Maneeruttanarungroj, Cherdsak

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School of Science

cherdsak.ma@kmitl.ac.th

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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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Response of green alga Tetraspora sp. CU2551 under potassium deprivation: a new promising strategy for hydrogen production
(2022-04-01)
Pewnual, Totsapon
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Jampapetch, Narirat
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Saladtook, Sathonkorn
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Raksajit, Wuttinun
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Klinsalee, Rachaneekorn
With increasing world energy consumption, renewable energy sources can fulfill the need and many have net zero carbon dioxide emissions. One of these is hydrogen, which is biologically produced molecular hydrogen from organisms. Although many studies have produced hydrogen from green algae and optimized conditions to obtain the highest yield, following our previous works on production optimization from Tetraspora sp. CU2551, we tested deprivation of Fe, Cu, Ca, S, Mg, or K, and surprisingly found that K deprivation showed another promising switch in increased H<inf>2</inf> yield, representing 9.2 ± 0.1 μmol mg−1 DW within 32 h incubation anaerobically. Moreover, longer cell adaptation by aerobic incubation, before production phase, resulted in higher hydrogen yield. Cells adapted in potassium deprivation (TAP-K medium) promoted hydrogen production to a yield of 14.8 ± 0.03 μmol mg−1 DW, or about 3.0 times higher than normal TAP (5.0 ± 0.82 μmol mg−1 DW). The increased yield was caused by (1) lowering PSII activity (direct biophotolysis), resulted in less oxygen being produced, lowering hydrogenase inhibitor levels, and (2) increasing accumulated starch degradation (indirect biophotolysis) (12.2% starch remaining or 7.2 ± 0.9 μg mg−1 DW), increased the flow of electrons to hydrogenase. Our finding makes potassium deprivation conditions as another the promising choice to enhance hydrogen production in biological systems.
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Bioethanol fermentation from acid/base-treated water hyacinth biomass using fermentation yeasts saccharomyces cerevisiae yrk 017 and candida shehatae ATCC 22984
(2019-01-01)
Ochaikul, Duangjai
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Jongmeesuk, Atcharaporn
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Maneeruttanarungroj, Cherdsak
Water hyacinth is considered as one of most abundant lignocellulosic feedstock in Thailand. It has been selected as a substrate for producing ethanol at an economically feasible manner. This study focuses on the bioethanol production from enzyme-treated hydrolysate of acid/base pretreatment water hyacinth. Two different pretreatment methods of water hyacinth were used including either 2 %(v/v) sulfuric acid or 2 %(w/v) sodium hydroxide at 121°C for 15 min. After acid/base pretreatments, the biomass residues in order to release glucose molecules were treated with a commercial cellulase enzyme complex (ACCELLERASE® 1500) at a concentration of 0.30 mL enzyme/g biomass residue. Two yeast species consisting of Saccharomyces cerevisiae YRK 017 and Candida shehatae ATCC 22984 were separately used as inoculum in fermentation process of the hydrolysate from enzymatic treatments. The results showed that ethanol yield obtained from acid pretreatment was slightly higher than the yield from base pretreatment. In acid pretreatment, S. cerevisiae YRK 017 could produce bioethanol in a yield of 3.82 ± 0.10 g/L, whereas C. shehatae ATCC 22984 could produce yielding 2.85 ± 0.04 g/L.
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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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Molecular detection and phylogeny of Ehrlichia canis and Anaplasma platys in naturally infected dogs in Central and Northeast Thailand
(2022-12-01)
Purisarn, Andaman
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Wichianchot, Sakulchit
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Maneeruttanarungroj, Cherdsak
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Mangkit, Bandid
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Raksajit, Wuttinun
Background and Aim: Ehrlichia canis and Anaplasma platys are tick-borne, Gram-negative bacteria that cause canine monocytic ehrlichiosis and canine cyclic thrombocytopenia, respectively. These diseases are of great importance and are distributed globally. This study aimed to create new primers for the identification of E. canis and A. platys in naturally infected dogs using polymerase chain reaction (PCR), DNA sequencing, and phylogenetic analysis using the 16S rDNA and gltA genes. Materials and Methods: In total, 120 blood samples were collected from dogs in three different locations (Saraburi, Buriram, and Nakhon Ratchasima provinces) in Central and Northeast Thailand. The molecular prevalence of E. canis and A. platys was assessed using PCR targeting the 16S rDNA and gltA genes. All positive PCR amplicons were sequenced, and phylogenetic trees were constructed based on the maximum likelihood method. Results: Ehrlichia canis had an overall molecular prevalence of 15.8% based on the 16S rDNA gene, compared to 8.3% based on the gltA gene. In addition, the overall molecular prevalence of A. platys using the 16S rDNA gene was 10.8%, while the prevalence rate was 5.8% using the gltA gene. Coinfection was 0.8% in Saraburi province. The partial sequences of the 16S rDNA and gltA genes of E. canis and A. platys in dogs in Central and Northeast Thailand showed 96.75%–100% identity to reference sequences in GenBank. Phylogenetic analysis of the 16S rDNA and gltA genes revealed that E. canis and A. platys sequences were clearly grouped into their own clades. Conclusion: This study demonstrated the molecular prevalence of E. canis and A. platys in Central and Northeast Thailand. The 16S rDNA and gltA genes were useful for the diagnosis of E. canis and A. platys. Based on the phylogenetic analysis, the partial sequences of the 16S rDNA and gltA genes in E. canis and A. platys were related to prior Thai strains and those from other countries.
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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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Proteomics identification of overexpressed serum proteins in dogs with Babesia canis infection
(2023-10-01)
Ritchoo, Sudpatchara
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Havanapan, Phattara Orn
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Sussadee, Metita
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Maneeruttanarungroj, Cherdsak
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Rucksaken, Rucksak
Background and Aim: Canine babesiosis, caused by the protozoan parasite Babesia canis, is characterized by clinical manifestations, including hemolytic anemia, thrombocytopenia, multiple organ failure, and may result in death. This disease is detected using conventional blood smears, which are time-consuming and have low sensitivity. This study aimed to investigate a more rapid and sensitive method for detecting B. canis infection in dogs by examining the expressed serum protein profiles using proteomics. Materials and Methods: We collected six sera samples from three healthy and three B. canis-infected dogs diagnosed using blood smear and polymerase chain reaction. We analyzed the proteins using two-dimensional gel electrophoresis. The candidate spots from the gel were subjected to protein identification using a nano-liquid chromatography system coupled to an ion-trap mass spectrometer equipped with an electrospray ionization nano-sprayer. Results: We found that 10 protein spots were overexpressed in the serum samples from infected dogs compared with healthy dogs, which corresponded to three proteins: serotransferrin, serotransferrin isoforms X1, and hemopexin. Furthermore, analysis of the protein-protein interaction network confirmed that they strongly interacted with each other. Conclusion: This study suggests that high levels of serotransferrin and hemopexin are related to B. canis infection, making these proteins potential candidates for the development of diagnostic molecules or vaccines.
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Exogenous Trehalose Improves Growth, Glycogen and Poly-3-Hydroxybutyrate (PHB) Contents in Photoautotrophically Grown Arthrospira platensis under Nitrogen Deprivation
(2024-02-01)
Mudtham, Nat Anong
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Promariya, Authen
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Duangsri, Chanchanok
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Maneeruttanarungroj, Cherdsak
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Ngamkala, Suchanit
Glycogen and poly-3-hydroxybutyrate (PHB) are excellent biopolymer products from cyanobacteria. In this study, we demonstrate that nitrogen metabolism is positively influenced by the exogenous application of trehalose (Tre) in Arthrospira platensis under nitrogen-deprived (−N) conditions. Cells were cultivated photoautotrophically for 5 days under −N conditions, with or without the addition of exogenous Tre. The results revealed that biomass and chlorophyll-a content of A. platensis experienced enhancement with the addition of 0.003 M and 0.03 M Tre in the −N medium after one day, indicating relief from growth inhibition caused by nitrogen deprivation. The highest glycogen content (54.09 ± 1.6% (w/w) DW) was observed in cells grown for 2 days under the −N + 0.003 M Tre condition (p < 0.05), while the highest PHB content (15.2 ± 0.2% (w/w) DW) was observed in cells grown for 3 days under the −N + 0.03 M Tre condition (p < 0.05). The RT-PCR analysis showed a significant increase in glgA and phaC transcript levels, representing approximately 1.2- and 1.3-fold increases, respectively, in A. platensis grown under −N + 0.003 M Tre and −N + 0.03 M Tre conditions. This was accompanied by the induction of enzyme activities, including glycogen synthase and PHA synthase with maximal values of 89.15 and 0.68 µmol min−1 mg−1 protein, respectively. The chemical structure identification of glycogen and PHB from A. platensis was confirmed by FTIR and NMR analysis. This research represents the first study examining the performance of trehalose in promoting glycogen and PHB production in cyanobacteria under nitrogen-deprived conditions.
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Maximizing Omega-3 production in Chlorella sp. KLSc59 through central composite design-driven cultivation and cellulase-assisted extraction
(2026-10-01)
Pansomsuay, Rawirat
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Vachiraroj, Nuttapon
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Pandhal, Jagroop
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Sirikwanpong, Sukrit
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Maneeruttanarungroj, Cherdsak
Chlorella sp. KLSc59 is a promising freshwater microalga for omega-3 oil production, but its application is limited by low biomass yield and inefficient lipid recovery. This study developed an integrated strategy to improve biomass production and omega-3 recovery. Carbon and nitrogen source screening in modified Tris-Acetate-Phosphate (TAP) medium identified acetic acid (CH<inf>3</inf>COOH) and ammonium nitrate (NH<inf>4</inf>NO<inf>3</inf>) as the best-performing carbon and nitrogen sources. Central Composite Design (CCD) was then applied to optimize CH<inf>3</inf>COOH and NH<inf>4</inf>NO<inf>3</inf> concentrations and pH. The resulting M2 formulation (33.9 mM CH<inf>3</inf>COOH, 16.1 mM NH<inf>4</inf>NO<inf>3</inf>, pH 6.6) gave the highest specific growth rate (0.20 h−1) and biomass concentration (1.16 g L−1) among the tested cultivation conditions. A second CCD was applied to optimize cell disruption by varying cellulase loading and sonication duration, identifying D6 (1,000 International Units (IU) g−1 DCW cellulase; 150 s sonication) as the best-performing extraction condition within the tested range for total and omega-3 fatty acid recovery. Among the tested comparative conditions, M2CaD6 yielded the highest omega-3 content and yield, increasing from 0.37 to 6.22 µg mg−1 DCW and from 0.74 to 14.90 mg L−1, respectively, corresponding to 17- and 20-fold increases over the previous TAP-CaCl<inf>2</inf> cultivation and extraction method (PD0). Reducing NH<inf>4</inf>NO<inf>3</inf> in M2 by half increased linoleic acid content, whereas higher light intensity (50 µmol m−2 s−1) reduced cellular total fatty acid accumulation. Overall, coordinated optimization of cultivation, CaCl<inf>2</inf> induction, and enzymatic–mechanical disruption improved omega-3 production from Chlorella sp. KLSc59 and provides a basis for further process development.
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Rapid method for DNA isolation from a tough cell wall green alga Tetraspora sp. CU2551
(2016-06-01)
Maneeruttanarungroj, Cherdsak
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Incharoensakdi, Aran
Genetic studies are important to understand the complex biological system of various organisms. Some eukaryotic green organisms have tough cell wall which precludes the efficient extraction of the genetic materials. Here, we developed the method for simple and rapid isolation of high quality DNA from a green alga Tetraspora sp. CU2551. The cell homogenization procedures were combined with physical force plus heat treatment to disrupt the cell envelope of Tetraspora sp. CU2551. Without protease treatment, vortexing with glass bead for 30–105 s at 70 °C led to the isolation of a high purity DNA which was suitable for downstream process. The improved method was successfully developed and could be applied for the rapid isolation of DNA from other unicellular and filamentous green microalgal strains.
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Maneeruttanarungroj, Cherdsak

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