wuttinun
Phycobiliproteins are colored water-soluble proteins found in cyanobacteria with potential applications in food and pharmaceuticals. Phycobiliproteins extracted from the Thai karstic cave cyanobacterium Nostoc sp. strain SW02 were purified, and their biological properties were characterized. Scale-up cultivation for 12 days revealed that maximum biomass and phycobiliproteins producing yield were 1.53 g/L and 31.92%, respectively. After purification, phycoerythrin was obtained as the principal component of phycobiliproteins, with a purity index up to 2.62, sufficient to be considered food and cosmetic grade. Native and SDS-PAGE analysis displayed that purified phycoerythrin and phycocyanin contained the αβ-subunits in hexamer form. Purified phycobiliproteins had a more stable structure and functionality in pH range 5.0–7.0, were resistant to strong oxidizers, and could withstand temperatures up to 60 °C. They also exhibited DPPH scavenging capabilities comparable to standard ascorbic acid. Pathogenic bacterial inhibition (MIC 250 μg/mL) and anticancer activity against the human HeLa cancer cell line (IC<inf>50</inf> 140 μg/mL) were also observed. With these prospective characteristics, phycobiliproteins obtained from Nostoc sp. SW02 have biotechnological value for biomedical research and also as a natural colorant for food and cosmetic applications.
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Highly effective reduction of phosphate and harmful bacterial community in shrimp wastewater using short-term biological treatment with immobilized engineering microalgae
(2023-01-01)
Krasaesueb, Nattawut
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Boonnorat, Jarungwit
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Maneeruttanarungroj, Cherdsak
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Khetkorn, Wanthanee
Shrimp farming wastewater includes high amounts of phosphate and microbiological contaminants, necessitating further treatment before release into receiving water bodies. After 24 h of shrimp wastewater treatment, alginate beads containing the blue-green algal Synechocystis strain lacking the phosphate regulator gene (mutant strain ΔSphU) at 150 mg L−1 reduced phosphate content from 17.5 mg L−1 to 5.0 mg L−1, representing 71.5% removal efficiency, with phosphate removal rate reaching 6.9 mg gDW−1 h−1 during photobioreactor operation. For short-term treatment, removal rates of nitrate, ammonium and nitrite were 42.7, 48.5 and 92.9%, respectively. Microalgal encapsulated beads also impacted the bacterial community composition dynamics in shrimp wastewater. Next-generation sequencing targeting the V3–V4 region of the 16S rDNA gene showed significant differences in bacterial community composition after 24 h of treatment. Proteobacteria are the most abundant phylum in shrimp wastewater. After 24 h of bioremediation, reductions of harmful bacteria in the Cellvibrionaceae and Pseudomonadaceae families were recorded at 5.85 and 3.18%, respectively. Engineered microalgal immobilization under optimal conditions can be applied as an alternative short-term bioremediation strategy to remove phosphate and other harmful microbial contamination from shrimp farming wastewater.
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Photobiohydrogen Production and Strategies for H2 Yield Improvements in Cyanobacteria
(2023-01-01)
Khetkorn, Wanthanee
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Raksajit, Wuttinun
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Maneeruttanarungroj, Cherdsak
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Lindblad, Peter
Hydrogen gas (H<inf>2</inf>) is one of the potential future sustainable and clean energy carriers that may substitute the use of fossil resources including fuels since it has a high energy content (heating value of 141.65 MJ/kg) when compared to traditional hydrocarbon fuels [1]. Water is a primary product of combustion being a most significant advantage of H<inf>2</inf> being environmentally friendly with the capacity to reduce global greenhouse gas emissions. H<inf>2</inf> is used in various applications. It generates electricity in fuel cells, including applications in transportation, and can be applied as fuel in rocket engines [2]. Moreover, H<inf>2</inf> is an important gas and raw material in many industrial applications. However, the high cost of the H<inf>2</inf> production processes requiring the use of other energy sources is a significant disadvantage. At present, H<inf>2</inf> can be prepared in many conventional ways, such as steam reforming, electrolysis, and biohydrogen production processes. Steam reforming uses high-temperature steam to produce hydrogen gas from fossil resources including natural gas. Electrolysis is an electrolytic process to decompose water molecules into O<inf>2</inf> and H<inf>2</inf>. However, both these two methods are energy-intensive and producing hydrogen from natural gas, which is mostly methane (CH<inf>4</inf>) and in steam reforming generates CO<inf>2</inf> and pollutants as by-products. On the other hand, biological hydrogen production is more environmentally sustainable and less energy intensive than thermochemical and electrochemical processes [3], but most concepts are not yet developed to production scale.
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Optimal conditions for maximized H2 yield from a new green algal strain Chlorella sp. KLSc61
(2022-08-01)
Laokua, Natwikar
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Rittiyan, Nutnicha
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Kornrawudaphikasama, Yosita
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Klinsalee, Rachaneekorn
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Tonawut, Yothawut
Hydrogen produced from microalgae is attracting the attention of scientists as a potential new ‘Bio-Circular-Green’ energy source. We screened a selection of naturally occurring algal strains for H<inf>2</inf> yield as a basis for high yield development. The best strain was a new strain of biohydrogen-producing green algae from a fresh water source at King Mongkut's Institute of Technology Ladkrabang, Thailand. Using morphological and 18S rDNA sequencing analyses, this alga was identified and classified as Chlorella sp. KLSc61. When adapted cells in TAP-K medium (with starting pH at 9.0) containing 25 mM ethanol, with 54 μmol photons m−2 s−1 light, and incubated at 35 °C, Chlorella sp. KLSc61 showed a maximum H<inf>2</inf> yield of 9,804 ± 469.5 μmol H<inf>2</inf> mg−1 Chl at day 7. Under these conditions, the cells produced ~ 12.2 times more H<inf>2</inf> gas than without optimization (753.0 μmol H<inf>2</inf> mg−1Chl). Our optimized system also produced H<inf>2</inf> at a rate 11–570 times greater than previously reported microalgae. Thus Chlorella sp. KLSc61 could be a viable strain for biohydrogen production as a clean future energy source.

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