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    INVESTIGATION OF ANTIFUNGAL ACTIVITY FROM SCENEDESMUS SP. AND CHLAMYDOMONAS SP. CRUDE EXTRACTS
    Scenedesmus sp. and Chlamydomonas sp. are green microalgae used for various biotechnology applications such as an alternative biofuel, antioxidant, human supplement, aquaculture feed etc. In a decade, finding a new antibiotic resource from a novel organism is significant for pharmaceutical research. Microalgae is a promising candidate since they consist of high-value biocompounds such as β-carotene, astaxanthin, flavonoids, polysaccharide. In this study, we aimed to preliminarily investigate antifungal activities of Scenedesmus and Chlamydomonas crude extracts against to Candida albicans, a fungal pathogen. We prepared both of intracellular and extracellular (culture media) crude extracts by using ethanol and methanol as a solvent and then tested C albicans inhibition performed by agar-well diffusion assay. From the preliminary results, we found that 50 mg/mL of Scenedesmus and Chlamydomonas ethanolic crude extracts from intracellular and extracellular could inhibit C. albicans. This preliminary finding could be a platform for a further study in some bio-based organic compounds that is significant for pharmaceutical application.
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    Genome Insights into the Plant Growth Promoting Features of a Newly Found Microbispora sp. SCL1-1
    Plant growth-promoting (PGP) actinobacteria can be used to promote plant growth. Their use is a promising strategy that can be employed instead of agricultural chemical fertilizers. An actinobacterium strain, designated SCL1-1, was collected and isolated from a soil sample in a herbal garden at Pathum Thani province, Thailand. Analysis revealed that the SCL1-1 strain was a Gram-positive bacterium that formed longitudinal paired spores that were borne directly on aerial mycelia. It contained meso-diaminopimelic acid in its cell wall peptidoglycan. Moreover, madurose, which is a diagnostic sugar, was present in its whole-cell hydrolysates. 16S rRNA gene analysis revealed that the SCL1-1 strain was a member of the Microbispora and showed a close relationship to Microbispora rosea ATCC 12950<sup>T</sup> (99.6%), followed by Microbispora hainanensis DSM 45428<sup>T</sup> (99.2%). However, a genome-based polyphasic study revealed that strain SCL1-1 had a low average nucleotide identity (ANI) (<95%), and digital DNA–DNA hybridization (dDDH) value (<70%) with M. rosea ATCC 12950<sup>T</sup> and M. hainanensis DSM 45428<sup>T</sup>, indicating that strain SCL1-1 was a different species to its close relatives. Genome mining of strain SCL1-1 showed the presence of genes related to the production of indole-3-acetic acid (IAA), and siderophore, which are agents that promote plant growth. In addition, the genome of strain SCL1-1 was found in several secondary metabolite biosynthetic gene clusters, which were possibly encoded for a broad range of remarkable natural products and antibiotics.
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    INVESTIGATION OF ANTIMICROBIAL CAPACITY AND ANTIOXIDANT EFFICIENCY FROM CHLORELLA SPP. CRUDE EXTRACTS
    Functional food, nutraceutical and pharmaceutical products are currently shared as an important part of the consumer market. Microalga plays a significant role in producing a source of good antioxidants including nutrient supplements such as chlorophyll, beta-carotene, astaxanthin etc. A Eukaryotic green microalga, Chlorella, is a unicellular microalga that itself is rich in a source of chlorophyll, carotenoids, the essential amino acids, bioactive compounds including fatty acids and these bioproducts make Chlorella as a great potential for human food and supplements, animal feed and biofuel feedstock. In this study, we aimed to test antimicrobial activity and antioxidant from four different Chlorella strains (Chlorella sp. KU11, Chlorella sp. V55, Chlorella sp. B2-59 and Chlorella sp. N11/59) for pharmaceutical and nutraceutical application. All four-ethanolic crude extracts of Chlorella were tested against six pathogens such as Streptococcus mutans ATCC25175, Bacillus subtilis ATCC6633, Staphylococcus aureus ATCC25923, Escherichia coli ATCC25922, Pseudomonas aeruginosa ATCC27853, and Candida albicans ATCC10231 using agar-well diffusion and the minimum inhibitory concentrating (MIC) assay. The result showed that 50 mg/mL of Chlorella sp. KU and Chlorella sp. V55 crude extracts could inhibit S. mutans, B. subtilis and S. aureus and 100 mg/mL of both crude extracts were able to inhibit E. coli, P. aeruginosa and C. albicans. From the minimum inhibitory concentrating (MIC) assay, we found that Chlorella sp. KU and Chlorella sp. V55 could inhibit S. mutans, B. subtilis, S. aureus E. coli, P. aeruginosa and C. albicans at 25, 50, 50, 100, 100 and 100 mg/mL, respectively and 50, 6.25, 25, 100, 100 and 100 mg/mL, respectively. The antioxidant activity of the four crude extracts were performed by DPPH radical scavenging assay and we found that the Chlorella sp. N11/59 crude extract showed the highest antioxidant efficiency with 63.73% and IC50 value was 2.8293 mg/mL. This finding suggested that microalgae can be a promising platform for pharmaceutical and nutraceutical application.
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    Lactobacillus plantarum JCM 1149 Growth Enhancement by using Chlorella sp. KLSc61-pretreated Cells
    (2025-06-01)
    Khanrin, Lalita
    ;
    Boonyakorn, Phonwimon
    ;
    ; ;
    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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    Using Phytoplankton as Bioindicators of Tourism Impact and Seasonal Eutrophication in the Andaman Sea (Koh Yaa, Thailand)
    (2026-01-01)
    Wongsnansilp, Tassnapa
    ;
    Khamcharoen, Manoch
    ;
    Boonrong, Jaran
    ;
    This study focuses on the diversity of phytoplankton in the Koh Yaa region of Thailand and their relationship with environmental variables, aiming to assess whether human activities (primarily tourism) pose potential threats to the marine ecosystem and provide scientific support for eco-sustainable tourism management decisions in the region. In April, August, and December 2024, corresponding to peak season, off-season, and shoulder season, a total of 156 discrete samples were collected from four coastal sites to analyze water quality parameters such as temperature, pH, total nitrogen (TN), and total phosphorus (TP), along with plankton diversity and abundance. Statistical analyses including two-way ANOVA with Duncan’s Multiple Range Test (DMRT), Pearson correlation analysis, and principal component analysis (PCA) were applied. The results showed a declining trend in plankton abundance over time, peaking at 1009 × 10<sup>6</sup> cells/m<sup>3</sup> in April and dropping to 281 × 10<sup>6</sup> cells/m<sup>3</sup> by December. A total of 15 types of phytoplankton were identified across four phyla: Bacillariophyta, Cyanobacteria, Dinoflagellata, and Chlorophyta. Notably, Chaetoceros from Bacillariophyta accounted for 47% of phytoplankton, while Oscillatoria from Cyanobacteria made up 29.6%. The diversity index and evenness index improved from 1.34 and 0.46 in April to 1.88 and 0.64 in December, respectively. Environmental factors like pH, temperature, and TP significantly affected phytoplankton abundance (p < 0.01), with TP levels ranging from 0.27 to 0.69 mg/L. These results indicate possible pollution in this region, and changes in phytoplankton abundance were linked to seasonal climate variations—especially during peak tourist seasons—which may exacerbate eutrophication affecting community structures.
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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
    ;
    Klinsalee, Rachaneekorn
    ;
    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<sup>−2</sup> s<sup>−1</sup> 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<sup>−1</sup> 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<sup>−1</sup>Chl). 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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    New Synthetic Operon Vectors for Expressing Multiple Proteins in the Chlamydomonas reinhardtii Chloroplast
    (2023-02-01)
    Yeon, Jihye
    ;
    Miller, Stephen M.
    ;
    Microalgae are a promising platform for generating valuable commercial products, including proteins that may not express well in more traditional cell culture systems. In the model green alga Chlamydomonas reinhardtii, transgenic proteins can be expressed from either the nuclear or chloroplast genome. Expression in the chloroplast has several advantages, but technology is not yet well developed for expressing multiple transgenic proteins simultaneously. Here, we developed new synthetic operon vectors to express multiple proteins from a single chloroplast transcription unit. We modified an existing chloroplast expression vector to contain intercistronic elements derived from cyanobacterial and tobacco operons and tested the ability of the resulting operon vectors to express two or three different proteins at a time. All operons containing two of the coding sequences (for C. reinhardtii FBP1 and atpB) expressed the products of those genes, but operons containing the other two coding sequences (C. reinhardtii FBA1 and the synthetic camelid antibody gene VHH) did not. These results expand the repertoire of intercistronic spacers that can function in the C. reinhardtii chloroplast, but they also suggest that some coding sequences do not function well in the context of synthetic operons in this alga.
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    Comprehensive Review of Strategies for Lactic Acid Bacteria Production and Metabolite Enhancement in Probiotic Cultures: Multifunctional Applications in Functional Foods
    (2025-05-01)
    Loo, Jiun Shen
    ;
    Oslan, Siti Nur Hazwani
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    Mokshin, Nur Anis Safiah
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    Othman, Rafidah
    ;
    Amin, Zarina
    Lactic acid bacteria (LAB) play a crucial role in probiotics, functional foods, and sustainable biotechnologies due to their ability to produce bioactive metabolites such as short-chain fatty acids, bacteriocins, vitamins, and exopolysaccharides. These metabolites aid in gut health, pathogen inhibition, and enhanced productivity in the food, pharmaceutical, and aquaculture industries. However, the high production cost remains a major challenge, necessitating cost-effective media formulations and bioprocess optimization. This review explores strategies for maximizing LAB yields and functionality through the precision control of key cultivation parameters, including temperature, pH, and agitation speed, ensuring probiotic viability in compliance with regulatory standards (≥10<sup>6</sup> CFU/g or mL). Furthermore, advances in metabolic engineering, synthetic biology, and the utilization of agro-industrial by-products are driving cost-efficient and eco-friendly LAB production. By integrating scalable fermentation technologies with sustainable resource management, LAB have the potential to bridge the gap between food security, environmental sustainability, and biotechnological innovation. This review provides a comprehensive overview of recent advances in LAB cultivation and bioprocess optimization, ensuring high-quality probiotic production for diverse industrial applications.
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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
    ;
    ;
    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<sup>−2</sup> s<sup>−1</sup> 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<sup>−1</sup> 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.